研究者詳細 - 香川　俊輔

論文 - 香川　俊輔

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Histone deacetylase inhibitor FR901228 enhances the antitumor effect of telomerase-specific replication-selective adenoviral agent OBP-301 in human lung cancer cells

T Watanabe, M Hioki, T Fujiwara, M Nishizaki, S Kagawa, M Taki, H Kishimoto, Y Endo, Y Urata, N Tanaka, T Fujiwara

EXPERIMENTAL CELL RESEARCH 312 ( 3 ) 256 - 265 2006年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER INC

Replication-competent oncolytic viruses are being developed for human cancer therapy. We previously reported that an attenuated adenovirus OBP-301 (Telomelysin), in which the human telomerase reverse transcriptase promoter element drives expression of EIA and El B genes linked with an internal ribosome entry site, could replicate in and causes selective lysis, of human cancer cells. Infection efficiency in target cancer cells is the most important factor that predicts the antitumor effects of OBP-301. The ob objectives of this study are to examine the effects of the histone deacetylase inhibitor FR901228 on the level of coxsackie and adenovirus receptor (CAR) expression and OBP-301-mediated oncolysis in human non-small cell lung cancer cell lines. Flow cytometric analysis revealed up-regulated CAR expression in A549 and H460 cells following treatment with 1 ng/ml of FR901228. which was associated with increased infection efficiency as confirmed by replication-deficient beta-galactosidase-expressing adenovirus vector. In contrast. neither CAR expression nor infection efficiency was affected by FR901228 in H1299 cells. To visualize and quantify viral replication in the presence of FR901228, we used 01311-401 (Telomelysin-GFP) that expresses the green fluorescent protein (GFP) reporter gene under the control of the cytomegalovirus promoter in the E3 region. Fluorescence microscopy and flow cytometry showed that FR901228 increased GFP expression in A549 and 11460 cells following OBP-401 infection in a dose-dependent manner, but this effect did not occur in H1299 cells. In addition. OBP-301 and FR901228 demonstrated a synergistic antitumor effect in A549 cells in vitro, as confirmed by isobologram analysis. Our data indicate that FR901228 preferentially increases adenovirus infectivity via up-regulation of CAR expression, leading to a profound oncolytic effect, which may have a significant impact on the outcome of adenovirus-based oncolytic virotherapy. (c) 2005 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.yexcr.2005.10.026

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テロメラーゼ特異的に増幅するGFP蛋白を用いたマウスの固形腫瘍の胸膜播種の可視化

梅岡達生, 川嶋健, 香川俊輔, 寺石文則, 滝正樹, 京哲, 永井勝幸, 浦田泰生, 田中紀章, 藤原俊義

岡山医学会雑誌 118 ( 1 ) 9 - 15 2006年

症例報告多発肝転移を伴う進行性胃癌に対するイリノテカンと低用量シスプラチン化学療法に併用したゲノシタビンに対する顕著な反応

寺石文則, 宇野太, 香川俊輔, 合地明, 藤原俊義, 田中紀章

癌と化学療法 2006年

CPT-11+Low-Dose5-FU+CDDP療法が著効した多発肝転移を有する噴門部胃癌の1例

香川俊輔, 藤原俊義, 徳永尚之, 西崎正彦, 宇野太, 寺石文則, 合地明, 松岡順治, 田中紀章

癌と化学療法 2006年

胃癌術後縦隔リンパ節再発による気管狭窄にステント+放射線治療が奏効した1例

香川俊輔, 藤原俊義, 西崎正彦, 徳永尚之, 合地明, 田中紀章

癌と化学療法 2006年

A case report--The marked response to gemcitabine combined with irinotecan and low-dose cisplatin chemotherapy for advanced gastric cancer with multiple liver metastases.

Fuminori Teraishi, Futoshi Uno, Shunsuke Kagawa, Akira Gochi, Toshiyoshi Fujiwara, Noriaki Tanaka

Gan to kagaku ryoho. Cancer & chemotherapy 33 ( 12 ) 1885 - 1887 2006年

記述言語：英語掲載種別：研究論文（学術雑誌）

A 69-year-old man with no sign of symptoms was admitted to our hospital for further examination and treatment of gastric cancer. Endoscopy revealed a Borrmann 3-type tumor in the cardia of the stomach. CT of the abdomen demonstrated a marked thickening of the stomach wall near the esophago-gastric junction, multiple liver metastases in bilateral liver lobes, and regional lymph node swelling around the cardia of the stomach. He consented and received systemic chemotherapy consisting of 0.5 h infusion of cisplatin (25 mg/body) followed by 0.5 h infusion of gemcitabine (800 mg/body) and 2.5 h infusion of irinotecan (60 mg/body) every 14 days as described below. Only the initial 1 course was administered with 5-FU (500 mg/body) as an inpatient, and further courses were performed as an outpatient with no severe adverse events. His tumors responded immediately to the chemotherapy, and restaging abdominal CT after 4-cycles of chemotherapy showed almost complete regression of liver metastases, and partial response to lymphadenopathy. The patient currently undergoes regular chemotherapy and remains in remission more than 6 months after the diagnosis of unresectable gastric cancer with liver metastases. It may be possible that the current chemotherapy will be neoadjuvant chemotherapy, and curative surgical resection can be performed.

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A case of advanced gastric cancer with liver metastasis completely responding to CPT-11+low-dose 5-FU+CDDP chemotherapy

Shunsuke Kagawa, Toshiyoshi Fujiwara, Naoyuki Tokunaga, Masahiko Nishizaki, Futoshi Uno, Fuminori Teraishi, Akira Gouchi, Junji Matsuoka, Noriaki Tanaka

Gan to kagaku ryoho. Cancer & chemotherapy 33 ( 9 ) 1317 - 1320 2006年

記述言語：日本語掲載種別：研究論文（学術雑誌）

The case was a 54-year-old man with type-3 gastric cancer in the cardia accompanied by multiple liver metastasis. He received combination chemotherapy consisting of CPT-11 (60 mg/body, day 1 and 8)+low-dose 5-FU and CDDP (5-FU 500 mg/body/day and CDDP 5 mg/body/day, day 1-5 and 8-12, continuous infusion) every 3 weeks. The initial 2 courses were administered on an inpatient basis,and further courses as an outpatient. After 7 courses of therapy without severe adverse events, not only primary lesion but also hepatic metastasis disappeared. He has been free from disease for 4 months, and chemotherapy was further continued with TS-1 (100 mg/body, day 1-14)+CPT-11 60 mg/body, day 1, 8), every 3 weeks. CPT-11 in combination with low-dose 5-FU+CDDP can be one of the most effective regimens for unresectable advanced gastric cancer.

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Stenting and radiotherapy for malignant tracheal stenosis due to mediastinal lymph node recurrence from gastric cancer--a case report

Shunsuke Kagawa, Toshiyoshi Fujiwara, Masahiko Nishizaki, Naoyuki Tokunaga, Akira Gochi, Noriaki Tanaka

Gan to kagaku ryoho. Cancer & chemotherapy 33 ( 1 ) 91 - 93 2006年

記述言語：日本語掲載種別：研究論文（学術雑誌）

The case is a 74-year-old woman,who underwent curative resection six years earlier for stomach cancer in the cardia extending to the esophagus (Stage IIIA). She noticed hoarseness and cough in September 2004, and CT revealed lymph node metastasis of the cancer in the upper mediastinum. The tumor pressured the SVC, the esophagus,and especially the trachea that narrowed up to a diameter of 5 mm.To secure the air way, first, a self-expandable metallic stent was placed in the narrowed trachea under general anesthesia. Irradiation to the upper mediastinal tumor was further applied for a total of 46 Gy from October to November, 2004.Symptom relief was obtained over the course by a marked reduction in the size of the metastatic tumor, and the disease remained stable for 5 months. In general,a stomach cancer that is mostly adenocarcinoma has been recognized to have low receptivity for radiotherapy. Nevertheless,this case suggests that radiotherapy can play an important role in local control and symptom relief for symptomatic stomach cancer patients.

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テロメラーゼ依存性GFP蛍光発現ウイルス製剤OBP-401を用いた微小癌組織ナビゲーション・システムの開発

藤原俊義, 岸本浩行, 香川俊輔, 宇野太, 田中紀章, 河村仁, 浦田泰生

日本癌治療学会誌 40 ( 2 ) 350 - 350 2005年9月

テロメラーゼ特異的腫瘍融解アデノウイルス(Telomelysin:OBP301)新たな抗腫瘍分子機構腫瘍免疫系への影響の解析

遠藤芳克, 徳永尚之, 池田義博, 酒井亮, 日置勝義, 岸本浩行, 藤原俊哉, 香川俊輔, 浦田泰生, 田中紀章, 藤原俊義

日本癌学会総会記事 64回 484 - 484 2005年9月

がん特異的増殖可能アデノウイルス製剤OBP-401を用いた生体内癌組織診断がん特異的GFP蛍光イメージング技術の開発

岸本浩行, 香川俊輔, 藤原俊哉, 遠藤芳克, 池田義博, 日置義勝, 酒井亮, 浦田泰生, 田中紀章, 藤原俊義

日本癌学会総会記事 64回 447 - 448 2005年9月

抗癌剤耐性細胞におけるテロメラーゼ特異的制限増殖型アデノウイルスTelomelysin(OBP-301)の抗腫瘍効果の検討

藤原俊哉, 藤原俊義, 香川俊輔, 岸本浩行, 遠藤芳克, 酒井亮, 日置勝義, 池田義博, 浦田泰生, 田中紀章

日本癌学会総会記事 64回 434 - 434 2005年9月

ZD1839 (Gefitinib, 'Iressa'), an epidermal growth factor receptor-tyrosine kinase inhibitor, enhances the anti-cancer effects of TRAIL in human esophageal squamous cell carcinoma

F Teralshi, S Kagawa, T Watanabe, Y Tango, T Kawashima, T Umeoka, M Nisizaki, N Tanaka, T Fujiwara

FEBS LETTERS 579 ( 19 ) 4069 - 4075 2005年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER SCIENCE BV

The EGF (epidermal growth factor) receptor-tyrosine kinase inhibitor ZD1839 (Getitinib, 'Iressa') blocks the cell signaling pathways involved in cell proliferation, survival, and angiogenesis in various cancer cells. TNF-related death apoptosis inducing ligand (TRAIL) acts as an anticancer agent. We investigated the antitumor effects of ZD-1839 alone or in combination with TRAIL against human esophageal squamous cell cancer (ESCC) lines. Although all ESCC cells expressed EGF receptor at a protein level, the effect of ZD1839 on cell growth did not correlate with the level of EGFR expression and phosphorylation of EGF receptor protein in ESCC lines. ZD1839 caused a dose-dependent growth arrest at GO-G, phase associated with increased p27 expression. As TE8 cells are resistant to TRAIL, we tested whether ZD1839 combined with TRAIL induced apoptosis of TE8 cells via the inhibition of EGF receptor signaling by ZD1839. ZD1839 inhibited the phosphorylation of Akt, and enhanced TRAIL-induced apoptosis via activation of caspase-3 and caspase-9, and inactivation of Bcl-xL. Our results indicated that ZD1839 has anti-cancer properties against human esophageal cancer cells. ZD1839 also augmented the anti-cancer activity of TRAIL, even in TRAIL-resistant tumors. These results suggest that treatment with ZD1839 and TRAIL may have potential in the treatment of ESCC patients. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

DOI： 10.1016/j.febslet.2005.06.031

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Enhanced oncolysis by a tropism-modified telomerase-specific replication-selective adenoviral agent OBP-405 ('Telomelysin-RGD)

M Taki, S Kagawa, M Nishizaki, H Mizuguchi, T Hayakawa, S Kyo, K Nagai, Y Urata, N Tanaka, T Fujiwara

ONCOGENE 24 ( 19 ) 3130 - 3140 2005年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：NATURE PUBLISHING GROUP

Replication-competent oncolytic viruses are being developed for human cancer therapy. We previously reported that an attenuated adenovirus (OBP-301, 'Telomelysin'), in which the hTERT promoter element drives expression of E1A and E1B genes linked with an IRES, could replicate in cancer cells, and causes selective lysis of cancer cells. We further constructed OBP-405 ('Telomelysin-RGD') that contains an RGD motif in the HI loop of the fiber knob. We examined whether OBP-405 could be effective in overcoming the limitations of OBP-301, specifically their inefficient infection into cells lacking the primary receptor, the coxsackievirus and adenovirus receptor ( CAR). By flow cytometric analysis, H1299 ( lung) and SW620 ( colorectal) tumor cells showed high levels of CAR expression, whereas LN444 ( glioblastoma), LNZ308 ( glioblastoma), and H1299-R5 ( lung) tumor cells were negative for CAR expression. A quantitative realtime PCR analysis demonstrated that fiber-modified OBP-405 infected more efficiently than OBP-301, although the intracellular replication rate of both viruses was consistent. The comparative antitumor effect of fiber-modified OBP-405 and unmodified OBP-301 for human cancer cells was evaluated in vitro by XTT assay as well as in vivo by using athymic mice carrying xenografts. OBP-405 had a profound oncolytic effect on human cancer cell lines compared to OBP-301, in particular on cells with low CAR expression. Intratumoral injection of 107 plaque-forming units of OBP-405 into CAR-negative H1299-R5 lung tumor xenografts in nu/nu mice resulted in a significant inhibition of tumor growth and long-term survival in all treated mice. Moreover, selective replication of OBP-405 in the distant, uninjected H1299-R5 tumors was demonstrated. Our results suggest that fiber-modified replication-competent adenovirus OBP-405 exhibits a broad target range by increasing infection efficiency, an outcome that has important implications for the treatment of human cancers.

DOI： 10.1038/sj.onc.1208460

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Human monocyte-derived dendritic cells pulsed with wild-type p53 protein efficiently induce CTLs against p53 overexpressing human cancer cells

N Tokunaga, T Murakami, Y Endo, M Nishizaki, S Kagawa, N Tanaka, T Fujiwara

CLINICAL CANCER RESEARCH 11 ( 3 ) 1312 - 1318 2005年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER ASSOC CANCER RESEARCH

Purpose: Dendritic cells are the most potent antigenpresenting cells for initiating cellular immune responses. Dendritic cells are attractive immunoregulatory cells for cancer immunotherapy, and their efficacy has been investigated in clinical trials. The tumor suppressor gene p53 is pivotal in the regulation of apoptosis, and p53-based immunization is an attractive approach to cancer immunotherapy because of the accumulation of p53 protein in malignant but not in normal cells. It has been shown that dendritic cells transduced with an adenoviral wild-type p53 (wt-p53) construct mediate the antitumor immune responses against p53-overexpressing tumor cells. We examined whether monocyte-derived human dendritic cells pulsed with the purified full-length wt-p53 protein were also capable of inducing the specific antitumor responses against p53-overexpressing tumors in vitro.
Experimental Design: Immature dendritic cells generated in the presence of interleukin4 and granulocyte/macrophage colony-stimulating factor from monocytes of HLA-A2- or HLA-A24-positive healthy individuals were pulsed with the purified p53 protein. Uptake of p53 protein by human dendritic cells was assessed by Western blotting and immunohistochemical staining using anti-p53 antibody. Induction of p53-specific CTL response was also evaluated by the cytotoxic assay against p53-overexpressing human tumor cells.
Results: Both Western blot and immunohistochemical analysis showed the accumulation of p53 protein in human immature dendritic cells. T cells obtained from HLA-A2- or HLA-A24-positive healthy donors were stimulated twice with p53 protein-pulsed dendritic cells and then applied to the cytotoxicity assay against p53-overexpressing target cells. The CTL activity was specific for p53-overexpressing tumor cells and MHC class I restricted. Moreover, the CTL activity generated by p53 protein-pulsed dendritic cells was nearly identical with that induced by adenoviral wt-p53-transduced dendritic cells.
Conclusions: Our results indicate that monocyte-derived human dendritic cells pulsed with the wt-p53 protein could induce the specific antitumor effect against p53-overexpressing tumors and that this in vitro model offers a new and more simple approach to the development of p53-based immunotherapy.

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Gene therapy for lung cancer

Toshiyoshi Fujiwara, Shunsuke Kagawa, Noriaki Tanaka

Nippon rinsho. Japanese journal of clinical medicine 63 507 - 511 2005年

癌に対する遺伝子治療：肺癌

藤原俊義, 香川俊輔, 田中紀章

遺伝子診療学（日本臨床増刊号） 63,507-511 2005年

Visualization of intrathoracically disseminated solid tumors in mice with optical imaging by telomerase-specific amplification of a transferred green fluorescent protein gene

T Umeoka, T Kawashima, S Kagawa, F Teraishi, M Taki, M Nishizaki, S Kyo, K Nagai, Y Urata, N Tanaka, T Fujiwara

CANCER RESEARCH 64 ( 17 ) 6259 - 6265 2004年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER ASSOC CANCER RESEARCH

Currently available methods for detection of tumors in vivo such as X-ray, computed tomography, and ultrasonography are noninvasive and have been well studied; the images, however, are not specific for tumors. Direct optical imaging of tumor cells in vivo that can clearly distinguish them from surrounding normal tissues may be clinically useful. Here, we describe a new approach to visualizing tumors whose fluorescence can be detected using tumor-specific replication-competent adenovirus (OBP-301, Telomelysin) in combination with Ad-GFP, a replication-deficient adenovirus expressing green fluorescent protein (GFP). Human telomerase reverse transcriptase is the catalytic subunit of telomerase, which is highly active in cancer cells but quiescent in most normal somatic cells. We constructed an adenovirus 5 vector in which the human telomerase reverse transcriptase promoter element drives expression of E1A and E1B genes linked with an internal ribosome entry site and showed that OBP-301 replicated efficiently in human cancer cells, but not in normal cells such as human fibroblasts. When the human lung and colon cancer cell lines were infected with Ad-GFP at a low multiplicity of infection, GFP expression could not be detected under a fluorescence microscope; in the presence of OBP-301, however, Ad-GFP replicated in these tumor cells and showed strong green signals. In contrast, coinfection with OBP-301 and Ad-GFP did not show any signals in normal cells such as fibroblasts and vascular endothelial cells. We also found that established subcutaneous tumors could be visualized after intraturnoral injection of OBP-301 and Ad-GFP. A549 human lung tumors and SW620 human colon tumors transplanted into BALB/c nu/nu mice were intraturnorally injected with 8 X 10(5) plaque-forming units of Ad-GFP in combination with 8 X 106 plaque-forming units of OBP-301. Within 3 days of treatment, the fluorescence of the expressed GFP became visible by a three-chip color cooled charged-coupled device camera in these tumors, whereas intraturnoral injection of Ad-GFP alone could not induce GFP fluorescence. Moreover, intrathoracic administration of Ad-GFP and OBP-301 could visualize disseminated A549 tumor nodules in mice after intrathoracic implantation. Our results indicate that intratumoral or intrathoracic injection of Ad-GFP in combination with OBP-301 might be a useful diagnostic method that provides a foundation for future clinical application.

DOI： 10.1158/0008-5472.CAN-04-1335

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GFP発現テロメラーゼ特異的制限増殖型ウイルスTelomelysin-GFPを用いた腫瘍組織ナビゲーション・システム

岸本浩行, 香川俊輔, 滝正樹, 藤原俊哉, 渡辺貴紀, 遠藤芳克, 京哲, 水口裕之, 田中紀章, 藤原俊義

日本癌学会総会記事 63回 454 - 454 2004年9月

テロメラーゼ特異的制限増殖型アデノウイルスTelomelysinと微小管作用型抗癌剤の併用効果の検討

藤原俊哉, 藤原俊義, 香川俊輔, 滝正樹, 岸本浩行, 渡邉貴紀, 遠藤芳克, 京哲, 水口裕之, 田中紀章

日本癌学会総会記事 63回 510 - 510 2004年9月

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