HGFによる形態形成および器官形成・再生の分子生物学的解析

2000 Fiscal Year Annual Research Report

• Project/Area Number: 11308025
• Research Institution: Osaka University
• Principal Investigator: 中村 敏一 大阪大学, 医学系研究科, 教授 (00049397)
• Co-Investigator(Kenkyū-buntansha): 船越 洋 大阪大学, 医学系研究科, 助手 (40273685) ; 小清水 右一 大阪大学, 医学系研究科, 助手 (50281126) ; 松本 邦夫 大阪大学, 医学系研究科, 助教授 (90201780)
• Keywords: HGF / c-Met / トランスジェニックマウス / 心筋梗塞 / 筋萎縮性側素硬化症 / 抗アポトーシス / 運動神経 / 再生・修復

Research Abstract

HGFの各種難治性疾患に対する治療効果
[心筋梗塞]心筋梗塞のラットモデルを作成し、心筋梗塞の進行におけるHGFの保護的役割を検討した。ラットの左心冠状動脈を結紮/解除すると、心筋梗塞の拡大に一致して血中HGF濃度は5-10倍に上昇する。そこでこの生理的機能を検証すべく心筋梗塞を発症したラットにHGF中和抗体を投与した結果、梗塞巣のさらなる拡大に一致して心機能が低下し、48時間以内に半数の動物が死亡した。逆に心筋梗塞発症に先立ってリコンビナントHGFを速やかに静脈内投与したところ、心筋細胞において抗アポトーシス分子であるBcl-xLの発現上昇に一致して心筋アポトーシスは抑制され、梗塞巣の拡大も強く抑制された。結果的にHGF投与によって心筋梗塞ラットでの心機能は著しく改善された。
[筋萎縮性側索硬化症]近年、HGFの神経栄養因子としての機能が注目されている。そこで神経変性疾患の中でも特に重篤で、治療が困難である筋萎縮性側索硬化症(ALS)に対するHGFの治療効果を検討した。まずALS発症モデルマウス(ALS-Tgマウス)では疾患の進行に伴い、脊髄組織におけるHGFおよびそのレセプター(c-Met)の発現が促進されることを見い出した。そこで神経特異的にHGFを発現するトランスジェニックマウス(NS-HGF-Tgマウス)を作成し、このマウスをALS-Tgマウスと交配させることにより、HGFのALS発症抑制効果を調べた。その結果、NS-HGF/ALS-TgマウスではALS-Tgマウスに比べて運動神経の軸索変性および細胞死が著明に改善されており、それに伴いALS発症時期の遅延ならびに寿命の延長が認められた。さらにHGFによるALS進行遅延効果は運動神経への直接作用に加えて、グリア細胞の特異蛋白質(グルタミン酸トランスポーター等)の発現制御や反応性増殖の阻害を介していることも明らかになった。

Research Products

• [Publications] S.Hiscox: "Inhibition of HGF/SF-induced breast cancer cell matility and invasion by the HGF/SF variant, NK4."Breast Cancer Res. & Treat.. 59. 245-254 (2000)
• [Publications] M.Nakano: "Hepatocyte growth factor is essential for amelioration of hyperglycemia in strepto zotocin-induced diabetic mice receiving a marginal mass of intrahepatic islet grafts."Transplantation. 69. 214-221 (2000)
• [Publications] S.Mizuno: "Reciprocal balance of hepatocyte growth factor and transforming growth factor-β1 in renal fibrosis in mice"Kidney Intern.. 57. 937-948 (2000)
• [Publications] K.Beppu: "Elevation of serum hepatocyte growth factor concentration in patients with gastric cancer is mediated by production by tumor tissue."Anticancer Res.. 20. 1263-1268 (2000)
• [Publications] M.Aoki: "Angiogenesis induced by hepatocyte growth factor in non-infarcted myocardium and infarcted myocardium : up-regalation of essential transcription factor for angiogenesis, ets."Gene Therapy. 7. 417-427 (2000)
• [Publications] T.Yamada: "Enhanced expression of hepatocyte growth factor by pulmonary ischemia reperfusion injury in the rat."Am.J.Resp.Crit Care Med.. 162. 707-715 (2000)
• [Publications] Y.Taniyama: "Potential contribution of a novel anti-fibrotic factor, hepatocyte growth factor, to prevention of myocardial fibrosis by angiotensin II blockade in cardiomyopathic hamster."Circulation. 102. 246-252 (2000)
• [Publications] T.Kunisada: "Keratinocyte expression of transgenic hepatocyte growth factor (HGF) affects melanocyte development, leading to dermal melanocytosis."Mech.Dev.. 94. 67-78 (2000)
• [Publications] S.Radaeva: "Unique epithelial cell production of hepatocyte growth factor/scatter factor by putative precancerous intestinal metaplasias and associated "intestinal-type" of biliary cancer chemically-induced in rat liver."Hepatolgy. 31. 1257-1265 (2000)
• [Publications] J.-M.Otte: "Functional expression of HGF and its receptor in human colorectol Cancer."Digestion. 61. 237-246 (2000)
• [Publications] O.Muto: "HGF/SF-induced spreading of MDCK cells corelates with disappearance of barmotin/7H6, a tight junction-associated protein, from the cell membrane."Cell Biol.Intern.. 24. 439-446 (2000)
• [Publications] T.Kikuchi: "Secretory leukoprotease inhibitor augments hepatocyte growth factor production in human lung fibroblasts."Am.J.Respir.Cell Mol.Biol.. 23. 364-370 (2000)
• [Publications] K.Hayashi: "In vivo transfer of human hepatocyte growth factor gene accelerates re-endothelization and inhibition neointimal formation after balloon injury in rat model."Gene Therapy. 7. 1664-1671 (2000)
• [Publications] H.Nakagami: "Anti-apoptotic action of hepatocyte growth factor through mitogen-activated protein kinase on human aortic endothelial cells."J.Hypertension. 18. 1411-1420 (2000)
• [Publications] T.Nakamura: "Myocardial protection from infarction by endogenous and exogenous hepatocyte growth factor."J.Clin.Invest.. 106. 1511-1519 (2000)
• [Publications] K.Kuba: "HGF/NK4, a four-kringle fragment of hepatocyte growth factor, is an angiogenesis inhibitor that suppresses tumor growth and metastasis in mice."Cancer Res.. 60. 6737-6743 (2000)
• [Publications] K.Kuba: "Kringle 1-4 of hepatocyte growth factor inhibits proliferation and migration of human microvascular endothelial cells."Biocem.Biophys.Res.Commun.. 279. 846-852 (2000)
• [Publications] W.Sun: "A sensitive quantification method for evaluating the lebel of hepatocyte growth factor and c-met/HGF receptor mRNAs in the nervous system using competitive RT-PCR."Brain Res.Protoc.. 5. 190-7 (2000)
• [Publications] G.H.Lai: "Unique epithelial cell production of hepatocyte growth factor/scatter factor by putative precancerous intestinal metaplasias and associated "intestinal-type" biliary cancer chemically induced in rat liver."Hepatology. 31. 1257-1265 (2000)
• [Publications] G.Shiota: "In vivo transfer of hepatocyte growth factor gene accelerates proliferation of hepatic oval cells in a 2-acetylaminofluorene/partial hepatectomy model in rats."FEBS Lett.. 470. 325-330 (2000)
• [Publications] K.Matsumoto: "Hepatocyte growth factor and met in tumor invasion-metastasis : Mechanisms related to cancer prevention In "Cancer metastasis, molecular and cellular biology and clinical aspects""Academic Publishers. 143-193 (2000)
• [Publications] K.Matsumoto: "Hepatocyte growth factor in renal regeneration, renal disease, and potential therapeutics."Hypertension. 9. 395-402 (2000)
• [Publications] T.Takahashi: "Functional involvement of LIM-kinase in Xenopus oocyte maturation."Develop.Biol.. 229. 554-567 (2001)
• [Publications] T.Sumi: "Specific activation of LIM-kinase 2 via threonine-505 phosphorylation by ROCK under the control of Rho."J.Biol.Chem.. 276. 670-676 (2001)
• [Publications] F.Tamura: "FK506 promotes liver regeneration by suppressing natural killer cell activity."Gastroenterol.and Hepatol.. (in press). (2001)
• [Publications] A.Uchiyama: "Significance of locally-secreted hepatocyte growth factor after thoracic cancer surgery as a stimulator for postoperative cancer invasion."Jpn.J.Clin.Oncol.. (in press). (2001)
• [Publications] H.Azuma: "Hepatocyte growth factor prevents development of chronic allograft nephropathy in an experimental rat transplant model."J.Am.Soc.Nephrol.. (in press). (2001)
• [Publications] S.Mizuno: "Hepatocyte growth factor suppresses interstitial fibrosis in a mouse model of obstructive nephropathy."Kidney Intern.. (in press). (2001)
• [Publications] K.Nakamura: "Molecular cloning and functional characterization of human scavenger receptor with ctype lectin (SRCL), a novel member of scavenger receptor family."Biochem.Biophys.Res.Commun.. (in press). (2001)
• [Publications] N.Maehara: "NK4, a four-kingle antagonist of HGF, inhibits spreading and invasion of human pancreatic cancer cells."Br.J.Cancer. (in press). (2001)
• [Publications] T.Nakamura: "Molecular cloning and characterization of Kremen, a novel Kringle-containing transmembrane protein."Biochem.Biophys.Acta. (in press). (2001)
• [Publications] N.Tsuzuki: "Hepatocyte growth fector reduces the infarct volume after transient focal cerebral ischemia."Neurol.Res.. (in press). (2001)
• [Publications] K.Matsumoto: "Hepatocyte growth factor : renotropic role and potential therapeutics for renal diseases."Kidney Intern.. (in press). (2001)
• [Publications] K.Matsumoto: "Cancer Metastasis : Molecular & Cellular Mechanisms and Clinical Interventions"Kluwer Academic Publisher. 50 (2000)
• [Publications] K.Matsumoto: "Growth Factors and Their Receptors in Cancer Metastasis"Kluwer Academic Publisher (in press). (2001)