Faculty Profiles - Otsuka Motoyuki

Papers - Otsuka Motoyuki

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PTBP1 protects Y RNA from cleavage leading to its apoptosis-specific degradation. Reviewed International journal

Takeshi Kamakura, Kazuaki Kameda, Masahiko Manabe, Kan Torii, Yuki Sugiura, Seiko Ito, Shunya Nakayama, Takanobu Shimizu, Etsuko Nagashima, Kosuke Kamiya, Masahiro Oka, Masafumi Tanaka, Motoyuki Otsuka, Masato Ohtsuka, Ai Kotani

Cell death discovery 10 ( 1 ) 322 - 322 2024.7

Language：English Publishing type：Research paper (scientific journal)

Some RNAs such as 28S rRNA, U1 small nuclear RNA (snRNA), and Y RNAs are known to be cleaved during apoptosis. The underlying mechanism, functions, and biological significance of RNA degradation in apoptosis remain elusive. Y RNAs are non-coding RNAs widely conserved from bacteria to mammals, and are major components of Ro ribonucleoprotein (RNP) complexes which contain the 60 kDa Ro protein (SS-A) and the 50 kDa La protein (SS-B). The autoantigenic Ro and La proteins were identified by autoantibodies present in the sera from patients with Systemic lupus erythematosus (SLE) and Sjögren's syndrome (SjS). We previously identified novel, functional small RNAs named AGO-taxis small RNAs (ASRs) that are specifically bound to Argonaute protein 1 (AGO1), which are processed from Y RNAs. Cell-free analysis combined with fractionation methods revealed that the apoptosis-specific biogenesis of ASRs or cleavage of Y RNA was induced by truncation of polypyrimidine tract-binding protein 1 (PTBP1), which is an endoribonuclease inhibitor of Y RNAs by caspase 3. Caspase 3-resistant PTBP1 mutant protected cleavage of Y RNAs in apoptosis induced by staurosporine. Furthermore, caspase 3-resistant PTBP1 mutant knock-in mice showed elevated cytokines, dysregulation of the germinal center formation compared to the wild-type mice at LPS stimulation, and high positivity of antinuclear antibody. Those results suggest that cleavage of Y RNAs or biogenesis of ASR during apoptosis has critical biological functions and their deregulation result in immune dysregulation and the formation of autoantibody, possibly leading to the development of autoimmune diseases.

DOI： 10.1038/s41420-024-02080-6

Virtual indigo carmine chromoendoscopy images: A novel modality for peroral cholangioscopy using artificial intelligence technology (with video). Reviewed International journal

Ryosuke Sato, Kazuyuki Matsumoto, Hideaki Kinugasa, Masahiro Tomiya, Takayoshi Tanimoto, Akimitsu Ohto, Kei Harada, Nao Hattori, Taisuke Obata, Akihiro Matsumi, Kazuya Miyamoto, Kosaku Morimoto, Hiroyuki Terasawa, Yuki Fujii, Daisuke Uchida, Koichiro Tsutsumi, Shigeru Horiguchi, Hironari Kato, Yoshiro Kawahara, Motoyuki Otsuka

Gastrointestinal endoscopy 2024.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: Accurately diagnosing biliary strictures is crucial for surgical decisions, and although peroral cholangioscopy (POCS) aids in visual diagnosis, diagnosing malignancies or determining lesion margins via this route remains challenging. Indigo carmine is commonly used to evaluate lesions during gastrointestinal endoscopy. We aimed to establish the utility of virtual indigo carmine chromoendoscopy (VICI) converted from POCS images using artificial intelligence. METHODS: This single-center, retrospective study analyzed 40 patients with biliary strictures who underwent POCS using white light imaging (WLI) and narrow-band imaging (NBI). A "cycle-consistent adversarial network" (CycleGAN) was used to convert the WLI into VICI of POCS images. Three experienced endoscopists evaluated WLI, NBI, and VICI via POCS in all patients. The primary outcome was the visualization quality of surface structures, surface microvessels, and lesion margins. The secondary outcome was diagnostic accuracy. RESULTS: VICI showed superior visualization of the surface structures and lesion margins compared with WLI (P<0.001) and NBI (P<0.001). The diagnostic accuracies were 72.5%, 87.5%, and 90.0% in WLI alone, WLI and VICI simultaneously, and WLI and NBI simultaneously, respectively. WLI and VICI simultaneously tended to result in higher accuracy than WLI alone (P=0.083) and the results were not significantly different from WLI and NBI simultaneously (P=0.65). CONCLUSIONS: VICI in POCS proved valuable for visualizing surface structures and lesion margins and contributed to higher diagnostic accuracy comparable to NBI. In addition to NBI, VICI may be a novel supportive modality for POCS.

DOI： 10.1016/j.gie.2024.06.013

Innovative submucosal injection solution for endoscopic resection with phosphorylated pullulan: a preclinical study. Reviewed International journal

Takuya Satomi, Yukari Ochi, Takumi Okihara, Hiroki Fujii, Yasuhiro Yoshida, Katsumi Mominoki, Haruko Hirayama, Junki Toyosawa, Yasushi Yamasaki, Seiji Kawano, Yoshiro Kawahara, Hiroyuki Okada, Motoyuki Otsuka, Akihiro Matsukawa

Gastrointestinal endoscopy 99 ( 6 ) 1039 - 1047 2024.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: A submucosal injection solution is used to assist in endoscopic surgery. The high viscosity of current solutions makes them difficult to inject. In the present study, we developed an extremely low-viscosity, easy-to-use submucosal injection solution using phosphorylated pullulan (PPL). METHODS: The PPL solutions were prepared at different concentrations, and their viscosities were measured. The mucosal elevation capacity was evaluated using excised porcine stomachs. Controls included 0.4% sodium hyaluronate (SH), 0.6% sodium alginate (SA), and saline. To evaluate the practicality, the catheter injectability of 0.7% PPL was measured, and EMR and endoscopic submucosal dissection (ESD) were performed using the stomach and colorectum of live pigs. As controls, 0.4% SH and saline were used. RESULTS: The PPL solutions were of extremely low viscosity compared to the solutions of 0.4% SH and 0.6% SA. Nevertheless, the mucosal elevation capacity of PPL solutions for up to 0.7% concentration was similar to that of 0.4% SH, and 0.7% PPL was less resistant to catheter infusion than 0.4% SH and 0.6% SA. In live pig experiments with endoscopic mucosal resection and ESD, snaring after submucosal injection of 0.7% PPL was easier than with 0.4% SH, ESD with 0.7% PPL produced less bubble formation than with 0.4% SH, and the procedure time tended to be shorter with 0.7% PPL than with 0.4% SH because of the shorter injection time. CONCLUSIONS: The PPL solution is an innovative and easy-to-use submucosal injection solution.

DOI： 10.1016/j.gie.2024.01.015

Methylation analysis of DCC gene in saliva samples is an efficient method for non-invasive detection of superficial hypopharyngeal cancer. Reviewed International journal

Ryosuke Hirai, Hideaki Kinugasa, Shumpei Yamamoto, Soichiro Ako, Koichiro Tsutsumi, Makoto Abe, Koji Miyahara, Masahiro Nakagawa, Motoyuki Otsuka

British journal of cancer 130 ( 10 ) 1725 - 1731 2024.3

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Advances in upper gastrointestinal endoscopic technology have enabled early detection and treatment of hypopharyngeal cancer. However, in-depth pharyngeal observations require sedation and are invasive. It is important to establish a minimally invasive and simple evaluation method to identify high-risk patients. METHODS: Eighty-seven patients with superficial hypopharyngeal cancer and 51 healthy controls were recruited. We assessed the methylation status of DCC, PTGDR1, EDNRB, and ECAD, in tissue and saliva samples and verified the diagnostic accuracy by methylation analyses of their promoter regions using quantitative methylation-specific PCR. RESULTS: Significant differences between cancer and their surrounding non-cancerous tissues were observed in the methylation values of DCC (p = 0.003), EDNRB (p = 0.001), and ECAD (p = 0.043). Using receiver operating characteristic analyses of the methylation values in saliva samples, DCC showed the highest area under the curve values for the detection of superficial hypopharyngeal cancer (0.917, 95% confidence interval = 0.864-0.970), compared with those for EDNRB (0.680) and ECAD (0.639). When the cutoff for the methylation values of DCC was set at ≥0.163, the sensitivity to detect hypopharyngeal cancer was 82.8% and the specificity was 90.2%. CONCLUSIONS: DCC methylation in saliva samples could be a non-invasive and efficient tool for early detection of hypopharyngeal cancer in high-risk patients.

DOI： 10.1038/s41416-024-02654-2

MicroRNA-34a-5p: A pivotal therapeutic target in gallbladder cancer. Reviewed International journal

Takashi Oda, Koichiro Tsutsumi, Taisuke Obata, Eijiro Ueta, Tatsuya Kikuchi, Soichiro Ako, Yuki Fujii, Tatsuhiro Yamazaki, Daisuke Uchida, Kazuyuki Matsumoto, Shigeru Horiguchi, Hironari Kato, Hiroyuki Okada, Ryota Chijimatsu, Motoyuki Otsuka

Molecular therapy. Oncology 32 ( 1 ) 200765 - 200765 2024.3

Language：English Publishing type：Research paper (scientific journal)

Gallbladder cancer incidence has been increasing globally, and it remains challenging to expect long prognosis with the current systemic chemotherapy. We identified a novel nucleic acid-mediated therapeutic target against gallbladder cancer by using innovative organoid-based gallbladder cancer models generated from KrasLSL-G12D/+; Trp53f/f mice. Using comprehensive microRNA expression analyses and a bioinformatics approach, we identified significant microRNA-34a-5p downregulation in both murine gallbladder cancer organoids and resected human gallbladder cancer specimens. In three different human gallbladder cancer cell lines, forced microRNA-34a-5p expression inhibited cell proliferation and induced cell-cycle arrest at the G1 phase by suppressing direct target (CDK6) expression. Furthermore, comprehensive RNA sequencing revealed the significant enrichment of gene sets related to the cell-cycle regulators after microRNA-34a-5p expression in gallbladder cancer cells. In a murine xenograft model, locally injected microRNA-34a-5p mimics significantly inhibited gallbladder cancer progression and downregulated CDK6 expression. These results provide a rationale for promising therapeutics against gallbladder cancer by microRNA-34a-5p injection, as well as a strategy to explore therapeutic targets against cancers using organoid-based models, especially for those lacking useful genetically engineered murine models, such as gallbladder cancer.

DOI： 10.1016/j.omton.2024.200765

Gut Bacteria-derived Membrane Vesicles Induce Colonic Dysplasia by Inducing DNA Damage in Colon Epithelial Cells. Reviewed International journal

Yu Miyakawa, Motoyuki Otsuka, Chikako Shibata, Takahiro Seimiya, Keisuke Yamamoto, Rei Ishibashi, Takahiro Kishikawa, Eri Tanaka, Takayuki Isagawa, Norihiko Takeda, Noriaki Kamio, Kenichi Imai, Mitsuhiro Fujishiro

Cellular and molecular gastroenterology and hepatology 17 ( 5 ) 745 - 767 2024.2

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND & AIMS: Colorectal cancer (CRC) is the third most common cancer in the world. Gut microbiota has recently been implicated in the development of CRC. Actinomyces odontolyticus is one of the most abundant bacteria in the gut of patients with very early stages of CRC. A odontolyticus is an anaerobic bacterium existing principally in the oral cavity, similar to Fusobacterium nucleatum, which is known as a colon carcinogenic bacterium. Here we newly determined the biological functions of A odontolyticus on colonic oncogenesis. METHODS: We examined the induction of intracellular signaling by A odontolyticus in human colonic epithelial cells (CECs). DNA damage levels in CECs were confirmed using the human induced pluripotent stem cell-derived gut organoid model and mouse colon tissues in vivo. RESULTS: A odontolyticus secretes membrane vesicles (MVs), which induce nuclear factor kappa B signaling and also produce excessive reactive oxygen species (ROS) in colon epithelial cells. We found that A odontolyticus secretes lipoteichoic acid-rich MVs, promoting inflammatory signaling via TLR2. Simultaneously, those MVs are internalized into the colon epithelial cells, co-localize with the mitochondria, and cause mitochondrial dysfunction, resulting in excessive ROS production and DNA damage. Induction of excessive DNA damage in colonic cells by A odontolyticus-derived MVs was confirmed in the gut organoid model and also in mouse colon tissues. CONCLUSIONS: A odontolyticus secretes MVs, which cause chronic inflammation and ROS production in colonic epithelial cells, leading to the initiation of CRC.

DOI： 10.1016/j.jcmgh.2024.01.010

CA19-9-Positive Extracellular Vesicle Is a Risk Factor for Cancer-Associated Thrombosis in Pancreatic Cancer. Reviewed International journal

Chikako Shibata, Motoyuki Otsuka, Kazunaga Ishigaki, Takahiro Seimiya, Takahiro Kishikawa, Mitsuhiro Fujishiro

Gastro hep advances 3 ( 4 ) 551 - 561 2024

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: Cancer-associated venous thromboembolism (VTE) is a frequent complication associated with high mortality in patients with cancer, particularly pancreatic cancer. While biological factors such as coagulation factors released from cancer cells may underlie the mechanisms of cancer-associated VTE, the detailed mechanisms have not been determined. Here, we aimed to determine whether extracellular vesicles carrying a glycan sialyl-Lewisa, known as carbohydrate antigen 19-9 (CA19-9), which is a clinically used serum tumor marker and selectin ligand, are a significant cause of cancer-associated VTE. METHODS: Risk factors for cancer-associated VTE were determined using clinical data. EVs derived from CA19-9-deficient or overexpressing pancreatic cancer cells were characterized. The protein levels of coagulation factors on the surface of the EVs were quantified using our newly developed sensitive method. RESULTS: Higher CA19-9 levels in the sera of patients were significantly associated with the occurrence of VTE. Using CA19-9-negative or overexpressing pancreatic cancer cells, we found that EVs derived from these cells interacted with E-selectin of endothelial cells in a CA19-9-dependent manner in cell-based assays and in vitro blood vessel models. EVs derived from cancer cells have higher tissue factor levels on their surfaces, and increased tissue factor activity is induced locally, where CA19-9-positive EVs bind to activated endothelial cells. CONCLUSION: These results suggest that the binding between CA19-9-positive EVs released from cancer cells and endothelial cell E-selectin explains the increased frequency of VTE in patients with pancreatic cancer.

DOI： 10.1016/j.gastha.2024.02.005

MicroRNA-451a inhibits gemcitabine-refractory biliary tract cancer progression by suppressing the MIF-mediated PI3K/AKT pathway. Reviewed International journal

Taisuke Obata, Koichiro Tsutsumi, Eijiro Ueta, Takashi Oda, Tatsuya Kikuchi, Soichiro Ako, Yuki Fujii, Tatsuhiro Yamazaki, Daisuke Uchida, Kazuyuki Matsumoto, Shigeru Horiguchi, Hironari Kato, Hiroyuki Okada, Motoyuki Otsuka

Molecular therapy. Nucleic acids 34 102054 - 102054 2023.12

Language：English Publishing type：Research paper (scientific journal)

Gemcitabine is an effective chemotherapeutic agent for biliary tract cancers (BTCs), including gallbladder cancer (GBC) and cholangiocarcinoma (CCA). However, few other effective agents are currently available, particularly for GEM-refractory BTCs. We previously identified microRNA-451a (miR-451a) as a potential therapeutic target in GBC. To elucidate the antineoplastic effects of miR-451a and its underlying mechanisms, we transfected miR-451a into GBC, gemcitabine-resistant GBC (GR-GBC), and gemcitabine-resistant CCA (GR-CCA) cell lines. Furthermore, mimicking in vivo conditions, tumorigenic GBC organoids and three-dimensional (3D) cell culture systems were employed to investigate the anti-proliferative effects of miR-451a on BTCs, and its effect on stem cell properties. We found that miR-451a significantly inhibited cell proliferation, induced apoptosis, and reduced chemoresistant phenotypes, such as epithelial-mesenchymal transition, in both GBC and GR-GBC. The principal mechanism is probably the negative regulation of the phosphatidylinositol 3-kinase/AKT pathway, partially accomplished by directly downregulating macrophage migration inhibitory factor. The Gene Expression Omnibus database revealed that miR-451a was the most significantly downregulated microRNA in CCA tissues. The introduction of miR-451a resulted in similar antineoplastic effects in GR-CCA. Furthermore, miR-451a reduced cell viability in 3D spheroid models and tumorigenic GBC organoids. These findings suggest that the supplementation of miR-451a is a potential treatment strategy for GEM-refractory BTCs.

DOI： 10.1016/j.omtn.2023.102054

Cabozantinib inhibits HBV-RNA transcription by decreasing STAT3 binding to the enhancer region of cccDNA. Reviewed International journal

Kazuyoshi Funato, Nozomi Miyake, Kazuma Sekiba, Yu Miyakawa, Takahiro Seimiya, Chikako Shibata, Takahiro Kishikawa, Motoyuki Otsuka

Hepatology communications 7 ( 11 ) 2023.11

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Precision medicine and customized therapeutics based on the features of each patient are important for maximizing therapeutic effects. Because most cases of HCC occur in the damaged liver through various etiologies, such as hepatitis virus infection, steatohepatitis, and autoimmune hepatitis, there should be a rationale for the choice of therapeutic options based on these etiologies. Although cabozantinib, an oral multikinase inhibitor, has demonstrated clinical effectiveness in advanced HCC, subgroup analyses showed a lower HR for death in HBV-related HCC. This study aimed to determine the therapeutic effects of cabozantinib in HBV-related HCC. METHODS: Using HBV infection models and gene knockout cells, we determined the crucial signaling axis responsible for the effects of cabozantinib on HBV. A chromatin immunoprecipitation assay was performed to determine the interaction between the signaling molecules and HBV DNA. Agonists and inhibitors were used for confirmation. RESULTS: Cabozantinib inhibited HBV replication through the HGF-mesenchymal-epithelial transition factor-signal transducer and activator of transcription 3 (MET-STAT3) signaling axis. The importance of STAT3 in viral replication has been confirmed using gene-edited STAT3 knockout cells. The chromatin immunoprecipitation assay revealed that the binding levels of phosphorylated STAT3 to enhancer region 1 of HBV covalently closed circular DNA were significantly increased by HGF stimulation. CONCLUSIONS: Cabozantinib has favorable therapeutic effects on HBV-related HCC because it inhibits HCC not only directly but also indirectly by means of inhibitory effects on HBV.

DOI： 10.1097/HC9.0000000000000313

Extracellular vesicle‑mediated RNA editing may underlie the heterogeneity and spread of hepatocellular carcinoma in human tissue and in vitro. Reviewed International journal

Chikako Shibata, Motoyuki Otsuka, Takayuki Shimizu, Takahiro Seimiya, Takahiro Kishikawa, Taku Aoki, Mitsuhiro Fujishiro

Oncology reports 50 ( 5 ) 2023.11

Language：English Publishing type：Research paper (scientific journal)

Extracellular vesicles (EVs) produced by various cells, including tumor cells, carry biomolecules to neighboring cells. In hepatocellular carcinoma (HCC), adenosine to inosine RNA editing of antizyme inhibitor 1 (AZIN1), specifically regulated by adenosine deaminase acting on RNA‑1 (ADAR1), promotes carcinogenesis. The present study examined if EVs and ADAR1 in the EVs released from HCC cells are transferred to neighboring cells in co‑culture systems and reporter assay. Distribution of the ADAR1 expression in human tissues were examined by immunohistochemistry. EVs released from HCC cells containing ADAR1 were delivered to neighboring HCC cells and non‑cancerous hepatocytes. The increased ADAR1 protein levels resulted in serine to glycine substitution at residue 367 of AZIN1, which augmented transformation potential and increased aggressive behavior of cancer cells. In clinically resected samples, ADAR1 distribution was highly heterogeneous within the tumor specimen and denser in non‑cancerous tissue surrounding the HCC tissue. These observations suggested that ADAR1 protein may be delivered from HCC cells to neighboring cells via EVs and that EV‑mediated RNA editing may serve a pivotal role in determining HCC heterogeneity and spread.

DOI： 10.3892/or.2023.8631

An unusual case of gastric polyposis. Reviewed International journal

Masaya Iwamuro, Seiji Kawano, Motoyuki Otsuka

Gastroenterology 165 ( 5 ) 1110 - 1113 2023.8

A rare case of severe colitis associated with a pancreatic neuroendocrine tumor. Reviewed International journal

Ryosuke Sato, Shigeru Horiguchi, Motoyuki Otsuka

Gastroenterology 166 ( 1 ) e13-e16 2023.7

Machine learning for individualized prediction of hepatocellular carcinoma development after the eradication of hepatitis C virus with antivirals. Reviewed International journal

Tatsuya Minami, Masaya Sato, Hidenori Toyoda, Satoshi Yasuda, Tomoharu Yamada, Takuma Nakatsuka, Kenichiro Enooku, Hayato Nakagawa, Hidetaka Fujinaga, Masashi Izumiya, Yasuo Tanaka, Motoyuki Otsuka, Takamasa Ohki, Masahiro Arai, Yoshinari Asaoka, Atsushi Tanaka, Kiyomi Yasuda, Hideaki Miura, Itsuro Ogata, Toshiro Kamoshida, Kazuaki Inoue, Ryo Nakagomi, Masatoshi Akamatsu, Hiroshi Mitsui, Hajime Fujie, Keiji Ogura, Koji Uchino, Hideo Yoshida, Kazuyuki Hanajiri, Tomonori Wada, Kiyohiko Kurai, Hisato Maekawa, Yuji Kondo, Shuntaro Obi, Takuma Teratani, Naohiko Masaki, Kayo Nagashima, Takashi Ishikawa, Naoya Kato, Hiroshi Yotsuyanagi, Kyoji Moriya, Takashi Kumada, Mitsuhiro Fujishiro, Kazuhiko Koike, Ryosuke Tateishi

Journal of hepatology 2023.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: Accurate risk stratification for hepatocellular carcinoma (HCC) after achieving a sustained viral response (SVR) is necessary for optimal surveillance. We aimed to develop and validate a machine learning (ML) model to predict the risk of HCC after achieving an SVR in individual patients. METHODS: In this multicenter cohort study, 1742 patients with chronic hepatitis C who achieved an SVR were enrolled. Five ML models were developed including DeepSurv, gradient boosting survival analysis, random survival forest (RSF), survival support vector machine, and a conventional Cox proportional hazard model. Model performance was evaluated using Harrel' c-index and was externally validated in an independent cohort (977 patients). RESULTS: During the mean observation period of 5.4 years, 122 patients developed HCC (83 in the derivation cohort and 39 in the external validation cohort). The RSF model showed the best discrimination ability using seven parameters at the achievement of an SVR with a c-index of 0.839 in the external validation cohort and a high discriminative ability when the patients were categorized into three risk groups (P <0.001). Furthermore, this RSF model enabled the generation of an individualized predictive curve for HCC occurrence for each patient with an app available online. CONCLUSIONS: We developed and externally validated an RSF model with good predictive performance for the risk of HCC after an SVR. The application of this novel model is available on the website. This model could provide the data to consider an effective surveillance method. Further studies are needed to make recommendations for surveillance policies tailored to the medical situation in each country. IMPACT AND IMPLICATIONS: A novel prediction model for HCC occurrence in patients after hepatitis C virus eradication was developed using machine learning algorithms. This model, using seven commonly measured parameters, has been shown to have a good predictive ability for HCC development and could provide a personalized surveillance system.

DOI： 10.1016/j.jhep.2023.05.042

Recent advances in extracellular vesicles in gastrointestinal cancer and lymphoma. Reviewed International journal

Motoyuki Otsuka, Ai Kotani

Cancer science 114 ( 6 ) 2230 - 2237 2023.2

Language：English Publishing type：Research paper (scientific journal)

Extracellular vesicles (EVs) are intercellular communication agents that transfer microRNAs (miRNAs), other non-coding RNAs (ncRNAs), messenger RNAs (mRNAs), proteins, lipids, metabolites, and other molecules from donor cells (e.g., cancer cells) to recipient cells (e.g., stromal cells). In 2007, miRNAs were reported to be abundant among the ncRNAs present in EVs. Since then, many studies have investigated the functions of miRNAs and have attempted to apply these molecules to aid in the diagnosis and treatment of cancer. Research on EVs has expanded, particularly in the field of cancer, because cancer cells heavily secrete EVs. The cargo of these EVs, especially those in small EVs such as exosomes, is assumed to work cooperatively and significantly in the tumor microenvironment and to promote metastasis. In this review, we first summarize recent studies on EVs in gastrointestinal cancer and highlight studies on human satellite II RNAs, which are a type of ncRNA found in EVs and possessing repetitive sequences. Second, since several recent studies have revealed that phospholipids, which are components of EV membranes, play important roles in intercellular communication and in the generation of lipid mediators in the tumor microenvironment, we review the reported roles of these molecules and discuss their potential use in the design of new cancer treatments.

DOI： 10.1111/cas.15771

Combination of serum human satellite RNA and miR-21-5p levels as a biomarker for pancreatic cancer. Reviewed International journal

Takahiro Seimiya, Tatsunori Suzuki, Takuma Iwata, Takahiro Kishikawa, Kazuma Sekiba, Chikako Shibata, Kazunaga Ishigaki, Hiroaki Fujiwara, Hiroki Oyama, Sachiko Kanai, Tatsuya Sato, Yousuke Nakai, Rei Ishibashi, Masaru Moriyama, Ryo Nakagawa, Hideaki Ijichi, Motoyuki Otsuka, Kazuhiko Koike

iScience 26 ( 2 ) 106021 - 106021 2023.2

Language：English Publishing type：Research paper (scientific journal)

Pancreatic ductal adenocarcinoma (PDAC) has a poor prognosis due to the difficulty of its diagnosis. Because human satellite II (HSATII) RNA, a satellite repeat RNA, is highly and specifically expressed in human PDAC, the serum HSATII RNA level may be a biomarker of PDAC. To measure the serum HSATII RNA level with high sensitivity and reproducibility, we previously developed a convenient method, tandem repeat amplification by nuclease protection (TRAP) combined with droplet digital PCR (ddPCR). Here, we refined the original method by simultaneously measuring the serum miR-21-5p level to enhance the detection of PDAC. The resulting PDAC-Index, constructed using serum HSATII RNA and miR-21-5p levels, discriminated patients with PDAC with high accuracy. We verified the clinical usefulness of the PDAC-Index as a supportive test in difficult-to-diagnose cases. The PDAC-Index has satisfactory diagnostic performance and may routinely be applied for detecting PDAC.

DOI： 10.1016/j.isci.2023.106021

Overcoming T-cell exhaustion: new therapeutic targets in HCC immunotherapy. Invited Reviewed International journal

Takahiro Seimiya, Motoyuki Otsuka, Mitsuhiro Fujishiro

Hepatology (Baltimore, Md.) 78 ( 4 ) 1009 - 1011 2023.1

Lipolysis by pancreatic cancer-derived extracellular vesicles in cancer-associated cachexia via specific integrins. Reviewed International journal

Chikako Shibata, Motoyuki Otsuka, Takahiro Seimiya, Takahiro Kishikawa, Kazunaga Ishigaki, Mitsuhiro Fujishiro

Clinical and translational medicine 12 ( 11 ) e1089 2022.11

Circulating Tumor DNA Guiding Adjuvant Therapy in Colon Cancer. Reviewed International journal

Motoyuki Otsuka

The New England Journal of Medicine 387 ( 8 ) 759 - 760 2022.8

MNX1-HNF1B Axis Is Indispensable for Intraductal Papillary Mucinous Neoplasm Lineages. Reviewed International journal

Hiroyuki Kato, Keisuke Tateishi, Hiroaki Fujiwara, Takuma Nakatsuka, Keisuke Yamamoto, Yotaro Kudo, Yoku Hayakawa, Hayato Nakagawa, Yasuo Tanaka, Hideaki Ijichi, Motoyuki Otsuka, Dosuke Iwadate, Hiroki Oyama, Sachiko Kanai, Kensaku Noguchi, Tatsunori Suzuki, Tatsuya Sato, Ryunosuke Hakuta, Kazunaga Ishigaki, Kei Saito, Tomotaka Saito, Naminatsu Takahara, Takahiro Kishikawa, Tsuyoshi Hamada, Ryota Takahashi, Koji Miyabayashi, Suguru Mizuno, Hirofumi Kogure, Yousuke Nakai, Yoshihiro Hirata, Atsushi Toyoda, Kazuki Ichikawa, Wei Qu, Shinichi Morishita, Junichi Arita, Mariko Tanaka, Tetsuo Ushiku, Kiyoshi Hasegawa, Mitsuhiro Fujishiro, Kazuhiko Koike

Gastroenterology 162 ( 4 ) 1272 - 1287 2022.4

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND & AIMS: Chromatin architecture governs cell lineages by regulating the specific gene expression; however, its role in the diversity of cancer development remains unknown. Among pancreatic cancers, pancreatic ductal adenocarcinoma (PDAC) and intraductal papillary mucinous neoplasms (IPMN) with an associated invasive carcinoma (IPMNinv) arise from 2 distinct precursors, and their fundamental differences remain obscure. Here, we aimed to assess the difference of chromatin architecture regulating the transcriptional signatures or biological features in pancreatic cancers. METHODS: We established 28 human organoids from distinct subtypes of pancreatic tumors, including IPMN, IPMNinv, and PDAC. We performed exome sequencing (seq), RNA-seq, assay for transposase-accessible chromatin-seq, chromatin immunoprecipitation-seq, high-throughput chromosome conformation capture, and phenotypic analyses with short hairpin RNA or clustered regularly interspaced short palindromic repeats interference. RESULTS: Established organoids successfully reproduced the histology of primary tumors. IPMN and IPMNinv organoids harbored GNAS, RNF43, or KLF4 mutations and showed the distinct expression profiles compared with PDAC. Chromatin accessibility profiles revealed the gain of stomach-specific open regions in IPMN and the pattern of diverse gastrointestinal tissues in IPMNinv. In contrast, PDAC presented an impressive loss of accessible regions compared with normal pancreatic ducts. Transcription factor footprint analysis and functional assays identified that MNX1 and HNF1B were biologically indispensable for IPMN lineages. The upregulation of MNX1 was specifically marked in the human IPMN lineage tissues. The MNX1-HNF1B axis governed a set of genes, including MYC, SOX9, and OLFM4, which are known to be essential for gastrointestinal stem cells. High-throughput chromosome conformation capture analysis suggested the HNF1B target genes to be 3-dimensionally connected in the genome of IPMNinv. CONCLUSIONS: Our organoid analyses identified the MNX1-HNF1B axis to be biologically significant in IPMN lineages.

DOI： 10.1053/j.gastro.2021.12.254

WWP1 inactivation enhances efficacy of PI3K inhibitors while suppressing their toxicities in breast cancer models. Reviewed International journal

Takahiro Kishikawa, Hiroshi Higuchi, Limei Wang, Nivedita Panch, Valerie Maymi, Sachem Best, Samuel Lee, Genso Notoya, Alex Toker, Lydia E Matesic, Gerburg M Wulf, Wenyi Wei, Motoyuki Otsuka, Kazuhiko Koike, John G Clohessy, Yu-Ru Lee, Pier Paolo Pandolfi

The Journal of clinical investigation 131 ( 24 ) 2021.12

Language：English Publishing type：Research paper (scientific journal)

Activation of the phosphatidylinositol 3-kinase (PI3K) signaling pathway is a pervasive event in tumorigenesis due to PI3K mutation and dysfunction of phosphatase and tensin homolog deleted on chromosome 10 (PTEN). Pharmacological inhibition of PI3K has resulted in variable clinical outcomes, however, raising questions regarding the possible mechanisms of unresponsiveness and resistance to treatment. WWP1 is an oncogenic HECT-type ubiquitin E3 ligase frequently amplified and mutated in multiple cancers, as well as in the germ lines of patients predisposed to cancer, and was recently found to activate PI3K signaling through PTEN inactivation. Here, we demonstrate that PTEN dissociated from the plasma membrane upon treatment with PI3K inhibitors through WWP1 activation, whereas WWP1 genetic or pharmacological inhibition restored PTEN membrane localization, synergizing with PI3K inhibitors to suppress tumor growth both in vitro and in vivo. Furthermore, we demonstrate that WWP1 inhibition attenuated hyperglycemia and the consequent insulin feedback, which is a major tumor-promoting side effect of PI3K inhibitors. Mechanistically, we found that AMPKα2 was ubiquitinated and, in turn, inhibited in its activatory phosphorylation by WWP1, whereas WWP1 inhibition facilitated AMPKα2 activity in the muscle to compensate for the reduction in glucose uptake observed upon PI3K inhibition. Thus, our identification of the cell-autonomous and systemic roles of WWP1 inhibition expands the therapeutic potential of PI3K inhibitors and reveals new avenues of combination cancer therapy.

DOI： 10.1172/JCI140436

HBx-induced degradation of Smc5/6 complex impairs homologous recombination-mediated repair of damaged DNA. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Kazuyoshi Funato, Yu Miyakawa, Eri Tanaka, Takahiro Seimiya, Mari Yamagami, Takeya Tsutsumi, Kazuya Okushin, Kei Miyakawa, Akihide Ryo, Kazuhiko Koike

Journal of hepatology 76 ( 1 ) 53 - 62 2021.8

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND & AIMS: Hepatitis B virus (HBV) causes hepatocellular carcinoma (HCC). While the need for HBV regulatory protein X (HBx) for viral transcription via impairment of the structural maintenance of chromosome 5/6 (Smc5/6) complex was recently demonstrated, HBx is also a potent driver of HCC. However, the mechanism by which HBx expression induces hepatocarcinogenesis is unclear. METHODS: Degradation of Smc5/6 complex and accumulation of DNA damage were observed in both in vivo and in vitro HBV infection models. Rescue experiments were performed using nitazoxanide (NTZ), which inhibits degradation of the Smc5/6 complex by HBx. RESULTS: Degradation of the Smc5/6 complex triggered by HBx impaired homologous recombination (HR) repair of DNA double-strand breaks (DSBs), leading to cellular transformation. We found that DNA damage accumulated in the liver tissue of HBV-infected humanized chimeric mice, HBx-transgenic mice, and human tissues. HBx suppressed the HR repair of DSBs, including that induced by the clustered regularly interspaced short palindromic repeats-Cas9 system, in an Smc5/6-dependent manner, which was rescued by restoring the Smc5/6 complex. NTZ restored HR repair in, and colony formation by, HBx-expressing cells. CONCLUSIONS: Degradation of the Smc5/6 complex by HBx increases viral transcription and promotes cellular transformation by impairing HR repair of DSBs. LAY SUMMARY: HBV regulatory protein X (HBx) degrades the structural maintenance of chromosome 5/6 (Smc5/6) complex, which is required for viral replication. We found that degradation of the Smc5/6 complex also plays a key role in the accumulation of DNA damage, as well as in HBx-mediated tumorigenesis.

DOI： 10.1016/j.jhep.2021.08.010

Impact of Obesity and Heavy Alcohol Consumption on Hepatocellular Carcinoma Development after HCV Eradication with Antivirals. Reviewed International journal

Tatsuya Minami, Ryosuke Tateishi, Naoto Fujiwara, Ryo Nakagomi, Takuma Nakatsuka, Masaya Sato, Koji Uchino, Kenichiro Enooku, Hayato Nakagawa, Hidetaka Fujinaga, Masashi Izumiya, Kazuyuki Hanajiri, Yoshinari Asaoka, Yuji Kondo, Yasuo Tanaka, Motoyuki Otsuka, Takamasa Ohki, Masahiro Arai, Atsushi Tanaka, Kiyomi Yasuda, Hideaki Miura, Itsuro Ogata, Toshiro Kamoshida, Kazuaki Inoue, Yukihiro Koike, Masatoshi Akamatsu, Hiroshi Mitsui, Hajime Fujie, Keiji Ogura, Hideo Yoshida, Tomonori Wada, Kiyohiko Kurai, Hisato Maekawa, Shuntaro Obi, Takuma Teratani, Naohiko Masaki, Kayo Nagashima, Takashi Ishikawa, Naoya Kato, Kyoji Moriya, Hiroshi Yotsuyanagi, Kazuhiko Koike

Liver cancer 10 ( 4 ) 309 - 319 2021.7

Language：English Publishing type：Research paper (scientific journal)

Background and Aims: It remains unclear whether obesity increases the risk of hepatocellular carcinoma (HCC) in patients with chronic hepatitis C who achieved a sustained virological response (SVR) with antiviral therapy. Methods: In this multicenter cohort study, we enrolled patients with chronic hepatitis C who achieved SVR with interferon (IFN)-based therapy (IFN group) or direct-acting antiviral (DAA) therapy (DAA group) between January 1, 1990, and December 31, 2018. The patients underwent regular surveillance for HCC. Cumulative incidence of and the risk factors for HCC development after SVR were assessed using the Kaplan-Meier method and Cox proportional hazard regression analysis, respectively. Results: Among 2,055 patients (840 in the IFN group and 1,215 in the DAA group), 75 developed HCC (41 in the IFN group and 34 in the DAA group) during the mean observation period of 4.1 years. The incidence rates of HCC at 1, 2, and 3 years were 1.2, 1.9, and 3.0%, respectively. Multivariate analysis revealed that in addition to older age, lower albumin level, lower platelet count, higher alpha-fetoprotein level, and absence of dyslipidemia, obesity (body mass index ≥25 kg/m2) and heavy alcohol consumption (≥60 g/day) were independent risk factors for HCC development, with adjusted hazard ratio (HR) of 2.53 (95% confidence interval [CI]: 1.51-4.25) and 2.56 (95% CI: 1.14-5.75), respectively. The adjusted HR was not significant between the 2 groups (DAA vs. IFN; HR 1.19, 95% CI: 0.61-2.33). Conclusions: Obesity and heavy alcohol consumption increased the risk of HCC development after SVR.

DOI： 10.1159/000513705

Aberrant expression of a novel circular RNA in pancreatic cancer. Reviewed International journal

Takahiro Seimiya, Motoyuki Otsuka, Takuma Iwata, Eri Tanaka, Kazuma Sekiba, Chikako Shibata, Masaru Moriyama, Ryo Nakagawa, Reo Maruyama, Kazuhiko Koike

Journal of human genetics 66 ( 2 ) 181 - 191 2021.2

Language：English Publishing type：Research paper (scientific journal)

Circular RNAs (circRNAs) are single-stranded, covalently closed RNA molecules that are produced from pre-mRNAs through a process known as back-splicing. Although circRNAs are expressed under specific conditions, current understanding of their comprehensive expression status is still limited. Here, we performed a large-scale circRNA profiling analysis in human pancreatic ductal adenocarcinoma (PDAC) tissues, using circular RNA-specific RNA sequencing. We identified more than 40,000 previously unknown circRNAs, some of which were upregulated in PDAC tissues, compared with normal pancreatic tissues. We determined the full-length sequence of a circRNA upregulated in PDAC, which was derived from two noncoding RNA loci on chromosome 12. The novel circRNA, named circPDAC RNA, was not expressed in normal human cells, but was expressed in PDAC and other carcinoma cells. While postulated biological functions, such as peptide production from the circPDAC RNA, were not detected, its aberrant expression was confirmed in other PDAC tissues and in serum from a PDAC patient. These results demonstrate that comprehensive studies are necessary to reveal the expression status of circRNAs and that the circPDAC RNA identified here might serve as a novel biomarker for cancers, including PDAC.

DOI： 10.1038/s10038-020-00826-5

Should Level of HBV RNA be Used to Determine When Patients Should Stop Treatment With Nucleos(t)ide Analogues. Reviewed International journal

Motoyuki Otsuka, Kazuhiko Koike

Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association 18 ( 3 ) 551 - 552 2020.3

Pevonedistat, a Neuronal Precursor Cell-Expressed Developmentally Down-Regulated Protein 8-Activating Enzyme Inhibitor, Is a Potent Inhibitor of Hepatitis B Virus. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Motoko Ohno, Mari Yamagami, Takahiro Kishikawa, Takahiro Seimiya, Tatsunori Suzuki, Eri Tanaka, Rei Ishibashi, Kazuyoshi Funato, Kazuhiko Koike

Hepatology (Baltimore, Md.) 69 ( 5 ) 1903 - 1915 2019.5

Language：English Publishing type：Research paper (scientific journal)

Hepatitis B virus (HBV) infection is a major health concern worldwide. To prevent HBV-related mortality, elimination of viral proteins is considered the ultimate goal of HBV treatment; however, currently available nucleos(t)ide analogs rarely achieve this goal, as viral transcription from episomal viral covalently closed circular DNA (cccDNA) is not prevented. HBV regulatory protein X was recently found to target the protein structural maintenance of chromosomes 5/6 (Smc5/6) for ubiquitination and degradation by DDB1-CUL4-ROC1 E3 ligase, resulting in enhanced viral transcription from cccDNA. This ubiquitin-dependent proteasomal pathway requires an additional ubiquitin-like protein for activation, neuronal precursor cell-expressed developmentally down-regulated protein 8 (NEDD8). Here, we show that pevonedistat, a NEDD8-activating enzyme inhibitor, works efficiently as an antiviral agent. Pevonedistat significantly restored Smc5/6 protein levels and suppressed viral transcription and protein production in the HBV minicircle system in in vitro HBV replication models and in human primary hepatocytes infected naturally with HBV. Conclusion: These results indicate that pevonedistat is a promising compound to treat chronic HBV infection.

DOI： 10.1002/hep.30491

Repression of MicroRNA Function Mediates Inflammation-associated Colon Tumorigenesis Reviewed

Takeshi Yoshikawa, Jianfeng Wu, Motoyuki Otsuka, Takahiro Kishikawa, Nobumi Suzuki, Akemi Takata, Motoko Ohno, Rei Ishibashi, Mari Yamagami, Ryo Nakagawa, Naoya Kato, Masaaki Miyazawa, Jiahuai Han, Kazuhiko Koike

Gastroenterology 152 ( 3 ) 631 - 643 2017.2

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO-ELSEVIER INC

BACKGROUND & AIMS: Little is known about the mechanisms by which chronic inflammation contributes to carcinogenesis, such as the development of colon tumors in patients with inflammatory bowel diseases. Specific microRNA (miRNAs) can function as suppressors or oncogenes, and widespread alterations in miRNA expression have been associated with tumorigenesis. We studied whether alterations in miRNA function contribute to inflammation-associated colon carcinogenesis. METHODS: We studied the effects of inflammatory cytokines, such as tumor necrosis factor, interleukin-1 alpha (IL1A), and IL1 beta (IL1B), on miRNA function, measured by activity of reporter constructs containing miRNA-binding sites in their 30 untranslated regions, in human 293T embryonic kidney, Caco-2, HT29, and HCT116 colon carcinoma cells, as well as dicer(+/+) and dicer(-/-), and Apobec3(+/+) and Apobec3(-/-) mouse embryonic fibroblasts. Cells were analyzed by immunoblots, immunohistochemistry, and flow cytometry. We generated transgenic mice expressing reporter constructs regulated by LET7B, MIR122, and MIR29b response elements; some mice were given injections of miRNA inhibitors (anti-MIR122 or anti-LET7B), a negative control, or tumor necrosis factor. Liver tissues were collected and analyzed by immunoblotting. Reporter mice were given azoxymethane followed by dextran sulfate sodium to induce colitis and colon tumors; some mice were given the ROCK inhibitor fasudil along with these agents (ROCK inhibitors increase miRNA function). Colon tissues were collected and analyzed by immunohistochemistry, immunoblots, and fluorescence microscopy. RESULTS: Incubation of cell lines with inflammatory cytokines reduced the ability of miRNAs to down-regulate expression from reporter constructs; dicer was required for this effect, so these cytokines relieve miRNA-dependent reductions in expression. The cytokines promoted degradation of APOBEC3G, which normally promotes miRNA loading into argonaute 2-related complexes. Mice with colitis had reduced miRNA function, based on increased expression of reporter genes. Administration of fasudil to mice did not reduce the severity of colitis that developed but greatly reduced the numbers of colon tumors formed (mean 2 tumors/colon in mice given fasudil vs 9 tumors/colon in mice given control agent). We made similar observations in IL10-deficient mice. CONCLUSIONS: We found inflammatory cytokines to reduce the activities of miRNAs. In mice with colitis, activities of miRNAs are reduced; administration of an agent that increases miRNA function prevents colon tumor formation in these mice. This pathway might be targeted to prevent colon carcinogenesis in patients with inflammatory bowel diseases.

DOI： 10.1053/j.gastro.2016.10.043

Satellite RNAs promote pancreatic oncogenic processes via the dysfunction of YBX1 Reviewed

Takahiro Kishikawa, Motoyuki Otsuka, Takeshi Yoshikawa, Motoko Ohno, Hideaki Ijichi, Kazuhiko Koike

Nature Communications 7 ( 1 ) 13006 2016.12

Language：English Publishing type：Research paper (scientific journal) Publisher：Springer Nature

Highly repetitive tandem arrays at the centromeric and pericentromeric regions in chromosomes, previously considered silent, are actively transcribed, particularly in cancer. This aberrant expression occurs even in K-ras-mutated pancreatic intraepithelial neoplasia (PanIN) tissues, which are precancerous lesions. To examine the biological roles of the satellite RNAs in carcinogenesis, we construct mouse PanIN-derived cells expressing major satellite (MajSAT) RNA and show increased malignant properties. We find an increase in frequency of chromosomal instability and point mutations in both genomic and mitochondrial DNA. We identify Y-box binding protein 1 (YBX1) as a protein that binds to MajSAT RNA. MajSAT RNA inhibits the nuclear translocation of YBX1 under stress conditions, thus reducing its DNA-damage repair function. The forced expression of YBX1 significantly decreases the aberrant phenotypes. These findings indicate that during the early stage of cancer development, satellite transcripts may act as 'intrinsic mutagens' by inducing YBX1 dysfunction, which may be crucial in oncogenic processes.

DOI： 10.1038/ncomms13006

Quantitation of circulating satellite RNAs in pancreatic cancer patients. Reviewed International journal

Takahiro Kishikawa, Motoyuki Otsuka, Takeshi Yoshikawa, Motoko Ohno, Keisuke Yamamoto, Natsuyo Yamamoto, Ai Kotani, Kazuhiko Koike

JCI insight 1 ( 8 ) e86646 2016.6

Language：English Publishing type：Research paper (scientific journal)

Pancreatic ductal adenocarcinoma (Pdac) is a malignancy with a poor prognosis due to difficulties in early detection. Although promising biomarkers are increasingly reported, such methods are not yet easy to apply clinically, mainly due to their low reproducibility or technical difficulties. In this study, we developed a convenient and sensitive method for quantifying aberrantly expressed satellite repeat RNAs in sera, which can be used to efficiently detect patients with Pdac. Here, we introduce a Tandem Repeat Amplification by nuclease Protection (TRAP) method combined with droplet digital PCR (ddPCR) to detect human satellite II (HSATII) RNAs, which are specifically expressed in human Pdacs at greater levels than normal tissues but are difficult to measure due to their repetitive sequences and irregularities. HSATII RNA core sequence levels in sera were significantly higher in Pdac patients compared with noncancer patients (median copy number: 14.75 and 3.17 per μl in the training set and 17.35 and 2.9 in the validation set, respectively). In addition, patients with intraductal papillary mucinous neoplasm (IPMN), a precancerous lesion of Pdac, could also be efficiently detected. This method can be routinely applied to screen patients with Pdac and high-risk patients, facilitating the development of preventive medicine for this disease.

DOI： 10.1172/jci.insight.86646

Regulation of NKT cell-mediated immune responses to tumours and liver inflammation by mitochondrial PGAM5-Drp1 signalling Reviewed

Young Jun Kang, Bo-Ram Bang, Kyung Ho Han, Lixin Hong, Eun-Jin Shim, Jianhui Ma, Richard A. Lerner, Motoyuki Otsuka

Nature Communications 6 ( 1 ) 8371 2015.12

The TNF Family Member 4-1BBL Sustains Inflammation by Interacting with TLR Signaling Components During Late-Phase Activation Reviewed

Jianhui Ma, Bo-Ram Bang, Jiawei Lu, So-Young Eun, Motoyuki Otsuka, Michael Croft, Peter Tobias, Jiahuai Han, Osamu Takeuchi, Shizuo Akira, Michael Karin, Hideo Yagita, Young Jun Kang

Science Signaling 6 ( 295 ) ra87 2013.10

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC ADVANCEMENT SCIENCE

Activation of Toll-like receptor (TLR)-dependent signaling leads to the expression of genes encoding proinflammatory factors, such as tumor necrosis factor-alpha (TNF-alpha), and this proinflammatory gene expression is sustained for the duration of the inflammatory response. TLR4-mediated inflammation, which occurs in two phases, depends on the TNF family member 4-1BB ligand (4-1BBL) to sustain TNF-alpha production during late-phase signaling. We showed that Toll-interleukin-1 receptor (TIR) domain-containing adaptor protein (TIRAP) and the kinase IRAK2 interacted with 4-1BBL to mediate late-phase TLR4 signaling. Expression of 4-1bbl depended on early TLR4 signaling that also induced Tnf expression, and 4-1BBL translocated to the plasma membrane, where it interacted with TLR4 to mediate late-phase signaling. TLR4-4-1BBL-mediated signaling depended on TIRAP and IRAK2, as well as a complex consisting of the E3 ubiquitin ligase TRAF6 (TNF receptor-associated factor 6), the kinase TAK1 (transforming growth factor-beta-activated kinase 1), and the adaptor protein TAB1 (TAK-binding protein 1). Inhibition of this late-phase pathway reduced the extent of TNF-alpha production by mouse macrophages exposed to the TLR4 ligand lipopolysaccharide (LPS) and ameliorated LPS-induced sepsis in mice. Together, these data suggest that TIRAP and IRAK2 are critical for the sustained inflammatory response that is mediated by late-phase signaling by the TLR-4-1BBL complex.

DOI： 10.1126/scisignal.2004431

Serum metabolomics reveals gamma-glutamyl dipeptides as biomarkers for discrimination among different forms of liver disease Reviewed

Tomoyoshi Soga, Masahiro Sugimoto, Masashi Honma, Masayo Mori, Kaori Igarashi, Kasumi Kashikura, Satsuki Ikeda, Akiyoshi Hirayama, Takehito Yamamoto, Haruhiko Yoshida, Motoyuki Otsuka, Shoji Tsuji, Yutaka Yatomi, Tadayuki Sakuragawa, Hisayoshi Watanabe, Kouei Nihei, Takafumi Saito, Sumio Kawata, Hiroshi Suzuki, Masaru Tomita, Makoto Suematsu

Journal of Hepatology 55 ( 4 ) 896 - 905 2011.10

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Background & Aims: We applied a metabolome profiling approach to serum samples obtained from patients with different liver diseases, to discover noninvasive and reliable biomarkers for rapid-screening diagnosis of liver diseases.
Methods: Using capillary electrophoresis and liquid chromatography mass spectrometry, we analyzed low molecular weight metabolites in a total of 248 serum samples obtained from patients with nine types of liver disease and healthy controls.
Results: We found that gamma-glutamyl dipeptides, which were bio-synthesized through a reaction with gamma-glutamylcysteine synthetase, were indicative of the production of reduced glutathione, and that measurement of their levels could distinguish among different liver diseases. Multiple logistic regression models facilitated the discrimination between specific and other liver diseases and yielded high areas under receiver-operating characteristic curves. The area under the curve values in training and independent validation data were 0.952 and 0.967 in healthy controls, 0.817 and 0.849 in drug-induced liver injury, 0.754 and 0.763 in asymptomatic hepatitis B virus infection, 0.820 and 0.762 in chronic hepatitis B, 0.972 and 0.895 in hepatitis C with persistently normal alanine transaminase, 0.917 and 0.707 in chronic hepatitis C, 0.803 and 0.993 in cirrhosis type C, and 0.762 and 0.803 in hepatocellular carcinoma, respectively. Several gamma-glutamyl dipeptides also manifested potential for differentiating between nonalcoholic steatohepatitis and simple steatosis.
Conclusions: gamma-Glutamyl dipeptides are novel biomarkers for liver diseases, and varying levels of individual or groups of these peptides have the power to discriminate among different forms of hepatic disease. (C) 2011 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jhep.2011.01.031

MicroRNA122 is a key regulator of alpha-fetoprotein expression and influences the aggressiveness of hepatocellular carcinoma Reviewed

Kentaro Kojima, Akemi Takata, Charles Vadnais, Motoyuki Otsuka, Takeshi Yoshikawa, Masao Akanuma, Yuji Kondo, Young Jun Kang, Takahiro Kishikawa, Naoya Kato, Zhifang Xie, Weiping J. Zhang, Haruhiko Yoshida, Masao Omata, Alain Nepveu, Kazuhiko Koike

Nature Communications 2 338 2011.6

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

alpha-fetoprotein (AFP) is not only a widely used biomarker in hepatocellular carcinoma (HCC) surveillance, but is also clinically recognized as linked with aggressive tumour behaviour. Here we show that deregulation of microRNA122, a liver-specific microRNA, is a cause of both AFP elevation and a more biologically aggressive phenotype in HCC. We identify CUX1, a direct target of microRNA122, as a common central mediator of these two effects. Using liver tissues from transgenic mice in which microRNA122 is functionally silenced, an orthotopic xenograft tumour model, and human clinical samples, we further demonstrate that a microRNA122/CUX1/microRNA214/ZBTB20 pathway regulates AFP expression. We also show that the microRNA122/CUX1/RhoA pathway regulates the aggressive characteristics of tumours. We conclude that microRNA122 and associated signalling proteins may represent viable therapeutic targets, and that serum AFP levels in HCC patients may be a surrogate marker for deregulated intracellular microRNA122 signalling pathways in HCC tissues.

DOI： 10.1038/ncomms1345

Genome-wide association study identifies a susceptibility locus for HCV-induced hepatocellular carcinoma Reviewed

Vinod Kumar, Naoya Kato, Yuji Urabe, Atsushi Takahashi, Ryosuke Muroyama, Naoya Hosono, Motoyuki Otsuka, Ryosuke Tateishi, Masao Omata, Hidewaki Nakagawa, Kazuhiko Koike, Naoyuki Kamatani, Michiaki Kubo, Yusuke Nakamura, Koichi Matsuda

Nature Genetics 43 ( 5 ) 455 - + 2011.5

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

To identify the genetic susceptibility factor(s) for hepatitis C virus-induced hepatocellular carcinoma (HCV-induced HCC), we conducted a genome-wide association study using 432,703 autosomal SNPs in 721 individuals with HCV-induced HCC (cases) and 2,890 HCV-negative controls of Japanese origin. Eight SNPs that showed possible association (P < 1 x 10(-5)) in the genome-wide association study were further genotyped in 673 cases and 2,596 controls. We found a previously unidentified locus in the 5' flanking region of MICA on 6p21.33 (rs2596542, P-combined = 4.21 x 10(-13), odds ratio = 1.39) to be strongly associated with HCV-induced HCC. Subsequent analyses using individuals with chronic hepatitis C (CHC) indicated that this SNP is not associated with CHC susceptibility (P = 0.61) but is significantly associated with progression from CHC to HCC (P = 3.13 x 10(-8)). We also found that the risk allele of rs2596542 was associated with lower soluble MICA protein levels in individuals with HCV-induced HCC (P = 1.38 x 10(-13)).

DOI： 10.1038/ng.809

Distinct Effects of p38 alpha Deletion in Myeloid Lineage and Gut Epithelia in Mouse Models of Inflammatory Bowel Disease Reviewed

Motoyuki Otsuka, Young Jun Kang, Jianlin Ren, Huiping Jiang, Yinbin Wang, Masao Omata, Jiahuai Han

Gastroenterology 138 ( 4 ) 1255 - U83 2010.4

Impaired microRNA processing causes corpus luteum insufficiency and infertility in mice Reviewed

Motoyuki Otsuka, Min Zheng, Masaaki Hayashi, Jing-Dwan Lee, Osamu Yoshino, Shengcai Lin, Jiahuai Han

Journal of Clinical Investigation 118 ( 5 ) 1944 - 1954 2008.5

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC CLINICAL INVESTIGATION INC

The microRNA (miRNA) processing enzyme Dicer1 is required for zygotic and embryonic development, but the early embryonic lethality of Dicer1 null alleles in mice has limited our ability to address the role of Dicer1 in normal mouse growth and development. To address this question, we used a mouse mutant with a hypomorphic Dicer1 allele (Dicer(d/d)) and found that Dicer1 deficiency resulted in female infertility. This defect in female Dicer(d/d) mice was caused by corpus luteum (CL) insufficiency and resulted, at least in part, from the impaired growth of new capillary vessels in the ovary. We found that the impaired CL angiogenesis in Dicer(d/d) mice was associated with a lack of miR17-5p and let7b, 2 miRNAs that participate in angiogenesis by regulating the expression of the antiangiogenic factor tissue inhibitor of metalloproteinase 1. Furthermore, injection of miR17-5p and let7b into the ovaries of Dicer(d/d) mice partially normalized tissue inhibitor of metalloproteinase 1 expression and CL angiogenesis. Our data indicate that the development and function of the ovarian CL is a physiological process that appears to be regulated by miRNAs and requires Dicer1 function.

DOI： 10.1172/JCI33680

Hypersusceptibility to vesicular stomatitis virus infection in Dicer1-deficient mice is due to impaired miR24 and miR93 expression Reviewed

Motoyuki Otsuka, Qing Jing, Philippe Georgel, Liguo New, Jianming Chen, Johann Mols, Young Jun Kang, Zhengfan Jiang, Xin Du, Ryan Cook, Subash C. Das, Asit K. Pattnaik, Bruce Beutler, Jiahuai Han

Immunity 27 ( 1 ) 123 - 134 2007.7

Language：English Publishing type：Research paper (scientific journal) Publisher：CELL PRESS

Dicer is essential for plant, Caenorhabditis elegans, and Drosophila antiviral responses because of its role in generating small interfering RNA (siRNA) from viral genomes. We show that because of impaired miRNA production, mice with a variant Dicer1 allele (Dicer1(d/d)) were more susceptible to vesicular stomatitis virus (VSV) infection. We did not detect VSV genome-derived siRNA in wild-type cells or any alteration of interferon-mediated antiviral responses by Dicer1 deficiency. Rather, we found that host miR24 and miR93 could target viral large protein (L protein) and phosphoprotein (P protein) genes, and a lack of miR24 and miR93 was responsible for increased VSV replication in Dicer1(d/d) cells. Our data suggest that host miRNA can play a role in host interactions with viruses.

DOI： 10.1016/j.immuni.2007.05.014

Cell surface 4-1BBL mediates sequential signaling pathways 'downstream' of TLR and is required for sustained TNF production in macrophages Reviewed

Young Jun Kang, Sung Ouk Kim, Shigeki Shimada, Motoyuki Otsuka, Alim Seit-Nebi, Byoung S. Kwon, Tania H. Watts, Jiahuai Han

Nature Immunology 8 ( 6 ) 601 - 609 2007.6

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

The stimulation of Toll-like receptors (TLRs) on macrophages triggers production of the cytokine tumor necrosis factor (TNF). TNF production occurs within 1 h of TLR stimulation and is sustained for 1 d. Here we document a function for the TNF family member 4-1BB ligand (4-1BBL) in sustaining TLR-induced TNF production. TLR signaling induced 4-1BBL, and 4-1BBL interacted with TLRs on the macrophage surface. The influence of 4-1BBL on TNF production was independent of its receptor (4-1BB) and did not require the adaptors MyD88 or TRIF. It did not influence TLR4-induced activation of transcription factor NF-kappa B (an early response) but was required for TLR4-induced activation of transcription factors CREB and C/EBP ( a late event). Transient TLR4-MyD88 complexes appeared during the first hour after lipopolysaccharide stimulation, and TLR4-4-1BBL interactions were detected between 2 h and 8 h after lipopolysaccharide stimulation. Our results indicate that two different TLR4 complexes sequentially form and selectively control early and late TNF production.

DOI： 10.1038/ni1471

Acetic Acid-Indigo Carmine Chromocolonoscopy for Proximal Serrated Lesions: A Randomized, Three-Arm Colonoscopy Study. International journal

Hideaki Kinugasa, Sakiko Hiraoka, Sayo Kobayashi, Minoru Matsubara, Teruya Nagahara, Reiji Higashi, Kensuke Takei, Masayasu Ohmori, Takashi Nakamura, Takao Tsuzuki, Shouichi Tanaka, Ryosuke Hirai, Junki Toyosawa, Yuki Aoyama, Yasushi Yamasaki, Toshihiro Inokuchi, Masahiro Takahara, Takehiro Tanaka, Toshiharu Mitsuhashi, Motoyuki Otsuka

The American journal of gastroenterology 2025.3

Language：English Publishing type：Research paper (scientific journal)

OBJECTIVES: Aggressive colorectal cancer (CRC) frequently originates from serrated lesions (SLs), particularly in the proximal colon, which are challenging to detect using standard screening colonoscopy. Although duplicate examinations or chromocolonoscopies are recommended for detecting proximal SLs, evidence from randomized trials is limited. We evaluated the effectiveness of tandem colonoscopy with an acetic acid-indigo carmine mixture (AIM) for detecting SLs in the proximal colon compared to white-light imaging (WLI) and indigo carmine (IC). METHODS: This three-arm, multicenter, randomized controlled trial involving nine institutions enrolled patients undergoing colonoscopy and assigned them randomly to the WLI, IC, or AIM group. The primary outcomes were the SL-detection rate (SDR) of proximal lesions during the second examination (SDR2nd) and SL additional rate (SAR). Secondary outcomes included the detection and additional rates of other polyps, factors contributing to SAR, and complications. RESULTS: Between 2021 and 2024, 1,319 participants with 1,267 polyps were included in the analysis. With AIM, the SDR2nd and SAR were significantly higher compared to WLI or IC (WLI vs. AIM: 2.7% vs. 14.0%, p<.001; IC vs. AIM: 7.9% vs. 14.0%, p=.002, and WLI vs. AIM: 22.4% vs. 69.3%, p<.001; IC vs. AIM: 45.8% vs. 69.3%, p=.001). AIM conferred a higher adenoma-detection rate (ADR)2nd than with WLI (10.5% vs. 24.7%; p<.001) and was an independent factor for SAR (odds ratio [95% confidence interval]: 7.79 [3.76-17.08]). No major adverse events were observed. CONCLUSIONS: AIM significantly improved proximal colon SDRs and outperformed WLI and IC. The relationship between SDR and CRC incidence warrants further investigation.

DOI： 10.14309/ajg.0000000000003411

Unraveling a Rare Case: Diarrhea, Alopecia, and Polyposis. International journal

Hidetaka Mizushima, Hideaki Kinugasa, Motoyuki Otsuka

Gastroenterology 2025.2

Changes of leucine-rich alpha 2 glycoprotein could be a marker of changes of endoscopic and histologic activity of ulcerative colitis. International journal

Yuki Aoyama, Sakiko Hiraoka, Eriko Yasutomi, Toshihiro Inokuchi, Takehiro Tanaka, Kensuke Takei, Shoko Igawa, Keiko Takeuchi, Masahiro Takahara, Junki Toyosawa, Yasushi Yamasaki, Hideaki Kinugasa, Jun Kato, Hiroyuki Okada, Motoyuki Otsuka

Scientific reports 15 ( 1 ) 5248 - 5248 2025.2

Language：English Publishing type：Research paper (scientific journal)

Leucine-rich alpha 2 glycoprotein (LRG) is one of the serum biomarkers for disease activity of ulcerative colitis (UC). We focused on the correlation between the changes of LRG and the changes of endoscopic and histologic activity of UC, in comparison to the changes of fecal calprotectin (Fcal), fecal immunochemical test (FIT), and C-reactive protein (CRP). Seventy-nine patients with two or more colonoscopies were enrolled, and 123 paired colonoscopies and 121 paired biopsies were examined. With regard to the change of endoscopic/histologic activity between the preceding and subsequent colonoscopy, there was improvement (n = 29/45), unchanging (n = 63/36), and worsening (n = 31/40). The correlations between the changes of marker levels and endoscopic/histologic activity were Fcal; r = 0.50/0.39 and FIT; r = 0.41/0.40, LRG; r = 0.42/0.40 and CRP; r = 0.22/0.17. Furthermore, when the correlation between the changes of LRG levels and the changes of endoscopic/histological activity was compared with those of other markers, the correlation of LRG tended to be superior to those of CRP (CRP vs. LRG; p = 0.08/0.01). LRG is equivalent to fecal markers and superior to CRP, when inferring changes in disease activity of UC based on changes in its level.

DOI： 10.1038/s41598-025-89615-8

Durable Response to Nivolumab Combined With Metformin in Advanced Pancreatic Cancer: A Case Report With Seven Years of Follow-Up. International journal

Ryosuke Sato, Katsuyuki Hotta, Toshio Kubo, Shigeru Horiguchi, Hironari Kato, Kazuyuki Matsumoto, Toshiyuki Kozuki, Heiichiro Udono, Katsuyuki Kiura, Motoyuki Otsuka

Cureus 17 ( 2 ) e79001 2025.2

Language：English Publishing type：Research paper (scientific journal)

We report a case of poorly differentiated pancreatic cancer that showed an exceptional response to combination therapy with nivolumab and metformin. A 58-year-old man presented with epigastric pain and was diagnosed with locally advanced pancreatic cancer with para-aortic lymph node metastasis. After disease progression following modified FOLFIRINOX therapy (a combination of fluorouracil, leucovorin, irinotecan, and oxaliplatin), the patient was enrolled in a phase Ib clinical trial of nivolumab (3 mg/kg biweekly) combined with metformin (750 mg/day). Post-treatment imaging showed marked tumor shrinkage with normalization of the tumor markers. During treatment, the patient was diagnosed with early-stage lung cancer and underwent successful left S1+S2 segmentectomy with temporary suspension of immunotherapy. The therapeutic response of pancreatic cancer has been sustained for seven years, with minimal residual disease. This unprecedented response duration is particularly noteworthy considering his microsatellite stability, which typically predicts a limited response to immune checkpoint inhibition. This case demonstrates an exceptional response to nivolumab and metformin combination therapy in poorly differentiated pancreatic cancer. The remarkable durability of the response suggests the need for further investigation to identify patients most likely to benefit from this therapeutic approach.

DOI： 10.7759/cureus.79001

WWP1 inhibition suppresses the proliferation of pancreatic cancer cells by regulating the PI3K-AKT pathway.

Genso Notoya, Takahiro Kishikawa, Kengo Yasugi, Takuma Iwata, Takahiro Seimiya, Koji Miyabayashi, Ryota Takahashi, Keisuke Yamamoto, Hideaki Ijichi, Motoyuki Otsuka, Mitsuhiro Fujishiro

Journal of gastroenterology 2024.12

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The proto-oncogene WWP1 is overexpressed in various cancers and contributes to tumor growth and poor prognosis. Recently, WWP1 inhibition was reported to suppress tumor development and cell proliferation by activating the PTEN function. However, the expression profiles and clinical significance of WWP1 in pancreatic ductal adenocarcinoma (PDAC) tissues remain undetermined. Therefore, this study aimed to evaluate the WWP1 expression in PDAC and investigate the therapeutic potential of WWP1 inhibition. METHODS: Cellular proliferation assays were performed using a doxycycline-inducible shWWP1 expression system. Transcriptome analyses were conducted to identify the altered pathways in WWP1-depleted cells. PTEN ubiquitination by WWP1 was confirmed using immunoprecipitation assays. In vivo xenograft and drug screening assays were performed to evaluate the clinical significance of WWP1 inhibition. RESULTS: WWP1 was significantly upregulated in PDAC tissues and associated with poor prognosis. WWP1 depletion significantly reduced the proliferation of PDAC cell lines, correlating with the suppression of the PI3K-AKT pathway. Mechanistically, as reported in other cancer types, PTEN is a target of WWP1 in PDAC cells. PTEN silencing abrogated the growth-inhibitory effects in WWP1-depleted cells, suggesting that the anti-tumor effects of WWP1 inhibition are mediated through PTEN activation. In vivo xenograft studies confirmed that WWP1 depletion substantially inhibited tumor growth. Moreover, drug screening assays revealed that WWP1 depletion had an additive effect with the PI3K-AKT pathway inhibitors on hindering tumor growth. CONCLUSION: WWP1 inhibition enhances the anti-tumor effects of PI3K-AKT pathway inhibitors through PTEN activation. Thus, WWP1 could be a potential therapeutic target in PDAC.

DOI： 10.1007/s00535-024-02192-x

Analysis of painful situations during unsedated esophagogastroduodenoscopy. International journal

Hiromitsu Kanzaki, Sakiko Kuraoka, Takuya Satomi, Shotaro Okanoue, Kenta Hamada, Yoshiyasu Kono, Masaya Iwamuro, Seiji Kawano, Yoshiro Kawahara, Hiroyuki Okada, Motoyuki Otsuka

Endoscopy international open 12 ( 11 ) E1267-E1276 2024.11

Language：English Publishing type：Research paper (scientific journal)

Background and study aims Although esophagogastroduodenoscopy (EGD) is a widely used technique, the procedure is often associated with discomfort. This study aimed to analyze painful situations, their frequency, and factors associated with patient discomfort during EGD. Patients and methods This prospective observational study included patients scheduled to undergo EGD. Seven endoscopists recruited patients scheduled for EGD screening or surveillance. Each endoscopist enrolled 20 patients, performing 10 EGD procedures using ultraslim endoscopes and 10 with standard-sized endoscopes. Data regarding painful situations and frequency were collected using specialized buttons pressed by the patients during EGD. A survey about overall discomfort was conducted after the procedure. Results We analyzed data from 140 patients. Esophageal insertion and duodenal observation were associated with the highest incidence of pressing the pain button, accounting for 59.3% and 40.7% of the cases, respectively. The factor associated with pressing the pain button during esophageal insertion was endoscopist experience (< 10 years). In contrast, younger age and female sex were the factors associated with pressing the pain button during duodenal observation. In the post-procedure survey, 63.6% of patients reported discomfort. Factors associated with patient discomfort included pressing the pain button during esophageal insertion (odds ratio [OR]: 2.84, P = 0.01) and previous painful EGD experience (OR: 2.41, P = 0.03). Concusions This study provides objective data on painful situations, their frequency, and related factors during EGD. Further research and interventions focusing on pain reduction during endoscopic procedures are warranted. The results of this study will help endoscopists manage painful situations and potentially improve skills.

DOI： 10.1055/a-2401-6804

Efficacy and Safety of Endoscopic Ultrasonography-Guided Ethanol Injections of Small Pancreatic Neuroendocrine Neoplasms: a prospective, multicenter study. International journal

Kazuyuki Matsumoto, Hironari Kato, Takao Itoi, Masayuki Kitano, Kazuo Hara, Masaki Kuwatani, Mamoru Takenaka, Reiko Ashida, Shuntaro Mukai, Nozomi Okuno, Kazumichi Kawakubo, Tatsuhiro Yamazaki, Jun Sakurai, Yuki Nakatsuka, Michihiro Yoshida, Motoyuki Otsuka

Endoscopy 2024.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND STUDY AIMS: Endoscopic ultrasonography (EUS)-guided ethanol injection (EI) has recently been introduced as one of the management strategies for pancreatic neuroendocrine neoplasms (PNENs). However, its role as a surgical alternative is unclear. We evaluated the efficacy and safety of EUS-EI in treating small PNENs through a prospective multicentre study. PATIENTS AND METHODS: Patients with Grade 1 tumours of ≤15 mm confirmed by pathology were included. The primary endpoint assessed efficacy and safety, measuring complete ablation using computed tomography at 1 and 6 months, prevention of adverse events (AEs) within 1 month, severe pancreatic fistula at 1 month, and diabetes mellitus (DM) incidence/worsening at 6 months. The composite endpoint of EUS-EI was compared with that of historical results of a study based on surgical treatment. RESULTS: Twenty-five patients with PNENs, with a median tumour size of 10.1 mm, were treated using EUS-EI. Seventy-six percent of the patients achieved the composite primary endpoint (19/25) (95% confidence interval [CI]=54.9%-90.6%), a proportion significantly higher than that of surgical treatment (P=0.0083). Regarding efficacy, 88% (22/25) of the patients achieved complete ablation at 1 and 6 months (95% CI=68.8%-97.5%). Regarding safety, 96% (24/25) of the patients had no severe AEs within 1 month (95% CI=79.7%-99.9%). No patients had severe pancreatic fistulas at 1 month, and 84% (21/25) of the patients had no incidence or exacerbation, or both, of DM at 6 months (95% CI=63.9%-95.5%). CONCLUSION: EUS-EI is safe and could be a potent treatment option for patients with small PNENs.

DOI： 10.1055/a-2452-4607

増大号炎症性腸疾患2024 主題炎症性腸疾患の内科治療基本治療薬(5-ASA製剤,副腎皮質ステロイド,チオプリン製剤)

平岡佐規子, 石黒美佳子, 豊澤惇希, 青山祐樹, 井川翔子, 竹内桂子, 山崎泰史, 井口俊博, 衣笠秀明, 髙原政宏, 大塚基之, 金谷信彦, 近藤喜太, 田中健大

胃と腸 59 ( 10 ) 1453 - 1461 2024.10

Short- and longer-term learning effects from virtual scale endoscopy videos: a useful tool for colorectal lesion size estimation (with video). International journal

Ryosuke Hirai, Hideaki Kinugasa, Mikako Ishiguro, Junki Toyosawa, Yuki Aoyama, Shoko Igawa, Yasushi Yamasaki, Toshihiro Inokuchi, Masahiro Takahara, Seiji Kawano, Sakiko Hiraoka, Motoyuki Otsuka

Gastrointestinal endoscopy 2024.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: Accurate assessment of colorectal polyp size is crucial for determining treatment and surveillance policies. However, visual estimation of lesion diameter is often inaccurate, making simple and effective educational tools essential. We aimed to evaluate the learning effects of virtual scale endoscopy (VSE). METHODS: Thirty-three endoscopists first watched pre-learning videos for SET1. They then estimated the diameters of 20 lesions and referred to instructional videos with VSE for self-study. Subsequently, they watched the post-learning videos for SET2 and estimated the lesion diameters. The error between the estimated and correct lesion sizes of both sets was compared. To evaluate longer-term learning effects, participants answered SET3 and SET4, which consisted of the same questions as SET2 and SET1, respectively, 2-3 months later without watching the instructional video for SET2. RESULTS: The error in the participants' estimation of the correct lesion diameter improved from SET1 to SET2 (34.7 mm ± 6.6 vs. 30.7 mm ± 7.7; p = 0.048), with a significant learning effect and error improvement specifically among non-experts (35.2 mm ± 5.3 vs. 30 mm ± 6.8; p = 0.028). In the SET3 and SET4, participants' errors indicated that the learning effect was well maintained (SET2 vs. SET3; 30.7 mm ± 7.7 vs. 28.6 mm ± 7.2; p = 0.1, and SET1 and SET4: 34.7 mm ± 6.6 vs. 31.7 mm ± 7.1; p = 0.025). CONCLUSIONS: VSE videos are a valuable learning tool for estimating lesion diameter, particularly for novice endoscopists, both in the short- and longer-term.

DOI： 10.1016/j.gie.2024.10.038

Characteristics of diabetes mellitus patients with nonviral chronic liver disease who developed hepatocellular carcinoma. International journal

Kyo Sasaki, Miwa Kawanaka, Yasuyuki Tomiyama, Akinobu Takaki, Motoyuki Otsuka, Fusao Ikeda, Naoko Yoshioka, Hideaki Kaneto, Jun Wada, Tetsuya Fukuda, Keisuke Hino, Sohji Nishina

Hepatology research : the official journal of the Japan Society of Hepatology 2024.10

Language：English Publishing type：Research paper (scientific journal)

AIM: Type 2 diabetes mellitus (T2DM) is a well-known risk factor for hepatocellular carcinoma (HCC). However, HCC is often diagnosed at an advanced stage in patients with diabetes because of the lack of the best criteria for surveillance candidates. The aim of this study was to identify risk factors for HCC development in patients with diabetes with nonviral chronic liver disease. METHOD: Three hundred thirty T2DM patients with nonviral chronic liver disease who underwent surveillance for HCC by imaging techniques between 2009 and 2020 were enrolled in this multicenter cross-sectional retrospective study. The clinical and laboratory parameters of patients with and without HCC were compared. RESULTS: Age ≥65 years, alcohol intake, lack of hepatic steatosis, triglyceride level <111 mg/dL, Mac2 binding protein glycosylation isomer (M2BPGi) ≥0.9 cut-off index (COI), α-fetoprotein concentration ≥5 ng/mL, and des-γ-carboxy prothrombin concentration ≥26 mAU/mL were independently associated with HCC development. When stratified by age, only alcohol intake (odds ratio [OR] 114.19, p < 0.001) was associated with HCC development in patients aged <65 years, and medication for diabetes mellitus (OR 5.72, p = 0.001), lack of hepatic steatosis (OR 4.47, p = 0.002), lactate dehydrogenase ≥198 IU/L (OR 2.751, p = 0.031), M2BPGi ≥1.18 COI (OR 9.05, p < 0.001), and FIB-4 index ≥2.59 (OR 3.22, p = 0.017) were associated with HCC development in patients aged ≥65 years. CONCLUSIONS: In addition to age and advanced liver fibrosis, alcohol intake in younger T2DM patients and medication for DM and lack of hepatic steatosis in older T2DM patients should be considered for HCC surveillance by imaging.

DOI： 10.1111/hepr.14124

Clinical Significance of Prior Ramucirumab Use on the Effectiveness of Nivolumab as the Third-Line Regimen in Gastric Cancer: A Multicenter Retrospective Study. International journal

Yuka Obayashi, Shoichiro Hirata, Yoshiyasu Kono, Makoto Abe, Koji Miyahara, Masahiro Nakagawa, Michihiro Ishida, Yasuhiro Choda, Kenta Hamada, Masaya Iwamuro, Seiji Kawano, Yoshiro Kawahara, Motoyuki Otsuka

Drugs - real world outcomes 2024.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND OBJECTIVE: Because vascular endothelial growth factor inhibition has been suggested to improve immune cell function in the cancer microenvironment, we examined whether using ramucirumab (RAM) before nivolumab usage is more effective in advanced gastric cancer. METHODS: This was a multicenter retrospective observational study. We analyzed patients who received nivolumab monotherapy as the third-line regimen for unresectable advanced or recurrent gastric cancer between October 2017 and December 2022. They were divided into the RAM (RAM-treated) group and the non-RAM (non-treated) group according to the RAM usage in the second-line regimen. The primary outcome was to compare the overall survival after nivolumab administration in the third-line regimen between the RAM and non-RAM groups. RESULTS: Fifty-two patients were included in the present study: 42 patients in the RAM group and ten patients in the non-RAM group. The median overall survival was significantly longer in the RAM group than in the non-RAM group (8.5 months vs 6.9 months, p < 0.05). In the RAM group, patients without peritoneal metastasis had significantly better median overall survival than those with peritoneal metastasis (23.8 months vs 7.7 months, p = 0.0033). Multivariate Cox-proportional hazards analyses showed that the presence of peritoneal metastasis (hazard ratio, 2.4; 95% confidence interval 1.0-5.7) alone was significantly associated with overall survival in the RAM group. CONCLUSIONS: The use of RAM prior to nivolumab monotherapy may contribute to prolonged survival in patients with gastric cancer, especially those without peritoneal metastasis.

DOI： 10.1007/s40801-024-00460-z

Long-term outcomes of endoscopic resection of superficial esophageal squamous cell carcinoma in late-elderly patients. International journal

Katsunori Matsueda, Seiji Kawano, Keisuke Fukui, Shoichiro Hirata, Takuya Satomi, Shoko Inoo, Kenta Hamada, Yoshiyasu Kono, Masaya Iwamuro, Yoshiro Kawahara, Motoyuki Otsuka

Journal of gastroenterology and hepatology 2024.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIM: As the population ages, the number of elderly patients with superficial esophageal squamous cell carcinoma (ESCC) is increasing. We aimed to clarify the indications for endoscopic resection (ER) in late-elderly patients with ESCC in terms of life expectancy. METHODS: Patients aged ≥75 years who underwent ER for ESCC at our institution from January 2005 to December 2018 were enrolled. Clinical data, including the Eastern Cooperative Oncology Group performance status, American Society of Anesthesiologists physical status (ASA-PS), Charlson comorbidity index, and prognostic nutritional index (PNI), were collected at the time of ER. The main outcome measure was overall survival (OS). RESULTS: Two hundred eight consecutive patients were enrolled. The patients' median age was 78 years (range, 75-89 years). The 5-year follow-up rate was 88.5% (median follow-up period, 6.6 years). The 5-year OS rate was 79.2% (95% confidence interval [CI], 72.2-84.8), and 5-year net survival standardized for age, sex, and calendar year was 1.04 (95% CI, 0.98-1.09). In the multivariate analysis, an ASA-PS of 3 (hazard ratio, 2.45; 95% CI, 1.16-5.17) and PNI of <44.0 (hazard ratio, 2.73; 95% CI, 1.38-5.40) were independent prognostic factors. When neither of these factors was met, the 5-year OS rate was 87.8% (95% CI, 80.0-92.9), and 5-year net survival was 1.08 (95% CI, 1.02-1.14). CONCLUSIONS: ER for ESCC in late-elderly patients may improve life expectancy. ER is recommended in patients with a good ASA-PS and PNI.

DOI： 10.1111/jgh.16764

Rates and risk factors of bleeding after gastric endoscopic submucosal dissection with continuous warfarin or 1-day withdrawal of direct oral anticoagulants. International journal

Shoichiro Hirata, Kenta Hamada, Masaya Iwamuro, Hirokazu Mouri, Koji Miyahara, Takao Tsuzuki, Kenji Yamauchi, Sayo Kobayashi, Sakuma Takahashi, Ryuta Takenaka, Shinichiro Hori, Masafumi Inoue, Tatsuya Toyokawa, Mamoru Nishimura, Shuhei Ishiyama, Jiro Miyaike, Ryo Kato, Minoru Matsubara, Naoko Yunoki, Hiromitsu Kanzaki, Yoshiro Kawahara, Hiroyuki Okada, Hideki Ishikawa, Motoyuki Otsuka

Journal of gastroenterology and hepatology 2024.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIM: The 2017 Japanese guidelines recommend continuing warfarin therapy during the perioperative period or discontinuing direct oral anticoagulants (DOACs) only on the day of endoscopic submucosal dissection for early gastric cancer. However, their safety has not been sufficiently explored. This study aimed to validate this management method. METHODS: This retrospective, multicenter study analyzed the characteristics and outcomes of patients who underwent gastric endoscopic submucosal dissection between July 2017 and June 2019. The patients were categorized according to the use of warfarin or DOACs. RESULTS: Among the 62 eligible patients, 53 (85%) were male (median age, 76 years). Warfarin was used in 10 patients (16%) and DOACs in 52 patients (84%). Fourteen patients taking DOACs (27%) used concomitant antiplatelet agents, with seven patients (13%) continuing treatment at the time of the endoscopic procedure. No postprocedural bleeding occurred in patients receiving warfarin (0%), whereas 10 cases (19%) of bleeding occurred in patients receiving DOACs: rivaroxaban, 0% (0/22); dabigatran, 0% (0/2); edoxaban, 43% (6/14); and apixaban, 29% (4/14). The type of anticoagulant (P < 0.01) and continuation of antiplatelet therapy (P = 0.02) were risk factors for postprocedural bleeding in patients receiving DOACs. Intraprocedural bleeding requiring transfusion or symptomatic thromboembolic events were not reported. CONCLUSIONS: Continuous warfarin therapy is preferred. DOAC withdrawal 1 day before a procedure is associated with a high bleeding rate, which may differ for different types of anticoagulants. The continuation of antiplatelet medications in patients receiving DOACs carries a high risk of bleeding and is a future challenge.

DOI： 10.1111/jgh.16757

Effectiveness of data-augmentation on deep learning in evaluating rapid on-site cytopathology at endoscopic ultrasound-guided fine needle aspiration. International journal

Yuki Fujii, Daisuke Uchida, Ryosuke Sato, Taisuke Obata, Matsumi Akihiro, Kazuya Miyamoto, Kosaku Morimoto, Hiroyuki Terasawa, Tatsuhiro Yamazaki, Kazuyuki Matsumoto, Shigeru Horiguchi, Koichiro Tsutsumi, Hironari Kato, Hirofumi Inoue, Ten Cho, Takayoshi Tanimoto, Akimitsu Ohto, Yoshiro Kawahara, Motoyuki Otsuka

Scientific reports 14 ( 1 ) 22441 - 22441 2024.9

Language：English Publishing type：Research paper (scientific journal)

Rapid on-site cytopathology evaluation (ROSE) has been considered an effective method to increase the diagnostic ability of endoscopic ultrasound-guided fine needle aspiration (EUS-FNA); however, ROSE is unavailable in most institutes worldwide due to the shortage of cytopathologists. To overcome this situation, we created an artificial intelligence (AI)-based system (the ROSE-AI system), which was trained with the augmented data to evaluate the slide images acquired by EUS-FNA. This study aimed to clarify the effects of such data-augmentation on establishing an effective ROSE-AI system by comparing the efficacy of various data-augmentation techniques. The ROSE-AI system was trained with increased data obtained by the various data-augmentation techniques, including geometric transformation, color space transformation, and kernel filtering. By performing five-fold cross-validation, we compared the efficacy of each data-augmentation technique on the increasing diagnostic abilities of the ROSE-AI system. We collected 4059 divided EUS-FNA slide images from 36 patients with pancreatic cancer and nine patients with non-pancreatic cancer. The diagnostic ability of the ROSE-AI system without data augmentation had a sensitivity, specificity, and accuracy of 87.5%, 79.7%, and 83.7%, respectively. While, some data-augmentation techniques decreased diagnostic ability, the ROSE-AI system trained only with the augmented data using the geometric transformation technique had the highest diagnostic accuracy (88.2%). We successfully developed a prototype ROSE-AI system with high diagnostic ability. Each data-augmentation technique may have various compatibilities with AI-mediated diagnostics, and the geometric transformation was the most effective for the ROSE-AI system.

DOI： 10.1038/s41598-024-72312-3

Gastric adenocarcinoma and proximal polyposis of the stomachに対する内視鏡診療の現状と今後の課題

川野誠司, 田中健大, 大塚基之

Gastroenterological Endoscopy 66 ( 9 ) 1663 - 1671 2024.9

Adverse events after different endoscopic resection procedures for small and intermediate-sized colorectal polyps. Reviewed International journal

Junki Toyosawa, Yasushi Yamasaki, Yuki Aoyama, Kensuke Takei, Shoko Igawa, Toshihiro Inokuchi, Hideaki Kinugasa, Masahiro Takahara, Sakiko Hiraoka, Hiroyuki Okada, Motoyuki Otsuka

Digestive diseases (Basel, Switzerland) 1 - 9 2024.8

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND STUDY AIMS: Cold snare polypectomy (CSP) and underwater endoscopic mucosal resection (UEMR) have been developed recently, in addition to conventional methods, but adverse events of each method have not been fully clarified. We compared outcomes of each method for the appropriate choice. PATIENTS AND METHODS: Patients who underwent CSP, endoscopic mucosal resection (EMR)/hot snare polypectomy (HSP) or UEMR for small and intermediate-sized colorectal polyps between April 2017 and June 2020 were retrospectively examined. The rate of adverse events and recurrences due to each method were determined as main outcomes. Clinical factors related with adverse events were examined. RESULTS: A total of 1,025 patients with 3,163 polyps underwent polypectomy using any of the methods. CSP, EMR/HSP and UEMR were performed for 704 (22.2%), 2,145 (67.8%) and 314 polyps (9.9%), and median size for each method was 4, 6 and 7 mm, respectively. Delayed bleeding for CSP, EMR/HSP and UEMR was 0%, 0.2% and 0.6% (P = 0.15), and perforation was 0%, 0.1% and 0%, respectively (P = 0.62). Recurrence after CSP, EMR/HSP and UEMR was 0.1%, 0.04% and 1.0%, respectively (P < 0.01). Recurrence for UEMR was significantly higher in the early stage of procedure introduction (P = 0.001). Oral anticoagulants were the risk factor for delayed bleeding (P < 0.01, respectively). CONCLUSIONS: There was no significant difference regarding adverse events among each method for small and intermediate-sized polyps, although recurrence rate after UEMR was higher than other methods.

DOI： 10.1159/000540365

Initial clinical experience with durvalumab plus tremelimumab in patients with unresectable hepatocellular carcinoma in real‑world practice. Reviewed International journal

Shigeo Shimose, Issei Saeki, Tetsu Tomonari, Takanori Ito, Joji Tani, Yasuto Takeuchi, Naoki Yoshioka, Takehito Naito, Mamiko Takeuchi, Satoru Kakizaki, Takeshi Hatanaka, Kyo Sasaki, Tetsuya Yasunaka, Masahiro Sakata, Hideki Iwamoto, Satoshi Itano, Tomotake Shirono, Norikazu Tanabe, Takafumi Yamamoto, Atsushi Naganuma, Soji Nishina, Motoyuki Otsuka, Hiroki Kawashima, Tetsuji Takayama, Taro Takami, Takumi Kawaguchi

Oncology letters 28 ( 2 ) 397 - 397 2024.8

Language：English Publishing type：Research paper (scientific journal)

Although durvalumab plus tremelimumab (Dur/Tre) has been approved as first-line therapy for patients with unresectable hepatocellular carcinoma (u-HCC), its outcomes in real-world clinical practice are unclear. The present study aimed to evaluate the efficacy and safety of Dur/Tre treatment. This multicenter study was conducted between March 2023 and January 2024, and included 120 patients with u-HCC treated with Dur/Tre. Among the patients, 44 had no history of systemic treatment. Progression-free survival (PFS), therapeutic response and adverse events (AEs) were assessed. The objective response rate (ORR) and disease control rates (DCR) were 15.8 and 53.3%, respectively. The median PFS was 3.9 months. The incidence rates of AEs of any grade and those grade 3 or higher were 83.3 and 36.7%, respectively. Liver injury was the most frequent AE of any grade and grade 3 or higher. Although there was no significant difference in ORR and PFS between the first and later line groups (ORR 15.8 vs. 15.7%, P=0.986; PFS 4.5 vs. 3.6 months, P=0.213), there was a significant difference in DCR between the two groups (65.8 vs. 45.9%, P=0.034). No significant differences were noted between the first- and later-line treatment groups regarding the incidence rate of AEs. Decision tree analysis revealed that poor liver function and advanced age were significant variables for discontinuation owing to AEs. In conclusion, Dur/Tre as first-line therapy had better disease control responses compared with later-line therapy; however, this regimen should be carefully administered to patients with deteriorating hepatic function or advanced age.

DOI： 10.3892/ol.2024.14530

Gastric syphilis displaying multiple annular lesions. Reviewed International journal

Masaya Iwamuro, Takehiro Tanaka, Motoyuki Otsuka

Gastrointestinal endoscopy 100 ( 2 ) 332 - 333 2024.8

【膵・胆道癌の発生・進展メカニズムを探る】胆道癌の薬剤耐性メカニズム

堤康一郎, 小幡泰介, 織田崇志, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 内田大輔, 松本和幸, 堀口繁, 大塚基之

胆と膵 45 ( 8 ) 857 - 862 2024.8

Language：Japanese Publisher：医学図書出版(株)

胆道癌の多くは根治切除が難しく,主に抗癌剤や分子標的薬を用いた薬物療法が行われるが,5年生存率は20%程度であり予後不良な疾患である。この一因として,癌細胞が抗腫瘍効果を有する薬剤から逃れて生存を維持しようとする機構,つまり癌の薬剤耐性保有やその獲得があげられる。これまでに胆道癌における七つの薬剤耐性メカニズム(mechanisms of chemoresistance:MOC)が報告されており,これらの複数が相互にまたは独立して関与していることが明らかとなっている。さまざまな遺伝子異常が基盤となって生じる薬剤耐性メカニズムの理解を深めることは,胆道癌に対する新規の治療戦略や治療法開発につながり,個々の患者における予後改善に寄与することが期待される。(著者抄録)

【今推しの肝胆膵研究手法と疾患モデル】新規モデルによる肝胆膵研究胆嚢オルガノイドを用いた新規治療の開発

織田崇志, 堤康一郎, 大塚基之

肝胆膵 89 ( 2 ) 249 - 254 2024.8

【膵・胆道癌の発生・進展メカニズムを探る】膵癌の薬剤耐性メカニズム

寺澤裕之, 堤康一郎, 原田圭, 服部直, 菊池達也, 小幡泰介, 佐藤亮介, 織田崇志, 松三明宏, 宮本和也, 藤井佑樹, 赤穂宗一郎, 内田大輔, 松本和幸, 堀口繁, 大塚基之

胆と膵 45 ( 8 ) 801 - 806 2024.8

Language：Japanese Publisher：医学図書出版(株)

膵癌は予後不良な難治癌であるが,その一因として薬剤耐性があげられる。薬剤耐性のメカニズムとして,標的治療が難しいKRAS変異の存在やエピジェネティック機構などの遺伝子変異や薬物代謝の変化,転移・浸潤を促進するEMT,膵癌に特徴的な間質成分の多い腫瘍微小環境,さらにnon-coding RNAによる遺伝子発現の調整などがあり,それらが複合的に影響していると考えられている。この複雑な薬剤耐性メカニズムのさらなる解明がその克服において重要である。(著者抄録)

【膵・胆道癌の発生・進展メカニズムを探る】胆道癌の薬剤耐性メカニズム

堤康一郎, 小幡泰介, 織田崇志, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 内田大輔, 松本和幸, 堀口繁, 大塚基之

胆と膵 45 ( 8 ) 857 - 862 2024.8

Language：Japanese Publisher：医学図書出版(株)

胆道癌の多くは根治切除が難しく,主に抗癌剤や分子標的薬を用いた薬物療法が行われるが,5年生存率は20%程度であり予後不良な疾患である。この一因として,癌細胞が抗腫瘍効果を有する薬剤から逃れて生存を維持しようとする機構,つまり癌の薬剤耐性保有やその獲得があげられる。これまでに胆道癌における七つの薬剤耐性メカニズム(mechanisms of chemoresistance:MOC)が報告されており,これらの複数が相互にまたは独立して関与していることが明らかとなっている。さまざまな遺伝子異常が基盤となって生じる薬剤耐性メカニズムの理解を深めることは,胆道癌に対する新規の治療戦略や治療法開発につながり,個々の患者における予後改善に寄与することが期待される。(著者抄録)

【膵・胆道癌の発生・進展メカニズムを探る】膵癌の薬剤耐性メカニズム

寺澤裕之, 堤康一郎, 原田圭, 服部直, 菊池達也, 小幡泰介, 佐藤亮介, 織田崇志, 松三明宏, 宮本和也, 藤井佑樹, 赤穂宗一郎, 内田大輔, 松本和幸, 堀口繁, 大塚基之

胆と膵 45 ( 8 ) 801 - 806 2024.8

Language：Japanese Publisher：医学図書出版(株)

膵癌は予後不良な難治癌であるが,その一因として薬剤耐性があげられる。薬剤耐性のメカニズムとして,標的治療が難しいKRAS変異の存在やエピジェネティック機構などの遺伝子変異や薬物代謝の変化,転移・浸潤を促進するEMT,膵癌に特徴的な間質成分の多い腫瘍微小環境,さらにnon-coding RNAによる遺伝子発現の調整などがあり,それらが複合的に影響していると考えられている。この複雑な薬剤耐性メカニズムのさらなる解明がその克服において重要である。(著者抄録)

【今推しの肝胆膵研究手法と疾患モデル】新規モデルによる肝胆膵研究胆嚢オルガノイドを用いた新規治療の開発

織田崇志, 堤康一郎, 大塚基之

肝胆膵 89 ( 2 ) 249 - 254 2024.8

Recent updates on treatment options for primary follicular lymphoma of the gastrointestinal tract. Reviewed International journal

Masaya Iwamuro, Takehiro Tanaka, Daisuke Ennishi, Motoyuki Otsuka

Expert review of gastroenterology & hepatology 18 ( 7 ) 1 - 9 2024.7

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Primary gastrointestinal follicular lymphoma is a subtype of follicular lymphoma that originates directly from the gastrointestinal tract. Pathologically, it exhibits substantial similarities with the secondary gastrointestinal involvement observed in nodal follicular lymphoma. However, primary gastrointestinal follicular lymphoma presents clinically distinct features, necessitating divergent considerations in treatment selection compared with nodal follicular lymphoma. AREAS COVERED: This narrative review focused on recent articles (2018-2023) regarding the long-term prognosis and treatment options for gastrointestinal follicular lymphoma. In addition, a brief overview of gastrointestinal follicular lymphomas is provided. EXPERT OPINION: Patients with primary gastrointestinal follicular lymphoma often present with a low tumor burden. Lymphoma lesions typically remain asymptomatic for several years or may undergo spontaneous regression without immediate treatment. Therefore, a 'watch and wait' approach is justified. Conversely, when large tumor masses are identified in the gastrointestinal tract, the potential for tumor bleeding or intestinal obstruction requires timely therapeutic interventions.

DOI： 10.1080/17474124.2024.2380337

Characteristics of Early Gastric Cancer in a Patient with a History of Helicobacter pylori Infection and No History of Eradication Therapy. Reviewed

Sakiko Kuraoka, Seiji Kawano, Shoko Ino, Takuya Satomi, Kenta Hamada, Yoshiyasu Kono, Masaya Iwamuro, Yoshiro Kawahara, Takehiro Tanaka, Hiroyuki Okada, Motoyuki Otsuka

Internal medicine (Tokyo, Japan) 2024.7

Language：English Publishing type：Research paper (scientific journal)

Objective The characteristics of gastric cancer in patients with atrophic mucosa and no apparent history of Helicobacter pylori eradication have not been thoroughly investigated. Therefore, this study examined the clinicopathological characteristics of gastric cancer in these patients. Methods We retrospectively examined the endoscopic and pathological characteristics of gastric cancer in patients who underwent endoscopic submucosal dissection. Patients or Materials We divided the patients into 2 groups: those with gastric atrophy and no history of eradication (group A; n =102) and those with a history of eradication (group B; n =161). In group A, patients were further divided into mild atrophy (group C) and severe atrophy (group D) groups, while group B was further divided into those who underwent eradication treatment >5 years ago (group E) and those who underwent eradication 1-5 years ago (group F). Results Group A comprised significantly older individuals (75±8.0 vs. 71±7.5 years old, p <0.001) with a higher frequency of elevated gastric cancer than group B (32.4% vs. 17.4%, p =0.006). Compared with group E, group A was older and had a greater incidence of elevated gastric cancer. The incidence of gastric cancer in the U or M region was lower in group C than in group D. Conclusion Gastric cancer in patients with gastric atrophy and no history of eradication was associated with an older age and higher frequency of elevated-type morphology than in those with a history of eradication.

DOI： 10.2169/internalmedicine.3617-24

Colorectal Ganglioneuromas Associated with Cowden Syndrome.

Toshiki Ozato, Yasushi Yamasaki, Toshihiro Inokuchi, Motoyuki Otsuka

Internal medicine (Tokyo, Japan) 63 ( 13 ) 1959 - 1960 2024.7

Response to the letter: "Predictive factors for transition to conversion therapy in HCC using atezolizumab plus bevacizumab". International journal

Tatsuya Kikuchi, Yasuto Takeuchi, Kazuhiro Nouso, Kazuya Kariyama, Kenji Kuwaki, Junichi Toshimori, Shota Iwado, Akio Moriya, Hiroaki Hagihara, Hiroyuki Takabatake, Toshifumi Tada, Tetsuya Yasunaka, Masahiro Sakata, Masahiko Sue, Nozomi Miyake, Takuya Adachi, Nozomu Wada, Hideki Onishi, Hidenori Shiraha, Akinobu Takaki, Motoyuki Otsuka

Liver international : official journal of the International Association for the Study of the Liver 44 ( 7 ) 1736 - 1737 2024.7

Small Intestinal Complications of Intestinal Transplant-Associated Microangiopathy Post-stem Cell Transplantation. International journal

Daisuke Kametaka, Masaya Iwamuro, Takehiro Tanaka, Ken-Ichi Matsuoka, Motoyuki Otsuka

Cureus 16 ( 7 ) e64111 2024.7

Language：English Publishing type：Research paper (scientific journal)

We present the case of a patient who underwent human leukocyte antigen-haploidentical transplantation for T-cell acute lymphoblastic leukemia. Seven weeks after transplantation, the patient developed intestinal transplant-associated microangiopathy (iTAM). Although the iTAM was resolved temporarily, it recurred. Video capsule enteroscopy revealed multiple erosions and shallow ulcers in the jejunum and ileum. To the best of our knowledge, this is the first report to present images of possible small intestinal lesions in iTAM. The small intestinal mucosal images presented herein may potentially aid in the management of similar patients.

DOI： 10.7759/cureus.64111

Risk factors and treatment strategies for cholecystitis after metallic stent placement for malignant biliary obstruction: a multicenter retrospective study. Reviewed International journal

Akihiro Matsumi, Hironari Kato, Taiji Ogawa, Toru Ueki, Masaki Wato, Masakuni Fujii, Tatsuya Toyokawa, Ryo Harada, Yuki Ishihara, Masahiro Takatani, Hirofumi Tsugeno, Naoko Yunoki, Takeshi Tomoda, Toshiharu Mitsuhashi, Motoyuki Otsuka

Gastrointestinal endoscopy 100 ( 1 ) 76 - 84 2024.7

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: Cholecystitis can occur after self-expandable metallic stent (SEMS) placement for malignant biliary obstruction (MBO), but the best treatment option for cholecystitis has not been determined. Here, we aimed to identify the risk factors of cholecystitis after SEMS placement and determine the best treatment option. METHODS: Incidence, treatments, and predictive factors of cholecystitis were retrospectively evaluated in 1084 patients with distal MBO (DMBO) and 353 patients with hilar MBO (HMBO) who underwent SEMS placement at 12 institutions from January 2012 to March 2021. RESULTS: Cholecystitis occurred in 7.5% of patients with DMBO and 5.9% of patients with HMBO. The recurrence rate was significantly lower (P = .043) and the recurrence-free period significantly longer (P = .039) in endoscopic procedures than in percutaneous procedures for cholecystitis treatment. EUS-guided gallbladder drainage (EUS-GBD) was better in terms of technical success, procedure time, and recurrence-free period than endoscopic transpapillary gallbladder drainage. Obstruction across the cystic duct orifice by tumor (P = .015) and by stent (P = .037) were independent risk factors for cholecystitis in DMBO. Cases with multiple SEMS placements (odds ratio [OR], 11; 95% confidence interval [CI], 0.68-190; P = .091) and with gallbladder stones (OR, 2.3; 95% CI ,0.92-5.6; P = .075) had a higher risk for cholecystitis in HMBO. CONCLUSIONS: The incidences of cholecystitis after SEMS placement for DMBO and HMBO were similar. EUS-GBD is the optimal treatment option for patients with cholecystitis after SEMS placement for MBO.

DOI： 10.1016/j.gie.2024.02.019

後期高齢者におけるIPMNの治療移行割合及び長期経過についての検討

宮本和也, 松本和幸, 服部直, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 森本光作, 寺澤裕之, 藤井佑樹, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 39 ( 3 ) A425 - A425 2024.7

膵EUS-FNA診断の功罪嚢胞・壊死を伴う膵管癌に対するFNA/B合併症リスクの検討

寺澤裕之, 松本和幸, 田中健大, 友田健, 小川泰司, 石原裕基, 菊池達也, 小幡泰介, 織田崇志, 松三明宏, 宮本和也, 森本光作, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 39 ( 3 ) A234 - A234 2024.7

Predictive factors for transition to conversion therapy in hepatocellular carcinoma using atezolizumab plus bevacizumab. International journal

Tatsuya Kikuchi, Yasuto Takeuchi, Kazuhiro Nouso, Kazuya Kariyama, Kenji Kuwaki, Junichi Toshimori, Shota Iwado, Akio Moriya, Hiroaki Hagihara, Hiroyuki Takabatake, Toshifumi Tada, Tetsuya Yasunaka, Masahiro Sakata, Masahiko Sue, Nozomi Miyake, Takuya Adachi, Nozomu Wada, Hideki Onishi, Hidenori Shiraha, Akinobu Takaki, Motoyuki Otsuka

Liver international : official journal of the International Association for the Study of the Liver 44 ( 6 ) 1456 - 1463 2024.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: To identify predictive factors associated with successful transition to conversion therapy following combination therapy with atezolizumab and bevacizumab in the treatment of unresectable hepatocellular carcinoma (HCC). METHODS: In total, 188 patients with HCC, who received atezolizumab plus bevacizumab combination therapy as the first-line chemotherapy, were studied. Patients who achieved complete response (CR) with systemic chemotherapy alone were excluded. Clinical factors possibly linked to successful transition to conversion therapy and the achievement of cancer-free status were identified. RESULTS: Fifteen (8.0%) patients underwent conversion therapy. In the conversion group, there was a significantly higher proportion of patients with Barcelona Clinic Liver Cancer (BCLC) stage A or B (73.3% versus [vs.] 45.1%; p = .03) and tended to have lower Child-Pugh scores and alpha-fetoprotein levels. Multivariate analysis revealed that BCLC stage was a predictive factor for the implementation of conversion therapy (A or B; odds ratio 3.7 [95% CI: 1.1-13]; p = .04). Furthermore, 10 (66.7%) patients achieved cancer-free status and exhibited a smaller number of intrahepatic lesions at the start of treatment (3.5 vs. 7; p < .01), and a shorter interval between systemic chemotherapy induction and conversion therapy (131 vs. 404 days; p < .01). In addition, the rate of achieving cancer-free status by undergoing surgical resection or ablation therapy was significantly higher (p = .03). CONCLUSION: BCLC stage was the sole predictive factor for successful transition to conversion therapy when using combination therapy with atezolizumab and bevacizumab to treat HCC. Furthermore, a small number of intrahepatic lesions and early transition to conversion therapy were associated with the achievement of cancer-free status.

DOI： 10.1111/liv.15907

A Duodenal Ulcer and Biliopancreatic Lesions: What is the Culprit? Reviewed International journal

Ryosuke Sato, Kazuyuki Matsumoto, Motoyuki Otsuka

Gastroenterology 2024.5

Current status of endoscopic ultrasound-guided antitumor treatment for pancreatic cancer. Reviewed International journal

Kazuyuki Matsumoto, Hironari Kato, Koichiro Tsutsumi, Motoyuki Otsuka

Digestive endoscopy : official journal of the Japan Gastroenterological Endoscopy Society 2024.5

Language：English Publishing type：Research paper (scientific journal)

Endoscopic ultrasound (EUS) was developed in the 1990s and has significantly transformed pancreatic tumor diagnosis. Subsequently, EUS has rapidly shifted from being a purely diagnostic procedure to being used in a wide range of interventional procedures. Recently, new therapeutic techniques, such as EUS-guided fine needle injection (EUS-FNI) or radiofrequency ablation (RFA), have been developed to deliver various antitumor agents. Despite technological advancements, pancreatic cancer (PC) has a poor prognosis and improvements in treatment outcomes are urgently required. One of the reasons for the limited response to antitumor agents in PC is the abundant desmoplasia and hypovascular nature of the tumor, complicating drug delivery into the tumor. Thus, changing the tumor microenvironment may be important to enhance the effectiveness of chemotherapy, and direct injection of antitumor agents into the tumor under EUS guidance can help overcome treatment challenges in PC. Treatment approaches using the EUS-FNI or RFA technique are expected to further improve the prognosis of PC. Therefore, this study reviewed the existing literature on EUS-guided antitumor therapy, specifically highlighting its application in PC to address the current challenges and to identify potential advancements in the field.

DOI： 10.1111/den.14815

Impact of Nutritional Status on Neutrophil-to-Lymphocyte Ratio as a Predictor of Efficacy and Adverse Events of Immune Check-Point Inhibitors. Reviewed International journal

Masahiko Sue, Yasuto Takeuchi, Shoichiro Hirata, Akinobu Takaki, Motoyuki Otsuka

Cancers 16 ( 10 ) 2024.5

Language：English Publishing type：Research paper (scientific journal)

The neutrophil -to-lymphocyte ratio (NLR) is useful for predicting the effectiveness of treatment with immune checkpoint inhibitors (ICIs) and immune-related adverse events (irAEs). Because a growing body of evidence has recently shown that the number of lymphocytes that comprise NLR fluctuates according to nutritional status, this study examined whether the usefulness of NLR varies in ICI treatment due to changes in nutritional status. A retrospective analysis was performed on 1234 patients who received ICI treatment for malignant tumors at our hospital. Progression-free survival (PFS) was significantly prolonged in patients with NLR < 4. Multivariate analysis revealed that the factors associated with the occurrence of irAE were NLR < 4 and the use of ipilimumab. However, when limited to cases with serum albumin levels <3.8 g/dL, lymphocyte counts significantly decreased, and the associations between NLR and PFS and between NLR and irAE occurrence disappeared. In contrast, when limited to the cases with serum albumin levels ≥3.8 g/dL, the associations remained, with significantly prolonged PFS and significantly increased irAE occurrence at NLR < 4. NLR may be a good predictive tool for PFS and irAE occurrence during ICI treatment when a good nutritional status is maintained.

DOI： 10.3390/cancers16101811

Association of screening tool scores with overall survival in older patients with unresectable gastrointestinal or biliary pancreatic cancer: A retrospective study. International journal

Shoko Inoo, Yoshiyasu Kono, Hiromitsu Kanzaki, Shigeru Horiguchi, Hironari Kato, Yasushi Yamasaki, Shoichiro Hirata, Sakiko Kuraoka, Katsunori Matsueda, Shotaro Okanoue, Takuya Satomi, Kenta Hamada, Masaya Iwamuro, Seiji Kawano, Yoshiro Kawahara, Hiroyuki Okada, Motoyuki Otsuka

Journal of geriatric oncology 15 ( 4 ) 101714 - 101714 2024.5

Drug-induced mucosal alterations observed during esophagogastroduodenoscopy. International journal

Masaya Iwamuro, Seiji Kawano, Motoyuki Otsuka

World journal of gastroenterology 30 ( 16 ) 2220 - 2232 2024.4

Language：English Publishing type：Research paper (scientific journal)

Several features of drug-induced mucosal alterations have been observed in the upper gastrointestinal tract, i.e., the esophagus, stomach, and duodenum. These include pill-induced esophagitis, desquamative esophagitis, worsening of gastroesophageal reflux, chemotherapy-induced esophagitis, proton pump inhibitor-induced gastric mucosal changes, medication-induced gastric erosions and ulcers, pseudomelanosis of the stomach, olmesartan-related gastric mucosal inflammation, lanthanum deposition in the stomach, zinc acetate hydrate tablet-induced gastric ulcer, immune-related adverse event gastritis, olmesartan-asso-ciated sprue-like enteropathy, pseudomelanosis of the duodenum, and lanthanum deposition in the duodenum. For endoscopists, acquiring accurate knowledge regarding these diverse drug-induced mucosal alterations is crucial not only for the correct diagnosis of these lesions but also for differential diag-nosis of other conditions. This minireview aims to provide essential information on drug-induced mucosal alterations observed on esophagogastroduodenoscopy, along with representative endoscopic images.

DOI： 10.3748/wjg.v30.i16.2220

Endoscopic manifestation of intestinal transplant-associated microangiopathy after stem cell transplantation. International journal

Masaya Iwamuro, Daisuke Ennishi, Nobuharu Fujii, Ken-Ichi Matsuoka, Takehiro Tanaka, Toshihiro Inokuchi, Sakiko Hiraoka, Motoyuki Otsuka

BMC gastroenterology 24 ( 1 ) 140 - 140 2024.4

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Endoscopic features of intestinal transplant-associated microangiopathy (iTAM) have not been comprehensively investigated. This study aimed to examine the endoscopic characteristics of patients diagnosed with iTAM. METHODS: This retrospective analysis included 14 patients pathologically diagnosed with iTAM after stem cell transplantation for hematolymphoid neoplasms (n = 13) or thalassemia (n = 1). The sex, age at diagnosis, endoscopic features, and prognosis of each patient were assessed. Serological markers for diagnosing transplant-associated thrombotic microangiopathy were also evaluated. RESULTS: The mean age at the time of iTAM diagnosis was 40.2 years. Patients diagnosed based on the pathognomonic pathological changes of iTAM presented with diverse symptoms at the times of endoscopic examinations, including diarrhea (n = 10), abdominal pain (n = 5), nausea (n = 4), appetite loss (n = 2), bloody stools (n = 2), abdominal discomfort (n = 1), and vomiting (n = 1). At the final follow-up, six patients survived, while eight patients succumbed, with a median time of 100.5 days (range: 52-247) post-diagnosis. Endoscopic manifestations included erythematous mucosa (n = 14), erosions (n = 13), ulcers (n = 9), mucosal edema (n = 9), granular mucosa (n = 9), and villous atrophy (n = 4). Erosions and/or ulcers were primarily observed in the colon (10/14, 71%), followed by the ileum (9/13, 69%), stomach (4/10, 40%), cecum (5/14, 36%), duodenum (3/10, 30%), rectum (4/14, 29%), and esophagus (1/10, 10%). Cytomegalovirus infection (n = 4) and graft-versus-host disease (n = 2) coexisted within the gastrointestinal tract. Patients had de novo prolonged or progressive thrombocytopenia (6/14, 43%), decreased hemoglobin concentration (4/14, 29%), reduced serum haptoglobin level (3/14, 21%), and a sudden and persistent increase in lactate dehydrogenase level (2/14, 14%). Peripheral blood samples from 12 patients were evaluated for schistocytes, with none exceeding 4%. CONCLUSIONS: This study provides a comprehensive exploration of the endoscopic characteristics of iTAM. Notably, all patients exhibited erythematous mucosa throughout the gastrointestinal tract, accompanied by prevalent manifestations, such as erosions (93%), ulcers (64%), mucosal edema (64%), granular mucosa (64%), and villous atrophy (29%). Because of the low positivity for serological markers of transplant-associated thrombotic microangiopathy in patients with iTAM, endoscopic evaluation and biopsy of these lesions are crucial, even in the absence of these serological features.

DOI： 10.1186/s12876-024-03221-y

Detection of residual stones by peroral direct cholangioscopy in patients with intrahepatic stones after hepaticojejunostomy: A prospective study (with video). International journal

Kazuyuki Matsumoto, Hironari Kato, Akihiro Matsumi, Kazuya Miyamoto, Kosaku Morimoto, Yuki Fujii, Ryosuke Sato, Nao Hattori, Taisuke Obata, Hiroyuki Terasawa, Daisuke Uchida, Shigeru Horiguchi, Koichiro Tsutsumi, Motoyuki Otsuka

Gastrointestinal endoscopy 2024.4

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND AND AIMS: The difficulty of radiographic confirmation of the presence of stones remains a challenge in the treatment of intrahepatic bile duct (IHBD) stones in patients after hepaticojejunostomy (HJ). Peroral direct cholangioscopy (PDCS) enables direct observation of the bile duct and is useful for detecting and removing residual stones; however, its effectiveness is not clearly established in this clinical context. METHODS: This single-center, single-arm, prospective study included 44 patients with IHBD who underwent bowel reconstruction with HJ during the study period. Stone removal was performed by short-type double-balloon enteroscopy (DBE). Following balloon-occluded cholangiography, the DBE was exchanged for an ultraslim endoscope through the balloon overtube for PDCS. The primary endpoint was the rate of residual stones detected by PDCS. Secondary endpoints were success rate of PDCS, residual stone removal with PDCS, procedure time for PDCS, procedure-related adverse events, and stone recurrence rate. RESULTS: PDCS was successful in 39/44 patients (89%), among whom residual stones were detected in 16 (41%) (95% CI: 28%-54%). Twelve patients (75%) had residual stones <5 mm. Stone removal was successful in 15 (94%) patients and median procedure time for PDCS was 16 (IQR: 10-26) min. The rate of procedure-related adverse events was 7% (3/44), all of which improved with conservative treatment. During median follow-up of 2.1 years (IQR: 1.4-3.3), the overall probability of recurrence-free status at 1, 2, and 3 years was 100%, 92%, and 86%, respectively. CONCLUSIONS: PDCS is a safe and effective procedure for complete stone removal in patients with IHBD stones after HJ.

DOI： 10.1016/j.gie.2024.04.014

UnderwaterとUnder-gelでの大腸腫瘍性病変に対する境界診断能の検討

豊澤惇希, 山崎泰史, 青山祐樹, 井川翔子, 井口俊博, 衣笠秀明, 高原政宏, 平岡佐規子, 大塚基之

Gastroenterological Endoscopy 66 ( Suppl.1 ) 969 - 969 2024.4

結節を有するIPMNの良悪性診断における新規TIC解析プログラムを用いた造影EUSの有用性

宮本和也, 内田大輔, 服部直, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 森本光作, 寺澤裕之, 藤井佑樹, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

Gastroenterological Endoscopy 66 ( Suppl.1 ) 977 - 977 2024.4

肝細胞癌の病態解明のためのトランスレーショナルリサーチ血清抗PD1自己抗体高値はアテゾリズマブ/ベバシズマブ治療予後不良の予測因子となる

高木章乃夫, 松本和幸, 佐々木裕貴, 須江真彦, 足立卓哉, 和田望, 竹内康人, 大西秀樹, 白羽英則, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A236 - A236 2024.3

経口胆道鏡検査におけるAIを用いた色素内視鏡画像への疑似変換

佐藤亮介, 松本和幸, 冨谷昌弘, 谷本太郁由, 大戸彰三, 原田圭, 服部直, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 内田大輔, 衣笠秀明, 堤康一郎, 堀口繁, 加藤博也, 河原祥朗, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A356 - A356 2024.3

膵癌の分子異常と個別化治療切除不能進行膵癌治療において生殖細胞BRCA病的変異は体細胞CGP結果によらず白金製剤治療効果予測因子となる

堀口繁, 加藤博也, 寺澤裕之, 藤井佑樹, 山崎辰洋, 松本和幸, 堤康一郎, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A87 - A87 2024.3

神経内分泌腫瘍の新しい治療戦略膵消化管神経内分泌腫瘍肝転移に対するPRRTの治療効果予測因子についての検討

堀口繁, 加藤博也, 寺澤裕之, 藤井佑樹, 山崎辰洋, 松本和幸, 堤康一郎, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A108 - A108 2024.3

神経線維腫症の消化管合併症に関する検討

本田真奈美, 岩室雅也, 山崎泰史, 田中健大, 川野誠司, 平岡佐規子, 河原祥朗, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A349 - A349 2024.3

Satellite double-stranded RNA induces mesenchymal transition in pancreatic cancer by regulating alternative splicing. Reviewed International journal

Takuma Iwata, Takahiro Kishikawa, Takahiro Seimiya, Genso Notoya, Tatsunori Suzuki, Chikako Shibata, Yu Miyakawa, Nariaki Odawara, Kazuyoshi Funato, Eri Tanaka, Mari Yamagami, Kazuma Sekiba, Motoyuki Otsuka, Kazuhiko Koike, Mitsuhiro Fujishiro

The Journal of biological chemistry 300 ( 3 ) 105742 - 105742 2024.3

Language：English Publishing type：Research paper (scientific journal)

Human satellite II (HSATII), composed of tandem repeats in pericentromeric regions, is aberrantly transcribed in epithelial cancers, particularly pancreatic cancer. Dysregulation of repetitive elements in cancer tissues can facilitate incidental dsRNA formation; however, it remains controversial whether dsRNAs play tumor-promoting or tumor-suppressing roles during cancer progression. Therefore, we focused on the double-stranded formation of HSATII RNA and explored its molecular function. The overexpression of double-stranded HSATII (dsHSATII) RNA promoted mesenchymal-like morphological changes and enhanced the invasiveness of pancreatic cancer cells. We identified an RNA-binding protein, spermatid perinuclear RNA-binding protein (STRBP), which preferentially binds to dsHSATII RNA rather than single-stranded HSATII RNA. The mesenchymal transition of dsHSATII-expressing cells was rescued by STRBP overexpression. Mechanistically, STRBP is involved in the alternative splicing of genes associated with epithelial-mesenchymal transition (EMT). We also confirmed that isoform switching of CLSTN1, driven by dsHSATII overexpression or STRBP depletion, induced EMT-like morphological changes. These findings reveal a novel tumor-promoting function of dsHSATII RNA, inducing EMT-like changes and cell invasiveness, thus enhancing our understanding of the biological significance of aberrant expression of satellite arrays in malignant tumors.

DOI： 10.1016/j.jbc.2024.105742

大腸鋸歯状病変の病態解明と診療戦略 Serrated polyposis syndromeの臨床病理学的特徴とその発癌ポテンシャルの検討

平井亮佑, 衣笠秀明, 平岡佐規子, 田中健大, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A215 - A215 2024.3

AIを用いた消化器疾患の診断・治療への応用内視鏡画像を用いた早期胃癌の深達度診断支援プログラムの性能

平田翔一郎, 濱田健太, 河原祥朗, 倉岡紗樹子, 内田大輔, 河野吉泰, 岩室雅也, 川野誠司, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A36 - A36 2024.3

免疫関連有害事象による胃炎の5例

岩室雅也, 田中健大, 平田翔一郎, 河野吉泰, 川野誠司, 河原祥朗, 大塚基之

Gastroenterological Endoscopy 66 ( 3 ) 266 - 272 2024.3

Language：Japanese Publisher：(一社)日本消化器内視鏡学会

免疫チェックポイント阻害薬投与後の免疫関連有害事象(immune-related adverse events:irAE)として胃病変を呈した5症例(男性1例,女性4例)を経験したので報告する.原疾患は全例が悪性黒色腫であった.内視鏡検査では粗そう粘膜,白色浸出物,発赤,易出血性をそれぞれ4例に認めたほか,浮腫が1例,小潰瘍が1例でみられ,拡大観察を行った4例では腺管構造が消失していた.CT検査では胃前庭部の壁肥厚を1例に認めたが,他の4例では上部消化管に異常所見はみられなかった.治療として全例でプレドニゾロンを用い,自覚症状の改善が得られ,うち3例では内視鏡所見の改善も確認した.上記の内視鏡所見に着目し,irAE胃炎と診断し治療にあたることが重要と考えられた.(著者抄録)

Other Link： https://search.jamas.or.jp/default/link?pub_year=2024&ichushi_jid=J00192&link_issn=&doc_id=20240405340004&doc_link_id=10.11280%2Fgee.66.266&url=https%3A%2F%2Fdoi.org%2F10.11280%2Fgee.66.266&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_2.gif

経口胆道鏡検査におけるAIを用いた色素内視鏡画像への疑似変換

佐藤亮介, 松本和幸, 冨谷昌弘, 谷本太郁由, 大戸彰三, 原田圭, 服部直, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 内田大輔, 衣笠秀明, 堤康一郎, 堀口繁, 加藤博也, 河原祥朗, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A356 - A356 2024.3

大腸鋸歯状病変の病態解明と診療戦略 Serrated polyposis syndromeの臨床病理学的特徴とその発癌ポテンシャルの検討

平井亮佑, 衣笠秀明, 平岡佐規子, 田中健大, 大塚基之

日本消化器病学会雑誌 121 ( 臨増総会 ) A215 - A215 2024.3

Long-term monitoring of gastric mucosa-associated lymphoid tissue lymphoma in patients with extra copies of the MALT1 gene. Reviewed International journal

Masaya Iwamuro, Ryuta Takenaka, Koji Miyahara, Shotaro Okanoue, Masao Yoshioka, Chihiro Sakaguchi, Kumiko Yamamoto, Yoshinari Kawai, Tatsuya Toyokawa, Takehiro Tanaka, Motoyuki Otsuka

Scientific reports 14 ( 1 ) 4953 - 4953 2024.2

Language：English Publishing type：Research paper (scientific journal)

The objective of this study was to clarify the long-term prognosis of patients with gastric mucosa-associated lymphoid tissue (MALT) lymphoma with additional copies of MALT1. In this multicenter retrospective study, we enrolled 145 patients with gastric MALT lymphoma who underwent fluorescence in situ hybridization (FISH) analysis to detect t(11;18) translocation. The patient cohort was divided into three groups: Group A (n = 87), comprising individuals devoid of the t(11;18) translocation or extra MALT1 copies; Group B (n = 27), encompassing patients characterized by the presence of the t(11;18) translocation; and Group C (n = 31), including patients with extra MALT1 copies. The clinical outcomes in each cohort were collected. Over the course of a mean follow-up of 8.5 ± 4.2 years, one patient died of progressive MALT lymphoma, while 15 patients died due to etiologies unrelated to lymphoma. The progression or relapse of MALT lymphoma was observed in 11 patients: three in Group A, two in Group B, and six in Group C. In Groups A, B, and C, the 10-year overall survival rates were 82.5%, 93.8%, and 86.4%, respectively, and the 10-year event-free survival rates were 96.1%, 96.0%, and 82.9%, respectively. The event-free survival rate in Group C was significantly lower than that in Group A. However, no differences were observed in the 10-year event-free survival rates among individuals limited to stage I or II1 disease (equivalent to excluding patients with stage IV disease in this study, as there were no patients with stage II2), with rates of 98.6%, 95.8%, and 92.3% for Groups A, B, and C, respectively. In conclusion, the presence of extra copies of MALT1 was identified as an inferior prognostic determinant of event-free survival. Consequently, trisomy/tetrasomy 18 may serve as an indicator of progression and refractoriness to therapeutic intervention in patients with gastric MALT lymphoma, particularly stage IV gastric MALT lymphoma.

DOI： 10.1038/s41598-024-55663-9

Endoscopic and clinical features of gastric emphysema. Reviewed International journal

Masaya Iwamuro, Ryuta Takenaka, Tatsuya Toyokawa, Masahide Kita, Takao Tsuzuki, Masao Yoshioka, Tatsuhiro Gotoda, Shotaro Okanoue, Minoru Matsubara, Chihiro Sakaguchi, Motoyuki Otsuka

Scientific reports 14 ( 1 ) 2202 - 2202 2024.1

Language：English Publishing type：Research paper (scientific journal)

Gastric emphysema is characterized by the presence of intramural gas in the stomach without bacterial infection. Due to its rarity, most reports on gastric emphysema have been limited to single-case studies, and this condition's clinical and endoscopic features have not been thoroughly investigated. In this study, we analyzed 45 patients with gastric emphysema from 10 institutions and examined their characteristics, endoscopic features, and outcomes. The mean age at diagnosis of gastric emphysema in our study population (35 males and 10 females) was 68.6 years (range, 14-95 years). The top five underlying conditions associated with gastric emphysema were the placement of a nasogastric tube (26.7%), diabetes mellitus (20.0%), post-percutaneous endoscopic gastrostomy (17.8%), malignant neoplasms (17.8%), and renal failure (15.6%). Among the 45 patients, 42 were managed conservatively with fasting and administration of proton pump inhibitors. Unfortunately, seven patients died within 30 days of diagnosis, and 35 patients experienced favorable recoveries. The resolution of gastric emphysema was confirmed in 30 patients through computed tomography (CT) scans, with a mean duration of 17.1 ± 34.9 days (mean ± standard deviation [SD], range: 1-180 days) from the time of diagnosis to the disappearance of the gastric intramural gas. There were no instances of recurrence. Endoscopic evaluation was possible in 18 patients and revealed that gastric emphysema presented with features such as redness, erosion, coarse mucosa, and ulcers, with fewer mucosal injuries on the anterior wall (72.2%), a clear demarcation between areas of mucosal injury and intact mucosa (61.1%), and predominantly longitudinal mucosal injuries on the stomach folds (50.0%). This study is the first English-language report to analyze endoscopic findings in patients with gastric emphysema.

DOI： 10.1038/s41598-024-52633-z

Anti-PD-1 Autoantibody Predicts Survival of Patients With Hepatocellular Carcinoma Receiving Atezolizumab/Bevacizumab

Yuki Sasaki, Kazuyuki Matsumoto, Akinobu Takaki, Takuya Adachi, Masahiro Takahara, Keita Ozato, Yasuto Takeuchi, Masahiko Sue, Nozomi Miyake, Nozomu Wada, Hideki Onishi, Hidenori Shiraha, Takashi Oda, Koichiro Tsutsumi, Kazuhiro Nouso, Kazuya Kariyama, Hiroaki Hagihara, Akio Moriya, Motoyuki Otsuka

Gastro Hep Advances 3 ( 8 ) 1138 - 1147 2024.1

Publishing type：Research paper (scientific journal)

Background and Aims: Methods for predicting therapeutic response to immune checkpoint inhibitors in cancer therapy are in high demand. In patients with advanced hepatocellular carcinoma (HCC), atezolizumab (anti-programmed cell death-ligand 1 [PD-L1]) and bevacizumab (anti-vascular endothelial growth factor) combination therapy (Atezo/Bev therapy) is a first-line treatment. However, no reliable biomarkers are currently available to predict its efficacy. Here, we examined serum anti-PD-1 autoantibody levels as candidate biomarkers. Methods: We prospectively enrolled 63 patients with advanced HCC who received Atezo/Bev therapy. Serum anti-PD-1 autoantibody levels were measured before treatment using an indirect enzyme-linked immunosorbent assay. The correlation between the titers and response to therapy was statistically examined. Results: Serum anti-PD-1 autoantibody levels were not significantly associated with the treatment response in any patient. However, when examining only patients who received the Atezo/Bev as their first-line therapy, higher anti-PD-1 autoantibody levels were significantly associated with worse overall survival rates. The titer was an independent risk factor for poor prognosis (odds ratio [OR] = 7.8, P = .013), in addition to a higher neutrophil-to-lymphocyte ratio (OR = 7.1, P = .009) and lower albumin levels (OR = 14.2, P = .003). Conclusion: Serum anti-PD-1 autoantibody levels correlated with the overall survival rate in patients who received Atezo/Bev as first-line therapy. Serum anti-PD-1 autoantibody levels may serve as new biomarkers for predicting the efficacy of immune checkpoint inhibitors in patients with HCC.

DOI： 10.1016/j.gastha.2024.07.018

Serum level of full-length connective tissue growth factor reflects liver fibrosis stage in patients with Fontan-associated liver disease. Reviewed International journal

Tomomi Kogiso, Kayo Takayanagi, Tsutomu Ishizuka, Motoyuki Otsuka, Kei Inai, Yuri Ogasawara, Kentaro Horiuchi, Makiko Taniai, Katsutoshi Tokushige

PloS one 19 ( 1 ) e0296375 2024

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Chronic liver disease leads to liver fibrosis, and an accurate diagnosis of the fibrosis stage is crucial for medical management. Connective tissue growth factor (CTGF) is produced by endothelial cells and platelets and plays a central role in inducing fibrosis in various organs. In the present study, we tested the validity of measuring the serum levels of two types of CTGF to estimate the biopsy-confirmed liver fibrosis stage. METHODS: We used two detection antibodies targeting the N- and C-terminal of CTGF to measure the serum levels of two forms of CTGF consisting of its full length and its N-terminal fragment. We analyzed the level of CTGF (via enzyme-linked immunosorbent assay) and the liver fibrosis stage in 38 patients with Fontan-associated liver disease (FALD) (26 cases of which were diagnosed pathologically). Correlations were determined by multivariate analysis and the area under the receiver operating characteristic curve. The 65 patients with nonalcoholic fatty liver disease (NAFLD) were included as a disease control group for examination. RESULTS: Full-length CTGF was significantly inversely correlated with liver fibrosis in patients with FALD. Although the platelet count was also associated with the liver fibrosis stage, full-length CTGF was more closely correlated with the fibrosis stage. Furthermore, the level of full-length CTGF was inversely associated with high central venous pressure. Conversely, the serum level of CTGF was not correlated with the fibrosis stage in NAFLD. CONCLUSION: The serum level of full-length CTGF may be useful for estimating the liver fibrosis stage in patients with FALD.

DOI： 10.1371/journal.pone.0296375

[Small intestinal perforation manifesting as post-transplant lymphoproliferative disorder occurring 23 years after living lung transplantation: a case report].

Daisuke Kametaka, Masaya Iwamuro, Shoko Inoo, Seiji Kawano, Takehiro Tanaka, Yoshiro Kawahara, Motoyuki Otsuka

Nihon Shokakibyo Gakkai zasshi = The Japanese journal of gastro-enterology 121 ( 11 ) 896 - 903 2024

Language：Japanese Publishing type：Research paper (scientific journal)

A Japanese female patient underwent a living lung transplant at age 29 and has been on immunomodulatory drug therapy since then. At age 52, she presented with sudden hematochezia. Despite abdominal computed tomography scan, esophagogastroduodenoscopy, and colonoscopy, no definitive source of bleeding was identified. Considering the possibility of small bowel bleeding, video capsule endoscopy was performed, which revealed a suspected ulcerative lesion in the jejunum. Subsequent per-oral double-balloon endoscopy confirmed the presence of an ulcerative lesion in the jejunum. A potential pathological diagnosis of post-transplant lymphoproliferative disorder (PTLD) was considered based on endoscopic biopsy specimens from the jejunal lesion. However, before the pathologic biopsy results were available, the patient experienced small intestinal perforation, necessitating emergency partial resection. Pathologic examination revealed a dense proliferation of medium-to-large atypical lymphocytes with neutrophilic infiltration in the perforated area of the small intestinal wall. Immunostaining showed lymphoid cells positive for CD20 but negative for CD3. Epstein-Barr virus (EBV) -encoded RNA was detected by in situ hybridization, and Ki-67 staining demonstrated a higher percentage of positive cells. Consequently, an EBV-positive diffuse large B-cell lymphoma, developed as PTLD, was diagnosed. Complete remission was achieved with a reduced immunomodulatory drug dosage and rituximab therapy. She has been alive for 8 months postoperatively without recurrence. This case suggests that PTLD should be considered in assessing patients presenting with abdominal symptoms following organ transplantation.

DOI： 10.11405/nisshoshi.121.896

Alpha-fetoprotein and des-gamma-carboxy prothrombin can predict the objective response of patients with hepatocellular carcinoma receiving durvalumab plus tremelimumab therapy. International journal

Issei Saeki, Shigeo Shimose, Tetsu Tomonari, Takanori Ito, Joji Tani, Yasuto Takeuchi, Naoki Yoshioka, Takehito Naito, Mamiko Takeuchi, Satoru Kakizaki, Takeshi Hatanaka, Kyo Sasaki, Tetsuya Yasunaka, Masahiro Sakata, Hideki Iwamoto, Satoshi Itano, Tomotake Shirono, Norikazu Tanabe, Takafumi Yamamoto, Yuki Kanayama, Atsushi Naganuma, Sohji Nishina, Motoyuki Otsuka, Hideki Kobara, Hiroki Kawashima, Tetsuji Takayama, Takumi Kawaguchi, Takahiro Yamasaki, Taro Takami

PloS one 19 ( 9 ) e0311084 2024

Language：English Publishing type：Research paper (scientific journal)

Durvalumab plus tremelimumab (Durva/Treme) combined immunotherapy is the first-line therapy recommended for unresectable hepatocellular carcinoma (HCC). Since sequential therapy is more effective in improving prognosis, tumor markers have been used as predictive biomarkers for response to systemic therapy. This study aimed to investigate the predictive ability of objective response (OR) by tumor markers for Durva/Treme therapy against HCC. In this multicenter study, 110 patients with HCC who received Durva/Treme therapy were retrospectively enrolled. The OR rate was 15.5%. To aid early decision-making regarding OR, we evaluated the predictors contributing to OR in two steps: before (first step) and 4 weeks after (second step) treatment induction. Changes in tumor markers (alpha-fetoprotein [AFP] and des-gamma-carboxy prothrombin [DCP]) from baseline to 4 weeks after treatment (ΔAFP/ΔDCP) were included as the input factors. In the first step, multivariable analysis identified only the baseline AFP level (odds ratio 3.497, p = 0.029) as a predictor of OR. Patients with AFP ≥ 400 ng/mL had a significantly higher OR rate than those with < 400 ng/mL (28.2 vs. 8.5%, p = 0.011), and there was no significant difference in progression-free survival (PFS) between the two groups. When AFP/DCP response was defined as a ≥10% reduction from baseline, multivariable analysis showed that AFP response (odds ratio 6.023, p = 0.042) and DCP response (odds ratio 11.657, p = 0.006) were both independent predictors of OR in the second step. The PFS of patients with AFP or DCP response was significantly longer than that of patients without AFP or DCP response. The study demonstrated that the use of AFP and DCP can predict the OR of patients with HCC receiving Durva/Treme therapy.

DOI： 10.1371/journal.pone.0311084

Prospective observational study of zinc deficiency symptoms during first-line chemotherapy for gastric and colorectal cancer. International journal

Chihiro Sakaguchi, Tomohiro Nishina, Yoshiyasu Kono, Kaori Hino, Akinori Asagi, Yoshinori Ohno, Takeshi Kajiwara, Seijin Nadano, Natsumi Yamashita, Mikiko Tohyama, Ichinosuke Hyodo, Hiroyuki Okada, Motoyuki Otsuka

Journal of gastrointestinal oncology 14 ( 6 ) 2384 - 2394 2023.12

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Zinc deficiency during long-term chemotherapy and its related symptoms, including skin rash, taste disorders, and oral mucositis, have not been sufficiently investigated. METHODS: This prospective observational study enrolled patients with gastric and colorectal cancer who underwent standard first-line chemotherapy. According to the Practice Guidelines for Zinc Deficiency, zinc deficiency is defined as a serum level of <60 µg/dL. Serum zinc levels were measured before and after (1, 3, and 6 months) chemotherapy, and symptoms were assessed using the Patient-Reported Outcomes version of the Common Terminology Criteria for Adverse Events version 1.0. Repeated measures were analyzed using a generalized linear mixed model. RESULTS: Of the 61 enrolled patients, 48 who underwent standard first-line chemotherapy with fluoropyrimidine plus oxaliplatin were analyzed. Zinc deficiency was observed in 18 patients (38%) before chemotherapy. The least-squares means of serum zinc levels significantly decreased at 3 and 6 months of chemotherapy in 30 patients without zinc deficiency at the start of chemotherapy (both P<0.01) but not in 18 with zinc deficiency at the beginning. Changes in serum zinc levels during chemotherapy negatively correlated with changes in taste, rash, and itching (all P<0.04) in patients without zinc deficiency before treatment initiation. CONCLUSIONS: Serum zinc levels decreased during chemotherapy in zinc-non-deficient patients at the beginning of chemotherapy and correlated with taste changes, skin rash, and itching. Therefore, investigating whether zinc supplementation ameliorates these symptoms is necessary.

DOI： 10.21037/jgo-23-517

A small molecule iCDM-34 identified by in silico screening suppresses HBV DNA through activation of aryl hydrocarbon receptor. Reviewed International journal

Yutaka Furutani, Yoshinori Hirano, Mariko Toguchi, Shoko Higuchi, Xian-Yang Qin, Kaori Yanaka, Yumi Sato-Shiozaki, Nobuaki Takahashi, Marina Sakai, Pornparn Kongpracha, Takehiro Suzuki, Naoshi Dohmae, Mutsuko Kukimoto-Niino, Mikako Shirouzu, Shushi Nagamori, Harukazu Suzuki, Kaoru Kobayashi, Takahiro Masaki, Hiroo Koyama, Kazuma Sekiba, Motoyuki Otsuka, Kazuhiko Koike, Michinori Kohara, Soichi Kojima, Hideaki Kakeya, Tomokazu Matsuura

Cell Death Discovery 9 ( 1 ) 467 - 467 2023.12

Language：English Publishing type：Research paper (scientific journal)

IFN-alpha have been reported to suppress hepatitis B virus (HBV) cccDNA via APOBEC3 cytidine deaminase activity through interferon signaling. To develop a novel anti-HBV drug for a functional cure, we performed in silico screening of the binding compounds fitting the steric structure of the IFN-alpha-binding pocket in IFNAR2. We identified 37 compounds and named them in silico cccDNA modulator (iCDM)-1-37. We found that iCDM-34, a new small molecule with a pyrazole moiety, showed anti-HCV and anti-HBV activities. We measured the anti-HBV activity of iCDM-34 dependent on or independent of entecavir (ETV). iCDM-34 suppressed HBV DNA, pgRNA, HBsAg, and HBeAg, and also clearly exhibited additive inhibitory effects on the suppression of HBV DNA with ETV. We confirmed metabolic stability of iCDM-34 was stable in human liver microsomal fraction. Furthermore, anti-HBV activity in human hepatocyte-chimeric mice revealed that iCDM-34 was not effective as a single reagent, but when combined with ETV, it suppressed HBV DNA compared to ETV alone. Phosphoproteome and Western blotting analysis showed that iCDM-34 did not activate IFN-signaling. The transcriptome analysis of interferon-stimulated genes revealed no increase in expression, whereas downstream factors of aryl hydrocarbon receptor (AhR) showed increased levels of the expression. CDK1/2 and phospho-SAMHD1 levels decreased under iCDM-34 treatment. In addition, AhR knockdown inhibited anti-HCV activity of iCDM-34 in HCV replicon cells. These results suggest that iCDM-34 decreases the phosphorylation of SAMHD1 through CDK1/2, and suppresses HCV replicon RNA, HBV DNA, and pgRNA formation.

DOI： 10.1038/s41420-023-01755-w

Successful removal of a proximally migrated pancreatic stent using a novel device delivery system. Reviewed International journal

Akihiro Matsumi, Kazuyuki Matsumoto, Daisuke Uchida, Shigeru Horiguchi, Koichiro Tsutsumi, Hironari Kato, Motoyuki Otsuka

Endoscopy 55 ( S 01 ) E641-E642 2023.12

Usefulness of the artificial intelligence-mediated virtual chromoendoscopy in peroral cholangioscopy. Reviewed International journal

Ryosuke Sato, Kazuyuki Matsumoto, Hideaki Kinugasa, Daisuke Uchida, Shigeru Horiguchi, Hironari Kato, Motoyuki Otsuka

Endoscopy 55 ( S 01 ) E971-E972 2023.12

高齢者におけるステント治療の現状と工夫高齢者の急性胆嚢炎におけるETGBDとEUS-GBDの比較検討

服部直, 宮本和也, 松本和幸, 加藤博也, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 62 - 62 2023.11

膵神経内分泌腫瘍の治療中にうっ血性左側結腸炎を発症した1例

佐藤亮介, 堀口繁, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 松本和幸, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 84 - 84 2023.11

ロボット支援下膵頭十二指腸切除術後の捻転を伴う輸入脚症候群に対して内視鏡的整復を行った1例

原田圭, 藤井佑樹, 小幡泰介, 佐藤亮介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 112 - 112 2023.11

短期間で急速に進行し、根治外科切除を行うも遠隔転移再発を来した十二指腸癌の一例

渋谷香苗, 河野吉泰, 神尾知宏, 平田翔一郎, 倉岡紗樹子, 稲生祥子, 里見拓也, 松枝克典, 濱田健太, 岩室雅也, 川野誠司, 河原祥朗, 高木弘誠, 楳田祐三, 田中健大, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 75 - 75 2023.11

特発性肺線維症に対する肺移植後に重症膵炎を発症し、診断に苦慮した膵頭部癌の一例

矢杉賢吾, 堀口しげる, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井祐樹, 山崎辰洋, 内田大輔, 松本和幸, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 85 - 85 2023.11

消化器診療の先端診断と治療の新展開下咽頭表在癌スクリーニングにおける唾液中DCC遺伝子メチル化解析の有用性

平井亮佑, 衣笠秀明, 大塚基之

日本消化器病学会中国支部例会・日本消化器内視鏡学会中国支部例会プログラム・抄録集 120回・131回 45 - 45 2023.11

Cysts or necrotic components in pancreatic ductal adenocarcinoma is associated with the risk of EUS-FNA/B complications including needle tract seeding. International journal

Hiroyuki Terasawa, Kazuyuki Matsumoto, Takehiro Tanaka, Takeshi Tomoda, Taiji Ogawa, Yuki Ishihara, Tatsuya Kikuchi, Taisuke Obata, Takashi Oda, Akihiro Matsumi, Kazuya Miyamoto, Kosaku Morimoto, Yuki Fujii, Tatsuhiro Yamazaki, Daisuke Uchida, Shigeru Horiguchi, Koichiro Tsutsumi, Hironari Kato, Motoyuki Otsuka

Pancreatology : official journal of the International Association of Pancreatology (IAP) ... [et al.] 23 ( 8 ) 988 - 995 2023.10

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: EUS-FNA/B for pancreatic ductal adenocarcinoma (PDAC) is generally considered to be safe; however, while the incidence is low, there are occurrences of complications. Among these complications, there are serious ones like needle tract seeding (NTS), and it is not known than which types of tumors have the risks of EUS-FNA/B complications. This study aimed to evaluate the risk of EUS-FNA/B complications in patients with PDAC, focusing on morphological features. METHODS: Overall, 442 patients who underwent EUS-FNA/B for solid pancreatic masses between January 2018 and May 2022 in four institutions were retrospectively surveyed. Finally, 361 patients histopathologically diagnosed with PDAC were analyzed. Among these patients, 79 tumors with cysts or necrotic components were compared with 282 tumors without cysts or necrotic components. The incidence and risk of EUS-FNA/B complications including NTS were evaluated. RESULTS: There were 9 (2.4 %) of total EUS-FNA/B complications and 3 (0.8 %) of NTS. The incidence of total complication rate and NTS in tumors with cysts or necrotic components were significantly higher than in those without cysts or necrotic components (total complication 6.3 % vs. 1.4 %, p = 0.026, NTS 3.7 % vs. 0 %, p = 0.01). The transgastric route of puncture (OR: 93.3, 95 % CI: 3.81-2284.23) and the existence of cysts or necrotic components (OR: 7.3, 95 % CI: 1.47-36.19) were risk factors for EUS-FNA/B complications identified by the multivariate analysis. CONCLUSIONS: We should pay attention to the risks of EUS-FNA/B complications, including NTS, when the tumor has cysts or necrotic components.

DOI： 10.1016/j.pan.2023.10.018

Optimal Bowel Preparation Method to Visualize the Distal Ileum via Small Bowel Capsule Endoscopy. International journal

Daisuke Kametaka, Mamoru Ito, Seiji Kawano, Shuhei Ishiyama, Akiko Fujiwara, Junichirou Nasu, Masao Yoshioka, Junji Shiode, Kazuhide Yamamoto, Masaya Iwamuro, Yoshiro Kawahara, Hiroyuki Okada, Motoyuki Otsuka

Diagnostics (Basel, Switzerland) 13 ( 20 ) 2023.10

Language：English Publishing type：Research paper (scientific journal)

Small bowel capsule endoscopy (SBCE) is a convenient and minimally invasive method widely used to evaluate the small intestine. However, especially in the distal ileum, visualization of the intestinal mucosa is frequently hampered by the remaining intestinal contents, making it difficult to detect critical lesions. Although several studies have reported on the efficacy of bowel preparation before SBCE, no standardized protocol has been established. Herein, we determined the optimal preparation method for better visualization of the distal ileum using SBCE. We retrospectively analyzed 259 consecutive patients who had undergone SBCE between July 2009 and December 2019, divided into three groups: Group A (no preparation except overnight fasting), Group B (ingestion of 1-2 L polyethylene glycol 4 h before colonoscopy after overnight fasting and performing SBCE immediately after colonoscopy), and Group C (ingestion of 0.9 L magnesium citrate [MC] before SBCE after overnight fasting). The visibility of the intestinal mucosa in the first 10 min and at the last 10 min during the period of observation of the distal ileum was examined using a scoring system and compared. The visibility of the images captured by SBCE was assessed based on the scoring of the degree of bile/chyme staining, residual fluid and debris, brightness, bubble reduction, and visualized mucosa. The status of intestinal collapse was also assessed. In the first 10 min of observation of the distal ileum, no significant differences were detected among the groups. In the last 10 min, significantly better images were acquired in Group C in terms of bile/chyme staining, brightness, bubble reduction, and visualized mucosa. Bowel preparation using a low-dose MC solution 2 h before SBCE provided significantly higher-quality images of the distal ileum. Further optimization, such as the timing of initiating the preparation, is necessary to determine the optimal regimen for bowel preparation prior to SBCE.

DOI： 10.3390/diagnostics13203269

Idiopathic Gastric Antral Ulcers.

Masaya Iwamuro, Takehiro Tanaka, Seiji Kawano, Yoshiro Kawahara, Motoyuki Otsuka

Internal medicine (Tokyo, Japan) 63 ( 10 ) 1367 - 1371 2023.10

Language：English Publishing type：Research paper (scientific journal)

A Japanese woman presented with gastric antral ulcers accompanied by erosion and edema, demonstrating a chronic pattern of improvement and recurrence for more than six years. The patient had no relevant treatment history, and Helicobacter pylori infection was ruled out. Other potential etiologies contributing to gastric ulcers were eliminated on the basis of endoscopic biopsy and blood laboratory findings. Consequently, the patient was diagnosed with idiopathic gastric antral ulcer. This disease is often overlooked, and the chronological endoscopic images provided in this report can be used as a reference.

DOI： 10.2169/internalmedicine.2554-23

Interventional EUSを用いた診療戦略-最適な低侵襲医療を目指して小病変かつ低悪性度の膵神経内分泌腫瘍に対するEUSガイド下低侵襲治療

松本和幸, 加藤博也, 大塚基之

Gastroenterological Endoscopy 65 ( Suppl.2 ) 1838 - 1838 2023.10

AIを用いた経口胆道鏡検査の良悪性診断支援システムの開発

佐藤亮介, 松本和幸, 加藤博也, 河原祥朗, 大塚基之

Gastroenterological Endoscopy 65 ( Suppl.2 ) 2028 - 2028 2023.10

Endoscopic Manifestations and Clinical Characteristics of Localized Gastric Light-Chain Amyloidosis.

Masaya Iwamuro, Shouichi Tanaka, Tatsuya Toyokawa, Mamoru Nishimura, Takao Tsuzuki, Koji Miyahara, Shin Negishi, Shogen Ohya, Takehiro Tanaka, Motoyuki Otsuka

Acta medica Okayama 77 ( 5 ) 545 - 552 2023.10

Language：English Publishing type：Research paper (scientific journal)

To determine the endoscopic and clinical features of localized gastric amyloid light-chain (AL) amyloidosis, we retrospectively examined the characteristics of nine patients (eight men and one woman) encountered by the hospitals in our network. Lesions were predominantly flat and depressed with surface vascular dilatation (n=5); others were characterized by subepithelial lesions (n=2), mucosal color change (n=1), and a mass-like morphology with swollen mucosal folds (n=1). Colonoscopy (n=7), video capsule enteroscopy (n=2), serum (n=5) and urine immunoelectrophoresis (n=4), and bone marrow examination (n=3) were performed to exclude involvement of organs other than the stomach. As treatment for gastric lesions of AL amyloidosis, one patient each underwent endoscopic submucosal dissection (n=1) and argon plasma coagulation (n=1), while the remaining seven patients underwent no specific treatment. During a mean follow-up of 4.2 years, one patient died 3.2 years after diagnosis, but the cause of death, which occurred in another hospital, was unknown. The remaining eight patients were alive at the last visit. In conclusion, although localized gastric AL amyloidosis can show various macroscopic features on esophagogastroduodenoscopy, flat, depressed lesions with vascular dilatation on the surface are predominant.

DOI： 10.18926/AMO/65978

高齢者の食道表在癌・早期胃癌に対する内視鏡治療の適応と問題点後期高齢表在食道癌患者に対する内視鏡治療後長期成績の検討

松枝克典, 川野誠司, 大塚基之

日本消化器病学会雑誌 120 ( 臨増大会 ) A586 - A586 2023.10

消化管領域における希少疾患に対する内視鏡診療消化管希少疾患 gastric adenocarcinoma and proximal polyposis of the stomach(GAPPS)の2家系の報告

平井亮佑, 川野誠司, 大塚基之

Gastroenterological Endoscopy 65 ( Suppl.2 ) 1906 - 1906 2023.10

The efficacy of non-anesthesiologist-administered propofol sedation with a target-controlled infusion system during double-balloon endoscopic retrograde cholangiopancreatography. International journal

Kazuya Miyamoto, Kazuyuki Matsumoto, Taisuke Obata, Ryosuke Sato, Akihiro Matsumi, Kosaku Morimoto, Taiji Ogawa, Hiroyuki Terasawa, Yuki Fujii, Tatsuhiro Yamazaki, Daisuke Uchida, Shigeru Horiguchi, Koichiro Tsutsumi, Hironari Kato, Motoyuki Otsuka

BMC gastroenterology 23 ( 1 ) 296 - 296 2023.9

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: The sedation method used during double-balloon endoscopic retrograde cholangiopancreatography (DB-ERCP) differs among countries and/or facilities, and there is no established method. This study aimed to evaluate the efficacy of non-anesthesiologist-administered propofol (NAAP) sedation using a target-controlled infusion (TCI) system during DB-ERCP. METHODS: This retrospective study was conducted between May 2017 and December 2020 at an academic center. One hundred and fifty-six consecutive patients who underwent DB-ERCP were sedated by gastroenterologists using diazepam (n = 77) or propofol with a TCI system (n = 79), depending on the period. The primary endpoint was a comparison of poor sedation rates between the two groups. Poor sedation was defined as a condition requiring the use of other sedative agents or discontinuation of the procedure. Secondary endpoints were sedation-related adverse events and risk factors for poor sedation. RESULTS: Poor sedation occurred significantly more often in the diazepam sedation group (diazepam sedation, n = 12 [16%] vs. propofol sedation, n = 1 [1%]; P = 0.001). Vigorous body movements (3 or 4) (diazepam sedation, n = 40 [52%] vs. propofol sedation, n = 28 [35%]; P = 0.038) and hypoxemia (< 85%) (diazepam sedation, n = 7 [9%] vs. propofol sedation, n = 1 [1%]; P = 0.027) occurred significantly more often in the diazepam sedation group. In the multivariate analysis, age < 70 years old (OR, 10.26; 95% CI, 1.57-66.98; P = 0.015), BMI ≥ 25 kg/m2 (OR, 11.96; 95% CI, 1.67-85.69; P = 0.014), and propofol sedation (OR, 0.06; 95% CI, 0.01-0.58; P = 0.015) were associated factors for poor sedation. CONCLUSIONS: NAAP sedation with the TCI system during DB-ERCP was safer and more effective than diazepam sedation.

DOI： 10.1186/s12876-023-02936-8

先天性胆道拡張症術後の肝内胆管狭窄に対し内視鏡的胆道拡張術が有効であった一例

宮本和也, 加藤博也, 小幡泰介, 佐藤亮介, 松三明宏, 森本光作, 藤井佑樹, 山崎辰洋, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 大塚基之

日本膵・胆管合流異常研究会プロシーディングス 46 76 - 76 2023.9

IBD診療におけるtreat to targetを考える Treat to targetを意識したクローン病におけるsurrogate markerの組み合わせ

井口俊博, 平岡佐規子, 青山祐樹, 竹井健介, 井川翔子, 高原政宏, 大塚基之

日本大腸肛門病学会雑誌 76 ( 9 ) A51 - A51 2023.9

胆膵癌ゲノムパネル検査における検査法についての検討(Investigation of test methods for pancreatobiliary cancer genomic panel test)

堀口繁, 加藤博也, 松本和幸, 大塚基之

日本癌学会総会記事 82回 828 - 828 2023.9

Biological Roles of Hepatitis B Viral X Protein in the Viral Replication and Hepatocarcinogenesis.

Motoyuki Otsuka

Acta medica Okayama 77 ( 4 ) 341 - 345 2023.8

Language：English Publishing type：Research paper (scientific journal)

Hepatitis B virus is a pathogenic virus that infects 300 million people worldwide and causes chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Hepatitis B virus encodes four proteins. Among them, the HBx protein plays a central role in the HBV pathogenesis. Because the HBx protein is considered to play a central role in the induction of viral replication and hepatocarcinogenesis, the regulation of its function could be a key factor in the development of new interventions against hepatitis B. In this review, HBx protein-related viral replication and hepatocarcinogenesis mechanisms are described, with a focus on the recently reported viral replication mechanisms related to degradation of the Smc5/6 protein complex. We also discuss our recent discovery of a compound that inhibits HBx protein-induced degradation of the Smc5/6 protein complex, and that exerts inhibitory effects on both viral replication and hepatocarcinogenesis. Finally, prospects for future research on the HBx protein are described.

DOI： 10.18926/AMO/65739

【胃良性疾患の近未来】[薬剤関連疾患]White globe appearance(WGA)

岩室雅也, 田中健大, 大塚基之

消化器内視鏡 35 ( 8 ) 1072 - 1074 2023.8

「術後胆管炎の診断と治療の現状」再発リスク因子からみた術後良性胆管空腸吻合部狭窄に対する最適な内視鏡治療戦略

松三明宏, 松本和幸, 大塚基之

胆道 37 ( 3 ) 417 - 417 2023.8

「難治性胆汁漏に対する治療」重症度からみた術後胆汁漏に対する内視鏡治療成績

小幡泰介, 松本和幸, 大塚基之

胆道 37 ( 3 ) 423 - 423 2023.8

Oxidative stress-related markers as prognostic factors for patients with primary sclerosing cholangitis in Japan. Reviewed International journal

Atsushi Oyama, Akinobu Takaki, Takuya Adachi, Nozomu Wada, Yasuto Takeuchi, Hideki Onishi, Hidenori Shiraha, Hiroyuki Okada, Motoyuki Otsuka

Hepatology international 17 ( 5 ) 1215 - 1224 2023.7

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND/PURPOSE: Primary sclerosing cholangitis (PSC) is a rare chronic liver disease. The mechanisms and prediction of PSC progression are unclear. Recent investigations have shown that general conditions, such as oxidative stress, affect the course of chronic diseases. We investigated the clinical course and oxidative stress-related condition of PSC to determine prognostic factors. METHODS: We recruited 58 patients with PSC (mean age; 37.4 years, mean observation period; 1382 days) who visited our department from 2003 to 2021. Clinical characteristics were investigated to define prognostic factors. Oxidative stress status was evaluated using two types of markers: an oxidative stress marker (serum reactive oxygen metabolite; dROM) and an antioxidant marker (serum OXY adsorbent test; OXY). RESULTS: The revised Mayo risk, Child-Pugh, model for end-stage liver disease-sodium (MELD-Na) scores or fibrosis-related FIB-4 index significantly predicted poor overall survival. High intestinal immunoglobulin A (IgA) levels predicted poor survival. Among patients with high and intermediate revised Mayo risk scores, those with physiologically high dROM levels showed better survival than those with lower dROM levels. In this population, dROM was negatively correlated with AST and IgA, which are both correlated with survival. CONCLUSIONS: High and intermediate revised Mayo risk score group predicted a poor clinical course in PSC. Additionally, the Child-Pugh score, MELD-Na score, FIB-4 index, and serum IgA were significantly correlated with survival. In patients with high and intermediate revised Mayo risk scores, physiologically high oxidative stress status correlated with low IgA levels and a good prognosis.

DOI： 10.1007/s12072-023-10557-2

Review of oral and pharyngolaryngeal benign lesions detected during esophagogastroduodenoscopy. Reviewed International journal

Masaya Iwamuro, Kenta Hamada, Seiji Kawano, Yoshiro Kawahara, Motoyuki Otsuka

World journal of gastrointestinal endoscopy 15 ( 7 ) 496 - 509 2023.7

Language：English Publishing type：Research paper (scientific journal)

Recent advancements in endoscopy equipment have facilitated endoscopists' detection of neoplasms in the oral cavity and pharyngolaryngeal regions. In particular, image-enhanced endoscopy using narrow band imaging or blue laser imaging play an integral role in the endoscopic diagnosis of oral and pharyngolaryngeal cancers. Despite these advancements, limited studies have focused on benign lesions that can be observed during esophagogastroduodenoscopy in the oral and pharyngolaryngeal regions. Therefore, this mini-review aimed to provide essential information on such benign lesions, along with representative endoscopic images of dental caries, cleft palate, palatal torus, bifid uvula, compression by cervical osteophytes, tonsil hyperplasia, black hairy tongue, oral candidiasis, oral and pharyngolaryngeal ulcers, pharyngeal melanosis, oral tattoos associated with dental alloys, retention cysts, papilloma, radiation-induced changes, skin flaps, vocal cord paresis, and vocal fold leukoplakia. Whilst it is imperative to seek consultation from otolaryngologists or dentists in instances where the diagnosis cannot be definitively ascertained by endoscopists, the merits of attaining foundational expertise pertaining to oral and pharyngolaryngeal lesions are unequivocal. This article will be a valuable resource for endoscopists seeking to enhance their understanding of oral and pharyngolaryngeal lesions.

DOI： 10.4253/wjge.v15.i7.496

当院における術後膵空腸吻合部狭窄に対する内視鏡治療成績の検討

森本光作, 山崎辰洋, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 38 ( 3 ) A337 - A337 2023.7

結節を有するIPMNの良悪性診断における新規TIC解析プログラムを用いた造影EUSの有用性

宮本和也, 松本和幸, 内田大輔, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 38 ( 3 ) A345 - A345 2023.7

結節を有するIPMNの良悪性診断における新規TIC解析プログラムを用いた造影EUSの有用性

宮本和也, 松本和幸, 内田大輔, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 38 ( 3 ) A345 - A345 2023.7

当院における術後膵空腸吻合部狭窄に対する内視鏡治療成績の検討

森本光作, 山崎辰洋, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

膵臓 38 ( 3 ) A337 - A337 2023.7

高齢者における膵癌術前化学療法の有効性と安全性の検討

藤井佑樹, 堀口繁, 佐藤亮介, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堤康一郎, 加藤博也, 大塚基之

膵臓 38 ( 3 ) A316 - A316 2023.7

Endoscopic evaluation by the Kyoto classification of gastritis combined with serum anti-Helicobacter pylori antibody testing reliably risk-stratifies subjects in a population-based gastric cancer screening program. Reviewed

Ryosuke Hirai, Mami Hirai, Motoyuki Otsuka, Toshiharu Mitsuhashi, Yuichi Shimodate, Hirokazu Mouri, Kazuhiro Matsueda, Hiroshi Yamamoto, Motowo Mizuno

Journal of gastroenterology 58 ( 9 ) 848 - 855 2023.6

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: We previously demonstrated that the Kyoto classification of gastritis was useful for judging the status of Helicobacter pylori infection in a population-based screening program, and that adding H. pylori antibody test improved its accuracy (UMIN000028629). Here, we tested whether our endoscopic diagnosis of H. pylori infection status reliably estimated gastric cancer risk in the program. METHODS: Data were collected from1345 subjects who underwent endoscopic follow-up 4 years after the end of the registration. We analyzed the association of three diagnostic methods of H. pylori infection with gastric cancer detection: (1) endoscopic diagnosis based on the Kyoto classification of gastritis; (2) serum diagnosis according to the ABC method (H. pylori antibody and pepsinogen I and II); and (3) endoscopic diagnosis together with H. pylori antibody test. RESULTS: During the follow-up, 19 cases of gastric cancer were detected. By Kaplan-Meier analysis, the detection rates of cancer were significantly higher in the past or current H. pylori infection groups than in the never-infected group with all 3 methods. By the Cox proportional hazards model, the hazard ratio for cancer detection was highest in evaluation with the combined endoscopic diagnosis and the antibody test (method 3; hazard ratio 22.6, 95% confidence interval 2.99-171) among the three methods (the endoscopic diagnosis (method 1); 11.3, 2.58-49.8, and the ABC method (method 2); 7.52, 2.49-22.7). CONCLUSIONS: Endoscopic evaluation of H. pylori status with the Kyoto classification of gastritis, especially combined with serum anti-Helicobacter pylori antibody testing, reliably risk-stratified subjects in a population-based gastric cancer screening program.

DOI： 10.1007/s00535-023-02010-w

Update in Molecular Aspects and Diagnosis of Autoimmune Gastritis. Reviewed International journal

Masaya Iwamuro, Takehiro Tanaka, Motoyuki Otsuka

Current issues in molecular biology 45 ( 7 ) 5263 - 5275 2023.6

Language：English Publishing type：Research paper (scientific journal)

Recent studies have advanced our understanding of the pathophysiology of autoimmune gastritis, particularly its molecular aspects. The most noteworthy recent advancement lies in the identification of several candidate genes implicated in the pathogenesis of pernicious anemia through genome-wide association studies. These genes include PTPN22, PNPT1, HLA-DQB1, and IL2RA. Recent studies have also directed attention towards other genes such as ATP4A, ATP4B, AIRE, SLC26A7, SLC26A9, and BACH2 polymorphism. In-depth investigations have been conducted on lymphocytes and cytokines, including T helper 17 cells, interleukin (IL)-17A, IL-17E, IL-17F, IL-21, IL-19, tumor necrosis factor-α, IL-15, transforming growth factor-β1, IL-13, and diminished levels of IL-27. Animal studies have explored the involvement of roseolovirus and H. pylori in relation to the onset of the disease and the process of carcinogenesis, respectively. Recent studies have comprehensively examined the involvement of autoantibodies, serum pepsinogen, and esophagogastroduodenoscopy in the diagnosis of autoimmune gastritis. The current focus lies on individuals demonstrating atypical presentations of the disease, including those diagnosed in childhood, those yielding negative results for autoantibodies, and those lacking the typical endoscopic characteristics of mucosal atrophy. Here, we discuss the recent developments in this field, focusing on genetic predisposition, epigenetic modifications, lymphocytes, cytokines, oxidative stress, infectious agents, proteins, microRNAs, autoantibodies, serum pepsinogen, gastrin, esophagogastroduodenoscopy and microscopic findings, and the risk of gastric neoplasm.

DOI： 10.3390/cimb45070334

肝内結石に対して内視鏡用イントロデューサーEndoSheatherが有用であった1例

佐藤亮介, 藤井佑樹, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堤康一郎, 堀口繁, 加藤博也, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 83 - 83 2023.6

悪性輸入脚症候群に対して超音波内視鏡下消化管ドレナージが有効であった1例

森本光作, 松本和幸, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 83 - 83 2023.6

EUS関連手技の言語化した教育法

松本和幸, 加藤博也, 堤康一郎, 大塚基之

日本消化器病学会四国支部例会プログラム・抄録集 119回 37 - 37 2023.6

長期経過を追えた膵Intraductal oncocytic papillary neoplasm(IOPN)の1例

藤井佑樹, 佐藤亮介, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 67 - 67 2023.6

進行胆道癌に対するゲムシタビン・シスプラチン・S-1併用療法の治療成績についての検討

小幡泰介, 松本和幸, 佐藤亮介, 菊池達也, 織田崇志, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 65 - 65 2023.6

長期経過を追えた膵Intraductal oncocytic papillary neoplasm(IOPN)の1例

藤井佑樹, 佐藤亮介, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 67 - 67 2023.6

進行胆道癌に対するゲムシタビン・シスプラチン・S-1併用療法の治療成績についての検討

小幡泰介, 松本和幸, 佐藤亮介, 菊池達也, 織田崇志, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 65 - 65 2023.6

肝内結石に対して内視鏡用イントロデューサーEndoSheatherが有用であった1例

佐藤亮介, 藤井佑樹, 小幡泰介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堤康一郎, 堀口繁, 加藤博也, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 83 - 83 2023.6

悪性輸入脚症候群に対して超音波内視鏡下消化管ドレナージが有効であった1例

森本光作, 松本和幸, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 宮本和也, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 83 - 83 2023.6

Helicobacter pylori除菌により寛解し、長期間経過後に再発した胃MALTリンパ腫の一例

稲生祥子, 岩室雅也, 平田翔一郎, 平井亮佑, 倉岡紗樹子, 小橋真由, 岡上昇太郎, 里見拓也, 松枝克典, 濱田健太, 河野吉泰, 川野誠司, 河原祥朗, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 71 - 71 2023.6

免疫関連有害事象(irAE:immune-related adverse events)による胃炎の5例

岩室雅也, 田中健大, 平田翔一郎, 河野吉泰, 川野誠司, 河原祥朗, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 70 - 70 2023.6

内反性増殖を示した十二指腸腺癌に対してUnderwater EMRにて切除し得た1例

松枝克典, 川野誠司, 平田翔一郎, 岡上昇太郎, 里見拓也, 稲生祥子, 濱田健太, 河野吉泰, 岩室雅也, 河原祥朗, 田中健大, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 76 - 76 2023.6

肝生検により診断された播種性bacillus Calmette-Guerin(BCG)症の1例

伊木道子, 竹内康人, 須江真彦, 三宅望, 足立卓哉, 和田望, 大西秀樹, 白羽秀則, 高木章乃夫, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 62 - 62 2023.6

血流豊富な粘膜下腫瘍の形態を呈した上行結腸血管周囲類上皮細胞腫の1例

青山祐樹, 井口俊博, 平岡佐規子, 豊澤惇希, 竹井健介, 井川翔子, 山崎泰史, 衣笠秀明, 高原政宏, 庄司良平, 近藤喜太, 柳井広之, 大塚基之

日本消化器内視鏡学会中国支部例会 130回 81 - 81 2023.6

免疫関連有害事象(irAE:immune-related adverse events)による胃炎の5例

岩室雅也, 田中健大, 平田翔一郎, 河野吉泰, 川野誠司, 河原祥朗, 大塚基之

日本消化器病学会中国支部例会プログラム・抄録集 119回 70 - 70 2023.6

Collagenous Colitis in a Patient With Gastric Cancer Who Underwent Chemotherapy. Reviewed International journal

Masaya Iwamuro, Takehiro Tanaka, Shunsuke Kagawa, Shoko Inoo, Motoyuki Otsuka

Cureus 15 ( 5 ) e39466 2023.5

Language：English

Herein, we present a case of collagenous colitis in a patient who underwent chemotherapy for gastric cancer, comprising five cycles of S-1 plus oxaliplatin and trastuzumab, followed by five cycles of paclitaxel and ramucirumab and seven cycles of nivolumab. The subsequent initiation of trastuzumab deruxtecan chemotherapy led to the development of grade 3 diarrhea after the second cycle of treatment. Collagenous colitis was diagnosed via colonoscopy and biopsy. The patient's diarrhea improved following the cessation of lansoprazole. This case highlights the importance of considering collagenous colitis as a differential diagnosis, in addition to chemotherapy-induced colitis and immune-related adverse event (irAE) colitis, in patients with similar clinical presentations.

DOI： 10.7759/cureus.39466

選択的胆管seekingのためのデバイス選択

小幡泰介, 内田大輔, 加藤博也, 佐藤亮介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 松本和幸, 堀口繁, 堤康一郎, 大塚基之

Gastroenterological Endoscopy 65 ( Suppl.1 ) 971 - 971 2023.4

選択的胆管seekingのためのデバイス選択

小幡泰介, 内田大輔, 加藤博也, 佐藤亮介, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 松本和幸, 堀口繁, 堤康一郎, 大塚基之

Gastroenterological Endoscopy 65 ( Suppl.1 ) 971 - 971 2023.4

EUS-HGSにおけるダブルガイドワイヤー法の有用性

松三明宏, 藤井佑樹, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 宮本和也, 森本光作, 寺澤裕之, 山崎辰洋, 内田大輔, 松本和幸, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会雑誌 120 ( 臨増総会 ) A321 - A321 2023.3

後期高齢者におけるIPMNの治療移行割合についての検討

宮本和也, 松本和幸, 小幡泰介, 佐藤亮介, 織田崇志, 菊池達也, 松三明宏, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 内田大輔, 堀口繁, 堤康一郎, 加藤博也, 大塚基之

日本消化器病学会雑誌 120 ( 臨増総会 ) A378 - A378 2023.3

胆嚢オルガノイドを用いた発癌モデルより同定したmiR-34a-5p補充による胆嚢癌治療の開発

織田崇志, 堤康一郎, 小幡泰介, 菊池達也, 松三明宏, 宮本和也, 森本光作, 寺澤裕之, 藤井佑樹, 山崎辰洋, 赤穂宗一郎, 内田大輔, 松本和幸, 堀口繁, 加藤博也, 大塚基之

日本消化器病学会雑誌 120 ( 臨増総会 ) A328 - A328 2023.3

Optimal liver drainage rate for survival in patients with unresectable malignant hilar biliary obstruction using 3D-image volume analyzer. International journal

Kosaku Morimoto, Kazuyuki Matsumoto, Taisuke Obata, Takashi Oda, Kazuya Miyamoto, Akihiro Matsumi, Hiroyuki Terasawa, Yuki Fujii, Tatsuhiro Yamazaki, Shigeru Horiguchi, Koichiro Tsutsumi, Hironari Kato, Motoyuki Otsuka

Therapeutic advances in gastroenterology 16 17562848231206980 - 17562848231206980 2023

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Drainage exceeding 50% of total liver volume is a beneficial prognostic factor in patients with unresectable malignant hilar biliary obstruction (UMHBO). However, it is unclear what threshold percentage of total liver volume drained ('liver drainage rate') significantly improves survival in patients with UMHBO who received systemic chemotherapy. OBJECTIVES: We aimed to assess the optimal liver drainage rate that improves survival in patients with UMHBO receiving chemotherapy using a three-dimensional (3D)-image volume analyzer. DESIGN: This study was a single-center retrospective cohort study. METHODS: Data from 90 patients with UMHBO who received chemotherapy after endoscopic biliary drainage using metal stents at Okayama University Hospital from January 2003 to December 2020 were reviewed. The liver drainage rate was calculated by dividing the drained liver volume by the total liver volume using a 3D-image volume analyzer. The primary endpoint was overall survival by liver drainage rate. The secondary endpoints were time to recurrent biliary obstruction (TRBO) and prognostic factors. RESULTS: The median total liver volume was 1172 (range: 673-2032) mL, and the median liver drainage rate was 83% (range: 50-100). Overall survival was 376 (95% CI: 271-450) days, and patients with >80% drainage (n = 67) had significantly longer survival than those with <80% drainage (n = 23) (450 days versus 224 days, p = 0.0033, log-rank test). TRBO was 201 (95% CI: 155-327) days and did not differ significantly by liver drainage rate. Multivariate Cox proportional hazards regression analysis revealed >80% liver drainage [hazard ratio (HR): 0.35, 95% CI: 0.20-0.62, p = 0.0003] and hilar cholangiocarcinoma (HR: 0.30, 95% CI: 0.17-0.50, p < 0.0001) as significant prognostic factors. CONCLUSION: In patients with UMHBO scheduled for chemotherapy, >80% drainage is associated with improved survival. Further prospective multicenter studies are needed to verify the results of this study. TRAIL REGISTRATION: Okayama University Hospital, IRB number: 2108-011.

DOI： 10.1177/17562848231206980

Smaller extracellular vesicles are released from pancreatic cancer cells by the alteration of the lipid composition under low glucose conditions. Reviewed International journal

Chikako Shibata, Motoyuki Otsuka, Takahiro Seimiya, Kazunaga Ishigaki, Yu Miyakawa, Takahiro Kishikawa, Mitsuhiro Fujishiro

Biochemical and biophysical research communications 637 314 - 321 2022.12

Language：English Publishing type：Research paper (scientific journal)

Extracellular vesicles (EVs) released from cells into the blood facilitate intercellular communication and serve as new biomarkers to understand the pathophysiology of several conditions. Although the importance of the cargo inside EVs has been extensively studied, the sizes of EVs that vary with different types of cancers are relatively poorly explored. Here, we show that pancreatic cancer cell-derived EVs are significantly smaller than non-cancer cell-derived EVs. The smaller size distribution of these EVs was confirmed by specifically isolating and examining tumor-derived EVs from the heterogeneous EV population isolated from the sera of patients with pancreatic ductal adenocarcinoma. In vitro analyses mimicking tumor microenvironment conditions revealed that low glucose conditions reduced the size distribution and increased the level of unsaturated fatty acids in the tumor-derived EVs. Because the lipid composition defines the fluidity of the membrane, the results suggest that the alterations in the size of EVs could be due to the alteration of the fluidity and stability of the membrane covering the EVs. Furthermore, the uptake of smaller EVs by recipient cells was increased, which may lead to enhanced functional results. These results provide fundamental insights into the factors defining the size of EVs, which may be important for developing cancer screening methods and understanding cancer-related pathophysiology.

DOI： 10.1016/j.bbrc.2022.11.040

HNF1B-driven three-dimensional chromatin structure for molecular classification in pancreatic cancers. Reviewed International journal

Hiroyuki Kato, Keisuke Tateishi, Dosuke Iwadate, Keisuke Yamamoto, Hiroaki Fujiwara, Takuma Nakatsuka, Yotaro Kudo, Yoku Hayakawa, Hideaki Ijichi, Motoyuki Otsuka, Takahiro Kishikawa, Ryota Takahashi, Koji Miyabayashi, Yousuke Nakai, Yoshihiro Hirata, Atsushi Toyoda, Shinichi Morishita, Mitsuhiro Fujishiro

Cancer science 114 ( 4 ) 1672 - 1685 2022.12

Language：English Publishing type：Research paper (scientific journal)

The molecular subtypes of pancreatic cancer (PC), either classical/progenitor-like or basal/squamous-like, are currently a major topic of research because of their direct association with clinical outcomes. Some transcription factors (TFs) have been reported to be associated with these subtypes. However, the mechanisms by which these molecular signatures of PCs are established remain unknown. Epigenetic regulatory processes, supported by dynamic changes in the chromatin structure, are essential for transcriptional profiles. Previously, we reported the importance of open chromatin profiles in the biological features and transcriptional status of PCs. Here, we aimed to analyze the relationships between three-dimensional (3D) genome structures and the molecular subtypes of human PCs using Hi-C analysis. We observed a correlation of the specific elements of 3D genome modules, including compartments, topologically associating domains, and enhancer-promoter loops, with the expression of related genes. We focused on HNF1B, a TF that is implicated in the progenitor subtype. Forced expression of HNF1B in squamous-type PC organoids induced the upregulation and downregulation of genes associated with progenitor and squamous subtypes, respectively. Long-range genomic interactions induced by HNF1B were accompanied by compartment modulation and H3K27ac redistribution. We also found that these HNF1B-induced changes in subtype-related gene expression required an intrinsically disordered region, suggesting a possible involvement of phase separation in compartment modulation. Thus, mapping of 3D structural changes induced by TFs, such as HNF1B, may become a useful resource for further understanding the molecular features of PCs.

DOI： 10.1111/cas.15690

The RNA-Binding Protein ELAVL1 Regulates Hepatitis B Virus Replication and Growth of Hepatocellular Carcinoma Cells. Reviewed International journal

Hiroaki Kanzaki, Tetsuhiro Chiba, Tatsuya Kaneko, Junjie Ao, Motoyasu Kan, Ryosuke Muroyama, Shingo Nakamoto, Tatsuo Kanda, Hitoshi Maruyama, Jun Kato, Yoh Zen, Ai Kotani, Kazuma Sekiba, Motoyuki Otsuka, Masayuki Ohtsuka, Naoya Kato

International journal of molecular sciences 23 ( 14 ) 2022.7

Language：English Publishing type：Research paper (scientific journal)

Previous RNA immunoprecipitation followed by proteomic approaches successfully demonstrated that Embryonic Lethal, Abnormal Vision, Drosophila-Like 1 (ELAVL1) interacts with hepatitis B virus (HBV)-derived RNAs. Although ELAVL family proteins stabilize AU-rich element (ARE)-containing mRNAs, their role in HBV transcription remains unclear. This study conducted loss-of-function assays of ELAVL1 for inducible HBV-replicating HepAD38 cells and HBx-overexpressed HepG2 cells. In addition, clinicopathological analyses in primary hepatocellular carcinoma (HCC) surgical samples were also conducted. Lentivirus-mediated short hairpin RNA knockdown of ELAVL1 resulted in a decrease in both viral RNA transcription and production of viral proteins, including HBs and HBx, probably due to RNA stabilization by ELAVL1. Cell growth of HepAD38 cells was more significantly impaired in ELAVL1-knockdown than those in the control group, with or without HBV replication, indicating that ELAVL1 is involved in proliferation by factors other than HBV-derived RNAs. Immunohistochemical analyses of 77 paired HCC surgical specimens demonstrated that diffuse ELAVL1 expression was detected more frequently in HCC tissues (61.0%) than in non-tumor tissues (27.3%). In addition, the abundant expression of ELAVL1 tended to affect postoperative recurrence in HBV-related HCC patients. In conclusion, ELAVL1 contributes not only to HBV replication but also to HCC cell growth. It may be a potent therapeutic target for HBV-related HCC treatment.

DOI： 10.3390/ijms23147878

Hepatitis B virus-associated hepatocellular carcinoma with Smc5/6 complex deficiency is susceptible to PARP inhibitors. Reviewed International journal

Kazuyoshi Funato, Motoyuki Otsuka, Kazuma Sekiba, Yu Miyakawa, Takahiro Seimiya, Chikako Shibata, Takahiro Kishikawa, Mitsuhiro Fujishiro

Biochemical and biophysical research communications 607 89 - 95 2022.6

Language：English Publishing type：Research paper (scientific journal)

DNA repair processes represent attractive synthetic lethal targets because many cancers exhibit impaired DNA repair pathways, which leads to dependence on specific repair proteins. The finding that poly (ADP-ribose) polymerase (PARP)-1 inhibitors are highly effective against cancers with deficient homologous recombination highlights the potential of this approach. In hepatitis B viral (HBV) infection, degradation of the structural maintenance of the chromosome 5/6 (Smc5/6) complex, which plays a key role in repairing double-stranded DNA breaks by homologous recombination, is induced by HBV regulatory protein X (HBx). Here, we hypothesized that a deficiency in the Smc5/6 complex in HBV-associated hepatocellular carcinoma (HCC) increases susceptibility to PARP inhibitors via a deficiency in homologous recombination. We confirmed impaired double-stranded DNA break repair in HBx-expressing HCC cells using a sensitive reporter to monitor homologous recombination. Treatment with a PARP inhibitor was significantly more effective against HBx-expressing HCC cells, and overexpression of Smc5/6 prevented these effects. Overall, our results suggest that homologous recombination deficiency in HBV-associated HCC leads to increased susceptibility to PARP inhibitors.

DOI： 10.1016/j.bbrc.2022.03.137

Roles of circular RNAs in the pathogenesis and treatment of pancreatic cancer. Reviewed International journal

Takahiro Seimiya, Motoyuki Otsuka, Mitsuhiro Fujishiro

Frontiers in cell and developmental biology 10 1023332 - 1023332 2022

Language：English Publishing type：Research paper (scientific journal)

Circular RNAs are single-stranded RNAs with a covalently closed structure formed by the process of back-splicing. Aberrant expression of circular RNAs contributes to the pathogenesis of a wide range of cancers. Pancreatic cancer is one of the most lethal cancers due to diagnostic difficulties and limited therapeutic options. Circular RNAs are emerging as novel diagnostic biomarkers and therapeutic targets for pancreatic cancer. Moreover, recent advances in the therapeutic application of engineered circular RNAs have provided a promising approach to overcoming pancreatic cancer. This review discusses the roles of circular RNAs in the pathogenesis of pancreatic cancer and in potential treatment applications and their usefulness as diagnostic biomarkers.

DOI： 10.3389/fcell.2022.1023332

Evidence-based clinical practice guidelines for nonalcoholic fatty liver disease/nonalcoholic steatohepatitis 2020. International journal

Motoyuki Otsuka

Hepatology research : the official journal of the Japan Society of Hepatology 51 ( 10 ) 1013 - 1025 2021.9

Language：English Publishing type：Research paper (scientific journal)

Nonalcoholic fatty liver disease (NAFLD) has become a serious public health issue not only in Western countries but also in Japan. Within the wide spectrum of NAFLD, nonalcoholic steatohepatitis (NASH) is a progressive form of disease that often develops into liver cirrhosis and increases the risk of hepatocellular carcinoma (HCC). While a definite diagnosis of NASH requires liver biopsy to confirm the presence of hepatocyte ballooning, hepatic fibrosis is the most important prognostic factor in NAFLD. With so many NAFLD patients, it is essential to have an effective screening method for NAFLD with hepatic fibrosis. As HCC with non-viral liver disease has increased markedly in Japan, effective screening and surveillance of HCC are also urgently needed. The most common death etiology in NAFLD patients is cardiovascular disease event. Gastroenterologists must, therefore, pay close attention to CVD when examining NAFLD patients. In the updated guidelines, we propose screening and follow-up methods for hepatic fibrosis, HCC, and CVD in NAFLD patients. Several drug trials are ongoing for NAFLD/NASH therapy, however, there is currently no specific drug therapy for NAFLD/NASH. In addition to vitamin E and thiazolidinedione derivatives, recent trials have focused on sodium glucose co-transporter 2 (SGLT2) inhibitors and glucagon-like peptide-1 (GLP-1) analogues, and effective therapies are expected to be developed. These practical guidelines for NAFLD/NASH were established by the Japanese Society of Gastroenterology in conjunction with the Japan Society of Hepatology. Clinical evidence reported internationally between 1983 and October 2018 was collected, and each clinical and background question was evaluated using the Grades of Recommendation Assessment, Development and Evaluation (GRADE) system. This English summary pro- vides the core essentials of these clinical practice guidelines, which include the definition and concept, screening systems for hepatic fibrosis, HCC and CVD, and current therapies for NAFLD/NASH in Japan.

DOI： 10.1111/hepr.13688

Evidence-based clinical practice guidelines for nonalcoholic fatty liver disease/nonalcoholic steatohepatitis 2020.

Motoyuki Otsuka

Journal of gastroenterology 56 ( 11 ) 951 - 963 2021.9

Language：English Publishing type：Research paper (scientific journal)

Nonalcoholic fatty liver disease (NAFLD) has become a serious public health issue not only in Western countries but also in Japan. Within the wide spectrum of NAFLD, nonalcoholic steatohepatitis (NASH) is a progressive form of disease that often develops into liver cirrhosis and increases the risk of hepatocellular carcinoma (HCC). While a definite diagnosis of NASH requires liver biopsy to confirm the presence of hepatocyte ballooning, hepatic fibrosis is the most important prognostic factor in NAFLD. With so many NAFLD patients, it is essential to have an effective screening method for NAFLD with hepatic fibrosis. As HCC with non-viral liver disease has increased markedly in Japan, effective screening and surveillance of HCC are also urgently needed. The most common death etiology in NAFLD patients is cardiovascular disease (CVD) event. Gastroenterologists must, therefore, pay close attention to CVD when examining NAFLD patients. In the updated guidelines, we propose screening and follow-up methods for hepatic fibrosis, HCC, and CVD in NAFLD patients. Several drug trials are ongoing for NAFLD/NASH therapy, however, there is currently no specific drug therapy for NAFLD/NASH. In addition to vitamin E and thiazolidinedione derivatives, recent trials have focused on sodium glucose co-transporter 2 (SGLT2) inhibitors and glucagon-like peptide-1 (GLP-1) analogues, and effective therapies are expected to be developed. These practical guidelines for NAFLD/NASH were established by the Japanese Society of Gastroenterology in conjunction with the Japan Society of Hepatology. Clinical evidence reported internationally between 1983 and October 2018 was collected, and each clinical and background question was evaluated using the Grades of Recommendation Assessment, Development and Evaluation (GRADE) system. This English summary provides the core essentials of these clinical practice guidelines, which include the definition and concept, screening systems for hepatic fibrosis, HCC and CVD, and current therapies for NAFLD/NASH in Japan.

DOI： 10.1007/s00535-021-01796-x

Humanized virus-suppressing factor inhibits hepatitis B virus infection by targeting viral cell entry. Reviewed International journal

Yu Miyakawa, Motoyuki Otsuka, Kazuma Sekiba, Kazuyoshi Funato, Kazuhiko Koike

Heliyon 7 ( 7 ) e07586 2021.7

Language：English Publishing type：Research paper (scientific journal)

Although nucleos(t)ide analogs and interferons suppress hepatitis B virus (HBV) replication, they must be taken continuously and have a low response rate. Therefore, therapeutics for HBV with novel modes of action are needed. Humanized virus-suppressing factor (hzVSF) is a monoclonal antibody against vimentin that exhibits broad-spectrum antiviral activity. Here, hzVSF significantly inhibited HBV infection. Although hzVSF inhibited HBV RNA production, it did not affect viral transcription from minicircle DNA mimicking covalently closed circular DNA. Additionally, hzVSF did not inhibit viral protein or DNA release from infected cells. Rather, hzVSF inhibited the cell entry of viral preS1 peptides, possibly by altering intracellular vimentin localization, which is important for HBV cell entry. These results suggest that hzVSF has therapeutic potential for HBV infection with a novel mode of action.

DOI： 10.1016/j.heliyon.2021.e07586

Potential of HBx Gene for Hepatocarcinogenesis in Noncirrhotic Liver. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Kazuhiko Koike

Seminars in liver disease 41 ( 2 ) 142 - 149 2021.5

Language：English Publishing type：Research paper (scientific journal)

Current treatments for hepatitis B virus (HBV) using nucleos(t)ide analogs cannot eliminate the risk of hepatocellular carcinoma (HCC) development. As HBV-associated HCC can develop even in the absence of liver cirrhosis, HBV is regarded to possess direct oncogenic potential. HBV regulatory protein X (HBx) has been identified as a primary mediator of HBV-mediated hepatocarcinogenesis. A fragment of the HBV genome that contains the coding region of HBx is commonly integrated into the host genome, resulting in the production of aberrant proteins and subsequent hepatocarcinogenesis. Besides, HBx interferes with the host DNA or deoxyribonucleic acid damage repair pathways, signal transduction, epigenetic regulation of gene expression, and cancer immunity, thereby promoting carcinogenesis in the noncirrhotic liver. However, numerous molecules and pathways have been implicated in the development of HBx-associated HCC, suggesting that the mechanisms underlying HBx-mediated hepatocarcinogenesis remain to be elucidated.

DOI： 10.1055/s-0041-1723033

Post-treatment cell-free DNA as a predictive biomarker in molecular-targeted therapy of hepatocellular carcinoma. Reviewed

Takuma Nakatsuka, Hayato Nakagawa, Yuki Hayata, Taijiro Wake, Tomoharu Yamada, Mizuki Nishibatake Kinoshita, Ryo Nakagomi, Masaya Sato, Tatsuya Minami, Koji Uchino, Kenichiro Enooku, Yotaro Kudo, Yasuo Tanaka, Takahiro Kishikawa, Motoyuki Otsuka, Ryosuke Tateishi, Kazuhiko Koike

Journal of gastroenterology 56 ( 5 ) 456 - 469 2021.5

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Liquid biopsies, particularly those involving circulating tumor DNA (ctDNA), are rapidly emerging as a non-invasive alternative to tumor biopsies. However, clinical applications of ctDNA analysis in hepatocellular carcinoma (HCC) have not been fully elucidated. METHODS: We measured the amount of plasma-derived cell-free DNA (cfDNA) in HCC patients before (n = 100) and a few days after treatment (n = 87), including radiofrequency ablation, transarterial chemoembolization, and molecular-targeted agents (MTAs), and prospectively analyzed their associations with clinical parameters and prognosis. TERT promoter mutations in cfDNA were analyzed using droplet digital PCR. Furthermore, we performed a comprehensive mutational analysis of post-treatment cfDNA via targeted ultra-deep sequencing (22,000× coverage) in a panel of 275 cancer-related genes in selected patients. RESULTS: Plasma cfDNA levels increased significantly according to HCC clinical stage, and a high cfDNA level was independently associated with a poor prognosis. TERT promoter mutations were detected in 45% of all cases but were not associated with any clinical characteristics. cfDNA levels increased significantly a few days after treatment, and a greater increase in post-treatment cfDNA levels was associated with a greater therapeutic response to MTAs. The detection rate of TERT mutations increased to 57% using post-treatment cfDNA, suggesting that the ctDNA was enriched. Targeted ultra-deep sequencing using post-treatment cfDNA after administering lenvatinib successfully detected various gene mutations and obtained promising results in lenvatinib-responsive cases. CONCLUSIONS: Post-treatment cfDNA analysis may facilitate the construction of biomarkers for predicting MTA treatment effects.

DOI： 10.1007/s00535-021-01773-4

Mutant KRAS drives metabolic reprogramming and autophagic flux in premalignant pancreatic cells. Reviewed International journal

Tatsunori Suzuki, Takahiro Kishikawa, Tatsuyuki Sato, Norihiko Takeda, Yuki Sugiura, Takahiro Seimiya, Kazuma Sekiba, Motoko Ohno, Takuma Iwata, Rei Ishibashi, Motoyuki Otsuka, Kazuhiko Koike

Cancer gene therapy 29 ( 5 ) 505 - 518 2021.4

Language：English Publishing type：Research paper (scientific journal)

Mutational activation of the KRAS gene occurs in almost all pancreatic ductal adenocarcinoma (PDAC) and is the earliest molecular event in their carcinogenesis. Evidence has accumulated of the metabolic reprogramming in PDAC, such as amino acid homeostasis and autophagic flux. However, the biological effects of KRAS mutation on metabolic reprogramming at the earlier stages of PDAC carcinogenesis are unclear. Here we report dynamic metabolic reprogramming in immortalized human non-cancerous pancreatic ductal epithelial cells, in which a KRAS mutation was induced by gene-editing, which may mimic early pancreatic carcinogenesis. Similar to the cases of PDAC, KRAS gene mutation increased the dependency on glucose and glutamine for maintaining the intracellular redox balance. In addition, the intracellular levels of amino acids were significantly decreased because of active protein synthesis, and the cells required greater autophagic flux to maintain their viability. The lysosomal inhibitor chloroquine significantly inhibited cell proliferation. Therefore, metabolic reprogramming is an early event in carcinogenesis initiated by KRAS gene mutation, suggesting a rationale for the development of nutritional interventions that suppress or delay the development of PDAC.

DOI： 10.1038/s41417-021-00326-4

The biological role of metabolic reprogramming in pancreatic cancer International journal

Tatsunori Suzuki, Motoyuki Otsuka, Takahiro Seimiya, Takuma Iwata, Takahiro Kishikawa, Kazuhiko Koike

MedComm 1 ( 3 ) 302 - 310 2020.12

Language：English Publishing type：Research paper (scientific journal) Publisher：Wiley

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease and highly resistant to all forms of therapy. PDAC cells reprogram their metabolism extensively to promote their survival and growth. Reflecting the vital role of altered metabolism, experimental and clinical trials targeting the rewired metabolism are currently underway. In this review, we summarize the vital role of metabolic reprogramming in the development of PDAC and the future of novel therapeutic applications.

DOI： 10.1002/mco2.37

The fatty-acid amide hydrolase inhibitor URB597 inhibits MICA/B shedding. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Takahiro Seimiya, Eri Tanaka, Kazuyoshi Funato, Yu Miyakawa, Kazuhiko Koike

Scientific reports 10 ( 1 ) 15556 - 15556 2020.9

Language：English Publishing type：Research paper (scientific journal)

MICA/B proteins are expressed on the surface of various types of stressed cells, including cancer cells. Cytotoxic lymphocytes expressing natural killer group 2D (NKG2D) receptor recognize MICA/B and eliminate the cells. However, cancer cells evade such immune recognition by inducing proteolytic shedding of MICA/B proteins. Therefore, preventing the shedding of MICA/B proteins could enhance antitumor immunity. Here, by screening a protease inhibitor library, we found that the fatty-acid amide hydrolase (FAAH) inhibitor, URB597, suppresses the shedding of MICA/B. URB597 significantly reduced the soluble MICA level in culture medium and increased the MICA level on the surface of cancer cells. The effect was indirect, being mediated by increased expression of tissue inhibitor of metalloproteinases 3 (TIMP3). Knockdown of TIMP3 expression reversed the effect of URB597, confirming that TIMP3 is required for the MICA shedding inhibition by URB597. In contrast, FAAH overexpression reduced TIMP3 expression and the cell-surface MICA level and increased the soluble MICA level. These results suggest that inhibition of FAAH could prevent human cancer cell evasion of immune-mediated clearance.

DOI： 10.1038/s41598-020-72688-y

Extracellular vesicles secreted by HBV-infected cells modulate HBV persistence in hydrodynamic HBV transfection mouse model. Reviewed International journal

Masatoshi Kakizaki, Yuichiro Yamamoto, Motoyuki Otsuka, Kouichi Kitamura, Masatoshi Ito, Hideki Derek Kawai, Masamichi Muramatsu, Tatehiro Kagawa, Ai Kotani

The Journal of biological chemistry 295 ( 35 ) 12449 - 12460 2020.8

Language：English Publishing type：Research paper (scientific journal)

Hepatitis B, a viral infection that affects the liver, is thought to affect over 257 million people worldwide, and long-term infection can lead to life-threatening issues such as cirrhosis or liver cancer. Chronic hepatitis B develops by the interaction between hepatitis B virus (HBV) and host immune response. However, questions of how HBV-infected cells thwart immune system defenses remain unanswered. Extracellular vesicles (EVs) are used for cellular communication, carrying cargoes such as RNAs, proteins, and lipids and delivering them intracellularly after being endocytosed by target cells. HBV-infected liver cells secrete several types of EVs into body fluids such as complete and incomplete virions, and exosomes. We previously demonstrated that monocytes that incorporated EVs moved to immunoregulatory phenotypes via up-regulation of PD-L1, an immunocheckpoint molecule, and down-regulation of CD69, a leukocyte activation molecule. In this study, we transfected mice with HBV using hydrodynamic injection and studied the effects of EVs secreted by HBV-infected liver cells. EVs secreted from cells with HBV replication strongly suppressed the immune response, inhibiting the eradication of HBV-replicating cells in the mice transfected with HBV. EVs were systemically incorporated in multiple organs, including liver, bone marrow (BM), and intestine. Intriguingly, the BM cells that incorporated EVs acquired intestinal tropism and the dendritic cell populations in the intestine increased. These findings suggest that the EVs secreted by HBV-infected liver cells exert immunosuppressive functions, and that an association between the liver, bone marrow, and intestinal tract exists through EVs secreted from HBV-infected cells.

DOI： 10.1074/jbc.RA120.014317

HBx increases EGFR expression by inhibiting miR129-5p function. Reviewed International journal

Masanori Ochi, Motoyuki Otsuka, Reo Maruyama, Kazuhiko Koike

Biochemical and biophysical research communications 529 ( 2 ) 198 - 203 2020.8

Language：English Publishing type：Research paper (scientific journal)

Despite the efficient suppression of hepatitis B virus (HBV) replication by nucelos(t)ide analogs, HBV RNA expression usually continues even during nucleots(t)ide analog therapy because episomal covalently closed circular DNA (ccDNA), which is the template for HBV RNA transcription, cannot be eliminated. Here, we found that the common sequences of all HBV RNAs and that encoding the X protein (HBx) have similarities with the sequences of a host cellular microRNA (miRNA), miR129-5p. HBx inhibits miR129-5p function, resulting in increased expression of ZBTB20, a target gene of miR129-5p. ZBTB20 activates transcription and increases cell-surface epidermal growth factor receptor (EGFR) levels, promoting the cell growth rate, and this effect was reversed through ZBTB20 knockdown. mir129-5p levels in Ago2-containing complexes were reduced by expression of HBx, suggesting that the viral RNA sequestered miR129-5p from Ago2-containing complexes. These results indicate the possibility that HBV RNA may maintain pathogenicity even through nucleos(t)ide analog therapy.

DOI： 10.1016/j.bbrc.2020.06.018

B型肝炎根治を目指した創薬研究宿主蛋白Smc5/6の発現回復によるHBV関連肝発癌制御の可能性

關場一磨, 大塚基之, 小池和彦

日本消化器病学会雑誌 117 ( 臨増総会 ) A93 - A93 2020.7

Emerging Roles of Exosomal Circular RNAs in Cancer. Reviewed International journal

Takahiro Seimiya, Motoyuki Otsuka, Takuma Iwata, Chikako Shibata, Eri Tanaka, Tatsunori Suzuki, Kazuhiko Koike

Frontiers in cell and developmental biology 8 568366 - 568366 2020

Language：English Publishing type：Research paper (scientific journal)

Circular RNA (circRNA) is a type of non-coding RNA that forms a covalently closed continuous loop. The expression pattern of circRNA varies among cell types and tissues, and many circRNAs are aberrantly expressed in various cancers. Aberrantly expressed circRNAs have been shown to play crucial roles in carcinogenesis, functioning as microRNA sponges or new templates for protein translation. Recent research has shown that circRNAs are enriched in exosomes. Exosomes are secretory vesicles that mediate intercellular communication through the delivery of cargo, including proteins, lipids, DNA, and RNA. Exosome-mediated crosstalk between cancer cells and the tumor microenvironment promotes the epithelial-mesenchymal transition, angiogenesis, and immune escape, and thus may contribute to cancer invasion and metastasis. In this review, we discuss the biological functions of exosomal circRNAs and their significance in cancer progression. Additionally, we discuss the potential clinical applications of exosomal circRNAs as biomarkers and in cancer therapy.

DOI： 10.3389/fcell.2020.568366

Deletion of Histone Methyltransferase G9a Suppresses Mutant Kras-driven Pancreatic Carcinogenesis

HIROYUKI KATO, KEISUKE TATEISHI, HIROAKI FUJIWARA, HIDEAKI IJICHI, KEISUKE YAMAMOTO, TAKUMA NAKATSUKA, MIWAKO KAKIUCHI, MAKOTO SANO, YOTARO KUDO, YOKU HAYAKAWA, HAYATO NAKAGAWA, YASUO TANAKA, MOTOYUKI OTSUKA, YOSHIHIRO HIRATA, MAKOTO TACHIBANA, YOICHI SHINKAI, KAZUHIKO KOIKE

Cancer Genomics - Proteomics 17 ( 6 ) 695 - 705 2020

Improved liver function in patients with cirrhosis due to chronic hepatitis C virus who achieve sustained virologic response is not accompanied by increased liver volume. Reviewed International journal

Taijiro Wake, Ryosuke Tateishi, Tsuyoshi Fukumoto, Ryo Nakagomi, Mizuki Nishibatake Kinoshita, Takuma Nakatsuka, Masaya Sato, Tatsuya Minami, Koji Uchino, Kenichiro Enooku, Hayato Nakagawa, Hidetaka Fujinaga, Yoshinari Asaoka, Yasuo Tanaka, Motoyuki Otsuka, Kazuhiko Koike

PloS one 15 ( 4 ) e0231836 2020

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND: Serum albumin level improves in patients with chronic hepatitis C virus (HCV) infection who achieve sustained virologic response (SVR) with antiviral therapy. However, it remains controversial whether liver volume increases along with SVR. METHODS: Patients with chronic HCV infection with a history of hepatocellular carcinoma (HCC) who achieved SVR with anti-HCV treatment from March 2003 to November 2017 were enrolled. Patients were followed up with periodic computed tomography (CT) scans to detect HCC recurrence. Patients who underwent treatment for HCC recurrence within 1 year after initiation of anti-HCV treatment were excluded. Laboratory data, including alanine aminotransferase (ALT) level, serum albumin level, and platelet count, were collected at baseline and timepoints after treatment initiation. Liver volume was evaluated at baseline and 24 and 48 weeks after treatment initiation using a CT volume analyzer. A linear mixed-effects model was applied to analyze the chronologic change in liver volume. The correlations between changes in ALT level, albumin level, and liver volume were also evaluated. RESULTS: Of 108 enrolled patients, 78 had cirrhosis. Serum albumin level continued to increase through 48 weeks after treatment initiation. A significant increase in liver volume was observed only in patients without cirrhosis (P = 0.005). There was a significant correlation between ALT level decrease and albumin level increase (P = 0.018). CONCLUSIONS: Improved liver albumin production with SVR was contributed by improved liver cell function rather than increased liver volume in patients with cirrhosis.

DOI： 10.1371/journal.pone.0231836

Identifying Inhibitors of the HBx-DDB1 Interaction Using a Split Luciferase Assay System. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Kazuhiko Koike

Journal of visualized experiments : JoVE ( 154 ) 2019.12

Language：English Publishing type：Research paper (scientific journal)

There is an urgent need for novel therapeutic agents for hepatitis B virus (HBV) infection. Although currently available nucleos(t)ide analogs potently inhibit viral replication, they have no direct effect on the expression of viral proteins transcribed from a viral covalently closed circular DNA (cccDNA). As high viral antigen load may play a role in this chronic and HBV-related carcinogenesis, the goal of HBV treatment is to eradicate viral proteins. HBV regulatory protein X (HBx) binds to the host DNA damage-binding protein 1 (DDB1) protein to degrade structural maintenance of chromosomes 5/6 (Smc5/6), resulting in activation of viral transcription from cccDNA. Here, using a split luciferase complementation assay system, we present a comprehensive compound screening system to identify inhibitors of the HBx-DDB1 interaction. Our protocol enables easy detection of interaction dynamics in real time within living cells. This technique may become a key assay to discover novel therapeutic agents for treatment of HBV infection.

DOI： 10.3791/60652

Detection of circulating colorectal cancer cells by a custom microfluid system before and after endoscopic metallic stent placement. Reviewed International journal

Rei Ishibashi, Shuntaro Yoshida, Nariaki Odawara, Takahiro Kishikawa, Ryo Kondo, Ayako Nakada, Ryunosuke Hakuta, Naminatsu Takahara, Eri Tanaka, Kazuma Sekiba, Takahiro Seimiya, Takashi Ohnaga, Motoyuki Otsuka, Kazuhiko Koike

Oncology letters 18 ( 6 ) 6397 - 6404 2019.12

Language：English Publishing type：Research paper (scientific journal)

Although the detection of circulating tumor cells (CTCs) should be crucial for future personalized medicine, no efficient and flexible methods have been established. The current study established a polymeric custom-made chip for capturing CTCs with a high efficiency and flexibility. As an example of clinical application, the effects of self-expandable metallic stent (SEMS) placement on the release of cancer cells into the blood of patients with colorectal cancer and bowel obstruction were analyzed. This was assessed as the placement of SEMS may cause mechanical damage and physical force to malignant tissue, increasing the risk of cancer cell release into the bloodstream. The present study examined the number of CTCs using a custom-made chip, before, at 24 h after and at 4 days after SEMS placement in patients with colorectal cancer. The results revealed that, among the 13 patients examined, the number of CTCs was increased in three cases at 24 h after SEMS placement. However, this increase was temporary. The number of CTCs also decreased at 4 days after stent placement in most cases. The CTC chip of the current study detected the number of CD133-positive cancer stem-like cells, which did not change, even in the patient whose total number of CTCs temporarily increased. The results indicated that this custom-made microfluid system can efficiently and flexibly detect CTCs, demonstrating its potential for obtaining information during the management of patients with cancer.

DOI： 10.3892/ol.2019.11047

Inhibition of HBV Transcription From cccDNA With Nitazoxanide by Targeting the HBx-DDB1 Interaction. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Motoko Ohno, Mari Yamagami, Takahiro Kishikawa, Tatsunori Suzuki, Rei Ishibashi, Takahiro Seimiya, Eri Tanaka, Kazuhiko Koike

Cellular and molecular gastroenterology and hepatology 7 ( 2 ) 297 - 312 2019.10

Language：English Publishing type：Research paper (scientific journal) Publisher：Elsevier {BV}

BACKGROUND & AIMS: Hepatitis B virus (HBV) infection is a major health concern worldwide. Although currently used nucleos(t)ide analogs efficiently inhibit viral replication, viral proteins transcribed from the episomal viral covalently closed circular DNA (cccDNA) minichromosome continue to be expressed long-term. Because high viral RNA or antigen loads may play a biological role during this chronicity, the elimination of viral products is an ultimate goal of HBV treatment. HBV regulatory protein X (HBx) was recently found to promote transcription of cccDNA with degradation of Smc5/6 through the interaction of HBx with the host protein DDB1. Here, this protein-protein interaction was considered as a new molecular target of HBV treatment. METHODS: To identify candidate compounds that target the HBx-DDB1 interaction, a newly constructed split luciferase assay system was applied to comprehensive compound screening. The effects of the identified compounds on HBV transcription and cccDNA maintenance were determined using HBV minicircle DNA, which mimics HBV cccDNA, and the natural HBV infection model of human primary hepatocytes. RESULTS: We show that nitazoxanide (NTZ), a thiazolide anti-infective agent that has been approved by the FDA for protozoan enteritis, efficiently inhibits the HBx-DDB1 protein interaction. NTZ significantly restores Smc5 protein levels and suppresses viral transcription and viral protein production in the HBV minicircle system and in human primary hepatocytes naturally infected with HBV. CONCLUSIONS: These results indicate that NTZ, which targets an HBV-related viral-host protein interaction, may be a promising new therapeutic agent and a step toward a functional HBV cure.

DOI： 10.1016/j.jcmgh.2018.10.010

Expression of circular RNA CDR1‑AS in colon cancer cells increases cell surface PD‑L1 protein levels. Reviewed International journal

Eri Tanaka, Yu Miyakawa, Takahiro Kishikawa, Takahiro Seimiya, Takuma Iwata, Kazuyoshi Funato, Nariaki Odawara, Kazuma Sekiba, Mari Yamagami, Tatsunori Suzuki, Rei Ishibashi, Motoyuki Otsuka, Kazuhiko Koike

Oncology reports 42 ( 4 ) 1459 - 1466 2019.10

Language：English Publishing type：Research paper (scientific journal)

The expression of CDR1‑AS, a representative circular RNA, is closely linked with poor prognosis in gastrointestinal cancers, such as colon, liver, and pancreatic cancers. Although it is well known that CDR1‑AS antagonizes microRNA‑7 function through its sequence similarities in the brain, its biological function and link with the malignant potential of cancer cells remain unclear, partly due to the difficulties of ectopic expression of circular RNAs. In the present study, SW620, a colon cancer cell line that stably expresses CDR1‑AS RNA circularized, was established using the laccase 2 gene cassette, and its biological function associated with malignant behavior was determined. In contrast to previous studies, cell growth or invasion ability was not altered by CDR1‑AS expression. However, the expression levels of CMTM4 and CMTM6, which were recently recognized as critical regulators of PD‑L1 protein expression at the cell surface, were significantly increased. Accordingly, the cell surface PD‑L1 protein levels were increased in CDR1‑AS‑expressing cells. Notably, the effects were not canceled out by overexpressing microRNA‑7, indicating that the increase in cell surface PD‑L1 in CDR1‑AS‑expressing cells was not dependent on microRNA‑7 function. These results indicated that expression of this circular RNA in cancer cells may lead to poor prognosis by increasing cell surface PD‑L1 levels through microRNA‑7‑independent mechanisms.

DOI： 10.3892/or.2019.7244

Inflammation and de-differentiation in pancreatic carcinogenesis. Reviewed International journal

Takahiro Seimiya, Motoyuki Otsuka, Takuma Iwata, Eri Tanaka, Tatsunori Suzuki, Kazuma Sekiba, Mari Yamagami, Rei Ishibashi, Kazuhiko Koike

World journal of clinical cases 6 ( 15 ) 882 - 891 2018.12

Language：English Publishing type：Research paper (scientific journal)

Pancreatic cancer is a malignancy with an extremely poor prognosis. Chronic pancreatitis is a well-known risk factor for pancreatic cancer. Inflammation is thought to influence carcinogenesis through DNA damage and activation of intracellular signaling pathways. Many transcription factors and signaling pathways co-operate to determine and maintain cell identity at each phase of pancreatic organogenesis and cell differentiation. Recent studies have shown that carcinogenesis is promoted through the suppression of transcription factors related to differentiation. Pancreatitis also demonstrates transcriptional changes, suggesting that multifactorial epigenetic changes lead to impaired differentiation. Taken together, these factors may constitute an important framework for pancreatic carcinogenesis. In this review, we discuss the role of inflammation and de-differentiation in the development of pancreatic cancer, as well as the future of novel therapeutic applications.

DOI： 10.12998/wjcc.v6.i15.882

Liver stiffness measurements in chronic hepatitis C: treatment evaluation and risk assessment. Reviewed

Nakagomi R, Tateishi R, Masuzaki R, Soroida Y, Iwai T, Kondo M, Fujiwara N, Sato M, Minami T, Uchino K, Enooku K, Nakagawa H, Asaoka Y, Kondo Y, Tanaka Y, Otsuka M, Kato N, Moriya K, Ikeda H, Koike K

Journal of gastroenterology and hepatology 2018.11

Circular RNA and exosomes in pancreatic cancer progression

Takahiro Seimiya, Motoyuki Otsuka, Hung-Yin Liu, Tatsunori Suzuki, Kazuma Sekiba, Mari Yamagami, Eri Tanaka, Rei Ishibashi, Kazuhiko Koike

Translational Cancer Research 7 S745 - S747 2018.8

Satellite RNA Increases DNA Damage and Accelerates Tumor Formation in Mouse Models of Pancreatic Cancer. Reviewed International journal

Takahiro Kishikawa, Motoyuki Otsuka, Tatsunori Suzuki, Takahiro Seimiya, Kazuma Sekiba, Rei Ishibashi, Eri Tanaka, Motoko Ohno, Mari Yamagami, Kazuhiko Koike

Molecular cancer research : MCR 16 ( 8 ) 1255 - 1262 2018.8

Language：English Publishing type：Research paper (scientific journal)

Highly repetitive tandem arrays such as satellite sequences in the centromeric and pericentromeric regions of chromosomes, which were previously considered to be silent, are actively transcribed in various biological processes, including cancers. In the pancreas, this aberrant expression occurs even in Kras-mutated pancreatic intraepithelial neoplasia (PanIN) tissues, which are precancerous lesions. To determine the biological role of satellite RNAs in carcinogenesis in vivo, we constructed mouse major satellite (MajSAT) RNA-expressing transgenic mice. However, these transgenic mice did not show spontaneous malignant tumor formation under normal breeding. Importantly, however, DNA damage was increased in pancreatic tissues induced by caerulein treatment or high-fat diet, which may be due to impaired nuclear localization of Y-Box Binding Protein 1 (YBX1), a component of the DNA damage repair machinery. In addition, when crossed with pancreas-specific Kras-mutant mice, MajSAT RNA expression resulted in an earlier increase in PanIN formation. These results suggest that aberrant MajSAT RNA expression accelerates oncogenesis by increasing the probability of a second driver mutation, thus accelerating cells to exit from the breakthrough phase to the expansion phase.Implications: Aberrant expression of satellite RNAs accelerates oncogenesis through a mechanism involving increased DNA damage. Mol Cancer Res; 16(8); 1255-62. ©2018 AACR.

DOI： 10.1158/1541-7786.MCR-18-0139

CPT2 downregulation adapts HCC to lipid-rich environment and promotes carcinogenesis via acylcarnitine accumulation in obesity. Reviewed International journal

Naoto Fujiwara, Hayato Nakagawa, Kenichiro Enooku, Yotaro Kudo, Yuki Hayata, Takuma Nakatsuka, Yasuo Tanaka, Ryosuke Tateishi, Yohko Hikiba, Kento Misumi, Mariko Tanaka, Akimasa Hayashi, Junji Shibahara, Masashi Fukayama, Junichi Arita, Kiyoshi Hasegawa, Hadassa Hirschfield, Yujin Hoshida, Yoshihiro Hirata, Motoyuki Otsuka, Keisuke Tateishi, Kazuhiko Koike

Gut 67 ( 8 ) 1493 - 1504 2018.8

Language：English Publishing type：Research paper (scientific journal)

OBJECTIVE: Metabolic reprogramming of tumour cells that allows for adaptation to their local environment is a hallmark of cancer. Interestingly, obesity-driven and non-alcoholic steatohepatitis (NASH)-driven hepatocellular carcinoma (HCC) mouse models commonly exhibit strong steatosis in tumour cells as seen in human steatohepatitic HCC (SH-HCC), which may reflect a characteristic metabolic alteration. DESIGN: Non-tumour and HCC tissues obtained from diethylnitrosamine-injected mice fed either a normal or a high-fat diet (HFD) were subjected to comprehensive metabolome analysis, and the significance of obesity-mediated metabolic alteration in hepatocarcinogenesis was evaluated. RESULTS: The extensive accumulation of acylcarnitine species was seen in HCC tissues and in the serum of HFD-fed mice. A similar increase was found in the serum of patients with NASH-HCC. The accumulation of acylcarnitine could be attributed to the downregulation of carnitine palmitoyltransferase 2 (CPT2), which was also seen in human SH-HCC. CPT2 downregulation induced the suppression of fatty acid β-oxidation, which would account for the steatotic changes in HCC. CPT2 knockdown in HCC cells resulted in their resistance to lipotoxicity by inhibiting the Src-mediated JNK activation. Additionally, oleoylcarnitine enhanced sphere formation by HCC cells via STAT3 activation, suggesting that acylcarnitine accumulation was a surrogate marker of CPT2 downregulation and directly contributed to hepatocarcinogenesis. HFD feeding and carnitine supplementation synergistically enhanced HCC development accompanied by acylcarnitine accumulation in vivo. CONCLUSION: In obesity-driven and NASH-driven HCC, metabolic reprogramming mediated by the downregulation of CPT2 enables HCC cells to escape lipotoxicity and promotes hepatocarcinogenesis.

DOI： 10.1136/gutjnl-2017-315193

Hepatitis B virus pathogenesis: Fresh insights into hepatitis B virus RNA. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Motoko Ohno, Mari Yamagami, Takahiro Kishikawa, Tatsunori Suzuki, Rei Ishibashi, Takahiro Seimiya, Eri Tanaka, Kazuhiko Koike

World journal of gastroenterology 24 ( 21 ) 2261 - 2268 2018.6

Language：English Publishing type：Research paper (scientific journal)

Hepatitis B virus (HBV) is still a worldwide health concern. While divergent factors are involved in its pathogenesis, it is now clear that HBV RNAs, principally templates for viral proteins and viral DNAs, have diverse biological functions involved in HBV pathogenesis. These functions include viral replication, hepatic fibrosis and hepatocarcinogenesis. Depending on the sequence similarities, HBV RNAs may act as sponges for host miRNAs and may deregulate miRNA functions, possibly leading to pathological consequences. Some parts of the HBV RNA molecule may function as viral-derived miRNA, which regulates viral replication. HBV DNA can integrate into the host genomic DNA and produce novel viral-host fusion RNA, which may have pathological functions. To date, elimination of HBV-derived covalently closed circular DNA has not been achieved. However, RNA transcription silencing may be an alternative practical approach to treat HBV-induced pathogenesis. A full understanding of HBV RNA transcription and the biological functions of HBV RNA may open a new avenue for the development of novel HBV therapeutics.

DOI： 10.3748/wjg.v24.i21.2261

Identification of PTBP1 responsible for caspase dependent YRNA cleavage Reviewed

Jun Ogata, Yuki Sugiura, Akinori Kanai, Masafumi Tanaka, Hirotaka Matsui, Masato Ohtsuka, Toshiya Inaba, Motoyuki Otsuka, Ai Kotani

BioRxiv doi: https://doi.org/10.1101/298851 2018.5

Publishing type：Research paper (other academic) Publisher：Cold Spring Harbor Laboratory

<title>ABSTRACT</title>Some RNAs such as 28S rRNA, U1 snRNA, and Y RNAs are known to be cleaved during apoptosis. As the underlying mechanism is yet unclear, the functions and biological significance of RNA degradation in apoptosis remain elusive. We previously identified novel, functional small RNAs named AGO-taxis small RNA (ASR) that are specifically bound to AGO1. Here, we investigated ASR biogenesis, which appears to be non-canonical. Y RNAs, non-coding RNAs degraded during apoptosis, were identified as the precursors of several ASRs. Cell-free analysis combined with fractionation methods revealed that the apoptosis-specific biogenesis of ASRs or Y RNA degradation was induced by PTBP1—an endoribonuclease inhibitor of Y RNAs. PTBP1, a splicing factor, was truncated by caspase 3, which subsequently activated endoribonuclease to induce biogenesis of ASRs and Y RNA cleavage.

DOI： 10.1101/298851

DHX9 regulates production of hepatitis B virus-derived circular RNA and viral protein levels. Reviewed International journal

Kazuma Sekiba, Motoyuki Otsuka, Motoko Ohno, Takahiro Kishikawa, Mari Yamagami, Tatsunori Suzuki, Rei Ishibashi, Takahiro Seimiya, Eri Tanaka, Kazuhiko Koike

Oncotarget 9 ( 30 ) 20953 - 20964 2018.4

Language：English Publishing type：Research paper (scientific journal)

Hepatitis B virus (HBV) infection, which is a major health concern worldwide, can lead to liver cirrhosis and hepatocellular carcinoma. Although current nucleos(t)ide analogs efficiently inhibit viral reverse transcription and viral DNA load clinically, episomal viral covalently closed circular DNA (cccDNA) minichromosomes and transcripts from cccDNA continue to be expressed over the long term. We hypothesized that, under these conditions, viral transcripts may have biological functions involved in pathogenesis. Here, we show that the host protein DExH-box helicase 9 (DXH9) is associated with viral RNAs. We also show that viral-derived circular RNA is produced during HBV replication, and the amount is increased by knockdown of the DHX9 protein, which, in turn, results in decreased viral protein levels but does not affect the levels of HBV DNA. These phenomena were observed in the HBV-producing cell culture model and HBV mini-circle model mimicking HBV cccDNA, as well as in human primary hepatocytes infected with HBV. Based on these results, we conclude that, in HBV infection, the RNA binding factor DHX9 is a novel regulator of viral circular RNA and viral protein levels.

DOI： 10.18632/oncotarget.25104

ISGF3 with reduced phosphorylation is associated with constitutive expression of interferon-induced genes in aging cells. Reviewed International journal

Mari Yamagami, Motoyuki Otsuka, Takahiro Kishikawa, Kazuma Sekiba, Takahiro Seimiya, Eri Tanaka, Tatsunori Suzuki, Rei Ishibashi, Motoko Ohno, Kazuhiko Koike

NPJ aging and mechanisms of disease 4 11 - 11 2018

Language：English Publishing type：Research paper (scientific journal)

During cellular aging, many changes in cellular functions occur. A hallmark of aged cells is secretion of inflammatory mediators, which collectively is referred to as the senescence-associated secretory phenotype (SASP). However, the mechanisms underlying such changes are unclear. Canonically, the expression of interferon (IFN)-stimulated genes (ISGs) is induced by IFNs through the formation of the tripartite transcriptional factor ISGF3, which is composed of IRF9 and the phosphorylated forms of STAT1 and STAT2. However, in this study, the constitutive expression of ISGs in human-derived senescent fibroblasts and in fibroblasts from a patient with Werner syndrome, which leads to premature aging, was mediated mainly by the unphosphorylated forms of STATs in the absence of INF production. Under homeostatic conditions, STAT1, STAT2, and IRF9 were localized to the nucleus of aged cells. Although knockdown of JAK1, a key kinase of STAT1 and STAT2, did not affect ISG expression or IFN-stimulated response element (ISRE)-mediated promoter activities in these senescent cells, knockdown of STAT1 or STAT2 decreased ISG expression and ISRE activities. These results suggest that the ISGF3 complex without clear phosphorylation is required for IFN-independent constitutive ISG transcription in senescent cells.

DOI： 10.1038/s41514-018-0030-6

Impact of histone demethylase KDM3A-dependent AP-1 transactivity on hepatotumorigenesis induced by PI3K activation Reviewed

T. Nakatsuka, K. Tateishi, Y. Kudo, K. Yamamoto, H. Nakagawa, H. Fujiwara, R. Takahashi, K. Miyabayashi, Y. Asaoka, Y. Tanaka, H. Ijichi, Y. Hirata, M. Otsuka, M. Kato, J. Sakai, M. Tachibana, H. Aburatani, Y. Shinkai, K. Koike

ONCOGENE 36 ( 45 ) 6262 - 6271 2017.11

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

Epigenetic gene regulation linked to oncogenic pathways is an important focus of cancer research. KDM3A, a histone H3 lysine 9 (H3K9) demethylase, is known to have a pro-tumorigenic function. Here, we showed that KDM3A contributes to liver tumor formation through the phosphatidylinositol 3-kinase (PI3K) pathway, which is often activated in hepatocellular carcinoma. Loss of Kdm3a attenuated tumor formation in Pik3ca transgenic (Tg) mouse livers. Transcriptome analysis of pre-cancerous liver tissues revealed that the expression of activator protein 1 (AP-1) target genes was induced by PI3K activation, but blunted upon Kdm3a ablation. Particularly, the expression of Cd44, a liver cancer stem marker, was regulated by AP-1 in a Kdm3a-dependent manner. We identified Cd44-positive hepatocytes with epithelial-mesenchymal transition-related expression profiles in the Pik3ca Tg liver and confirmed their in vivo tumorigenic capacity. Notably, the number and tumor-initiating capacity of Cd44-positive hepatocytes were governed by Kdm3a. As a mechanism in Kdm3a-dependent AP-1 transcription, Kdm3a recruited c-Jun to the AP-1 binding sites of Cd44, Mmp7 and Pdgfrb without affecting c-Jun expression. Moreover, Brg1, a component of the SWI/SNF chromatin remodeling complex, interacted with c-Jun in a Kdm3a-dependent manner and was bound to the AP-1 binding site of these genes. Finally, KDM3A and c-JUN were co-expressed in 33% of human premalignant lesions with PI3K activation. Our data suggest a critical role for KDM3A in the PI3K/AP-1 oncogenic axis and propose a novel strategy for inhibition of KDM3A against liver tumor development under PI3K pathway activation.

DOI： 10.1038/onc.2017.222

膵発癌過程における反復配列RNA異常発現の生物学的意義の検討 Reviewed

岸川孝弘, 大塚基之, 大野元子, 山上まり, 石橋嶺, 關場一磨, 鈴木辰典, 小池和彦

日本消化器病学会雑誌 114 ( 臨増大会 ) A743 - A743 2017.9

RASAL1 is a potent regulator of hepatic stellate cell activity and liver fibrosis Reviewed

Akemi Takata, Motoyuki Otsuka, Takahiro Kishikawa, Mari Yamagami, Rei Ishibashi, Kazuma Sekiba, Tatsunori Suzuki, Motoko Ohno, Yui Yamashita, Takaya Abe, Ryota Masuzaki, Tsuneo Ikenoue, Kazuhiko Koike

ONCOTARGET 8 ( 39 ) 64840 - 64852 2017.9

Language：English Publishing type：Research paper (scientific journal) Publisher：IMPACT JOURNALS LLC

Liver fibrosis, leading to cirrhosis and liver failure, can occur after chronic liver injury. The transition of hepatic stellate cells (HSCs) from quiescent cells into proliferative and fibrogenic cells is a central event in liver fibrosis. Here, we show that RAS protein activator like-1 (RASAL1), a RAS-GTPase-activating protein, which switches off RAS activity, is significantly decreased during HSC activation, and that HSC activation can be antagonized by forced expression of the RASAL1 protein. We demonstrate that RASAL1 suppresses HSC proliferation by regulating the Ras-MAPK pathway, and that RASAL1 suppresses HSC fibrogenic activity by regulating the PKA-LKB1-AMPK-SRF pathway by interacting with angiotensin II receptor, type 1. We also show that RASAL1-deficient mice are more susceptible to liver fibrosis. These data demonstrate that deregulated RASAL1 expression levels and the affected downstream intracellular signaling are central mediators of perpetuated HSC activation and fibrogenesis in the liver.

DOI： 10.18632/oncotarget.17609

Biliary epithelial injury-induced regenerative response by IL-33 promotes cholangiocarcinogenesis from peribiliary glands Reviewed

Hayato Nakagawa, Nobumi Suzuki, Yoshihiro Hirata, Yohko Hikiba, Yoku Hayakawa, Hiroto Kinoshita, Sozaburo Ihara, Koji Uchino, Yuji Nishikawa, Hideaki Ijichi, Motoyuki Otsuka, Junichi Arita, Yoshihiro Sakamoto, Kiyoshi Hasegawa, Norihiro Kokudo, Keisuke Tateishi, Kazuhiko Koike

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 114 ( 19 ) E3806 - E3815 2017.5

Language：English Publishing type：Research paper (scientific journal) Publisher：NATL ACAD SCIENCES

The carcinogenic mechanism of extrahepatic cholangiocarcinoma (ECC) is unclear, due at least in part to the lack of an appropriate mouse model. Because human studies have reported frequent genetic alterations in the Ras-and TGF beta/SMAD-signaling pathways in ECC, mice with tamoxifen-inducible, duct-cell-specific Kras activation and a TGF beta receptor type 2 (TGF beta R2) deletion were first generated by crossing LSL-Kras(G12D), Tgfbr2(flox/flox), and K19(CreERT) mice (KT-K19(CreERT)). However, KT-K19CreERT mice showed only mild hyperplasia of biliary epithelial cells (BECs) in the extrahepatic bile duct (EHBD) and died within 7 wk, probably a result of lung adenocarcinomas. Next, to analyze the additional effect of E-cadherin loss, KT-K19CreERT mice were crossed with CDH1flox/flox mice (KTCK19CreERT). Surprisingly, KTC-K19CreERT mice exhibited a markedly thickened EHBD wall accompanied by a swollen gallbladder within 4 wk after tamoxifen administration. Histologically, invasive periductal infiltrating-type ECC with lymphatic metastasis was observed. Time-course analysis of EHBD revealed that recombined BECs lining the bile duct lumen detached due to E-cadherin loss, whereas recombined cells could survive in the peribiliary glands (PBGs), which are considered a BEC stem-cell niche. Detached dying BECs released high (l)evels of IL-33, as determined by microarray analysis using biliary organoids, and stimulated inflammation and a regenerative response by PBGs, leading eventually to ECC development. Cell lineage tracing suggested PBGs as the cellular origin of ECC. IL-33 cooperated with Kras and TGF beta R2 mutations in the development of ECC, and anti-IL-33 treatment suppressed ECC development significantly. Thus, this mouse model provided insight into the carcinogenic mechanisms, cellular origin, and potential therapeutic targets of ECC.

DOI： 10.1073/pnas.1619416114

Transcriptional activation of the MICA gene with an engineered CRISPR-Cas9 system Reviewed

Kazuma Sekiba, Mari Yamagami, Motoyuki Otsuka, Tatsunori Suzuki, Takahiro Kishikawa, Rei Ishibashi, Motoko Ohno, Masaya Sato, Kazuhiko Koike

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 486 ( 2 ) 521 - 525 2017.4

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Major histocompatibility complex class I polypeptide-related sequence A (MICA) is a prototypical NKG2D ligand. Because immune cells, such as natural killer (NK) cells, recognize virally infected or transformed cells and eliminate them through the interaction between NKG2D receptors on NK cells and NKG2D ligands on pathogenic cells, MICA expression levels are associated with NK cell-mediated immunity. Here, we report that an engineered clustered regularly interspaced short palindromic repeats-Cas9-related complex targeting MICA gene promoter sequences activates transcription of the MICA gene from its endogenous locus. Inhibiting microRNA function, which targets the 3' untranslated region of the MICA gene, enhances this activation. These results demonstrate that the combination of Cas9-based transcriptional activators and simultaneous modulation of microRNA function may be a powerful tool for enhancing MICA protein expression and efficient anti-pathogenic cell immunity. (C) 2017 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2017.03.076

MicroRNAs and liver disease Reviewed

Motoyuki Otsuka, Takahiro Kishikawa, Takeshi Yoshikawa, Mari Yamagami, Motoko Ohno, Akemi Takata, Chikako Shibata, Rei Ishibashi, Kazuhiko Koike

JOURNAL OF HUMAN GENETICS 62 ( 1 ) 75 - 80 2017.1

Language：English Publisher：NATURE PUBLISHING GROUP

The biological roles of microRNAs (miRNAs) have been extensively studied. miRNA122 represents more than half of the miRNAs expressed in the liver and has various physiological and pathological functions, which include enhancing hepatitis virus replication, regulating lipid metabolism and suppressing hepatocellular carcinoma. miRNAs, whether globally or individually, have been linked with hepatocarcinogenesis. Furthermore, some miRNAs have been shown to be involved in the pathogenesis of nonalcoholic steatohepatitis. Using nucleotide-based strategies, these miRNAs may be developed as potential therapeutic targets. Because changes in miRNA expression can be measured in sera, they may be used as non-invasive biomarkers if they correctly reflect the pathological state of the liver. In this review, we show the biological roles of representative miRNAs in liver disease and discuss the current issues that remain to be clarified for future clinical applications.

DOI： 10.1038/jhg.2016.53

Mutual antagonism between hepatitis B viral mRNA and host microRNA let-7 Reviewed

Akemi Takata, Motoyuki Otsuka, Motoko Ohno, Takahiro Kishikawa, Takeshi Yoshikawa, Kazuhiko Koike

SCIENTIFIC REPORTS 6 23237 2016.3

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

The interplay between viral and host factors plays a major role in viral pathogenesis. Hepatitis B virus (HBV) infection is a global health problem that leads to liver cirrhosis and hepatocellular carcinoma (HCC). Although HBV proteins have been studied extensively about their implication in hepatocarcinogenesis, the molecular mechanisms of oncogenesis are still largely unknown. A recent concept in gene regulation, in which competitive endogenous RNAs compete for common microRNAs (miRNAs), suggests that mRNA targets are key elements in the regulation of miRNA availability. Here, we show that HBV mRNA in the preS2 region can be targeted by host miRNA let-7 g. This leads to the sequestration of let-7 g and inhibition of let-7 g function. The expression of HBV transcripts, including the preS2 region, de-repressed let-7 g targets, which may contribute to long-term oncogenesis. HBV transcript-expressing transgenic mice, but not non-targeted transcript-expressing mice, were more prone to chemically induced hepatoocarcinogenesis. Let-7 target protein expression was upregulated in human HCC tissues derived from HBV-infected patients. On the other hand, let-7 g inhibited HBV preS2 protein expression and viral products. These results suggest that the interplay between viral intermediate transcripts during HBV replication and host miRNAs is crucial to the pathogenesis of chronic viral infection.

DOI： 10.1038/srep23237

Development of a screening method to identify regulators of MICA shedding Reviewed

Takahiro Kishikawa, Motoyuki Otsuka, Motoko Ohno, Takeshi Yoshikawa, Masaya Sato, Kazuhiko Koike

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 465 ( 4 ) 764 - 768 2015.10

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Immune cells, such as natural killer (NK) cells, recognize virally infected and transformed cells, and eliminate them through the interaction between NKG2D receptors on NK cells and NKG2D ligands on pathogenic cells. Shedding of NKG2D ligands is thought to be a type of counter-mechanism employed by pathogenic cells to evade from NKG2D-mediated immune surveillance. MHC class I polypeptide-related sequence A (MICA) is a prototypical NKG2D ligand. We previously reported that, in soluble form, MICA expression levels are significantly associated with hepatitis virus-induced hepatocellular carcinoma. Here, we report a MICA shedding assay that utilizes membrane-bound MICA tagged at its N-terminus with a nano-luciferase reporter to quantify MICA shedding into culture media. Using this method, we screened a compound library and identified putative regulators of MICA shedding that have the potential to enhance the immune reaction by simultaneously increasing cell surface MICA levels and decreasing soluble MICA levels. This shedding assay may be useful for screening regulators of cell surface molecule shedding. (C) 2015 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2015.08.081

ROCK inhibition enhances microRNA function by promoting deadenylation of targeted mRNAs via increasing PAIP2 expression Reviewed

Takeshi Yoshikawa, Jianfeng Wu, Motoyuki Otsuka, Takahiro Kishikawa, Motoko Ohno, Chikako Shibata, Akemi Takata, Felicia Han, Young Jun Kang, Chyi-Ying A. Chen, Ann-Bin Shyu, Jiahuai Han, Kazuhiko Koike

NUCLEIC ACIDS RESEARCH 43 ( 15 ) 7577 - 7589 2015.9

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

The reduced expression levels and functional impairment of global miRNAs are related to various human diseases, including cancers. However, relatively little is known about how global miRNA function may be upregulated. Here, we report that global miRNA function can be enhanced by Rho-associated, coiled-coil-containing protein kinase (ROCK) inhibitors. The regulation of miRNA function by ROCK inhibitors is mediated, at least in part, by poly(A)-binding protein-interacting protein 2 (PAIP2), which enhances poly(A)-shortening of miRNA-targeted mRNAs and leads to global upregulation of miRNA function. In the presence of a ROCK inhibitor, PAIP2 expression is enhanced by the transcription factor hepatocyte nuclear factor 4 alpha (HNF4A) through increased ROCK1 nuclear localization and enhanced ROCK1 association with HNF4A. Our data reveal an unexpected role of ROCK1 as a cofactor of HNF4A in enhancing PAIP2 transcription. ROCK inhibitors may be useful for the various pathologies associated with the impairment of global miRNA function.

DOI： 10.1093/nar/gkv728

Circulating RNAs as new biomarkers for detecting pancreatic cancer Reviewed

Takahiro Kishikawa, Motoyuki Otsuka, Motoko Ohno, Takeshi Yoshikawa, Akemi Takata, Kazuhiko Koike

WORLD JOURNAL OF GASTROENTEROLOGY 21 ( 28 ) 8527 - 8540 2015.7

Language：English Publisher：BAISHIDENG PUBLISHING GROUP INC

Pancreatic cancer remains difficult to treat and has a high mortality rate. It is difficult to diagnose early, mainly due to the lack of screening imaging modalities and specific biomarkers. Consequently, it is important to develop biomarkers that enable the detection of early stage tumors. Emerging evidence is accumulating that tumor cells release substantial amounts of RNA into the bloodstream that strongly resist RNases in the blood and are present at sufficient levels for quantitative analyses. These circulating RNAs are upregulated in the serum and plasma of cancer patients, including those with pancreatic cancer, compared with healthy controls. The majority of RNA biomarker studies have assessed circulating microRNAs (miRs), which are often tissue-specific. There are few reports of the tumor-specific upregulation of other types of small non-coding RNAs (ncRNAs), such as small nucleolar RNAs and Piwi-interacting RNAs. Long ncRNAs (lncRNAs), such as HOTAIR and MALAT1, in the serum/plasma of pancreatic cancer patients have also been reported as diagnostic and prognostic markers. Among tissue-derived RNAs, some miRs show increased expression even in pre-cancerous tissues, and their expression profiles may allow for the discrimination between a chronic inflammatory state and carcinoma. Additionally, some miRs and lncRNAs have been reported with significant alterations in expression according to disease progression, and they may thus represent potential candidate diagnostic or prognostic biomarkers that may be used to evaluate patients once detection methods in peripheral blood are well established. Furthermore, recent innovations in high-throughput sequencing techniques have enabled the discovery of unannotated tumor-associated ncRNAs and tumor-specific alternative splicing as novel and specific biomarkers of cancers. Although much work is required to clarify the release mechanism, origin of tumor-specific circulating RNAs, and selectivity of carrier complexes, and technical advances must also be achieved, such as creating a consensus normalization protocol for quantitative data analysis, circulating RNAs are largely unexplored and might represent novel clinical biomarkers.

DOI： 10.3748/wjg.v21.i28.8527

Novel therapeutic approaches for hepatitis B virus covalently closed circular DNA Reviewed

Motoko Ohno, Motoyuki Otsuka, Takahiro Kishikawa, Takeshi Yoshikawa, Akemi Takata, Kazuhiko Koike

WORLD JOURNAL OF GASTROENTEROLOGY 21 ( 23 ) 7084 - 7088 2015.6

Language：English Publisher：BAISHIDENG PUBLISHING GROUP INC

Hepatitis B virus (HBV) infection is a major global health problem. Although current therapies, such as the use of nucleos(t) ide analogs, inhibit HBV replication efficiently, they do not eliminate covalently closed circular DNA (cccDNA), which persists in hepatocyte nuclei. As HBV cccDNA is a viral transcription template, novel therapeutic approaches to directly target HBV cccDNA are necessary to completely eradicate persistent HBV infections. HBV cccDNA levels in HBV-infected human liver cells are extremely low; thus, more reliable and simple measurement methods are needed to correctly monitor their levels during therapeutic treatment. Although reverse transcription-polymerase chain reaction or Southern blot procedures are currently used in research studies, these methods are not completely reliable and are also time-consuming and labor-intensive. Genome editing technologies, such as zinc finger nucleases, transcription activator-like effector nucleases, and the clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9) system, which are designed to target specific DNA sequences, represent highly promising potential therapeutic tools. In particular, the CRISPR/Cas9 system is an easily customizable sequences-pecific nuclease with high flexibility and may be the most feasible approach to target HBV cccDNA. Further research to develop easier, safer, and more effective protocols should be pursued.

DOI： 10.3748/wjg.v21.i23.7084

Tissue-Specific Regulation of p38 alpha-Mediated Inflammation in Con A-Induced Acute Liver Damage Reviewed

Young Jun Kang, Bo-Ram Bang, Motoyuki Otsuka, Kinya Otsu

JOURNAL OF IMMUNOLOGY 194 ( 10 ) 4759 - 4766 2015.5

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC IMMUNOLOGISTS

Because p38 alpha plays a critical role in inflammation, it has been an attractive target for the development of anti-inflammation therapeutics. However, p38 alpha inhibitors showed side effects, including severe liver toxicity, that often prevailed over the benefits in clinical studies, and the mechanism of toxicity is not clear. In this study, we demonstrate that p38 alpha regulates the inflammatory responses in acute liver inflammation in a tissue-specific manner, and liver toxicity by p38 alpha inhibitors may be a result of the inhibition of protective activity of p38 alpha in the liver. Genetic ablation of p38 alpha in T and NKT cells protected mice from liver injury in Con A-induced liver inflammation, whereas liver-specific deletion of p38 alpha aggravated liver pathology. We found that p38 alpha deficiency in the liver increased the expression of chemokines to recruit more inflammatory cells, indicating that p38 alpha in the liver plays a protective anti-inflammatory role during acute liver inflammation. Therefore, our results suggest that p38 alpha regulates the inflammatory responses in a tissue-specific manner, and that the tissue-specific p38 alpha targeting strategies can be used for the development of an effective anti-inflammation treatment with an improved side-effect profile.

DOI： 10.4049/jimmunol.1402954

Decreased miR122 in hepatocellular carcinoma leads to chemoresistance with increased arginine Reviewed

Takahiro Kishikawa, Motoyuki Otsuka, Poh Seng Tan, Motoko Ohno, Xiaochen Sun, Takeshi Yoshikawa, Chikako Shibata, Akemi Takata, Kentaro Kojima, Kenji Takehana, Maki Ohishi, Sana Ota, Tomoyuki Noyama, Yuji Kondo, Masaya Sato, Tomoyoshi Soga, Yujin Hoshida, Kazuhiko Koike

Oncotarget 6 ( 10 ) 8339 - 8352 2015

Language：English Publishing type：Research paper (scientific journal) Publisher：Impact Journals LLC

Reduced expression of microRNA122 (miR122), a liver-specific microRNA, is frequent in hepatocellular carcinoma (HCC). However, its biological significances remain poorly understood. Because deregulated amino acid levels in cancers can affect their biological behavior, we determined the amino acid levels in miR122- silenced mouse liver tissues, in which intracellular arginine levels were significantly increased. The increased intracellular arginine levels were through upregulation of the solute carrier family 7 (SLC7A1), a transporter of arginine and a direct target of miR122. Arginine is the substrate for nitric oxide (NO) synthetase, and intracellular NO levels were increased in miR122-silenced HCC cells, with increased resistance to sorafenib, a multikinase inhibitor. Conversely, maintenance of the miR122-silenced HCC cells in arginine-depleted culture media, as well as overexpression of miR122 in miR122-low-expressing HCC cells, reversed these effects and rendered the cells more sensitive to sorafenib. Using a reporter knock-in construct, chemical compounds were screened, and Wee1 kinase inhibitor was identified as upregulators of miR122 transcription, which increased the sensitivity of the cells to sorafenib. These results provide an insight into sorafenib resistance in miR122-low HCC, and suggest that arginine depletion or a combination of sorafenib with the identified compound may provide promising approaches to managing this HCC subset.

DOI： 10.18632/oncotarget.3234

Diagnostic and therapeutic application of noncoding RNAs for hepatocellular carcinoma Reviewed

Chikako Shibata, Motoyuki Otsuka, Takahiro Kishikawa, Motoko Ohno, Takeshi Yoshikawa, Akemi Takata, Kazuhiko Koike

World Journal of Hepatology 7 ( 1 ) 1 - 6 2015

Language：English Publishing type：Research paper (scientific journal) Publisher：Baishideng Publishing Group Co

MicroRNAs (miRNAs) are small, noncoding RNA molecules that regulate gene expression posttranscriptionally, targeting thousands of messenger RNAs. Long noncoding RNAs (lncRNAs), another class of noncoding RNAs, have been determined to be also involved in transcription regulation and translation of target genes. Since deregulated expression levels or functions of miRNAs and lncRNAs in hepatocellular carcinoma (HCC) are frequently observed, clinical use of noncoding RNAs for novel diagnostic and therapeutic applications in the management of HCCs is highly and emergently expected. Here, we summarize recent findings regarding deregulated miRNAs and lncRNAs for their potential clinical use as diagnostic and prognostic biomarkers of HCC. Specifically, we emphasize the deregulated expression levels of such noncoding RNAs in patients' sera as noninvasive biomarkers, a field that requires urgent improvement in the clinical surveillance of HCC. Since nucleotide-based strategies are being applied to clinical therapeutics, we further summarize clinical and preclinical trials using oligonucleotides involving the use of miRNAs and small interfering RNAs against HCC as novel therapeutics. Finally, we discuss current open questions, which must be clarified in the near future for realistic clinical applications of these new strategies.

DOI： 10.4254/wjh.v7.i1.1

Loss of histone demethylase KDM6B enhances aggressiveness of pancreatic cancer through downregulation of C/EBP alpha Reviewed

Keisuke Yamamoto, Keisuke Tateishi, Yotaro Kudo, Tomohiko Sato, Shinzo Yamamoto, Koji Miyabayashi, Keisuke Matsusaka, Yoshinari Asaoka, Hideaki Ijichi, Yoshihiro Hirata, Motoyuki Otsuka, Yousuke Nakai, Hiroyuki Isayama, Tsuneo Ikenoue, Mineo Kurokawa, Masashi Fukayama, Norihiro Kokudo, Masao Omata, Kazuhiko Koike

CARCINOGENESIS 35 ( 11 ) 2404 - 2414 2014.11

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

Genetic mutations in pancreatic ductal adenocarcinoma (PDAC) with critical roles have been well examined The recent discovery of alterations in genes encoding histone modifiers suggests their possible roles in the complexity of cancer development. We previously reported loss of heterozygosity of the KDM6B gene, which encodes a histone demethylase for trimethylated histone 113 lysine 27, a repressive chromatin mark, in PDAC cells. In this study, we demonstrated that loss of KDM6B enhanced aggressiveness of PDAC cells. KDM6B has been regarded as a tumor suppressor that mediates oncogenic KRAS-induced senescence. Consistently, KDM6B was highly expressed in pancreatic precancerous lesions (pancreatic intraepithelial neoplasms); then, the expression decreased as the malignant grade progressed. We found that knockdown of KDM6B in PDAC cells promoted tumor sphere formation and increased peritoneal dissemination and liver metastasis in vivo. Microarray and chromatin immunoprecipitation analysis implicated CEBPA for aggressiveness induced by KDM6B knockdown. CEBPA knockdown recapitulated the phenotypic change of PDAC cells after KDM6B knockdown, which was reversed by forced expression of C/EBP alpha. Moreover, similar protein expression patterns of KDM6B and C/EBP alpha in human PDAC emphasized their functional correlation. Notably, pharmacological inhibition of the H3K27 methylase EZH2 in PDAC cells inhibited tumor sphere formation along with the upregulation of CEBPA expression, and this effect was impaired in KDM6B knockdown cells, highlighting the role for KDM6B in the activation of CEBPA. Together, our results propose a significant role for the KDM6B-C/EBP alpha axis in the PDAC phenotype.

DOI： 10.1093/carcin/bgu136

Impact of PNPLA3 polymorphisms on the development of hepatocellular carcinoma in patients with chronic hepatitis C virus infection Reviewed

Masaya Sato, Naoya Kato, Ryosuke Tateishi, Ryosuke Muroyama, Norie Kowatari, Wenwen Li, Kaku Goto, Motoyuki Otsuka, Shuichiro Shiina, Haruhiko Yoshida, Masao Omata, Kazuhiko Koike

HEPATOLOGY RESEARCH 44 ( 10 ) E137 - E144 2014.10

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

Aim: The PNPLA3 rs738409 C>G polymorphism (encoding for I148M) has recently been identified as a susceptibility factor for steatosis-mediated liver damage. We evaluated the influence of this polymorphism on hepatocarcinogenesis in patients with chronic hepatitis C (CHC) virus infection.
Methods: We genotyped the rs738409 single nucleotide polymorphism in 358 hepatitis C-associated hepatocellular carcinoma (HCC) patients and correlated the age at onset of HCC and the interval between hepatitis C virus (HCV) infection and the development of HCC in patients with each genotype.
Results: The frequencies of CC, CG and GG genotypes were 27.9% (100/358), 49.2% (176/358) and 22.9% (82/358), respectively, and were in Hardy-Weinberg equilibrium. The median age at onset of HCC for the GG genotype was significantly younger compared to for non-GG genotypes (67.81 vs 69.87 years, P < 0.001), and the median interval between HCV infection and the development of HCC was significantly shorter in patients with the GG genotype (39.96 vs 40.85 years, P = 0.008). PNPLA3 GG genotype was also associated with a higher aspartate aminotransferase level (69.5 vs 59.0 IU/L, P = 0.02), lower prothrombin time (73.0% vs 78.0%, P = 0.008) and a higher prevalence of histological steatosis (40.0% vs. 22.2%, P = 0.01) at the time of HCC onset.
Conclusion: The PNPLA3 genotype GG may be associated with accelerated hepatocarcinogenesis in CHC patients through increased steatosis in the liver.

DOI： 10.1111/hepr.12258

The flavonoid apigenin inhibits hepatitis C virus replication by decreasing mature microRNA122 levels Reviewed

Chikako Shibata, Motoko Ohno, Motoyuki Otsuka, Takahiro Kishikawa, Kaku Goto, Ryosuke Muroyama, Naoya Kato, Takeshi Yoshikawa, Akemi Takata, Kazuhiko Koike

VIROLOGY 462 42 - 48 2014.8

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Despite recent progress in the development of direct-acting antivirals against hepatitis C virus (HCV), chronic HCV infection remains an important health burden worldwide. MicroRNA122 (miR122), a liver-specific microRNA (miRNA), positively regulates HCV replication, and systemic application of antisense oligonucleotides against miR122 led to the long-lasting suppression of HCV viremia in human clinical trials. Here, we report that apigenin, a flavonoid and an inhibitor of maturation of a subset of miRNAs, inhibits HCV replication in vitro. Apigenin decreased the expression levels of mature miR122 without significantly affecting cell growth. Because supplementation of synthesized miR122 oligonucleotides or overexpression of constitutively active TRBP blocked these effects, the inhibitory effects of apigenin on HCV replication seemed to be dependent on the reduction of mature miR122 expression levels through inhibition of TRBP phosphorylation. Thus, apigenin intake, either through regular diet or supplements, may decrease HCV replication in chronically infected patients. (C) 2014 The Authors. Published by Elsevier Inc.

DOI： 10.1016/j.virol.2014.05.024

Specific delivery of microRNA93 into HBV-replicating hepatocytes downregulates protein expression of liver cancer susceptible gene MICA Reviewed

Motoko Ohno, Motoyuki Otsuka, Takahiro Kishikawa, Chikako Shibata, Takeshi Yoshikawa, Akemi Takata, Ryosuke Muroyama, Norie Kowatari, Masaya Sato, Naoya Kato, Shun'ichi Kuroda, Kazuhiko Koike

ONCOTARGET 5 ( 14 ) 5581 - 5590 2014.7

Language：English Publishing type：Research paper (scientific journal) Publisher：IMPACT JOURNALS LLC

Chronic hepatitis B virus (HBV) infection is a major cause of hepatocellular carcinoma (HCC). To date, the lack of efficient in vitro systems supporting HBV infection and replication has been a major limitation of HBV research. Although primary human hepatocytes support the complete HBV life cycle, their limited availability and difficulties with gene transduction remain problematic. Here, we used human primary hepatocytes isolated from humanized chimeric uPA/SCID mice as efficient sources. These hepatocytes supported HBV replication in vitro. Based on analyses of mRNA and microRNA (miRNA) expression levels in HBV-infected hepatocytes, miRNA93 was significantly downregulated during HBV infection. MiRNA93 is critical for regulating the expression levels of MICA protein, which is a determinant for HBV-induced HCC susceptibility. Exogenous addition of miRNA93 in HBV-infected hepatocytes using bionanocapsules consisted of HBV envelope L proteins restored MICA protein expression levels in the supernatant. These results suggest that the rescued suppression of soluble MICA protein levels by miRNA93 targeted to HBV-infected hepatocytes using bionanocapsules may be useful for the prevention of HBV-induced HCC by altering deregulated miRNA93 expression.

IL28B minor allele is associated with a younger age of onset of hepatocellular carcinoma in patients with chronic hepatitis C virus infection Reviewed

Masaya Sato, Naoya Kato, Ryosuke Tateishi, Ryosuke Muroyama, Norie Kowatari, Wenwen Li, Kaku Goto, Motoyuki Otsuka, Shuichiro Shiina, Haruhiko Yoshida, Masao Omata, Kazuhiko Koike

JOURNAL OF GASTROENTEROLOGY 49 ( 4 ) 748 - 754 2014.4

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER JAPAN KK

IL28B polymorphisms were shown to be associated with a response to peg-interferon-based treatment in chronic hepatitis C (CHC) and spontaneous clearance. However, little is known about how this polymorphism affects the course of CHC, including the development of hepatocellular carcinoma (HCC). We evaluated the influence of IL28B polymorphisms on hepatocarcinogenesis in CHC patients.
We genotyped the rs8099917 single-nucleotide polymorphism in 351 hepatitis C-associated HCC patients without history of IFN-based treatment, and correlated the age at onset of HCC in patients with each genotype.
Frequencies of TT, TG, and GG genotypes were 74.3 % (261/351), 24.8 % (87/351), and 0.9 % (3/351), respectively. The mean ages at onset of HCC for TT, TG, and GG genotypes were 69.9, 67.5 and 66.8, respectively. In multivariate analysis, IL28B minor allele (TG and GG genotypes) was an independent risk factor for younger age at onset of HCC (P = 0.02) in males (P < 0.001) with higher body mass index (BMI; P = 0.009). The IL28B minor allele was also associated with a lower probability of having aspartate aminotransferase-to-platelet ratio index (APRI) > 1.5 (minor vs. major, 46.7 vs. 58.6 %; P = 0.01), lower AST (69.1 vs. 77.7 IU/L, P = 0.02), lower ALT (67.8 vs. 80.9 IU/L, P = 0.002), higher platelet count (12.8 vs. 11.2 x 10(4)/mu L, P = 0.002), and higher prothrombin time (79.3 vs. 75.4 %, P = 0.002).
The IL28B minor allele was associated with lower inflammatory activity and less progressed fibrosis of the liver; however, it constituted a risk factor for younger-age onset of HCC in CHC patients.

DOI： 10.1007/s00535-013-0826-x

The role of microRNAs in hepatocarcinogenesis: current knowledge and future prospects Reviewed

Motoyuki Otsuka, Takahiro Kishikawa, Takeshi Yoshikawa, Motoko Ohno, Akemi Takata, Chikako Shibata, Kazuhiko Koike

JOURNAL OF GASTROENTEROLOGY 49 ( 2 ) 173 - 184 2014.2

Language：English Publisher：SPRINGER JAPAN KK

MicroRNAs (miRNAs) are small, noncoding RNA molecules that regulate gene expression post-transcriptionally through complementary base pairing with thousands of messenger RNAs. Although the precise biological functions of individual miRNAs are still unknown, miRNAs are speculated to play important roles in diverse biological processes through fine regulation of their target gene expression. A growing body of data indicates the deregulation of miRNAs during hepatocarcinogenesis. In this review, we summarize recent findings regarding deregulated miRNA expression and their possible target genes in hepatocarcinogenesis, with emphasis on inflammation-related hepatocarcinogenesis. Because miRNA-based strategies are being applied to clinical therapeutics, precise knowledge of miRNA functions is crucial both scientifically and clinically. We discuss the current open questions from these points of view, which must be clarified in the near future.

DOI： 10.1007/s00535-013-0909-8

Histone demethylase KDM4C regulates sphere formation by mediating the cross talk between Wnt and Notch pathways in colonic cancer cells Reviewed

Shinzo Yamamoto, Keisuke Tateishi, Yotaro Kudo, Keisuke Yamamoto, Takayuki Isagawa, Genta Nagae, Takuma Nakatsuka, Yoshinari Asaoka, Hideaki Ijichi, Yoshihiro Hirata, Motoyuki Otsuka, Tsuneo Ikenoue, Hiroyuki Aburatani, Masao Omata, Kazuhiko Koike

CARCINOGENESIS 34 ( 10 ) 2380 - 2388 2013.10

Language：English Publishing type：Research paper (scientific journal) Publisher：OXFORD UNIV PRESS

Alterations in genes coding for histone modifiers are found in human cancers, suggesting that histone modification is involved in malignant features of neoplastic cells. This study showed that a histone demethylase KDM4C is significant for colonosphere formation by mediating the cross talk between oncogenic pathways through a feed-forward mechanism. The expression of KDM4C gene was increased in spheres from colorectal cancer (CRC) cells and the knockdown (KD) of KDM4C eliminated colonosphere formation. We found that the KD of -catenin, an important oncogenic factor in CRC, resulted in not only decreased sphere formation but also impaired upregulation of KDM4C gene in spheres. -Catenin bound to the KDM4C promoter, suggesting that KDM4C is involved in the sphere-forming ability downstream of -catenin in CRC cells. Microarray analysis identified the JAG1 gene that codes for a notch ligand Jagged1 responsible for sphere formation as a target of KDM4C. KDM4C KD decreased the expression of JAG1 gene, and the downregulation of JAG1 gene recapitulated the impaired colonosphere formation. JAG1 is also a target of -catenin, and chromatin immunoprecipitation analysis showed the binding of -catenin and KDM4C onto the JAG1 promoter during colonosphere formation. Importantly, KDM4C KD ruined the recruitment of -catenin onto the JAG1 promoter independently of the H3-K9 methylation status and blunted JAG1 expression during sphere formation. These data indicate that KDM4C maintains the sphere-forming capacity in CRCs by mediating the -catenin-dependent transcription of JAG1 in a feed-forward manner.

DOI： 10.1093/carcin/bgt174

Activation of p38α in T cells regulates the intestinal host defense against attaching and effacing bacterial infections Reviewed

Eun-Jin Shim, Bo-Ram Bang, Seung-Goo Kang, Jianhui Ma, Motoyuki Otsuka, Jiman Kang, Martin Stahl, Jiahuai Han, Changchun Xiao, Bruce A. Vallance, Young Jun Kang

Journal of Immunology 191 ( 5 ) 2764 - 2770 2013.9

Language：English Publishing type：Research paper (scientific journal) Publisher：5

Intestinal infections by attaching and effacing (A/E) bacterial pathogens cause severe colitis and bloody diarrhea. Although p38α in intestinal epithelial cells (IEC) plays an important role in promoting protection against A/E bacteria by regulating T cell recruitment, its impact on immune responses remains unclear. In this study, we show that activation of p38α in T cells is critical for the clearance of the A/E pathogen Citrobacter rodentium. Mice deficient of p38α in T cells, but not in macrophages or dendritic cells, were impaired in clearing C. rodentium. Expression of inflammatory cytokines such as IFN-γ by p38α-deficient T cells was reduced, which further reduced the expression of inflammatory cytokines, chemokines, and antimicrobial peptide by IECs and led to reduced infiltration of T cells into the infected colon. Administration of IFN-γ activated the mucosal immunity to C. rodentium infection by increasing the expression of inflammation genes and the recruitment of T cells to the site of infection. Thus, p38α contributes to host defense against A/E pathogen infection by regulating the expression of inflammatory cytokines that activate host defense pathways in IECs. Copyright © 2013 by The American Association of Immunologists, Inc.

DOI： 10.4049/jimmunol.1300908

Unique Haploinsufficient Role of the MicroRNA-Processing Molecule Dicer1 in a Murine Colitis-Associated Tumorigenesis Model Reviewed

Takeshi Yoshikawa, Motoyuki Otsuka, Takahiro Kishikawa, Akemi Takata, Motoko Ohno, Chikako Shibata, Young Jun Kang, Haruhiko Yoshida, Kazuhiko Koike

PLOS ONE 8 ( 9 ) e71969 2013.9

Language：English Publishing type：Research paper (scientific journal) Publisher：PUBLIC LIBRARY SCIENCE

A widespread downregulated expression of microRNAs (miRNAs) is commonly observed in human cancers. Similarly, deregulated expression of miRNA-processing pathway components, which results in the reduction of global miRNA expression, may also be associated with tumorigenesis. Here, we show that specific ablation of Dicer1 in intestinal epithelial cells accelerates intestinal inflammation-associated tumorigenesis. This effect was apparent only when a single copy of Dicer1 was deleted, but not with complete Dicer1 ablation. DICER expression and subsequent mature miRNA levels were inversely correlated with the number of intact Dicer1 alleles. Because the expression levels of DICER were retained in tumors and its surrounding tissues even after induction of colitis-associated tumors, the effects of Dicer1 deletion were cell-autonomous. Although the expression levels of representative oncogenes and tumor suppressor genes were in most cases inversely correlated with the expression levels of DICER, some genes were not affected by Dicer1 deletion. Thus, deregulating the delicate balance between the expression levels of tumor-promoting and - suppressive genes may be crucial for tumorigenesis in this unique haploinsufficient case.

DOI： 10.1371/journal.pone.0071969

Inhibition of microRNA122 decreases SREBP1 expression by modulating suppressor of cytokine signaling 3 expression Reviewed

Chikako Shibata, Takahiro Kishikawa, Motoyuki Otsuka, Motoko Ohno, Takeshi Yoshikawa, Akemi Takata, Haruhiko Yoshida, Kazuhiko Koike

Biochemical and Biophysical Research Communications 438 ( 1 ) 230 - 235 2013.8

Language：English Publishing type：Research paper (scientific journal) Publisher：1

While inhibition of microRNA122 (miR122) function in vivo results in reduced serum cholesterol and fatty acid levels, the molecular mechanisms underlying the link between miR122 function and lipid metabolism remains unclear. Because the expression of SREBP1, a central transcription factor involved in lipid metabolism, is known to be increased by suppressor of cytokine signaling 3 (SOCS3) expression, and because we previously found that SOCS3 expression is regulated by miR122, in this study, we examined the correlation between miR122 status and the expression levels of SOCS3 and SREBP1. SREBP1 expression decreased when SOCS3 expression was reduced by miR122 silencing in vitro. Conversely, SREBP1 expression in miR122-silenced cells was restored by enforced expression of SOCS3. Such correlations were observed in human liver tissues with different miR122 expression levels. These signaling links may explain one of the molecular mechanisms linking inhibition of miR122 function or decreased expression of miR122 to decreased fatty acid and cholesterol levels, in the inhibition of miR122 function, or in pathological status in chronic liver diseases. © 2013 Elsevier Inc.

DOI： 10.1016/j.bbrc.2013.07.064

The flavonoid apigenin improves glucose tolerance through inhibition of microRNA maturation in miRNA103 transgenic mice Reviewed

Motoko Ohno, Chikako Shibata, Takahiro Kishikawa, Takeshi Yoshikawa, Akemi Takata, Kentaro Kojima, Masao Akanuma, Young Jun Kang, Haruhiko Yoshida, Motoyuki Otsuka, Kazuhiko Koike

SCIENTIFIC REPORTS 3 2553 2013.8

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

Polyphenols are representative bioactive substances with diverse biological effects. Here, we show that apigenin, a flavonoid, has suppressive effects on microRNA (miRNA) function. The effects were mediated by impaired maturation of a subset of miRNAs, probably through inhibition of the phosphorylation of TRBP, a component of miRNA-generating complexes via impaired mitogen-activated protein kinase (MAPK) Erk activation. While glucose intolerance was observed in miRNA103 (miR103)-overexpressing transgenic mice, administration of apigenin improved this pathogenic status likely through suppression of matured miR103 expression levels. These results suggest that apigenin may have favorable effects on the pathogenic status induced by overexpression of miRNA103, whose maturation is mediated by phosphorylated TRBP.

DOI： 10.1038/srep02553

Transdifferentiation of human fibroblasts into hepatocyte-like cells by defined transcriptional factors Reviewed

Tomomi Kogiso, Hikaru Nagahara, Motoyuki Otsuka, Keiko Shiratori, Steven F. Dowdy

HEPATOLOGY INTERNATIONAL 7 ( 3 ) 937 - 944 2013.7

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

Liver transplantation is currently the only curative therapeutic option for end-stage liver cirrhosis. However, due to the limitations of donor liver availability and occasional rejection, it cannot always be successfully applied. In this study, we determined whether fibroblasts can be transdifferentiated into hepatocyte-like cells by transcription factors that initiate and maintain hepatocyte differentiation.
Fibroblasts were transduced with retrovirus vectors carrying FOXA2, HNF4 alpha, and C/EBP beta. To enhance the efficiency of transdifferentiation, cMyc was also expressed.
Transdifferentiation was successful using both neonatal fibroblasts and human forehead fibroblasts. The transdifferentiated cells produced hepatocyte-specific proteins such as albumin and cytochrome, and had important hepatocyte-specific functions, such as glycogen storage and indocyanine green uptake, suggesting that the cells function at least as partial hepatocytes.
These results provide a novel method of generating differentiated hepatocyte-like cells, and may represent an alternative source of cells for future cell-based therapeutics for end-stage liver diseases.

DOI： 10.1007/s12072-013-9432-5

A genome-wide association study of HCV-induced liver cirrhosis in the Japanese population identifies novel susceptibility loci at the MHC region Reviewed

Yuji Urabe, Hidenori Ochi, Naoya Kato, Vinod Kumar, Atsushi Takahashi, Ryosuke Muroyama, Naoya Hosono, Motoyuki Otsuka, Ryosuke Tateishi, Paulisally Hau Yi Lo, Chizu Tanikawa, Masao Omata, Kazuhiko Koike, Daiki Miki, Hiromi Abe, Naoyuki Kamatani, Joji Toyota, Hiromitsu Kumada, Michiaki Kubo, Kazuaki Chayama, Yusuke Nakamura, Koichi Matsuda

Journal of Hepatology 58 ( 5 ) 875 - 882 2013.5

Language：English Publishing type：Research paper (scientific journal)

Background &amp
Aims: We performed a genome-wide association study (GWAS) of hepatitis C virus (HCV)-induced liver cirrhosis (LC) to identify predictive biomarkers for the risk of LC in patients with chronic hepatitis C (CHC). Methods: A total of 682 HCV-induced LC cases and 1045 CHC patients of Japanese origin were genotyped by Illumina Human Hap 610-Quad bead Chip. Results: Eight SNPs which showed possible associations (p &lt
1.0 × 10-5) at the GWAS stage were further genotyped using 936 LC cases and 3809 CHC patients. We found that two SNPs within the major histocompatibility complex (MHC) region on chromosome 6p21, rs910049 and rs3135363, were significantly associated with the progression from CHC to LC (pcombined = 9.15 × 10 -11 and 1.45 × 10-10, odds ratio (OR) = 1.46 and 1.37, respectively). We also found that HLA-DQA1*0601 and HLA-DRB1*0405 were associated with the progression from CHC to LC (p = 4.53 × 10-4 and 1.54 × 10-4 with OR = 2.80 and 1.45, respectively). Multiple logistic regression analysis revealed that rs3135363, rs910049, and HLA-DQA1*0601 were independently associated with the risk of HCV-induced LC. In addition, individuals with four or more risk alleles for these three loci have a 2.83-fold higher risk for LC than those with no risk allele, indicating the cumulative effects of these variations. Conclusions: Our findings elucidated the crucial roles of multiple genetic variations within the MHC region as prognostic/predictive biomarkers for CHC patients. © 2013 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jhep.2012.12.024

MicroRNA-140 acts as a liver tumor suppressor by controlling NF-kappa B activity by directly targeting DNA methyltransferase 1 (Dnmt1) expression Reviewed

Akemi Takata, Motoyuki Otsuka, Takeshi Yoshikawa, Takahiro Kishikawa, Yohko Hikiba, Shuntaro Obi, Tadashi Goto, Young Jun Kang, Shin Maeda, Haruhiko Yoshida, Masao Omata, Hiroshi Asahara, Kazuhiko Koike

HEPATOLOGY 57 ( 1 ) 162 - 170 2013.1

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

MicroRNAs (miRNAs) are small RNAs that regulate the expression of specific target genes. While deregulated miRNA expression levels have been detected in many tumors, whether miRNA functional impairment is also involved in carcinogenesis remains unknown. We investigated whether deregulation of miRNA machinery components and subsequent functional impairment of miRNAs are involved in hepatocarcinogenesis. Among miRNA-containing ribonucleoprotein complex components, reduced expression of DDX20 was frequently observed in human hepatocellular carcinomas, in which enhanced nuclear factor-?B (NF-?B) activity is believed to be closely linked to carcinogenesis. Because DDX20 normally suppresses NF-?B activity by preferentially regulating the function of the NF-?B-suppressing miRNA-140, we hypothesized that impairment of miRNA-140 function may be involved in hepatocarcinogenesis. DNA methyltransferase 1 (Dnmt1) was identified as a direct target of miRNA-140, and increased Dnmt1 expression in DDX20-deficient cells hypermethylated the promoters of metallothionein genes, resulting in decreased metallothionein expression leading to enhanced NF-?B activity. MiRNA-140-knockout mice were prone to hepatocarcinogenesis and had a phenotype similar to that of DDX20 deficiency, suggesting that miRNA-140 plays a central role in DDX20 deficiency-related pathogenesis. Conclusion: These results indicate that miRNA-140 acts as a liver tumor suppressor, and that impairment of miRNA-140 function due to a deficiency of DDX20, a miRNA machinery component, could lead to hepatocarcinogenesis. (HEPATOLOGY 2013; 57: 162-170)

DOI： 10.1002/hep.26011

MicroRNA-140 acts as a liver tumor suppressor by controlling NF-κB activity by directly targeting DNA methyltransferase 1 (Dnmt1) expression. International journal

Akemi Takata, Motoyuki Otsuka, Takeshi Yoshikawa, Takahiro Kishikawa, Yohko Hikiba, Shuntaro Obi, Tadashi Goto, Young Jun Kang, Shin Maeda, Haruhiko Yoshida, Masao Omata, Hiroshi Asahara, Kazuhiko Koike

Hepatology (Baltimore, Md.) 57 ( 1 ) 162 - 70 2013.1

Language：English Publishing type：Research paper (scientific journal)

UNLABELLED: MicroRNAs (miRNAs) are small RNAs that regulate the expression of specific target genes. While deregulated miRNA expression levels have been detected in many tumors, whether miRNA functional impairment is also involved in carcinogenesis remains unknown. We investigated whether deregulation of miRNA machinery components and subsequent functional impairment of miRNAs are involved in hepatocarcinogenesis. Among miRNA-containing ribonucleoprotein complex components, reduced expression of DDX20 was frequently observed in human hepatocellular carcinomas, in which enhanced nuclear factor-κB (NF-κB) activity is believed to be closely linked to carcinogenesis. Because DDX20 normally suppresses NF-κB activity by preferentially regulating the function of the NF-κB-suppressing miRNA-140, we hypothesized that impairment of miRNA-140 function may be involved in hepatocarcinogenesis. DNA methyltransferase 1 (Dnmt1) was identified as a direct target of miRNA-140, and increased Dnmt1 expression in DDX20-deficient cells hypermethylated the promoters of metallothionein genes, resulting in decreased metallothionein expression leading to enhanced NF-κB activity. MiRNA-140-knockout mice were prone to hepatocarcinogenesis and had a phenotype similar to that of DDX20 deficiency, suggesting that miRNA-140 plays a central role in DDX20 deficiency-related pathogenesis. CONCLUSION: These results indicate that miRNA-140 acts as a liver tumor suppressor, and that impairment of miRNA-140 function due to a deficiency of DDX20, a miRNA machinery component, could lead to hepatocarcinogenesis.

DOI： 10.1002/hep.26011

Current status of miRNA-targeting therapeutics and preclinical studies against gastroenterological carcinoma. Reviewed

Shibata C, Otsuka M, Kishikawa T, Yoshikawa T, Ohno M, Takata A, Koike K

Molecular and cellular therapies 1 5 2013

Regulation of the expression of the liver cancer susceptibility gene MICA by microRNAs Reviewed

Takahiro Kishikawa, Motoyuki Otsuka, Takeshi Yoshikawa, Motoko Ohno, Akemi Takata, Chikako Shibata, Yuji Kondo, Masao Akanuma, Haruhiko Yoshida, Kazuhiko Koike

Scientific Reports 3 2739 2013

Language：English Publishing type：Research paper (scientific journal)

Hepatocellular carcinoma (HCC) is a threat to public health worldwide. We previously identified the association of a single nucleotide polymorphism (SNP) at the promoter region of the MHC class I polypeptide-related sequence A (MICA) gene with the risk of hepatitis-virus-related HCC. Because this SNP affects MICA expression levels, regulating MICA expression levels may be important in the prevention of HCC. We herein show that the microRNA (miR) 25-93-106b cluster can modulate MICA levels in HCC cells. Overexpression of the miR 25-93-106b cluster significantly suppressed MICA expression. Conversely, silencing of this miR cluster enhanced MICA expression in cells that express substantial amounts of MICA. The changes in MICA expression levels by the miR25-93-106b cluster were biologically significant in an NKG2D-binding assay and an in vivo cell-killing model. These data suggest that the modulation of MICA expression levels by miRNAs may be a useful method to regulate HCCs during hepatitis viral infection.

DOI： 10.1038/srep02739

A genome-wide association study of HCV-induced liver cirrhosis in the Japanese population identifies novel susceptibility loci at the MHC region Reviewed International journal

Urabe Y, Ochi H, Kato N, Kumar V, Takahashi A, Muroyama R, Hosono N, Otsuka M, Tateishi R, Lo P. H, Tanikawa C, Omata M, Koike K, Miki D, Abe H, Kamatani N, Toyota J, Kumada H, Kubo M, Chayama K, Nakamura Y, Matsuda K

J Hepatol 58 ( 5 ) 875 - 82 2013

Language：English Publishing type：Research paper (scientific journal)

BACKGROUND & AIMS: We performed a genome-wide association study (GWAS) of hepatitis C virus (HCV)-induced liver cirrhosis (LC) to identify predictive biomarkers for the risk of LC in patients with chronic hepatitis C (CHC). METHODS: A total of 682 HCV-induced LC cases and 1045 CHC patients of Japanese origin were genotyped by Illumina Human Hap 610-Quad bead Chip. RESULTS: Eight SNPs which showed possible associations (p<1.0 × 10(-5)) at the GWAS stage were further genotyped using 936 LC cases and 3809 CHC patients. We found that two SNPs within the major histocompatibility complex (MHC) region on chromosome 6p21, rs910049 and rs3135363, were significantly associated with the progression from CHC to LC (pcombined=9.15 × 10(-11) and 1.45 × 10(-10), odds ratio (OR)=1.46 and 1.37, respectively). We also found that HLA-DQA1(*)0601 and HLA-DRB1(*)0405 were associated with the progression from CHC to LC (p=4.53 × 10(-4) and 1.54 × 10(-4) with OR=2.80 and 1.45, respectively). Multiple logistic regression analysis revealed that rs3135363, rs910049, and HLA-DQA1(*)0601 were independently associated with the risk of HCV-induced LC. In addition, individuals with four or more risk alleles for these three loci have a 2.83-fold higher risk for LC than those with no risk allele, indicating the cumulative effects of these variations. CONCLUSIONS: Our findings elucidated the crucial roles of multiple genetic variations within the MHC region as prognostic/predictive biomarkers for CHC patients.

DOI： 10.1016/j.jhep.2012.12.024

Silencing of microRNA-122 enhances interferon-alpha signaling in the liver through regulating SOCS3 promoter methylation Reviewed

Takeshi Yoshikawa, Akemi Takata, Motoyuki Otsuka, Takahiro Kishikawa, Kentaro Kojima, Haruhiko Yoshida, Kazuhiko Koike

SCIENTIFIC REPORTS 2 637 2012.9

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

Hepatitis C virus (HCV) infection is a major cause of chronic liver disease worldwide. Although novel drugs against HCV are under development, the current standard therapy consists principally of interferon (IFN). To improve the response to IFN treatment by enhancing interferon-stimulated response element (ISRE)-mediated gene transcription, we screened 75 microRNAs highly expressed in hepatocytes for their ability to modulate ISRE activity. Overexpression of microRNA-122 (miR122) significantly suppressed ISRE activity. Conversely, silencing of miR122 function enhanced IFN-induced ISRE activity, by decreasing expression of suppressor of cytokine signaling 3 (SOCS3). This decrease in SOCS3 level was not mediated by microRNA target gene suppression, but rather by enhanced methylation at SOCS3 gene promoter. Taken together, our data, along with the fact that antisense oligonucleotides of miR122 also directly inhibit HCV replication, suggest that a combination therapy comprising IFN and silencing of miR122 function may be a promising therapeutic option in the near future.

DOI： 10.1038/srep00637

A miRNA machinery component DDX20 controls NF-kappa B via microRNA-140 function Reviewed

Akemi Takata, Motoyuki Otsuka, Takeshi Yoshikawa, Takahiro Kishikawa, Yotaro Kudo, Tadashi Goto, Haruhiko Yoshida, Kazuhiko Koike

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 420 ( 3 ) 564 - 569 2012.4

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Hepatocellular carcinoma is the third leading cause of cancer mortality worldwide, but the molecular mechanisms in tumorigenesis remain largely unknown. Previously, a DEAD-box protein DDX20, a component of microRNA-containing ribonucleoprotein complexes, was identified as a liver tumor suppressor candidate in an oncogenomics-based in vivo RNAi screen. However, the molecular mechanisms were unknown. Here, we show that deficiency of DDX20 results in the enhancement of NF-kappa B activity, a crucial intracellular signaling pathway closely linked with hepatocarcinogenesis. While DDX20 normally suppresses NF-kappa B activity by regulating NF-kappa B-suppressing miRNA-140 function, this suppressive effect was lost in DDX20-deficient cells. The impairment of miRNA function due to DDX20 deficiency appears to be miRNA species-specific at the point of loading miRNAs into the RNA-induced silencing complex. These results indicate that DDX20 deficiency enhances NF-kappa B activity by impairing the NF-kappa B-suppressive action of microRNAs, and suggest that dysregulation of the microRNA machinery components may also be involved in pathogenesis in various human diseases. (C) 2012 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2012.03.034

Altered composition of fatty acids exacerbates hepatotumorigenesis during activation of the phosphatidylinositol 3-kinase pathway Reviewed

Yotaro Kudo, Yasuo Tanaka, Keisuke Tateishi, Keisuke Yamamoto, Shinzo Yamamoto, Dai Mohri, Yoshihiro Isomura, Motoko Seto, Hayato Nakagawa, Yoshinari Asaoka, Motohisa Tada, Miki Ohta, Hideaki Ijichi, Yoshihiro Hirata, Motoyuki Otsuka, Tsuneo Ikenoue, Shin Maeda, Shuichiro Shiina, Haruhiko Yoshida, Osamu Nakajima, Fumihiko Kanai, Masao Omata, Kazuhiko Koike

JOURNAL OF HEPATOLOGY 55 ( 6 ) 1400 - 1408 2011.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Background & Aims: Some clinical findings have suggested that systemic metabolic disorders accelerate in vivo tumor progression. Deregulation of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway is implicated in both metabolic dysfunction and carcinogenesis in humans; however, it remains unknown whether the altered metabolic status caused by abnormal activation of the pathway is linked to the protumorigenic effect.
Methods: We established hepatocyte-specific Pik3ca transgenic (Tg) mice harboring N1068fs*4 mutation.
Results: The Tg mice exhibited hepatic steatosis and tumor development. PPAR gamma-dependent lipogenesis was accelerated in the Tg liver, and the abnormal profile of accumulated fatty acid (FA) composition was observed in the tumors of Tg livers. In addition, the Akt/mTOR pathway was highly activated in the tumors, and in turn, the expression of tumor suppressor genes including Pten, Xpo4, and Dlc1 decreased. Interestingly, we found that the suppression of those genes and the enhanced in vitro colony formation were induced in the immortalized hepatocytes by the treatment with oleic acid (OA), which is one of the FAs that accumulated in tumors.
Conclusions: Our data suggest that the unusual FA accumulation has a possible role in promoting in vivo hepato-tumorigenesis under constitutive activation of the PI3K pathway. The Pik3ca Tg mice might help to elucidate molecular mechanisms by which metabolic dysfunction contributes to in vivo tumor progression. (C) 2011 European Association for the Study of the Liver. Published by Elsevier B. V. All rights reserved.

DOI： 10.1016/j.jhep.2011.03.025

Direct differentiation of hepatic cells from human induced pluripotent stem cells using a limited number of cytokines Reviewed

Akemi Takata, Motoyuki Otsuka, Tomomi Kogiso, Kentaro Kojima, Takeshi Yoshikawa, Ryosuke Tateishi, Naoya Kato, Shuichiro Shiina, Haruhiko Yoshida, Masao Omata, Kazuhiko Koike

HEPATOLOGY INTERNATIONAL 5 ( 4 ) 890 - 898 2011.12

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

Purpose Development of improved protocols for differentiating induced pluripotent stem (iPS) cells into hepatic cells is an important step toward their use in the field of hepatology. Specifically, the number of different cytokines should be reduced to limit undesired effects and to reduce the cost of the process. In this report, we describe a simple method for directing human iPS cells to differentiate into hepatic cells using only two cytokines and a short incubation time.
Methods A two-step protocol for differentiating iPS cells into hepatic cells was developed. A high dose of activin A was applied for 3 days to induce definitive endoderm formation. Subsequently, cells were treated with hepatocyte growth factor (HGF) for 5 days to generate hepatic cells. Differentiation was confirmed by immunostaining for differentiation markers. Albumin mRNA levels in differentiated hepatic cells generated using a previously tested three-step protocol that uses activin A, fibroblast growth factor (FGF)/bone morphogenetic protein (BMP), and HGF, and our new protocol were compared to determine the efficiency of differentiation.
Results Our two-step protocol induced the differentiation of iPS cells into hepatic cells and required a shorter differentiation period than the previous three-step protocol. The differentiation efficiencies of the two protocols were comparable and the induced hepatic cells were functional.
Conclusions Developing efficient induction and culture methods to generate more highly matured hepatocytes is essential for regenerative cell-based therapies. Our protocol provides a simple, cost-effective, and time-saving approach for generating hepatic cells from iPS cells.

DOI： 10.1007/s12072-011-9251-5

Receptor for Activated Protein Kinase C: Requirement for Efficient MicroRNA Function and Reduced Expression in Hepatocellular Carcinoma Reviewed

Motoyuki Otsuka, Akemi Takata, Takeshi Yoshikawa, Kentaro Kojima, Takahiro Kishikawa, Chikako Shibata, Mutsuhiro Takekawa, Haruhiko Yoshida, Masao Omata, Kazuhiko Koike

PLOS ONE 6 ( 9 ) 2011.9

Language：English Publishing type：Research paper (scientific journal) Publisher：PUBLIC LIBRARY SCIENCE

MicroRNAs (miRNAs) are important regulators of gene expression that control physiological and pathological processes. A global reduction in miRNA abundance and function is a general trait of human cancers, playing a causal role in the transformed phenotype. Here, we sought to newly identify genes involved in the regulation of miRNA function by performing a genetic screen using reporter constructs that measure miRNA function and retrovirus-based random gene disruption. Of the six genes identified, RACK1, which encodes "receptor for activated protein kinase C" (RACK1), was confirmed to be necessary for full miRNA function. RACK1 binds to KH-type splicing regulatory protein (KSRP), a member of the Dicer complex, and is required for the recruitment of mature miRNAs to the RNA-induced silencing complex (RISC). In addition, RACK1 expression was frequently found to be reduced in hepatocellular carcinoma. These findings suggest the involvement of RACK1 in miRNA function and indicate that reduced miRNA function, due to decreased expression of RACK1, may have pathologically relevant roles in liver cancers.

DOI： 10.1371/journal.pone.0024359

MicroRNA-22 and microRNA-140 suppress NF-kappa B activity by regulating the expression of NF-kappa B coactivators Reviewed

Akemi Takata, Motoyuki Otsuka, Kentaro Kojima, Takeshi Yoshikawa, Takahiro Kishikawa, Haruhiko Yoshida, Kazuhiko Koike

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 411 ( 4 ) 826 - 831 2011.8

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Nuclear factor kappa B (NF-kappa B) is a transcription factor that regulates a set of genes that are critical to many biological phenomena, including liver tumorigenesis. To identify microRNAs (miRNAs) that regulate NF-kappa B activity in the liver, we screened 60 miRNAs expressed in hepatocytes for their ability to modulate NF-kappa B activity. We found that miRNA-22 and miRNA-140-3p significantly suppressed NF-kappa B activity by regulating the expression of nuclear receptor coactivator 1 (NCOA1) and nuclear receptor-interacting protein 1 (NRIP1), both of which are NF-kappa B coactivators. Our results provide new information about the roles of miRNAs in the regulation of NF-kappa B activity. (C) 2011 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.bbrc.2011.07.048

HCV関連肝細胞癌患者の高齢化と非硬変肝からの発癌の増加

吉田晴彦, 建石良介, 佐藤雅哉, 三神信太郎, 内野康史, 新野徹, 榎奥健一郎, 後藤絵里子, 中川隼人, 増崎良太, 浅岡良成, 大塚基之, 山敷宣代, 五藤忠, 椎名秀一朗, 小俣政男, 小池和彦

肝臓 52 ( Suppl.1 ) A411 - A411 2011.4

Reduced expression of RAS protein activator like-1 in gastric cancer Reviewed

Motoko Seto, Miki Ohta, Tsuneo Ikenoue, Takafumi Sugimoto, Yoshinari Asaoka, Motohisa Tada, Dai Mohri, Yotaro Kudo, Hideaki Ijichi, Keisuke Tateishi, Motoyuki Otsuka, Yoshihiro Hirata, Shin Maeda, Kazuhiko Koike, Masao Omata

INTERNATIONAL JOURNAL OF CANCER 128 ( 6 ) 1293 - 1302 2011.3

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-BLACKWELL

RAS signaling is frequently deregulated in human neoplasms. However, RAS mutations have been found in only a small proportion of human gastric cancers, implicating other mechanisms in the activation of RAS signaling in gastric tumorigenesis. We have previously reported that decreased expression of RAS protein activator like-1 (RASAL1), a member of the RAS-GTPase-activating proteins that switch off RAS activity, contributes to colon tumor progression. In our study, we explored the involvement of decreased RASAL1 expression in gastric tumorigenesis. RASAL1 expression was reduced in 6 of 10 gastric cancer cell lines examined by immunoblotting. Knockdown of RASAL1 increased mitogen-activated protein kinase signaling in response to growth factor stimulation, and the forced expression of RASAL1 reduced proliferation of gastric cancer cells. Immunohistochemical analyses in primary gastric tumors showed that RASAL1 expression was reduced in 23 of 48 (48%) of the gastric cancers but in none of the adenomas (0/10). Methylation of the RASAL1 promoter region and loss of heterozygosity (LOH) at the RASAL1 locus were examined to investigate the causes of RASAL1 silencing. All cell lines with reduced RASAL1 had RASAL1 methylation, and two had LOH. In primary gastric cancers, methylation or LOH was detected in 50% (6/12) of those with reduced RASAL1. Furthermore, RASAL1 expression was restored in some cell lines by histone deacetylase inhibitor treatment. Our findings demonstrate that reduced RASAL1 expression, partly due to genetic and epigenetic changes, contributes to gastric carcinogenesis, and also re-emphasize the importance of RAS signaling in gastric cancer development.

DOI： 10.1002/ijc.25459

Epithelial p38 alpha Controls Immune Cell Recruitment in the Colonic Mucosa Reviewed

Young Jun Kang, Motoyuki Otsuka, Arjen van den Berg, Lixin Hong, Zhe Huang, Xiurong Wu, Duan-Wu Zhang, Bruce A. Vallance, Peter S. Tobias, Jiahuai Han

PLOS PATHOGENS 6 ( 6 ) e1000934 2010.6

Language：English Publishing type：Research paper (scientific journal) Publisher：PUBLIC LIBRARY SCIENCE

Intestinal epithelial cells (IECs) compose the first barrier against microorganisms in the gastrointestinal tract. Although the NF-kappa B pathway in IECs was recently shown to be essential for epithelial integrity and intestinal immune homeostasis, the roles of other inflammatory signaling pathways in immune responses in IECs are still largely unknown. Here we show that p38 alpha in IECs is critical for chemokine expression, subsequent immune cell recruitment into the intestinal mucosa, and clearance of the infected pathogen. Mice with p38 alpha deletion in IECs suffer from a sustained bacterial burden after inoculation with Citrobacter rodentium. These animals are normal in epithelial integrity and immune cell function, but fail to recruit CD4(+) T cells into colonic mucosal lesions. The expression of chemokines in IECs is impaired, which appears to be responsible for the impaired T cell recruitment. Thus, p38 alpha in IECs contributes to the host immune responses against enteric bacteria by the recruitment of immune cells.

DOI： 10.1371/journal.ppat.1000934

DDX20 deficiency enhances NF-kappa B by impairing NF-kappa B suppressive-microRNA function and leads to hepatocarcinogenesis Reviewed

Takata Akemi, Otsuka Motoyuki, Kudo Yotaro, Kojima Kentaro, Maeda Shin, Tateishi Keisuke, Ikenoue Tsuneo, Ijichi Hideaki, Hirata Yoshihiro, Yoshida Haruhiko, Omata Masao, Koike Kazuhiko

CANCER RESEARCH 70 2010.4

Apoptosis Signal-Regulating Kinase 1 Regulates Colitis and Colitis-Associated Tumorigenesis by the Innate Immune Responses Reviewed

Yoku Hayakawa, Yoshihiro Hirata, Hayato Nakagawa, Kei Sakamoto, Yohko Hikiba, Motoyuki Otsuka, Hideaki Ijichi, Tsuneo Ikenoue, Keisuke Tateishi, Masao Akanuma, Keiji Ogura, Haruhiko Yoshida, Hidenori Ichijo, Masao Omata, Shin Maeda

GASTROENTEROLOGY 138 ( 3 ) 1055 - U334 2010.3

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO-ELSEVIER INC

BACKGROUND & AIMS: Mitogen-activated protein kinase (MAPK) signaling pathways regulate multiple cellular functions and are implicated in the pathogenesis of inflammatory bowel disease and colitis-associated cancer (CAC). Apoptosis signal-regulating kinase 1 (ASK1) is a MAPK kinase kinase; little is known about the role of ASK1 in colonic disease. We assessed the involvement of ASK1 in the development of intestinal inflammation and CAC. METHODS: Dextran sodium sulfate (DSS) or Citrobacter rodentium was used to induce colitis in wildtype (WT) and ASK1 knock-out (ASK1(-/-)) mice; CAC was induced by azoxymethane injection followed by repeated intake of DSS by the mice. Primary macrophages were isolated from WT and ASK1(-/-) mice and used to investigate the involvement of ASK1 in innate immune responses. Bone marrow chimeric mice were used to study the contribution of myeloid cells to colitis activity. RESULTS: ASK1 deficiency increased susceptibility to colonic inflammation in both models of colitis. In vitro, ASK1(-/-) macrophages were impaired in their ability to kill bacteria and had increased susceptibility to bacterial-induced apoptosis, because p38 was inactivated. Expression of antiapoptotic genes was greatly reduced in ASK1(-/-) macrophages. WT mice given transplants of ASK1(-/-) mouse-derived bone marrow cells developed more severe DSS-induced colitis than mice with WT-derived bone marrow cells. In the CAC model, ASK1(-/-) mice developed more numerous and larger tumors than WT mice through increased colonic inflammation. CONCLUSIONS: ASK1 controls the development of intestinal inflammation and CAC through the regulation of innate immunity.

DOI： 10.1053/j.gastro.2009.11.015

TNF-alpha stimulation inhibits siRNA-mediated RNA interference through a mechanism involving poly-(A) tail stabilization. Reviewed

Mols J, van den Berg A, Otsuka M, Zheng M, Chen J, Han J

Biochimica et biophysica acta 1779 712 - 719 2008.11

TNF-alpha stimulation inhibits siRNA-mediated RNA interference through a mechanism involving poly-(A) tail stabilization Reviewed

Johann Mols, Arjen van den Berg, Motoyuki Otsuka, Min Zheng, Jianming Chen, Jiahuai Han

BIOCHIMICA ET BIOPHYSICA ACTA-GENE REGULATORY MECHANISMS 1779 ( 11 ) 712 - 719 2008.11

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

The control of mRNA stability is a complex biological process that involves numerous factors, including rnicroRNA (miRNA) and short interfering RNA (siRNA). Here, we show that short interfering RNA (siRNA) and rnicroRNA share some similarities in their response to cellular stress. rniR16 expedites the degradation of mRNAs containing AU-rich elements (ARE) in their 31 untranslated region (UTR). si20 is an siRNA designed to target a non-ARE sequence in the TNF 3'UTR. We found that both si20 and miR16/ARE-mediated degradation of mRNAs can be inhibited by stimulating cells with different stresses. By analyzing TNF-alpha stimulation-mediated stabilization of si20- and miR16-targeted mRNA, we show that this stabilization is not caused by modifying si20 and miR16 loading into Ago2 complexes, or mRNA targeting to Ago2, but by inhibiting mRNA deadenylation. This is the first report showing that a specific siRNA-mediated mRNA degradation can be regulated by inflammatory stimuli, and that deadenylation is involved in this siRNA-mediated mRNA decay. (C) 2008 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.bbagrm.2008.03.007

Potential contribution of tumor suppressor p53 in the host defense against hepatitis C virus Reviewed

Narayan Dharel, Naoya Kato, Ryosuke Muroyama, Hiroyoshi Taniguchi, Motoyuki Otsuka, Yue Wang, Amarsanaa Jazag, Run-Xuan Shao, Jin-Hai Chang, Mark K. Adler, Takao Kawabe, Masao Omata

HEPATOLOGY 47 ( 4 ) 1136 - 1149 2008.4

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS INC

Infection by hepatitis C virus (HCV) usually results into chronic hepatitis that can ultimately lead to cirrhosis and hepatocellular carcinoma. Type 1 interferons (IFN-alpha/beta) constitute the primary cellular defense against viral infection including HCV. IFN binding to their receptors activates associated Jak1 and Tyk2 kinases, which ultimately leads to phosphorylation and assembly of a signal transducer and activator of transcription protein (STAT)1-STAT2-interferon regulatory factor (IRF)9 trimetric complex called interferon-stimulated gene factor 3 that translocates into the nucleus and binds to the interferon-stimulated response elements (ISRE), leading to transcriptional induction of several antiviral genes, including double-stranded RNA-activated protein kinase (PKR), 2',5'-oligoadenylate synthetase (OAS), and myxovirus resistance protein A (MxA). Understanding the mechanisms of how the virus evades this cellular innate defense and establishes a chronic infection is the key for the development of better therapeutics against HCV infection. Here, we demonstrate that p53 could have a crucial role in the cellular innate defense against HCV. We observed significantly higher levels of HCV RNA replication and viral protein expression in the Huh7 cells when their p53 expressions were knocked down. Moreover, IFN treatment was less effective in inhibiting the HCV RNA replication in the p53-knocked-down (p53kd) Huh? cells. In fact, the activation of the ISRE and the induction of ISGs were significantly attenuated in the p53kd Huh? cells and p53 was found to directly interact with IRF9. Conclusion: These observations underscore the potential contributions of the tumor suppressor p53 in cellular antiviral immunity against HCV with possible therapeutic implications.

DOI： 10.1002/hep.22176

Macrophage deletion of p38 alpha partially impairs lipopolysaccharide-induced cellular activation Reviewed

Young Jun Kang, Jianming Chen, Motoyuki Otsuka, Johann Mols, Shuxun Ren, Yinbin Wang, Jiahuai Han

JOURNAL OF IMMUNOLOGY 180 ( 7 ) 5075 - 5082 2008.4

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC IMMUNOLOGISTS

The activation of p38 alpha, a MAPK family member, is associated with macrophage activation by microbial pattern molecules, such as LPS. The requirement of p38 alpha in inflammatory responses has been shown in a number of studies using chemical inhibitors, though the inhibitors also inhibit p38 beta and perhaps some other enzymes. In this study, we used conditional knockout of p38 alpha in macrophages to address the role of p38 alpha in macrophage activation. We found that p38 alpha deficiency causes a significant inhibition in the production of LPS-induced TNF-alpha, IL-12, and IL-18, but it has little or no effect on IL-6 or IFN-beta production. Knockout of p38a in macrophages did not affect LPS-induced activation of the other major signaling pathways (NF-kappa B, Jnk, and Erk), nor did it affect the transcriptional activity of NF-kappa B. It had little inhibitory effect on LPS-induced AP-1 activity, but it significantly inhibited LPS-induced C/EBP-beta and CREB activation, indicating that the role of p38 alpha in cytokine production in macrophages is at least in part through its regulation of C/EBP-beta and CREB activation. In addition, we also confirmed that p38 alpha is important for phagocytosis of bacteria by macrophages. Our in vivo studies with two murine models showed that p38 alpha is involved in sepsis. Collectively, our data demonstrate that p38 alpha is an important player in inflammatory responses.

Calcineurin negatively regulates TLR-Mediated activation pathways Reviewed

Young Jun Kang, Brenda Kusler, Motoyuki Otsuka, Michael Hughes, Nobutaka Suzuki, Shinobu Suzuki, Wen-Chen Yeh, Shizuo Akira, Jiahuai Han, Patricia P. Jones

JOURNAL OF IMMUNOLOGY 179 ( 7 ) 4598 - 4607 2007.10

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC IMMUNOLOGISTS

In innate immunity, microbial components stimulate macrophages to produce antimicrobial substances, cytokines, other proinflammatory mediators, and IFNs via TLRs, which trigger signaling pathways activating NF-kappa B, MAPKs, and IFN response factors. We show in this study that, in contrast to its activating role in T cells, in macrophages the protein phosphatase calcineurin negatively regulates NF-kappa B, MAPKs, and IFN response factor activation by inhibiting the TLR-mediated signaling pathways. Evidence for this novel role for calcineurin was provided by the findings that these signaling pathways are activated when calcineurin is inhibited either by the inhibitors cyclosporin A or FK506 or by small interfering RNA-targeting calcineurin, and that activation of these pathways by TLR ligands is inhibited by the overexpression of a constitutively active form of calcineurin. We further found that I kappa B-alpha degradation, MAPK activation, and TNF-alpha production by FK506 were reduced in macrophages from mice deficient in MyD88, Toll/IL-1R domain-containing adaptor-inducing IFN-beta (TRIF), TLR2, or TLR4, whereas macrophages from TLR3-deficient or TLR9 mutant mice showed the same responses to FK506 as those of wild-type cells. Biochemical studies indicate that calcineurin interacts with MyD88, TRIF, TLR2, and TLR4, but not with TLR3 or TLR9. Collectively, these results suggest that calcineurin negatively regulates TLR-mediated activation pathways in macrophages by inhibiting the adaptor proteins MyD88 and TRIF, and a subset of TLRs.

Interferon-beta is activated by hepatitis C virus NS5B and inhibited by NS4A, NS4B, and NS5A Reviewed

Masaru Moriyama, Naoya Kato, Motoyuki Otsuka, Run-Xuan Shao, Hiroyoshi Taniguchi, Takao Kawabe, Masao Omata

HEPATOLOGY INTERNATIONAL 1 ( 2 ) 302 - 310 2007.6

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER

Innate immunity is part of the antiviral response. Interferon (IFN)-beta plays a leading role in this system. To investigate the influence of hepatitis C virus (HCV) on innate immunity, we examined the effect of viral proteins on IFN-beta induction. HepG2 cells were co-transfected with plasmids for seven HCV proteins (core protein, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) and the IFN-beta promoter luciferase. Toll-like receptor (TLR) 3 and Toll/IL-1 receptor domain-containing adapter inducing IFN-beta (TRIF) play key roles in dsRNA-mediated activation of interferon regulatory factor (IRF)-3 and IFN-beta; therefore, the participation of TLR3/TRIF in NS5B-mediated IFN induction was examined. Among seven HCV proteins, only NS5B, a viral RNA-dependent RNA polymerase (RdRp), activated the IFN-beta promoter. However, mutant NS5B without RdRp activity or template/primer association did not activate the IFN-beta promoter. Activation of the IFN-beta promoter by NS5B required the positive regulatory domain III, a binding sequence for IRF-3. Moreover, IRF-3 was phosphorylated by NS5B. Both inhibition of TLR3 expression by small interfering RNA and expression of the dominant negative form of TRIF significantly reduced NS5B-induced activation of IFN-beta. Of the six other HCV proteins, NS4A, NS4B, and NS5A efficiently inhibited this activation. HCV NS5B is a potent activator of the host innate immune system, possibly through TLR3/TRIF and synthesis of dsRNA. Meanwhile, NS4A, NS4B, and NS5A block IFN-beta induction by NS5B, which may contribute toward the persistence of this virus.

DOI： 10.1007/s12072-007-9003-8

Gene expressions associated with chemosensitivity in human hepatoma cells Reviewed

Yujin Hoshida, Masaru Moriyama, Motoyuki Otsuka, Naoya Kato, Hiroyoshi Taniguchi, Yasushi Shiratori, Naohiko Seki, Masao Omata

HEPATO-GASTROENTEROLOGY 54 ( 74 ) 489 - 492 2007.3

Language：English Publishing type：Research paper (scientific journal) Publisher：H G E UPDATE MEDICAL PUBLISHING S A

Background/Aims: Only limited patients with hepatoma. benefit from chemotherapy without a clear explanation. We aimed to identify genes associated with chemosensitivity using transcriptional profiles. Methodology: In 8 hepatoma cells (HLE, HLF, Huh7, Hep3B, PLC/PRF/5, SK-Hep1, Huh6, and HepG2) transcriptional profiles were obtained using cDNA microarray including 2,300 genes. Chemosensitivities to 8 anticancer drugs (nimustine, mitomycin C, cisplatin, carboplatin, doxorubicin, epirubicin, mitoxantrone, and 5-fluorouracil) were measured by obtaining 50% growth inhibitory concentrations (GI50) using MTT assay. Genes having drug-specific association with chemosensitivity were selected. Results: Up-regulation of topoisomerase II beta was associated with chemo-resistance, the target of doxorubicin. Platinum-specific resistance was associated with superoxide dismutase 2 expression. Antigen peptide transporter I expression correlated with nimustine and mitoxantrone-specific susceptibility. These results were verified by semi-quantitative RT-PCR. Drug inactivators reported in non-liver cancers such as multidrug transporters and drug metabolizers showed less diversity of chemosensitivity in hepatoma cells. Conclusions: To evaluate these gene expressions may be useful to select anticancer drugs, and possibly to consider new therapeutic target to modify drug action.

Absence of tyrosine kinase mutations in Japanese colorectal cancer patients Reviewed

R-X Shao, N. Kato, L-J Lin, R. Muroyama, M. Moriyama, T. Ikenoue, H. Watabe, M. Otsuka, B. Guleng, M. Ohta, Y. Tanaka, S. Kondo, N. Dharel, J-H Chang, H. Yoshida, T. Kawabe, M. Omata

ONCOGENE 26 ( 14 ) 2133 - 2135 2007.3

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

Tyrosine kinases, which are important regulators of intracellular signal-transduction pathways, have mutated forms that are often associated with oncogenesis and are attractive targets for therapeutic intervention. Recently, systematic mutational analyses of tyrosine kinases revealed that a minimum of 30% of colorectal cancer contain at least one mutation in the tyrosine kinases. To further explore these mutations, we examined all reported mutations of NTRK3, FES, KDR, EPHA3, NTRK2, JAK1, PDGFRA, EPHA7, EPHA8, ERBB4, FGFR1, MLK4 and GUCY2F genes in the 24 colorectal cancer cell lines. Unexpectedly, among 24 colorectal cancer cell lines, only two cell lines (LoVo and CaR1) harbored mutation C1408T (R470C) in MLK4 gene. The mutation rate was extremely low compared to that previously reported. Therefore, we analyzed mutations in 46 colorectal cancer samples resected from the same number of Japanese patients. Surprisingly, none of the 46 samples contained any of the mutations reported. Based on our study, we advise that a more comprehensive tyrosine kinase gene mutation assay is necessary in the future.

DOI： 10.1038/sj.onc.1210007

Nucleotide change of codon 38 in the X gene of hepatitis B virus genotype C is associated with an increased risk of hepatocellular carcinoma Reviewed

Ryosuke Muroyama, Naoya Kato, Haruhiko Yoshida, Motoyuki Otsuka, Masaru Moriyama, Yue Wang, Run-Xuan Shao, Narayan Dharel, Yasuo Tanaka, Miki Ohta, Ryosuke Tateishi, Shuichiro Shiina, Masashi Tatsukawa, Kenichi Fukai, Fumio Imazeki, Osamu Yokosuka, Yasushi Shiratori, Masao Omata

JOURNAL OF HEPATOLOGY 45 ( 6 ) 805 - 812 2006.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Background/Aims: The hepatitis B virus (HBV) genotype C is associated with the development of hepatocellular carcinoma (HCC). In addition, the HBV X gene, which encodes the pleiotropic transactivator HBx, has also been associated with the development of HCC. In this study, we investigated whether nucleotide changes in the X gene of genotype C are associated with the development of HCC.
Methods/Results: We sequenced the X gene in age- and sex-matched 39 HBV-infected patients with HCC and 36 HBV-infected patients without HCC. A novel nucleotide change that resulted in a proline to serine substitution at codon 38 in HBx (codon-38 change) was preferentially found in patients with HCC. Then, sera were collected from a new group of age- and sex-matched 52 patients with HCC and 51 patients without HCC. In this cohort also, the codon-38 change was associated with HCC. Multiple logistic regression analysis showed the prevalence of the codon-38 change was significantly associated with HCC in all patients (P = 0.001, odds ratio: 4.89).
Conclusion: The codon-38 change in genotype C is an independent risk factor for the development of HCC and may serve as a useful molecular marker for predicting the clinical outcomes in patients infected with HBV. (c) 2006 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.jhep.2006.07.025

Hepatitis C virus core protein is a potent inhibitor of RNA silencing-based antiviral response Reviewed

Y Wang, N Kato, A Jazag, N Dharel, M Otsuka, H Taniguchi, T Kawabe, M Omata

GASTROENTEROLOGY 130 ( 3 ) 883 - 892 2006.3

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO-ELSEVIER INC

Background & Aims: Persistent infection with hepatitis C virus (HCV) leads to chronic hepatitis and hepatocellular carcinoma (HCC). RNA interference (RNAi) may act as a host antiviral response against viral RNA. Methods: The effects of RNAi on both the replicative intermediates and the internal ribosome entry site (IRES) of HCV were studied by using HCV-related short interfering RNA (siRNA) detection assay. The mechanism that permits HCV to escape RNAi was studied by using RNAi assay materials. Results: These studies demonstrate that the Dicer, an RNase enzyme that generates short siRNA, can target and digest both the IRES and the replicative intermediate of HCV into siRNA of similar to 22 nucleotides. Further studies also show that Dicer can inhibit the replication of the HCV subgenomic replicon. However, the HCV core protein inhibits this RNAi and rescues the replication of the HCV subgenomic replicon through a direct interaction with Dicer. Conclusions: RNAi is a limiting factor for HCV infection, and the core protein suppresses the RNA silencing-based antiviral response. This ability of the core protein to counteract the host defense may lead to a persistent viral infection and may contribute to the pathogenesis of HCV.

DOI： 10.1053/j.gastro.2005.12.028

Identifying genes with differential expression in gemcitabine-resistant pancreatic cancer cells using comprehensive transcriptome analysis Reviewed

Y Nakai, M Otsuka, Y Hoshida, M Tada, Y Komatsu, T Kawabe, M Omata

ONCOLOGY REPORTS 14 ( 5 ) 1263 - 1267 2005.11

Language：English Publishing type：Research paper (scientific journal) Publisher：PROFESSOR D A SPANDIDOS

Pancreatic cancer is often unresectable at diagnosis, and chemotherapy using gemcitabine is now the standard treatment for advanced pancreatic cancer. However, acquired resistance to gemcitabine resulting in therapeutic failure is often encountered. Therefore, we sought to identify genes that determine gemcitabine resistance by evaluating the relationship between gene expression profiles and gemcitabine sensitivity to provide molecular targets for overcoming gemcitabine resistance. First, the gemcitabine concentration needed for 50% growth inhibition was examined in six pancreatic cancer cell lines. By exposing MIA PaCa-2 cells to long-term gemcitabine, we established gemcitabine-resistant cells. The gene expression profiles of the six pancreatic cancer cell lines and gemcitabine-resistant cells were determined using cDNA microarray analysis. By comparing the results, 30 genes were identified as differentially expressed genes correlated with gemcitabine sensitivity. Differentially expressed genes in the parental cell lines were also examined, and six overlapping genes were identified as genes correlated with gemcitabine sensitivity in both assays. Of these genes, the down-regulated expression of TNFSF6 protein, also known as Fas ligand, was confirmed in the gemcitabine-resistant cell line. These results should provide therapeutic molecular targets for overcoming gemcitabine resistance.

Large-scale search of single nucleotide polymorphisms for hepatocellular carcinoma susceptibility genes in patients with hepatitis C Reviewed

N Kato, GJ Ji, Y Wang, M Baba, Y Hoshida, M Otsuka, H Taniguchi, M Moriyama, N Dharel, T Goto, RX Shao, T Matsuura, K Ishii, S Shiina, T Kawabe, M Muramatsu, M Omata

HEPATOLOGY 42 ( 4 ) 846 - 853 2005.10

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS INC

Hepatitis C virus (HCV) infection is a major risk factor for developing hepatocellular carcinoma (HCC). The host genetic factors that are involved in the development of HCC in patients with HCV infection remain to be investigated. To search for single nucleotide polymorphisms (SNPs) in HCC susceptibility genes, 393 SNPs in 171 candidate genes were examined in 188 Japanese patients with chronic HCV infection, including 77 patients with HCC. HCC-related SNPs were then examined in another 188 patients (including 93 patients with HCC) with chronic HCV infection. Haplotype analyses of HCC-related genes were performed in a total of 376 patients. Of the 393 SNPs, 31 SNPs in 29 genes were significantly associated with HCC based on an initial screening (P < .05). Of these 31 SNPs, 3 SNPs of 3 genes (SCYB14, GFRAI, and CRHR2) were significantly associated with HCC in a secondary screening. Haplotype analyses of these 3 genes identified 2 haplotype blocks associated with HCC. In conclusion, these SNPs and haplotypes located in the SCBY14, CRHR2, and GFRA1 genes will be used as markers to identify a subgroup of Japanese patients with chronic HCV infection who are at high risk of developing HCC.

DOI： 10.1002/hep.20860

NF-kappa B and ERK-signaling pathways contribute to the gene expression induced by cag PAI-positive-Helicobacter pylori infection Reviewed

Wataru Shibata, Yoshihiro Hirata, Haruhiko Yoshida, Motoyuki Otsuka, Yujin Hoshida, Keiji Ogura, Shin Maeda, Tomoya Ohmae, Ayako Yanai, Yuzo Mitsuno, Naohiko Seki, Takao Kawabe, Masao Omata

WORLD JOURNAL OF GASTROENTEROLOGY 11 ( 39 ) 6134 - 6143 2005.10

Language：English Publishing type：Research paper (scientific journal) Publisher：BAISHIDENG PUBL GRP CO LTD

AIM: To elucidate the sequential gene expression profile in AGS cells co-cultured with wild-type Helicobacter pylori (H pylori) as a model of H pylori-infected gastric epithelium, and to further examine the contribution of cag-pathogenicity islands (cagPAI)-coding type IV secretion system and the two pathways, nuclear factor kappa B (NF-kappa B) and extracellular signal-regulated kinases (ERK) on wild-type H pylori-induced gene expression.
METHODS: Gene expression profiles induced by H pylori were evaluated in AGS gastric epithelial cells using cDNA microarray, which were present in the 4 600 independent clones picked up from the human gastric tissue. We also analyzed the contribution of NF-kappa B and ERK signaling on H pylori-induced gene expression by using inhibitors of specific signal pathways. The isogenic mutant with disrupted cagE (Delta cagE) was used to elucidate the role of cagPAI-encoding type IV secretion system in the gene expression profile.
RESULTS: According to the expression profile, the genes were classified into four clusters. Among them, the clusters characterized by continuous upregulation were most conspicuous, and it contained many signal transducer activity-associated genes. The role of cagPAI on cultured cells was also investigated using isogenic mutant cagE, which carries non-functional cagPAI. Then the upregulation of more than 80% of the induced genes (476/566) was found to depend on cagPAI. Signal transducer pathway through NF-kappa B or ERK are the major pathways which are known to be activated by cagPAI-positive H pylori. The role of these pathways in the whole signal activation H pylori was analyzed. The specific inhibitors against NF-kappa B or ERK pathway blocked the activation of gene expression in 65% (367/566) or 76% (429/566) of the genes whose activation appealed to depend on cagPAI.
CONCLUSION: These results suggest that more than half of the genes induced by cagPAI-positive H pylori depend on NF-kappa B and ERK signaling activation, and these pathways may play a role in the gene expression induced by host-bacterial interaction which may associate with H pylori-related gastro-duodenal diseases. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.

No mutations in the tyrosine kinases of human hepatic, pancreatic, and gastric cancer cell lines Reviewed

RX Shao, M Otsuka, N Kato, JH Chang, R Muroyama, H Taniguchi, M Moriyama, Y Wang, T Kawabe, M Omata

JOURNAL OF GASTROENTEROLOGY 40 ( 9 ) 918 - 918 2005.9

Hepatitis C virus core protein and hepatitis activity are associated through transactivation of interleukin-8 Reviewed

Y Hoshida, N Kato, H Yoshida, Y Wang, M Tanaka, T Goto, M Otsuka, H Taniguchi, M Moriyama, F Imazeki, O Yokosuka, T Kawabe, Y Shiratori, M Omata

JOURNAL OF INFECTIOUS DISEASES 192 ( 2 ) 266 - 275 2005.7

Language：English Publishing type：Research paper (scientific journal) Publisher：UNIV CHICAGO PRESS

Background. We evaluated the association between variations in hepatitis C virus (HCV) core protein and hepatitis severity in patients with chronic HCV infection who achieved remission without viral eradication and had a biochemical response to interferon (IFN) therapy, to evaluate the effect of HCV core sequence in the absence of the influence of host factors.
Methods. Using serum from 10 patients with a biochemical response and 10 patients with no response, we measured serum levels of interleukin (IL)-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IFN-gamma, and tumor necrosis factor-alpha before and after IFN therapy. Expression vectors with the core region were transfected into Huh7 cells, and cytokine induction was evaluated by reporter assay.
Results. In biochemical responders, only IL-8 levels decreased after IFN therapy (P = .04). Changes in the C-terminal hydrophobic region were observed more frequently in biochemical responders. Activation of the IL-8 promoter by HCV core protein was significantly decreased in biochemical responders after IFN therapy (P = .04). When 69 C-terminal amino acids from before IFN therapy were replaced with those from after IFN therapy in 3 biochemical responders, their ability to transactivate IL-8 decreased.
Conclusions. Differences in amino acids in the HCV core protein correlates with hepatitis activity through the modulation of IL-8 induction in HCV-infected patients.

DOI： 10.1086/430924

Interaction between the HCVNS3 protein and the host TBK1 protein leads to inhibition of cellular antiviral responses Reviewed

M Otsuka, N Kato, M Moriyama, H Taniguchi, Y Wang, N Dharel, T Kawabe, M Omata

HEPATOLOGY 41 ( 5 ) 1004 - 1012 2005.5

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS INC

The persistent nature of hepatitis C virus (HCV) infection suggests that HCV encodes proteins that enable it to overcome host antiviral responses. Toll-like receptor 3 (TLR3)-mediated signaling, which recognizes the double-stranded RNA that is produced during viral replication and induces type I interferons, including interferon beta (IFN-beta), is crucial to the host defense against viruses. Recent studies suggest that a TIR domain-containing adaptor protein, TRIF, and two protein kinases, TANK-binding kinase-1 (TBK1) and I kappa B kinase-epsilon (IKK epsilon), play essential roles in TLR3-mediated IFN-beta production through the activation of the transcriptional factor interferon regulatory factor 3 (IRF-3). We report that the HCV NS3 protein interacts directly with TBK1, and that this binding results in the inhibition of the association between TBK1 and IRF-3, which leads to the inhibition of IRF-3 activation. In conclusion, these results suggest the mechanisms of the inhibition of the innate immune responses of HCV infection by NS3 protein.

DOI： 10.1002/hep.20666

Hepatic gene expression profiles associated with fibrosis progression and hepatocarcinogenesis in hepatitis C patients Reviewed

Run-Xuan Shao, Yujin Hoshida, Motoyuki Otsuka, Naoya Kato, Ryosuke Tateishi, Takuma Teratani, Shuichiro Shiina, Hiroyoshi Taniguchi, Masaru Moriyama, Takao Kawabe, Masao Omata

WORLD JOURNAL OF GASTROENTEROLOGY 11 ( 13 ) 1995 - 1999 2005.4

Language：English Publishing type：Research paper (scientific journal) Publisher：BAISHIDENG PUBL GRP CO LTD

AIM: To determine fibrosis progression and hepatocellular carcinoma (HCC), using simultaneous gene expression analysis.
METHODS: Total RNA samples were extracted from liver biopsies from 19 patients with hepatitis C virus (HCV) infection and 3 patients without HCV infection. Among the 19 HCV-infected patients, 7 and 12 patients had grade F1-2 and F3-4 fibrosis, respectively. Of the 12 patients with F3-4 fibrosis, 8 had HCC. Gene expression in the liver samples was determined using an oligonucleotide microarray. The following comparisons were performed: normal livers vs HCV-infected livers; F1-2 vs F3-4; and F3-4 with HCC vs F3-4 without HCC. Genes that were differentially expressed between these groups were identified based on signal-to-noise ratios.
RESULTS: In the HCV-infected livers, genes involved in immune responses were highly expressed. Expression levels of genes for plasma proteins and drug-metabolizing enzymes were decreased and those of genes involved in the cell cycle and oncogenesis were increased in the F3-4 cases as compared to the F1-2 cases. Among the F3-4 cases, genes involved in carbohydrate metabolism tended to be more highly expressed in patients with HCC than in patients without HCC.
CONCLUSION: We identified genes that are associated with fibrosis progression and hepatocarcinogenesis. This information may be used to detect increased carcinogenic potential in the livers of patients with HCV infection. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.

Vitamin K2 binds 17 beta-hydroxysteroid dehydrogenase 4 and modulates estrogen metabolism Reviewed

M Otsuka, N Kato, T Ichimura, S Abe, Y Tanaka, H Taniguchi, Y Hoshida, M Moriyama, Y Wang, RX Shao, D Narayan, R Muroyama, F Kanai, T Kawabe, T Isobe, M Omata

LIFE SCIENCES 76 ( 21 ) 2473 - 2482 2005.4

Language：English Publishing type：Research paper (scientific journal) Publisher：PERGAMON-ELSEVIER SCIENCE LTD

Vitamin K is a cofactor for gamma-glutamyl carboxylase, an enzyme that is important for blood coagulation. Recent studies have shown that vitamin K has other roles, in addition to post-transcriptional modification, such as bone metabolism and antitumoral actions; these findings have indicated that there might be unknown intracellular binding proteins that are specific for vitamin K. In this study, vitamin K-binding proteins were characterized by pull-down experiment using a chemically synthesized biotynylated vitamin K followed by mass spectrometric identification of the pull-downed components. The results indicated that 17 beta hydroxy steroid dehydrogenase 4, apolipoportein E, and 40S ribosomal proteins S7 and S13 might be the candidates of the vitamin K-binding proteins. Subsequent experiments showed that vitamin K2 binds 17 beta hydroxysteroid dehydrogenase 4 and decreases the intracellular estradiol:estrone ratio, which resulted in the inhibition of the amount of estrogen receptor alpha-binding to its target DNA. These results suggest a. possible novel role for vitamin K in modulating estrogen function. (c) 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.lfs.2004.12.020

Acyclic retinoid inhibits human hepatoma cell growth by suppressing fibroblast growth factor-mediated signaling pathways Reviewed

RX Shao, M Otsuka, N Kato, H Taniguchi, Y Hoshida, M Moriyama, T Kawabe, M Omata

GASTROENTEROLOGY 128 ( 1 ) 86 - 95 2005.1

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO

Background& Aims: Hepatocellular carcinoma (HCC) is one of the most common human malignancies. Its high mortality rate is mainly a result of high intrahepatic recurrence. The novel synthetic retinoid acyclic retinoid (ACR) has been reported to prevent the recurrence of human HCC after surgical resection of primary tumors, but the molecular mechanisms underlying its effects remain to be elucidated. In this study, we clarified the molecular targets of ACR. Methods: The inhibitory effects by ACR on growth were examined. Intracellular signaling induced by ACR was comprehensively studied by a reporter assay. Gene expression changes by ACR were examined using a microarray. From these results, a candidate signaling pathway modulated by ACR was determined and whether antagonizing this pathway reverses the effect was examined. Results: We show that ACR inhibits the growth of HCC cells through the down-regulation of fibroblast growth factor (FGF) receptor 3 expression and FGF-mediated signaling, which in turn suppresses the activity of Rho and serum response factor-mediated transcription. Conversely, overexpression of the active form of FGF receptor 3 or the addition of FGF reverses the ACR-mediated inhibition of growth. In addition, silencing the FGF receptor 3 gene by RNA interference inhibits cell growth. Conclusions: These studies show that ACR is a potent inhibitor of FGF signaling and that selective blocking of the FGFmediated pathway could be a promising therapeutic approach for the management of patients with HCC.

DOI： 10.1053/j.gastro.2004.09.077

A simple combination of serum type IV collagen and prothrombin time to diagnose cirrhosis in patients with chronic active hepatitis C Reviewed

Y Qiu, YJ Hoshida, N Kato, M Moriyama, M Otsuka, H Taniguchi, T Kawabe, M Omata

HEPATOLOGY RESEARCH 30 ( 4 ) 214 - 220 2004.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI IRELAND LTD

Background/aim: Cirrhosis in chronic hepatitis C is a major cause of mortality. The components of reported diagnostic indices of cirrhosis based on biochemical markers may be modified by therapies for hepatic inflammation. We aimed to construct index of cirrhosis in patients treated for chronic active hepatitis.
Methods: Using sera of consecutive 140 patients with chronic hepatitis C, routine blood tests including fibrosis markers, type IV collagen and procollagen type III peptide (PIIIP), were performed. Diagnosis of cirrhosis was determined by biopsy. Using multivariate analyses, diagnostic indices of cirrhosis were constructed.
Results: Fifty-eight patients were diagnosed to have cirrhosis. Platelet count, prothrombin time, and albumin were lower, and type IV collagen and PIIIP were higher in patients with cirrhosis (p < 0.05). There was no difference in aspartate and alanine aminotransferases (AST, ALT) and gamma-glutamyl-transpeptidase (GGT) (p > 0.3). Our diagnostic indices I (prothrombin time and platelet count) and II (prothrombin time and type IV collagen) of cirrhosis showed the area under the ROC curves (AUC) of 0.77 and 0.81, respectively. The index II was relatively superior to the index I.
Conclusions: Using combination of type IV collagen and prothrombin time, efficient diagnosis of cirrhosis can be performed in patients with chronic active hepatitis C. (C) 2004 Elsevier B.V. All rights reserved.

DOI： 10.1016/j.hepres.2004.10.006

Genes associated with human hepatocellular carcinoma cell chemosensitivity to 5-fluorouracil plus interferon-alpha combination chemotherapy Reviewed

M Moriyama, Y Hoshida, N Kato, M Otsuka, H Yoshida, T Kawabe, M Omata

INTERNATIONAL JOURNAL OF ONCOLOGY 25 ( 5 ) 1279 - 1287 2004.11

Language：English Publishing type：Research paper (scientific journal) Publisher：PROFESSOR D A SPANDIDOS

Recently, combined chemotherapy with 5-fluorouracil (5-FU) and interferon (IFN)-alpha has been reported to show marked effects in patients with advanced hepatocellular carcinoma. We investigated the genes associated with susceptibility to this combination therapy. The gene expression profiles of eight human hepatocellular carcinoma cells (HepG2, Hep3B, Huh7, Huh6, PLC/PRF/5, HLE, HLF, and SK-Hep1) were evaluated using an oligonucleotide microarray that consisted of 3,800 genes. The 50% growth inhibitory concentration (GI50) values for 5-FU, IFN-alpha, and the combination of 5-FU plus IFN-a were determined by the MTT assay. We selected genes that were expressed differentially between the cells with increased susceptibility to the combination therapy and the remaining cells. Relevance networks of the gene expression patterns and GI50 values of the susceptible cells were constructed to find genes associated with susceptibility to the combination therapy. Of the eight cells tested, five showed increased susceptibility to 5-FU plus IFN-alpha compared with 5-FU treatment alone. Among the 3,800 genes, 25 were expressed differentially between susceptible cells and resistant cells. The relevance networks revealed that sensitivity to 5-FU plus IFN-a involved the expression of 27 independent genes, which included 10 genes that are commonly associated with sensitivity to 5-FU alone. We selected a set of genes to predict susceptibility to 5-FU plus IFN-alpha combination therapy. We also selected genes that play key roles in the synergistic effect of this combination therapy. These gene sets should prove useful in evaluations of the efficacy and underlying molecular mechanisms of this combination therapy.

Vitamin K-2 inhibits the growth and invasiveness of hepatocellular carcinoma cells via protein kinase A activation Reviewed

M Otsuka, N Kato, RX Shao, Y Hoshida, H Ijichi, Y Koike, H Taniguchi, M Moriyama, Y Shiratori, T Kawabe, M Omata

HEPATOLOGY 40 ( 1 ) 243 - 251 2004.7

Language：English Publishing type：Research paper (scientific journal) Publisher：JOHN WILEY & SONS INC

Hepatocellular carcinoma (HCC) is a common human malignancy. Its high mortality rate is mainly a result of high intrahepatic recurrence and portal venous invasion (PVI). We previously reported that the development of PVI is related to levels of des-gamma-carboxy prothrombin (DCP), a serum protein that increases at a notably higher rate in patients with HCC. Because DCP is produced by a vitamin K shortage, we examined the biological effects of extrinsic supplementation of vitamin K-2 in HCC cells in vitro and in vivo. Consequently, vitamin K-2 inhibits the growth and invasion of HCC cells through the activation of protein kinase A, which modulates the activities of several transcriptional factors and inhibits the small GTPase Rho, independent of suppression of DCP. In addition, administration of vitamin K-2 to nude mice inoculated with liver tumor cells reduced both tumor growth and body weight loss. In conclusion, similar to an acyclic retinoid-which was previously reported to prevent the recurrence of HCC-vitamin K-2, another lipid-soluble vitamin, may be a promising therapeutic means for the management of HCC.

DOI： 10.1002/hep.20260

UDP-glucuronosyltransferase 1A7 genetic polymorphisms are associated with hepatocellular carcinoma in Japanese patients with hepatitis C virus infection Reviewed

Y Wang, N Kato, Y Hoshida, M Otsuka, H Taniguchi, M Moriyama, S Shiina, T Kawabe, YM Ito, M Omata

CLINICAL CANCER RESEARCH 10 ( 7 ) 2441 - 2446 2004.4

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC CANCER RESEARCH

Purpose: Genetic polymorphisms of UDP-glucuronosyl-transferase 1A7 (UGT1A7), which detoxifies endogenous and environmental carcinogens, have been reported to be associated with hepatocellular carcinoma (HCC) in German populations. On the other hand, we reported that interleukin-1beta (IL-1beta) gene polymorphisms were associated with hepatitis C virus (HCV)-related HCC. In this study, we evaluated the association of both genes with the risk of HCC in Japanese HCV-infected patients.
Experimental Design: Genetic polymorphisms of UGT1A7 and IL-1beta were investigated in 280 Japanese patients (122 with HCC and 158 without HCC) with chronic HCV infections, by use of standard PCR-based genotyping techniques.
Results: We designated the UGT1A7*1 allele (a haplotype conferring higher activity) as H and the *2, *3, and *4 alleles (haplotypes conferring lower activity) as L. The proportions of UGT1A7 L/L and H/L alleles (genotypes) in patients with HCC (25% and 45%, respectively) were higher than those in patients without HCC (15% and 39%, respectively) with odds ratios of 2.73 (95% confidence interval, 1.40-5.35) and 1.80 (95% confidence interval, 1.05-3.09), respectively, compared with the UGT1A7 H/H alleles. Multivariate analyses revealed that UGT1A7 L/L and IL-1beta/-31T/T-511C/C genotypes, the presence of cirrhosis, age 60 years, male sex, and alpha-fetoprotein >20 mug/ml were associated with the presence of HCC (odds ratios, 2.33, 2.67, 4.20, 3.12, 3.09, and 2.90, respectively).
Conclusion: The UGT1A7 polymorphisms together with IL-1beta were associated with the presence of HCC in Japanese HCV-infected patients.

DOI： 10.1158/1078-0432.CCR-1187-3

Smad4-independent regulation of p21/WAF1 by transforming growth factor-beta Reviewed

H Ijichi, M Otsuka, K Tateishi, T Ikenoue, T Kawakami, F Kanai, Y Arakawa, N Seki, K Shimizu, K Miyazono, T Kawabe, M Omata

ONCOGENE 23 ( 5 ) 1043 - 1051 2004.2

Language：English Publishing type：Research paper (scientific journal) Publisher：NATURE PUBLISHING GROUP

The transforming growth factor-beta (TGF-beta)-Smad signaling pathway inhibits the growth of human epithelial cells and plays a role in tumor suppression. The Smad4 gene is mutated or deleted in 50% of pancreatic cancers. In this study, the Smad4-null pancreatic cancer cell line BxPC-3 was transfected with either the Smad4 expression vector or the empty vector and incubated in the presence or absence of TGF-beta. The cells were analysed using a cDNA microarray, which included 2280 named genes to screen for target genes regulated by TGF-beta in either a Smad4-dependent or -independent manner. The microarray and subsequent quantitative RT-PCR analysis demonstrated that the Smad4-independent and -dependent signaling pathways driven by TGF-beta upregulated only one of the 2280 genes, respectively, suggesting that Smad4-independent signaling downstream of TGF-beta might be as widespread as Smad4-dependent signaling. In this study, we demonstrated that the cyclin-dependent kinase inhibitor p21/WAF1, which has been considered the major effector of the Smad-dependent growth inhibitory signal of TGF-beta, is upregulated in a Smad4-independent manner. The upregulation occurs through Smad2/3-dependent transcriptional activation of the p21/WAF1 promoter region. These results suggest a novel mechanism of gene regulation, that is, a novel signal mediator other than Smad4.

DOI： 10.1038/sj.onc.1207222

Hepatitis C virus core protein upregulates transforming growth factor-beta 1 transcription Reviewed

H Taniguchi, N Kato, M Otsuka, T Goto, H Yoshida, Y Shiratori, M Omata

JOURNAL OF MEDICAL VIROLOGY 72 ( 1 ) 52 - 59 2004.1

Language：English Publishing type：Research paper (scientific journal) Publisher：WILEY-LISS

The majority of persons with chronic hepatitis C virus (HCV) infection develop liver fibrosis. Transforming growth factor (TGF)-beta1 plays a pivotal role in the pathogenesis of post-inflammatory liver scarring. To clarify the influence of HCV infection on liver fibrosis, a reporter assay was used to investigate the effect of viral proteins on TGF-beta1 expression in human hepatoma cells. Of all HCV proteins investigated (core, E1/E2/p7, NS2, NS3, NS4A, NS4B, NS5A, and NS5B), only the core protein activated the TGF-beta1 promoter and upregulated TGF-beta1 expression measured by an RNase protection assay. Bases -376 to -331 by in the promoter region of TGF-beta1 are responsible for upregulation by HCV core protein, and the nuclear protein that binds to this region increased with the stimulation of HCV core protein. Blocking the mitogen-activated protein kinase pathway prevented upregulation of TGF-beta1 by HCV core protein. The immunological response is supposed to be a major factor to cause the secretion of TGF-beta1 from non-parenchymal cells, but the results suggest that the HCV core protein expression may upregulate directly TGF-beta1 transcription in parenchymal cells and suggest a new paradigm for exacerbation of liver fibrosis by HCV infection.

DOI： 10.1002/jmv.10545

Antiapoptotic regulation by hepatitis C virus core protein through up-regulation of inhibitor of caspase-activated DNase Reviewed

R Sacco, T Tsutsumi, R Suzuki, M Otsuka, H Aizaki, S Sakamoto, M Matsuda, N Seki, Y Matsuura, T Miyamura, T Suzuki

VIROLOGY 317 ( 1 ) 24 - 35 2003.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

The hepatitis C virus (HCV) core protein is considered to influence multiple cellular processes. We developed a human hepatoblastoma HepG2-derived inducible cell line, Hep191, which allows tightly regulated expression of the core protein at relatively low but physiological levels under control of the ecdysone-regulated promoter. By transcriptional profiling, we identified differentially expressed genes, some of which are involved in cell growth or apoptosis such as inhibitor of caspase-activated DNase (ICAD), defender against cell death 1, tumor necrosis factor (TNF) receptor 1, and cytochrome c oxidase subunit VIII. Furthermore, we found that core protein expression increases a steady-state level of ICAD protein, possibly through enhancing its promoter activity, and inhibits caspase-3 activity induced by anti-Fas antibody. Since Fas- or TNF-mediated DNA fragmentation is suppressed in the core-induced Hep191 cells, these findings suggest that expression of HCV core at physiological levels confers blocking activity of caspase-activated DNase and consequently inhibiting apoptotic cell death. (C) 2003 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.virol.2003.08.028

Comparing gene expression profiles in human liver, gastric, and pancreatic tissues using full-length-enriched cDNA libraries Reviewed

M Otsuka, M Arai, M Mori, M Kato, N Kato, O Yokosuka, T Ochiai, M Takiguchi, M Omata, N Seki

HEPATOLOGY RESEARCH 27 ( 1 ) 76 - 82 2003.9

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCI IRELAND LTD

In the post-genome-sequencing era, full-length cDNA-sequence resources are extremely useful for functional analyses of genes. In addition, comprehensive gene profiling of human tissues at the mRNA level is also useful in understanding the molecular mechanisms of tissue-specific functions and disease pathogenesis. In this study, to obtain a wide variety of full-length cDNA clones derived from digestive tissues, numerous expressed sequence tags were generated from libraries enriched with full-length cDNAs. In total, 13 575 sequences were obtained from three cDNA libraries, which were constructed from tissues and cell lines of human liver, stomach, and pancreas. The integration of overlapping clones categorized the sequences into 5936 clusters (1666, 2746, and 2222 clusters in the liver, stomach, and pancreas, respectively). Of these, 1138 clones were scored as full-length cDNAs. Surprisingly, the redundant clones from all three tissues were assembled to show that only 101 genes (1.7% of the assembled 5936 genes) were shared. These results suggest that functional differences between tissues are probably related to their divergent gene expression profiles, and form a basis for understanding the molecular mechanisms underlying tissue-specific pathogenesis that are expressed in different organs. In addition, the full-length cDNAs obtained in this study should prove useful for future functional analyses of the genes expressed in digestive tissues. (C) 2003 Elsevier B.V. All rights reserved.

DOI： 10.1016/S1386-6346(03)0161-X

肝細胞癌患者血清のプロテオミクス解析 Reviewed

星田有人, 川上高幸, 大塚基之, 谷口寿章, 西村信一郎, 小尾俊太郎, 椎名秀一朗, 川邊隆夫, 小俣政男

肝臓 44 ( Suppl.1 ) A174 - A174 2003.4

Liver chip and gene shaving Reviewed

M Otsuka, Y Hoshida, N Kato, M Moriyama, H Taniguchi, M Arai, M Mori, N Seki, M Omata

JOURNAL OF GASTROENTEROLOGY 38 89 - 92 2003.3

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG TOKYO

A comprehensive profile of genes expressed at the mRNA level in various human tissues is considered to be important for understanding the molecular mechanisms of the tissue-specific function and the pathogenesis of related diseases. Here, the gene expression profiling in three human digestive tissues, liver, stomach, and pancreas, was catalogued by generating a large number of expressed sequence tags, and clarified how quantitatively the gene expressions are different. After assembling the redundant clones among three tissues, the results showed that only 1.7% among the assembled genes was expressed commonly in the investigated tissues. These results suggest that the significant functional divergences in different tissues must be related to the divergence of the gene expression profiles. Recently, microarray technologies are widely used. Considering the results that different genes express in different tissues, however, it is important to spot the cDNAs derived from the same tissues or cells examined to acquire information efficiently. For the study of digestive diseases, we constructed an in-house microarray by using the cDNA sets derived from the digestive tissues (liver and gastric chip). In addition, because the amount of information acquired by the microarray analyses is huge, the power of bioinformatics for unifying the obtained data is indispensable. Some examples of the strategies for handling the microarray data obtained by our in-house microarrays are shown in this article.

Synergistic activation of the serum response element-dependent pathway by hepatitis B virus X protein and large-isoform hepatitis delta antigen Reviewed

T Goto, N Kato, H Yoshida, M Otsuka, M Moriyama, Y Shiratori, K Koike, M Matsumura, M Omata

JOURNAL OF INFECTIOUS DISEASES 187 ( 5 ) 820 - 828 2003.3

Language：English Publishing type：Research paper (scientific journal) Publisher：UNIV CHICAGO PRESS

Hepatitis delta virus (HDV) is a naturally occurring satellite of hepatitis B virus (HBV). There are few studies of the effects of the combination of HBV and HDV proteins (HDV antigens [HDAgs]) on intracellular signaling pathways. To understand the influence of HBV and HDV coinfection on hepatocytes, we investigated the effect of HBV proteins and HDAgs on the serum response element (SRE)-dependent pathway. Reporter assays revealed that only HBV X protein (HBx), alone or with the large isoform of HDAg (LHDAg), synergistically activated the SRE-dependent pathway. The effect of HBx and LHDAg on Elk1 or serum response factor (SRF) was examined, because both proteins bind to the SRE. HBx activated the transcriptional ability of Elk1, whereas LHDAg activated the transcriptional ability of SRF. Thus, HBx and LHDAg synergistically activated the SRE-dependent pathway. These results may help us understand clinical phenomena in patients coinfected with HBV and HDV.

DOI： 10.1086/368389

Study of digestive diseases in post-genome era Reviewed

Masao Omata, Motoyuki Otsuka, Takayuki Kawakami, Yujin Hoshida

Japanese Journal of Gastroenterology 100 ( 2 ) 135 - 143 2003.2

Relevance network between chemosensitivity and transcriptome in human hepatoma cells Reviewed

M Moriyama, Y Hoshida, M Otsuka, S Nishimura, N Kato, T Goto, H Taniguchi, Y Shiratori, N Seki, M Omata

MOLECULAR CANCER THERAPEUTICS 2 ( 2 ) 199 - 205 2003.2

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER ASSOC CANCER RESEARCH

Generally, hepatoma is not a chemosensitive tumor, and the mechanism of resistance to anticancer drugs is not fully elucidated. We aimed to comprehensively evaluate the relationship between chemosensitivity and gene expression profile in human hepatoma cells, by using microarray analysis, and analyze the data by constructing relevance networks.
In eight hepatoma cell lines (HLE, HLF, Huh7, Hep3B, PLC/PRF/5, SK-Hep1, Huh6, and HepG2), the baseline expression levels of 2300 genes were measured by cDNA microarray. The concentrations of eight anticancer drugs (nimustine, mitomycin C, cisplatin, carboplatin, doxorubicin, epirubicin, mitoxantrone, and 5-fluorouracil) needed for 50% growth inhibition were examined and used as a measure of chemosensitivity. These data were combined and comprehensive pair-wise correlations between gene expression levels and the 50% growth inhibition values were calculated. Significant correlations with significance were used to construct networks of similarity.
Fifty-two relations, including 42 genes, were selected. Among them, nearly 20% were various types of transporters, and most of them negatively correlated with chemosensitivity. Transporter associated with antigen processing 1 was associated with resistance to mitoxantrone, consistent with previous reports. Other transporters were not reported previously to associate with chemosensitivity. Resistance to doxorubicin and its analogue, epirubicin, were positively correlated with topoisomerase II beta expression, whereas it negatively correlated with expression of carboxypeptidases A3 and Z. Response to nimustine was associated with expression of superoxide dismutase 2.
Relevance networks identified several negative correlations between gene expression and resistance, which were missed by hierarchical clustering. Our results suggested the necessity of systematically evaluating the transporting systems that may play a major role in resistance in hepatoma. This may provide useful information to modify anticancer drug action in hepatoma.

Interieukin-1 beta gene polymorphisms associated with hepatocellular carcinoma in hepatitis C virus infection Reviewed

Y Wang, N Kato, Y Hoshida, H Yoshida, H Taniguchi, T Goto, M Moriyama, M Otsuka, S Shiina, Y Shiratori, Y Ito, M Omata

HEPATOLOGY 37 ( 1 ) 65 - 71 2003.1

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO

Hepatitis C virus (HCV) infection is a major risk factor for developing hepatocellular carcinoma (HCC), a life-threatening sequel. However, the factors that affect disease progression to HCC have not been thoroughly elucidated. Genetic polymorphisms in proinflammatory cytokines, the interleukin 1 (IL-1) family (IL-1beta and IL-1ra) and tumor necrosis factor-alpha (TNF-alpha), were studied in 274 Japanese patients with chronic HCV infection and 55 healthy individuals using standard polymerase chain reaction-based genotyping techniques. The association between these polymorphisms and disease status was evaluated while controlling for confounding clinical variables. The proportion of patients with HCC in the IL-1beta-31 T/T (55%, odds ratio to C/C was 2.63, P =.009) genotype was higher than in the T/C (44%, odds ratio to C/C was 1.64, P =.149) and C/C genotypes (35%). The IL-1beta-31 and -511 loci were in near complete linkage disequilibrium, and the IL-1beta-511/-31 haplotype C-T was significantly associated with the presence of HCC (odds ratio of 1.5 1, P =.02). Polymorphisms in the TNF-alpha gene were not associated with disease. A multivariate analysis revealed that the IL-1beta-31 T/T genotype, alpha-fetoprotein >20 mug/L, presence of cirrhosis, male sex, and age > 60 years were associated with the presence of HCC at odds ratios of 3.73 (T/T vs. C/C), 4.12, 4.03, 3.89, and 3.27, respectively. In conclusion, the IL-1beta-31 genotype T/T or the IL-1beta-511/-31 haplotype C-T is associated with the presence of HCC in Japanese patients with chronic HCV infection.

DOI： 10.1053/jhep.2003.50017

Differential cellular gene expression induced by hepatitis B and C viruses Reviewed

M Otsuka, H Aizaki, N Kato, T Suzuki, T Miyamura, M Omata, N Seki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 300 ( 2 ) 443 - 447 2003.1

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Hepatitis B virus (HBV) is a hepatotropic virus that causes acute and chronic hepatocellular injury and hepatocellular carcinoma. To clarify how HBV proteins regulate host cellular gene expression, we used our in-house cDNA microarray and HepG2.2.15 cells, which are derived from HepG2 cells and produce all HBV proteins. Of 2304 genes investigated, several genes were differentially expressed in HepG2.2.15 cells compared with HepG2 cells. These genes included insulin-like growth factor II and alpha-fetoprotein, consistent with previous reports. Furthermore. we previously performed similar microarray analyses to clarify the effects of hepatitis C virus (HCV) proteins on host cells, using a HepG2-derivative cell line, which produces all HCV proteins. Using these two microarray results, we compared the differences in cellular gene expression induced by HBV and HCV proteins. The expression of the majority of genes investigated differed only slightly between HBV and HCV protein-producing cells. However, HBV and HCV proteins clearly regulated several genes in a reciprocal manner. Combined, these microarray results shed new light on the effects of HBV proteins on cellular gene expression and on the differences in the pathogenic activities of these two hepatitis viruses. (C) 2002 Elsevier Science (USA). All rights reserved.

DOI： 10.1016/S0006-291X(02)02861-9

Signals induced by HCV proteins Reviewed

M Otsuka, N Kato, H Taniguchi, H Yoshida, Y Shiratori, M Omata

HCV/OXIDATIVE STRESS AND LIVER DISEASE 32 - 47 2003

Language：English Publishing type：Research paper (international conference proceedings) Publisher：SPRINGER-VERLAG TOKYO

Hepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, cirrhosis, and hepatocellular carcinoma. To explore the influence of HCV infection on hepatocytes, the effects of HCV proteins on intracellular signal transduction pathways, especially those related to apoptosis, fibrosis, and cell growth, were investigated. The effects of nine HCV proteins (core, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) on the CRE-, SRE-, NFB-, AP-1-, SRF-, ISRE-, HSE-, GRE-, and p53-associated pathways were investigated by use of a reporter assay. The effects of core protein on apoptosis were examined by DNA laddering and Western blotting after induction of apoptosis by Fas stimulation. The possible mechanisms of anti-apoptotic effects of core protein were investigated by RNase protection assay and a reporter assay. The effects of HCV proteins on TGF beta production were determined by a reporter assay. Among seven HCV proteins investigated, core protein had the strongest influence on intracellular signaling, especially for the SRE-, AP-1-, NFkappaB-, and p53-associated pathways. Core protein promoted Bcl-x(L) expression through the mitogen-activated protein kinase (MAPK) pathway and inhibited apoptosis. Among HCV proteins, only core protein caused TGF beta promoter activity through the MAPK pathway. From these results, core protein was considered to regulate cell growth exquisitely in hepatocytes, inhibit apoptosis, and promote liver fibrosis directly without any inflammation or the mediation of any other cells. These functions may reflect the direct action of HCV proteins on the intracellular signal transduction pathways related to chronic hepatitis pathogenesis.

[HCV-related hepatocellular carcinoma]. Reviewed

Otsuka M, Kato N, Yoshida H, Yoshida H, Shiratori Y, Omata M

Uirusu 52 ( 2 ) 295 - 300 2002.12

Expression profiling of liver cell lines expressing entire or parts of hepatitis C virus open reading frame Reviewed

H Aizaki, T Harada, M Otsuka, N Seki, M Matsuda, YW Li, H Kawakami, Y Matsuura, T Miyamura, T Suzuki

HEPATOLOGY 36 ( 6 ) 1431 - 1438 2002.12

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO

Although hepatitis C virus (HCV) is a causative agent of liver diseases, its mechanism of pathogenesis is still unclear, mainly because of the lack of adequate cell culture systems to support HCV infection and replication. In this report, we describe development and characterization of human hepatoma cell lines constitutively expressing entire (Hep394) or parts (Hep352, Hep3294) of the HCV open reading frame (ORF). The viral and cellular proteolytic machinery involved in the viral precursor processing was consistently functional, and processed HCV proteins were synthesized in these established cell lines. By using a cDNA microarray analysis coupled with semiquantitative reverse-transcription polymerase chain reaction (RT-PCR), we identified 12 genes up-regulated and 4 genes down-regulated in Hep394 cells. With regard to genes related to cell growth regulation, we found up-regulation of forkhead transcription factor FREAC-1, poly (A) binding protein PABP2,, and Ras suppressor Rsu-1. Another category of changes in gene expression includes MHC antigens, which play an important role in the T-cell-mediated immune reaction in the liver. In conclusion, functional genomic approaches comparing expression among the different cell lines expressing parts of the HCV genome may promote our understanding of the molecular basis of pathogenicity of HCV infection.

DOI： 10.1053/jhep.2002.36937

Identification of candidate genes for Sjogren's syndrome using MRL/lpr mouse model of Sjogren's syndrome and cDNA microarray analysis Reviewed

T Azuma, M Takei, T Yoshikawa, Y Nagasugi, M Kato, M Otsuka, H Shiraiwa, S Sugano, K Mitamura, S Sawada, Y Masuho, N Seki

IMMUNOLOGY LETTERS 81 ( 3 ) 171 - 176 2002.5

Language：English Publishing type：Research paper (scientific journal) Publisher：ELSEVIER SCIENCE BV

Sjogren's syndrome is a chronic autoimmune disease characterized by focal lymphocytic infiltration of lacrimal and salivary Glands, but the precise mechanism of this syndrome is poorly understood. To clarify the mechanism of onset and progression of Sjogren's syndrome, it is necessary to identify Sjogren's syndrome-related genes. For this purpose, we used MLR/MpJ-1pr/lpr(MRL/lpr) mouse as a model of human Secondary Sjogren's syndrome and analyzed specific mRNA expression pattern in MRL/lpr mouse salivary glands by in-house cDNA microarray. Among arrayed 2304 genes, 13 genes were isolated as highly expressed genes in MRL/lpr mouse salivary gland in comparison with MRL/MpJ-(+/+) (MRL/+) mouse tissue. Subsequently, we performed RTPCR analysis and confirmed the high expression level of nine genes; caspase3, Ly-6C.2, vimentin, Mel-14 antigen, cathepsin B, mpt1, Laptm5, Gnai2 and UCP2. Five of the nine genes have already been identified in patients with Sjogren's syndrome or mice models of the syndrome, but the remaining four genes; mpt1, Laptin5, Gnai2, and UCP2 have not been reported previously as Sjogren's syndrome-related Genes. Although, further experiments are necessary to examine the relationship between these four genes and Sjogren's syndrome, our system of mouse model of Sjogren's syndrome combined with in-house cDNA microarray is suitable for the isolation of Sjogron's syndronic-related genes. (C) 2002 Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S0165-2478(01)00344-3

Hepatitis C virus core protein inhibits apoptosis via enhanced Bcl-x(L) expression Reviewed

M Otsuka, N Kato, H Taniguchi, H Yoshida, T Goto, Y Shiratori, M Omata

VIROLOGY 296 ( 1 ) 84 - 93 2002.4

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

Previous studies indicated that hepatitis C virus core protein influences cellular apoptosis. However, the precise mechanisms of the effects are not fully understood. Therefore, in this study, we examined the mechanisms of the effects on cell apoptosis by core protein, using transiently transfected and magnetically collected core-producing HepG2 cells. First, to elucidate the target site of core protein in the apoptotic pathway, we examined the activation of caspases after anti-Fas antibody stimulation. Core protein inhibited the apoptotic cascade downstream from caspase 8 and upstream from caspase 3. Next, to clarify more direct mechanisms of this effect, mRNA levels of several bcl-2-related genes were examined. An RNase protection assay showed that the mRNA of bcl-xl increased in the core-producing cells. We showed that this increase was mediated by the enhancement of bcl-x promoter activity by core protein through an extracellular-regulated kinase pathway. These results suggest that core protein inhibits apoptosis at the mitochondria level through augmentation of Bcl-x expression, resulting in an inhibition of caspase 3 activation. (C) 2002 Elsevier Science (USA).

DOI： 10.1006/viro.2002.1371

The evaluation of putative virulence factors of Helicobacter pylori for gastroduodenal disease by use of a short-term Mongolian gerbil infection model Reviewed

M Akanuma, S Maeda, K Ogura, Y Mitsuno, Y Hirata, T Ikenoue, M Otsuka, T Watanabe, Y Yamaji, H Yoshida, T Kawabe, Y Shiratori, M Omata

JOURNAL OF INFECTIOUS DISEASES 185 ( 3 ) 341 - 347 2002.2

Language：English Publishing type：Research paper (scientific journal) Publisher：UNIV CHICAGO PRESS

Few virulence determinants of Helicobacter pylori have been tested in vivo. We conducted this study to establish an animal model for their screening. Six-week-old male Mongolian gerbils were inoculated with wild-type H. pylori (TN2) or its isogenic mutant with deletion of cagE (TN2DeltacagE), total cag pathogenicity island (TN2Deltacag PAI), HP0499 (TN2DeltaHP499), or HP0638 (TN2DeltaHP638) (n = 5 each). The animals were killed 3 weeks later, and the density of bacteria and the degree of inflammation in the stomach were compared. Infection was established in all animals except those inoculated with TN2DeltaHP638. TN2 and TN2DeltaHP499, but not TN2DcagE and TN2Deltacag PAI, induced intense inflammation, although the densities of bacteria were similar. The Mongolian gerbil model was useful for the screening of virulence determinants in vivo, which confirmed the importance of cag PAI while questioning that of HP0499.

DOI： 10.1086/338772

Identification of genes associated with sensitivity to 5-fluorouracil and cisplatin in hepatoma cells Reviewed

Y Hoshida, M Moriyama, M Otsuka, N Kato, T Goto, H Taniguchi, Y Shiratori, N Seki, M Omata

JOURNAL OF GASTROENTEROLOGY 37 92 - 95 2002

Language：English Publishing type：Research paper (scientific journal) Publisher：SPRINGER-VERLAG TOKYO

Background. The prognosis of patients with advanced hepatoma is grim. Although chemotherapy is adapted to such patients, the efficacy is low and the outcome cannot be predicted before therapy. In this study, we aimed to identify genes associated with sensitivity to 5-fluorouracil and cisplatin, drugs widely used in treatment, using gene expression profiles. Methods: Gene expression was evaluated in eight human hepatoma cell lines using an in-house cDNA microarray including 2300 known genes. The 50% growth inhibitory concentrations (Gl(50)) of 5-fluorouracil and cisplatin were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and designated as chemosensitivity. Genes with expression ratios associated with Gl(50) were selected using the permutation test. Results: For 5-fluorouracil and cisplatin, 21 and 40 genes, respectively, were selected. From among the genes associated with 5-fluorouracil and cisplatin, several encoding metabolic enzymes were selected. In addition, several genes involved in the cell cycle and transcription were identified. Conclusions: We identified genes that may be associated with sensitivity to 5-fluorouracil and cisplatin. A list of these genes may be useful to elucidate how these drugs work on human hepatoma.

Induction of proliferation-related signals by hepatitis C virus Reviewed

N Kato, H Yoshida, M Otsuka, Y Shiratori, M Omata

GROWTH, PROLIFERATION, AND APOPTOSIS IN HEPATOCYTES 81 - 98 2002

Language：English Publishing type：Research paper (international conference proceedings) Publisher：SPRINGER-VERLAG TOKYO

Hepatitis C virus (HCV) causes persistent infection, chronic hepatitis, cirrhosis, and hepatocellular carcinoma. To explore the influence of HCV infection on hepatocytes, the effects of HCV proteins on intracellular signal transduction pathways were investigated. The effects of seven HCV proteins (core, nonstructural [NS]2, NS3, NS4A, NS4B, NS5A, and NS5B) on cyclic AMP response element (CRE)-, serum response element (SRE)-, nuclear factor (NF)-kappaB-, activator protein (AP)-1-, serum response factor (SRF)-, and p53-associated pathways were investigated by use of a reporter assay. The activation of signals by HCV proteins was examined using a reporter plasmid with an interleukin (IL)-8 or p21(waf1) promoter. The possible mechanisms by which HCV proteins activate these pathways were investigated. Among the seven HCV proteins investigated, core protein had the strongest influence on intracellular signaling, especially SRE-, AP-1-, NF-kappaB-, and p53-associated pathways. Core protein activated IL-8 promoter through NF-kappaB and AP-1 and activated p21 promoter having a p53-binding site. Core protein activated the NF-kappaB pathway mainly through IKKbeta and tumor necrosis factor receptor-associated factor 2/6 and augmented p53 function by increasing both p53-DNA binding affinity and transcriptional ability itself. Direct interaction between core protein and the C-terminus of p53 was detected. In addition, the interaction between core protein and human TBP-associated factor 1128, a component of the transcriptional factor complex, was also demonstrated. Core protein may directly promote cell proliferation and induce an inflammatory reaction by activating SRE, AP-1-, and NF-kappaB-associated pathways. On the other hand, core protein could enhance p53 function. These opposing functions may result in exquisitely balancing the proliferation of hepatocytes infected with HCV.

Differential expression of the L-plastin gene in human colorectal cancer progression and metastasis Reviewed

M Otsuka, M Kato, T Yoshikawa, H Chen, EJ Brown, Y Masuho, M Omata, N Seki

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 289 ( 4 ) 876 - 881 2001.12

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC ELSEVIER SCIENCE

To identify molecular alterations in the progression of colorectal carcinoma, we analyzed gene expression profiles of colon cancer cell lines derived from primary and metastatic tumors from a single patient. Of 2280 cDNAs investigated using our in-house microarray, the expression of 6 genes (tumor-associated antigen L6, L-plastin, the human homologue of yeast ribosomal protein S28, the B-cell translocation gene, mitochondrial aspartate-aminotransferase, and HLA-A) increased, while that of 2 genes (keratin 5 and phosphoglucomutase) decreased in metastatic-tumor-derived cells compared with primary-tumor-derived cells. Of these genes, we assessed the L-plastin gene, an actin-bundling protein, at the protein level using a tissue microarray consisting of 58 clinically stratified colorectal cancer specimens. Consistent with our microarray results, the expression of L-plastin was significantly correlated with the progression of cancer staging. Therefore, our results suggest that the L-plastin gene is a potential metastatic marker. In addition, combining cDNA microarrays and tissue arrays, as shown here, is thought to facilitate the rapid characterization of candidate biomarkers. (C) 2001 Elsevier Science.

DOI： 10.1006/bbrc.2001.6047

cDNA microarray analysis of Helicobacter pylori-mediated alteration of gene expression in gastric cancer cells Reviewed

S Maeda, M Otsuka, Y Hirata, Y Mitsuno, H Yoshida, Y Shiratori, Y Masuho, M Muramatsu, N Seki, M Omata

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 284 ( 2 ) 443 - 449 2001.6

Language：English Publishing type：Research paper (scientific journal) Publisher：ACADEMIC PRESS INC

Helicobacter pylori infection stimulates several intracellular signaling pathways and is accompanied by increased gene expression in gastric epithelial cells. High-density cDNA microarray was used to characterize the mRNA expression profile of genes in human gastric cancer cells (MKN45, AGS) cocultured with H. pylori. Coculture with cag pathogenicity island (PAI)-positive H. pylori (wild-type) significantly upregulated mRNA expression in 8 of 2304 genes tested. In 6 (interleukin-8, I kappaB alpha, A20, ERF-1, keratin K7, glutathione peroxidase) of the 8 genes, up-regulation was confirmed by RT-PCR. In coculture with isogenic cagE-negative mutant (Delta cagE), which encodes a type TV secretion system with other genes in the cag PAI, no significant up-regulation was found. We further analyzed the role of A20. Transfection of expression vector encoding A20 resulted in an inhibition of H. pylori-mediated NF-KB activation, indicating that H, pylori-mediated A20 expression could be a negative regulator of NF-KB activation. Taken together, these results indicate the importance of microarray technology as a tool for analyzing the complex interplay between H. pylori and the host. (C) 2001 Academic Press.

DOI： 10.1006/bbrc.2001.5006

Hepatitis C virus core protein activates nuclear factor kappa B-dependent signaling through tumor necrosis factor receptor-associated factor Reviewed

H Yoshida, N Kato, Y Shiratori, M Otsuka, S Maeda, J Kato, M Omata

JOURNAL OF BIOLOGICAL CHEMISTRY 276 ( 19 ) 16399 - 16405 2001.5

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Hepatitis C virus (HCV) core protein, a viral nucleocapsid, has been shown to affect various intracellular events including the nuclear factor kappaB (NF-kappaB) signaling supposedly associated with inflammatory response, cell proliferation, and apoptosis. In order to elucidate the effect of HCV core protein on the NF-kappaB signaling in HeLa and HepG2 cells, a reporter assay was utilized. HCV core protein significantly activated NF-kappaB signaling in a dose-dependent manner not only in HeLa and HepG2 cells transiently transfected with core protein expression plasmid, but also in HeLa cells induced to express core protein under the control of doxycycline. HCV core protein increased the DNA binding affinity of NF-kappaB in the electrophoretic mobility shift assay. Acetyl salicylic acid, an IKK beta -specific inhibitor, and dominant negative form of IKK beta significantly blocked NF-kappaB activation by HCV core protein, suggesting HCV core protein activates the NF-kappaB pathway mainly through IKK beta. Moreover, the dominant negative forms of TRAF2/6 significantly blocked activation of the pathway by HCV core protein, suggesting HCV core protein mimics proinflammatory cytokine activation of the NF-kappaB pathway through TRAF2/6. In fact, HCV core protein activated interleukin-lp promoter mainly through NF-kappaB pathway. Therefore, this function of HCV core protein may play an important role in the inflammatory reaction induced by this hepatotropic virus.

Improvement of Chemosensitivity Prediction by Transcriptional Profiling in Hepatoma Cells

Hoshida Yujin, Moriyama Masaru, Otsuka Motoyuki, Kato Naoya, Shiratori Yasushi, Seki Naohiko, Omata Masao

GI 12 257 - 258 2001

Large isoform of hepatitis delta antigen activates serum response factor-associated transcription Reviewed

T Goto, N Kato, SK Ono-Nita, H Yoshida, M Otsuka, Y Shiratori, M Omata

JOURNAL OF BIOLOGICAL CHEMISTRY 275 ( 48 ) 37311 - 37316 2000.12

Language：English Publishing type：Research paper (scientific journal) Publisher：AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Hepatitis delta virus infection sometimes causes severe and fulminant hepatitis as a coinfection or superinfection along with the hepatitis B virus. To elucidate the underlying mechanism of injury caused by hepatitis delta virus, we examined whether two isoforms of the hepatitis delta antigen (HDAg) had any effect on five well defined intracellular signal transduction pathways: serum response factor (SRF)-, serum response element (SRE)-, nuclear factor kappaB-, activator protein 1-, and cyclic AMP response element-dependent pathways. Reporter assays revealed that large HDAg (LIDDAg) activated the SRF- and SRE-dependent pathways. In contrast, small HDAg (LHDAg) did not activate any of five pathways. LHDAg enhanced the transcriptional ability of SRF without changing its DNA binding affinity in an electrophoretic mobility shift assay. In addition, LHDAg activated a rat SM22 alpha promoter containing SRF binding site and a human c-fos promoter containing SRE. In conclusion, LHDAg, but not SHDAg, enhances SRF-associated transcriptions. Despite structural similarities be tween the two HDAgs, there are significant differences in their effects on intracellular signal transduction pathways. These results may provide clues that will aid in the clarification of functional differences between LHDAg and SHDAg and the pathogenesis of delta hepatitis.

Hepatitis C virus core protein enhances p53 function through augmentation of DNA binding affinity and transcriptional ability Reviewed

Motoyuki Otsuka, Naoya Kato, Keng-Hsin Lan, Hideo Yoshida, Jun Kato, Tadashi Goto, Yasushi Shiratori, Masao Omata

Journal of Biological Chemistry 275 ( 44 ) 34122 - 34130 2000.11

Language：English Publishing type：Research paper (scientific journal) Publisher：American Society for Biochemistry and Molecular Biology Inc.

Hepatitis C virus (HCV) causes a persistent infection, chronic hepatitis, and hepatocellular carcinoma. Since there are several reports indicating that some viruses influence the tumor suppressor p53 function, we determined the effects of HCV proteins on p53 function and its mechanism determined by use of a reporter assay. Among seven HCV proteins investigated (core, NS2, NS3, NS4A, NS4B, NS5A, and NS5B), only core protein augmented the transcriptional activity of p53 and increased the expression of p21(waf1) protein, which is a major target of p53. Core protein increased both DNA-binding affinity of p53 in electrophoretic morbidity shift assay and transcriptional ability of p53 itself in a reporter assay. The direct interaction between core protein and C terminus of p53 was also shown by glutathione S-transferase fusion protein binding assay. In addition, core protein interacted with hTAF(II)28, a component of the transcriptional factor complex in vivo and in vitro. These results suggest that HCV core protein interacts with p53 and modulates p53-dependent promoter activities during HCV infection.

DOI： 10.1074/jbc.M000578200

Activation of intracellular signaling by hepatitis B and C viruses: C-viral core is the most potent signal inducer Reviewed

N Kato, H Yoshida, SK Ono-Nita, J Kato, T Goto, M Otsuka, KH Lan, K Matsushima, Y Shiratori, M Omata

HEPATOLOGY 32 ( 2 ) 405 - 412 2000.8

Language：English Publishing type：Research paper (scientific journal) Publisher：W B SAUNDERS CO

To clarify the effects of hepatitis C virus (HCV) infection on hepatocytes, we analyzed and compared the induction of intracellular signals by HCV and hepatitis B virus (HBV) proteins. We examined the influence of 7 HCV (core, NS2, NS3, NS4A, NS4B, NS5A, and NS5B) and 4 HBV (precore, core, polymerase, and X) proteins on 5 well-defined intracellular signaling pathways associated with cell proliferation, differentiation, and apoptosis by use of a reporter assay. Viral protein-expression vectors were cotransfected into mammalian cells with reporter vectors having a luciferase gene driven by the following inducible cis-enhancer elements: the cyclic adenosine monophosphate response element, the serum response element (SRE), and the binding sites for nuclear factor kappa B (NF-kappa B), activator protein 1 (AP-1), and serum response factor (SRF). In addition, the activation of signals by HCV proteins was examined in a reporter plasmid having a natural interleukin-8 (IL-8) promoter upstream of a luciferase gene. Of 11 HCV and HBV proteins, HCV core had the strongest influence on intracellular signals, especially NF-kappa B-, AP-1-, and SRE-associated pathways. HCV core's activation level exceeded that of HBV X protein, a well-characterized transactivator of these signals. Moreover, HCV core activated the IL-8 promoter through NF-kappa B and AP-1. For the other proteins, HCV NS4B showed signal activation, but signals were activated at a lesser extent. The luciferase reporter assay, a recently introduced technique, helped in the elucidation of molecular events underlying the inflammatory and proliferation process in the liver induced by HCV.

超音波内視鏡下吸引細胞診(EUS-FNA)による膵腫瘍性病変の診断

小松裕, 多田稔, 平田喜裕, 大塚基之, 笹平直樹, 山形道子, 戸田信夫, 大橋誠, 川邊隆夫, 白鳥康史

超音波医学 27 ( 5 ) 816 - 816 2000.5

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