Grant amount：\4290000 （ Direct expense: \3300000 、 Indirect expense：\990000 ）

For animal experiments, aortic dissection model was made by subcutaneous infusion of lysyl oxidase inhibitor and angiotensin II. The incidence of aortic dissection in integrin α1 Knockout (Itga1KO) mice was significantly reduced compared with control mice. These results suggests that Itga1 play an important role in the development of aortic dissection. Next, the differences in mRNA expressions between Itga1 KO mice and control mice of aortic dissection model were evaluated by real-time quantitative PCR. The mRNA expressions of inflammatory genes and matrix matrix metalloproteinase in Itga1 KO mice were significantly lower than those in control mice. Taken together, Itga1 may be crucial for the development of aortic dissection.

researchmap

Grant amount：\3900000 （ Direct expense: \3000000 、 Indirect expense：\900000 ）

We investigated the mechanism of macular hole (MH) closure following the inverted internal limiting membrane (ILM) technique. We performed the inverted ILM flap surgical technique as an experimental MH model in monkeys, and investigated the process of MH closure immunohistochemically. We then investigated the effects of type IV collagen, fibronectin, and laminin, which are constituent proteins of the ILM, on the proliferation and migration of cultivated Muller cells(MIO-M1). We also investigated the expression of neurotrophic factors in human ILM and MIO-M1 cells. From these results, during MH closure, the ILM functioned as a scaffold for the proliferation and migration of Muller cells, and may promote Muller cell activation. Neurotrophic factors produced by activated Muller cells and present on the surface of the ILM may contribute to MH closure.

researchmap

Grant amount：\4680000 （ Direct expense: \3600000 、 Indirect expense：\1080000 ）

Abdominal aortic aneurysm is a silent disease, but lethal once ruptured. The prevention of AAA expansion and rupture is clinically difficult. This study investigated whether alpha1 integrin, an adhesion molecule, is involved in the development of AAA in murine model. Genetic deficiency of alpha1 integrin suppressed the rupture of AAA in angiotensin II infusion model. These findings suggest that alpha1 integrin may be a promising target to treat AAA.

researchmap

Grant amount：\4940000 （ Direct expense: \3800000 、 Indirect expense：\1140000 ）

In this study, with the aim of clarifying the mechanism by which xanthophyll is incorporated into the retina and the role played by retinal function, we investigated the uptake into the cell, its pathway, and the effect on cell migration by using retinal pigment epithelial cells and retinal glial cells which constitute the retina. It was revealed that lutein is taken up into cells in concentration and time dependence. Inhibition of the known lutein transport pathway did not cause a significant change in lutein uptake. Also, lutein had no significant effect on cell migration. It is important to investigate the metabolism of intracellular lutein and the role of lutein further in the near future.

researchmap

Grant amount：\4810000 （ Direct expense: \3700000 、 Indirect expense：\1110000 ）

To identify the molecules which would compose "the tran-plasma membrane molecular machinery" of collagen types XV/XVIII and mitochondria, we have screened thousands of genes expressed in muscular tissues and identified possible candidate G-protein coupled receptors. In addition, our histological analyses on collagen types XV/XVIII mutant animals revealed abnormal structures of cytoskeletons, which would potentially compromise the mitochondrial and cellular functions. Moreover, we had successfully optimized the immunohistochemical conditions for the anti-collagen XV/XVIII monoclonal antibodies, which enabled us to use variously processed tissue samples to diagnose human diseases caused by defective collagen XV/XVIII.

researchmap

Grant amount：\4940000 （ Direct expense: \3800000 、 Indirect expense：\1140000 ）

Immunohistochemical analysis of human abdominal aortic aneurysm revealed that the increased expression of CD44 and the degradation of hyaluronic acid were possibly involved in pathogenesis of abdominal aortic aneurysm. Experimental abdominal aortic aneurysm using CaCl2-induced model, the incidence of abdominal aortic aneurysm was significantly less in CD44 knockout mice compared to wild type mice. Liposome targeting CD44 was developed and the utility of its liposome is being studied.

researchmap

Grant amount：\5330000 （ Direct expense: \4100000 、 Indirect expense：\1230000 ）

Retinal pigment epithelial cells (RPE) has a variety of physiological functions to maintain homeostasis of the retina. In the age-related macular degeneration, homeostasis of the retina collapses by decreased function of the RPE associated with aging, and leads to visual disturbance. In this study, for the ARPE-19 and human iPS-derived RPE, we induced senescence of RPE by chronic oxidative stress, and revealed a change in the functions of the RPE associated with aging. In addition, using models of inflammation to induce epithelial-mesenchymal transition in the RPE, we revealed that the activation of AMPK inhibits epithelial-mesenchymal transformation in the RPE.

researchmap

Grant amount：\19240000 （ Direct expense: \14800000 、 Indirect expense：\4440000 ）

Abnormalities of the basement membranes in brain small vessels are considered to cause the break-down of blood-brain barrier (BBB) and intracerebral hemorrhage. We established the mouse encephalopathy model by the administration of TNF-alpha intravenously, in which BBB permeability increase transiently. To know the change of type IV collagen, as major component of the basement membranes, in the encephalopathy model, we performed Western blot and immunohistochemistry. The results suggested that the degradation of type IV collagen was associated with the break-down of BBB.

researchmap

Grant amount：\4420000 （ Direct expense: \3400000 、 Indirect expense：\1020000 ）

Astroglia form a glia limitans around the injured tissue after spinal cord injury. We used a mouse spinal cord injury model and found that the glia limitans was associated with the suppression of infiltration of inflammatory cells and the perivascular space provides a conduit for inflammatory cells to the expansion from the injury core. In addition, we found that type IV collagen induced expression of thrombospondin-1 in astrocytes. Our results suggest that glia limitans play an important role in the repair process following injured central nervous tissue.

researchmap

Grant amount：\3300000 （ Direct expense: \3300000 ）

整形外科において骨や手術時に挿入した人工物におこる感染症はしばしば難治性である。この難治性にはバイオフィルムが関与すると言われており、バイオフィルムは抗生剤の効果を減少させ、残存すると感染を再燃させる。マイクロバブルは下水管の洗浄や半導体におけるシリコンウエハーの洗浄に利用されている。マイクロバブルの粒径はバイオフィルムの狭間よりも小さく、バイオフィルムの除去に有用とされる。
昨年度明らかとなったバイオフィルム形成能が高い黄色ブドウ球菌を用いて、バイオフィルムの作成と洗浄の実験を行った。整形外科感染症のモデルとしてポリフィラメントの繊維成分を当初は用いていたが、慢性骨髄炎など生体骨内の環境を再現するためには不十分であったため、連通孔を有する人工骨にバイオフィルムを作成するモデルの検討を行った。連通孔を介して深部にまでバイオフィルムを作成するために陰圧下で培地を浸潤させること、培地にグルコースを混ぜることにより安定したバイオフィルムが作成可能であった。このバイオフィルムを作用させた人工骨を動物に移植することで容易に慢性骨髄炎モデルとすることができた。
また,昨年度から開発を進めてきたマイクロバブル発生装置については、条件の最適化や効率的な発生方法の検討を行ったが、マイクロバブルの持続時間が短かったため洗浄、滅菌が困難であった。そこで人工骨に作成したバイオフィルムのモデルに対してマイクロバブルの洗浄が有用かを検討するために、滅菌状態は維持できないがマイクロバブルを長時間安定して供給できる器械をレンタルし、洗浄実験を行い一定の効果を得た。
今後はオゾンを含有させたマイクロバブルでのin vitroでの洗浄実験と、in vivoでの洗浄実験を行い、臨床応用への研究を継続していく予定である。

researchmap

Grant amount：\14400000 （ Direct expense: \14400000 ）

We analyzed molecular components of the new basement membrane-like structure of capillaries in the brain, fractone. We also tried identification and separation of adult neural stem cells existing subventricular zone, and investigated mechanisms of stem cell differentiation.
[1]Molecular and cellular analysis of the new basement membrane like structure fractone.
We detected the basement mambrane molecules such as laminin and type IV collagen in the mouse fractone by immunohistochemistry. Especially, several laminin chains and type IV collagen chains were present in the fractone, but fibronectin was not expressed there. Moreover, the fractone was distributed around almost all of subventricular zone in the brain and spinal code but it was not present in some areas of the third and forth ventricle. Developmental studies showed that the fractone started its expression around subventricular zone at postnatal day seven, and then expression pattern was limited to lateral part of ventricle at postnatal day 14. Furthermore, we recognized the fractone structure more precisely using immunoelectron-microscopic analysis.
[2]Identification and separation of neural stem cells from subventricular zone
We tried to separate neural stem cells from adult mouse subventricular zones and allowed them to further differentiate into neurons and astrocytes in various conditions. To set-up differentiation conditions, we prepared substrates rich in matrix macromolecules.
[3]Control of stem cell differentiation by extracellular matrix
We successfully culture neurospheres prepared from subventricular zone of the lateral ventricles and set-up conditions to further differentiate into neurons and astrocytes.

researchmap

Grant amount：\3400000 （ Direct expense: \3400000 ）

中枢神経系疾患では血液脳関門(BBB)の破綻を伴うがその分子機構はよく分かっていない.本研究は,中枢神経系疾患動物モデルを作製し,グリア境界膜の破綻とBBB破綻との関連性,さらにその破綻によって発現変化をきたす分子について解析し,グリア境界膜のBBBに対する重要性およびその形成と維持の分子メカニズムを明らかにすることを目的とした.本研究の成果によってグリア境界膜の働きが血液脳関門に重要であると示唆され,外傷モデルにおける血液脳関門不全が生じるメカニズムの一つを見出した.
1,中枢神経系疾患モデルの作製
外傷モデルとして「マウス脳凍結損傷モデル」及び「マウス脊髄損傷モデル」を確立した.
2,脊髄損傷モデルにおけるグリア境界膜の組織学的な解析
損傷組織の経時的な変化を組織染色,免疫組織染色によって解析した.limitrinは損傷後に速やかに消失し,壊死組織周辺の組織修復に伴いグリア境界膜への局在が回復した.さらに他のグリア境界膜構成分子についても観察した.これらの結果から,損傷によってグリア境界膜は破綻し,その後,修復されると示唆された.またグリア境界膜の構成単位である基底膜構築も変化し,血液脳関門不全の兆候を伴うことが明らかとなった.
3,limitrin遺伝子破壊マウスの作製と表現型解析
上記研究の結果からグリア境界膜の機能に重要と示唆された分子の一つである,limitrinの遺伝子破壊マウスを作製し,表現型解析を行った.

researchmap

Grant amount：\15000000 （ Direct expense: \15000000 ）

1 Expression analysis of novel aggrecanase
RNA was extracted from IL-1 b stimulated chondrosarcoma cells and chondrocytes and the aggrecanase expressions were analyzed by quantitative RT-PCR. When compared with fibroblasts, chondrosarcoma cells and chondrocytes showed higher induction of novel aggrecanase, thus indicating this novel aggrecanase is IL-1 induced specifically in cartilage cells. When chondrosarcoma cells were stimulated with IL-1b and TNFa, novel aggrecanase induced synergistically.
2 Antibody against novel aggrecanase
We raised a polyclonal antibody against novel aggrecanase, and checked its specificity by Western blotting. A novel aggrecanase was induced by IL-1b stimulation by Western blot analysis.
3 KO mouse
We got hetero mice
4 Signal mechanism for novel aggrecanase induction
MAP kinase inhibitor(PD98059,SB203580) were added to IL- 1b stimulated chondrosarcoma cells and the signaling pathway was determined.

researchmap

Grant amount：\2800000 （ Direct expense: \2800000 ）

血管新生を抑制するマルチプレキシンコラーゲンの性質を調べる意味で、以下の実験を行い、結果を得ることが出来た。マルチプレキシンコラーゲンの一つであるXVIII型コラーゲンは、そのカルボキシル末端の約250アミノ酸残基のNCドメインの中にはエンドスタチンとよばれ、腫瘍の周囲の血管新生を抑制することが1997年に初めて報告されてから、有望な抗がん作用を示す薬剤になりうる可能性を秘めていると思われて来た。しかしながら、いくつかの研究グループはその部分のペプチドに再現性を認めず、その作用機序を含め未解決の部分は多い。そこで、今回私どもは、XVIII型コラーゲンを多く産生する肝臓細胞株を探し出し、それをヌードマウスに打つことによって作られる腫瘍塊を作成した。そこで、XVIII型コラーゲンに特異的モノクローン抗体を担癌マウスに接種することにより、癌塊が増悪腫大するかどうかを観察することとした。その結果、この特異的モノクローン抗体はがん細胞をアポトーシスに導くことなく、肝癌の容積の増大を認めた。しかしながら、この特異抗体だけでは腫瘍細胞の増殖を促進することはなさそうであった。これらの結果は、エンドスタチンの抗腫瘍効果を支持する重要な所見であり、それだけでなく今回使用したものクローン抗体は、特異的にXVIII型コラーゲンに反応し、エンドスタチンの抗腫瘍効果を抑制することにまで至ったものであり、重要な知見であると思われる。

researchmap

Grant amount：\14100000 （ Direct expense: \14100000 ）

We report the molecular cloning of a new member of the transmembrane-type immunoglobulin superfamily and designate the encoded protein as limitrin since it selectively localized to glia limitans in mouse brain. Limitrin cDNA was obtained using a subtractive hybridization procedure designed to identify molecules responsible for blood-brain barrier function. Western blots using a limitrin-specific antibody demonstrated that the gene product is expressed significantly in mouse brain and primary murine astrocytes, and is distributed in the plasma membrane. Immunohistochemical studies using confocal and electron microscopy clearly demonstrated highly polarized localization in astroglial endfeet in the perivascular region and under the pia mater in vivo. Limitrin is expressed in spinal cord and many areas of the brain but not in the median eminence or subfornical organ (the circumventricular organs) where the blood-brain barrier is lacking. Disruption of the blood-brain barrier by cold injury resulted in a drastic reduction in limitrin expression. Furthermore, during retrieval from cold injury the increased expression of limitrin in perivascular endfeet correlated with the recovery of angiogenesis in capillaries within the lesion margins. Our results suggest that limitrin is physically and functionally associated with the blood-brain barrier, implying that this protein may be useful as a diagnostic determinant of barrier integrity.

researchmap

Grant amount：\14000000 （ Direct expense: \14000000 ）

To investigate new aggrecanase, we have done the following experiments and got some data.
1) Various ADAMTS-specific primers were designed and RT-PCR was performed. We cultured human chondrosarcoma cell lines OUMS-27 (Okayama University Medical School-27) and stimulated with interleukine-1 beta (IL-1β). To determine gene expression quantitatively, we employed real-time RT-PCR method. Briefly, total RNA was extracted and then DNAse treatmeat was done to eliminate contaminating genomic DNA. After reverse transcribed with random primers and enzymes, cDNA was served as a template for RT-PCR. GAPDH was used for the internal control.
2) The expression and gene regulation by IL-1β was different amonf the ADAMTS-1,4, and -5, which were reported to have aggrecan cleaving property in vitro. We also investigated other ADAMTS gene expressions.
3) We identified another up-regulating ADAMTS gene by IL-1β in OUMS-27 cells. We raised polyclonal antibody against this new ADAMTS gene using peptide sequence of this ADAMTS.
4) We also started to making knock-out mouse for this gene. We screened mouse genomic library and identified several clones including this new ADAMTS gene. Under the collaboration with Dr. Apte's lab in the USA, we started to put our clones to ES cells.

researchmap

Grant amount：\2000000 （ Direct expense: \2000000 ）

ADAMファミリーのうち、ADM-10とADAM-17はαセクレターゼ機能を持つことが知られているが、ADAMTSが同様な機能を持つか検討した。
まず、基礎的実験としてADAMTS-1はその遺伝子発現がリポポリサッカライド刺激により、各臓器において発現が上昇することから炎症において何らかの役割を持つことが示唆されている。そこで、ラット実験的心筋梗塞モデルを用いてADAMTS-1の発現形式をノーザンブロット法にて検討した。ADAMTS-1は非梗塞心臓では弱い発現しか認めなかったが、梗塞心においては、梗塞後6時間でその発現が大きく上昇していた。
マウス脳におけるADAMTSの発現をADAMTS-1〜7において検討したが、いずれもそれほど強くなかった。海外共同研究として、アミロイド前駆体蛋白を強制発現し、恒常的に発現する細胞株を樹立している、アラバマ大学Fukuchi教授らと共同実験を開始した。同細胞株は、通常の神経系細胞よりも過剰にアミロイド前駆体蛋白を発現している。これまでの検討によって過剰なアミロイド蛋白前駆体が細胞表面及び培養上清中に存在することが確認された。この実験系において過剰なアミロイド前駆体の切断がαセクレターゼによって制御されているかどうかを検討する目的でこの細胞系におけるαセクレターゼ発現の検討を開始した。実験の条件検討が複雑であり、切断の確認は困難であった。今後は、In vivoの系における実験系の確立およびアルツハイマー脳におけるADAMTSの発現解析が重要と考えられた。

researchmap

Grant amount：\14700000 （ Direct expense: \14700000 ）

To investigate the function of ten-m2/Neurestin gene for the olfactory sensory neuron projection, we have done the following experiments and got some data.
Ten-m2 specific antibodies were raised using the C-terminal unique amino acid sequence among the ten-m family. The specific staining was confirmed in the mouse section and confirmed its embryonic expression in combination with in situ hybridization.
Detection of Ten-m2 ligand: The extracellular domain of Ten-m2 protein was expressed in HEK293 cell culture system. We could detect specific protein band in the SDS-PAGE which bound to the ecto-domain. We are now identifying the protein.
Chromosomal mapping of human ten-m2 gene. The ten-m2, 3, and 4 genes were mapped to the chromosome 5, 4and 11, respectively. There was no genetic disorder reported to the locus.
Phenotypic analysis of the ten-m2 knockout mouse: By introducing a neomycin expression cassette in to the transmembrane exon of type II transmembrane protein, ten-m2, ten-m2 null mice were generated. There was no obvious phenotype so far. Ten-m2 null mice were viable and fertile, had a normal life span. There could be some compensation by another ten-m family members. The olfactory neuronal axon is now stained with MBP, marker of myelin and observed carefully.
Another approach to look for the ligand for ten-m2: The ten-m protein seems to be quite unique for its protein structure. Structural analysis will become a very important information to search the ligands. Ultrastructural analysis of recombinants revealed the dimerfomation through the disufide-bond. The dimer could be formed between the different members of ten-m family in our reconstitution experiments. Since CSPGs are known to act for the axon guidance, and the versican is expressed in the olfactory bulb, we tried to clone some related genes. The Brain link protein is cloned and the expression pattern was analyzed. The data for the novel link protein is published (see references).

researchmap

Grant amount：\8200000 （ Direct expense: \8200000 ）

By analyzing Col4a knockout mice, it is now possible to predict the biological function of α(IV) chains in vivo. We have now knockout mice of two strains: col4a3 and col4a6. Since we have evidence that there are major three molecular forms in basement membranes: (α1)2α2, α3α4α5, and (α5)2α6, it is now possible to set-up experiments to presume in vivo function of several α chains. We also set-up international collaboration to create another new project of gene targeting for col4α1 or col4α2, which makes it possible to make mice without type IV collagen.
We have solved problems of which of the 6 α(IV) chains can make molecules. We raised monoclonal antibodies specific for α(IV) chains. By using these 6 antibodies, we came to the conclusion that there are at least three molecular forms as mentioned above: (α1)2α2, α3α4α5, and (α5)2α6. New biochemical analysis using antibodies that recognize native chains made possible to immunoprecipitate specific molecules when they are digested by pure bacterial collagenase. The products run on SDS-PAGE were analyzed by other monoclonal antibodies that recognize now denatured α-chains by Western blots. This now method concluded that our prediction from immunocytochemistry was indeed true. In other words, there were three (α1)2α2, α3α4α5, and (α5)2α6 molecules. By this method, we analyzed basement membranes from renal glomeruli and bladder and aortic smooth muscle cells.
Specialized basement membrane from glomeruli was analyzed to contain supramolecular aggregates of (α1)2α2 and α3α4α5 molecules. The function of it is to filter blood to create urine. The similar phenomenon is found in choroid plexus in the brain. We analyzed it and found that it contains also the same compositions as glomerulus.

researchmap

Grant amount：\13900000 （ Direct expense: \13900000 ）

Under low oxygen condition, expression of collagen XVIII gene by cultured vascular endothelial cells reduced significantly. When it was analyzed by specific monoclonal antibodies, the collagen XVIII protein level was also reduced.
Endostatin induced regression of chondrosarcoma growth and tumor angiogenesis in vivo. However, endostatin showed no effects on the proliferation and migration of chondrosarcoma cells in vitro. Next, we investigated the interactions between endostatin and endothelial cells in detail. Endostatin inhibited the migration and attachment on collagen I but did not affect proliferation of endothelial cells. Although the migration of endothelial cells was stimulated with angiogenic factors such as basic fibroblast growth factor and vascular endothelial growth factor, endostatin showed similar inhibitory effects on it in the presence or absence of stimulants.
We came to the conclusion that non-vascular BMs contain predominantly one of the two types; I.e., subepithelial basement membranes contained type XVIII in general, whereas skeletal and cardiac muscles harbored prominently type XV. But basement membranes surrounding smooth muscle cells in vascular tissues contained one or both of them, depending on their locations. Interestingly, continuous or somatic capillaries contained both type XV and type XVIII collagens in their basement membranes; however, fenestrated or specialized capillaries such as glomeruli, liver sinusoids, lung alveoli, and splenic sinusoids expressed only type XVIII chains in their basement membranes, lacking type XV chain. This observation could imply different functions of basement membranes in various tissues and organs that use different mechanisms for the endogenous control of angiogenesis.

researchmap

Grant amount：\2000000 （ Direct expense: \2000000 ）

1 血管系特に毛細血管全体におけるXVIIIコラーゲンおよびXVコラーゲン遺伝子の発現
両コラーゲンは内皮細胞直下基底膜と血管壁に存在する平滑筋細胞周囲の基底膜であることがわかった。毛細血管内皮細胞下基底膜ではXVIII型とXV型を共通に発現するものが認められる.即ち腎や小腸粘膜、皮膚の毛細血管の内皮細胞下基底膜はXVIII型/XV型両者が共存する.しかしこれは組織臓器により異なり、肺胞壁、肝類洞、糸球体基底膜はXVIII型が主であり、他方胎盤、心臓、骨格筋、小腸筋層ではXV型が主であった.しかし、XVIII型/XV型両者の発現のない毛細血管は認められなかった.このように、毛細血管内皮細胞下基底膜のXVIII型とXV型の発現には多様性がある、XVIII型とXV型ともに発現するものが基本だが、類洞や糸球体基底膜のように特殊化した毛細血管ではXVIII型が優位に発現された.
2 基底膜IV型コラーゲンの新しい機能
IV型コラーゲンのα鎖は6種類あり、中央にコラーゲン部分、N末(NC2ドメイン)とC末(NC1ドメイン)に非コラーゲンドメインを有する。私達は、これらの6種類のα鎖のNC1ドメインをリコンビナントで作製し、これらを用いて血管内皮細胞の接着性と走化性をみたところ、α2、α3、α6鎖NC1ドメインにそれを抑制する活性が認められた。さらに、血管内皮細胞の接着と走化性はインテグリンαvβ1依存性であることが分かった。鶏胚の系(CAMアッセイ)でbFGFによって誘導された血管新生が、明らかにコントロールとくらべて優位に、リコンビナントα2、α3、α6鎖NC1ドメインによって抑制されるという結果を得た。このことは初めての報告であり、IV型コラーゲンの断片に新しい機能があることが明らかになったという意味で意義のある発見である。

researchmap

Grant amount：\12900000 （ Direct expense: \12900000 ）

Ultimate goal of the research project is to make artificial organs to insert basement membrane materials between epithelial layers and extracellular matrix. In order to make type IV collagen molecules in vitro, we we did the following investigations :
* Molecular composition of basement membranes underneath smooth muscle cells varies in different organs, which could be related to specific function of the individual organs.
* We first isolated and characterized genomic DNA fragments that cover the 5'flanking sequences of COL4A3 and COL4A4 genes, which provided information to delineate the promoter activity for the tissue-specific expression of the six type IV collagen genes.
* Molecular forms of collagen IV in follicle basement membranes are different in development.
* We identified interactions between the α2(IV) and ProMMP-9 by immunoprecipitations and blotting.
* Basement membrane collagen IV molecules diminish in parallel with malignancy and invasiveness when analyzed in tumor progression.
* Although mRNA expression level of α5 decreased in kidney of a canine Alport case, that of α6 resulted in unchanged. But the protein level of both chains cannot be found any. These results suggest a possibility of a molecular form of α5/α6.
* Differential expression of collagen IV genes in epithelial basement membranes was analyzed in detail.
* Here we define a novel function for soluble non-collagenous (NC1) domains of the α2(IV), α3(IV), and α6(IV) chains of human collagen type IV in the regulation of angiogenesis and tumor growth. These NCI domains were shown to regulate endothelial cell adhesion and migration by distinct αv and β1 integrin-dependent mechanisms.
* In Lmx1b(-/-) mice, expression of both α3 and α4 is strongly diminished in GBM, whereas that of α1, α2 and α5 is unchanged. Moreover, LMX1B binds specifically to a putative enhancer in intron 1 of mouse/human COL4A4 and upregulates reporter constructs containing the enhancer-like sequence.

researchmap