HGFによる形態形成機構の分子細胞生物学的研究

1997 Fiscal Year Annual Research Report

• Project/Area Number: 08408027
• Research Institution: Osaka University
• Principal Investigator: 中村 敏一 大阪大学, 医学部, 教授 (00049397)
• Co-Investigator(Kenkyū-buntansha): 宮本 一政 大阪大学, 医学部, 助手 (60273679) ; 小清水 右一 大阪大学, 医学部, 助手 (50281126) ; 船越 洋 大阪大学, 医学部, 助手 (40273685) ; 松本 邦夫 大阪大学, 医学部, 助教授 (90201780)
• Keywords: HGF / c-Met / 劇症肝炎 / 肝細胞アポトーシス / Bcl-xl / LIMキナーゼ / Sky / Gas6

Research Abstract

今年度の主な研究成果を以下に記す。
1.HGFによる肺の器官形成機構:ラット13日目胎仔肺においてHGFとそのレセプターc-Met mRNAの発現はそれぞれ間葉組織および気管支皮組織に限局して認められる。肺原基より間葉組織を取り除いた上皮単独培養系にHGFを添加すると上皮組織の形態を維持し、細胞死を抑制する″histoprotective″な活性が認められた。この培養系にHGFとaFGFあるいはKGFを同時添加すると、活発なbuddingと共に管腔組織の伸長が促進された。次に肺原基の培養系にHGFの中和抗体ならびにFGFレセプターのアンチセンスDNAを添加すると細気管支の枝分れが強く抑制された。以上の結果より肺上皮組織の分枝管腔形成にはHGFとFGFシグナルの両者の存在が必須であることが明らかになった。
2.HGFによる劇症肝炎の防止作用:マウスに抗Fas抗体を投与すると投与8時間後には大量肝細胞死による劇症肝炎を発症し、80%以上のマウスが肝不全死する。これに対してHGFを投与したマウスでは抗Fas抗体による肝細胞アポトーシスがほぼ完全に抑制されマウスは全例生存した。同様な結果はエンドトキシンとD-ガラクトサミンで誘起した劇症肝炎モデルでも認められた。両モデルにおいてHGFは肝臓でのBcl-xlの発現を著明に誘導し、この作用がHGFによる強力な劇症肝炎抑制効果の1つの機作と考えられた。
3.LIMキナーゼ(LIMK)相互作用分子の同定:LIMK1ならびにLIMK2と相互作用する分子をyeast two hybrid法により約50種類同定した。そのうち新規分子は23種類であり、細胞骨格、細胞周期、転写調節に関係する分子と考えられた。さらに精巣において特異的に発現するLIMK2のバリアント(LIMドメイン欠損型)を見い出した。
4.脳神経特異的レセプター型キナーゼSkyの機能:私達の研究室で発見したSkyと、そのリガンドGas6の脳神経系に於ける役割を解析した結果、Gas6がSkyを介して大脳皮質、海馬などの多くの神経細胞の生存を促す新しいneurotrophic factorとして機能することを見い出した。

Research Products

• [Publications] K.Takai et al.: "Hepatocyte growth factor is constitutively produced by human bone marrow stromal cells and promotes erythropoiesis." Blood. 89. 1560-1565 (1997)
• [Publications] R,Morishita et.al.: "Potential role of endothelium-specific growth factor hepatocyte growth factor,on endothelial damage in diabetes militus." Diabetes. 46. 138-142 (1997)
• [Publications] I.S.Weimar et al.: "Hepatocyte growth factor/scatter factor(HGF/SF)promotes adhesion of lymphoma cells to extra-cellular matrix molecules via a4b,and a5b,integrens." Blood. 89. 990-1000 (1997)
• [Publications] N.Kawakami-Kimura et al.: "Involvement of hepatocyte growth factor in increased integrin expression on Hep G2 cells triggered by adthesion to endothelial cells." Br.J.Cancer. 75. 47-53 (1997)
• [Publications] T.Sakai et al.: "Hepatocyte growth factor in bronchoalveolar lavage fluid and cells in patients with in flammatory chest diseases of the lower respitatory tract:Defection by RIA and in situ hybridization." Am.J.Respir.Cell Mole Biol.16. 388-397 (1997)
• [Publications] T.Takahashi et al.: "Xenopus LIM motif-containing protein kinase,XLIMK,is expressed in the developing head structure of the embryo." Develop.Dyn.209. 196-205 (1997)
• [Publications] S.Aoki et al.: "Activation of Met tyrosine kinase by hepatocyte growth factor is essential for intemal ganogenesis in Xenopus embryo." Biochem.Biophys.Res.Commun.234. 8-14 (1997)
• [Publications] Y.Ikeda et al.: "Bile ductular proliferation in the granulation tissue of rat liver.:Histological and serial reconstruction studies." Biomed.Res.18. 113-127 (1997)
• [Publications] Y.Matsuda et al.: "Prevential and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis." Hepatology. 26. 81-89 (1997)
• [Publications] K.Furugori et al.: "Level of Hepatocyte growth factor and its messenger ribonucleic acid in uncomplecated pregnancies and those complicated by preeclamptia." J.Clin.Endocrinol.&Metabo.82. 2726-2730 (1997)
• [Publications] T.Nakamura et al.: "Induction of hepatocyte growth factor in fibroblasts by tumor-derived factors affects invasive growth of tumor cells:In vitro analysis of tumor-stromal interaction." Cancer Res.57. 3305-3313 (1997)
• [Publications] T.Imai et al.: "Sparse distribution of hepatocyte growth factor-producing cells inside hepatocellular foci in rats treated with hepatocarcinogens." Jpn.J.Cancer Res.88. 615-618 (1997)
• [Publications] T.Kaido et al.: "Continous hepatocyte growth factor sypply prevents lipopolysacharide-induccd liver injury in rats." FEBS Lett.411. 378-382 (1997)
• [Publications] S.E.Hiscox et al.: "Expression of the HGF/SF receptor c-met,and its ligand in human colon cancers." Cancer Investigations. 15. 513-521 (1997)
• [Publications] R.Morishita et al.: "Role of hepatocyte growth factor in endothelial regulation:Prevention of high D-glucose induced endothelia lcell death by prostglandins and phosphodiesterase type 3 inhibitor." Diabetologia. 40. 1053-1061 (1997)
• [Publications] H.Li et al.: "Hepatocyte gorwht factor stimuolates invasion of gallbladder carcinoma cell lines in vitro." Clin.Exp.Metastasis.15. 484-493 (1997)
• [Publications] K.Liu et al.: "Existence of two nonliner elimination mechanisms for hepatocyte growth factor in rats." Am.J.Physiol. 273. E891-E897 (1997)
• [Publications] S.Wakitani et al.: "Hepatocyte growth factor faulitates Cartilage repair:full thickness articular cartilage defect studied in rabbit knee." Acta Orthop.Stand.68. 474-480 (1997)
• [Publications] T.Adachi et al.: "Possible involvement of pertussis toxin-sensitive G protein in hepatocyte growth factor(HGF)-induced signal transduction in cultured rat hepatocytes." Hepatology.26. 295-300 (1997)
• [Publications] V.R.Wallenius et al.: "Chromosomal localization of rat hepatocyte growth factor(Hgf)and HGF receptor(Met)and chractrization of HGF receptor cDNA." Mammalian Genome. 8. 661-667 (1997)
• [Publications] N.Nakano et al.: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor(HGF),in experimental hypertensive rats." Hypertension. 30. 1448-1454 (1997)
• [Publications] K.Date et al.: "HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor." FEBS Lett.420. 1-6 (1997)
• [Publications] A.Schmassman et al.: "Role of antagotensin II in the reguration of a novel vascular modulator,hepatocyte growth factor(HGF),in experiemental hypertensive rats." Gastroenterolgy. 113. 1858-1872 (1997)
• [Publications] U.Koshimizu et al.: "cDNA cloning,genomic organization and chromosomal localization of the mouse Lim motif-containing kinase,Limk 2 gene." Biochem.Biophys.Res.Commun.241. 243-250 (1997)
• [Publications] M.Yaekashiwa et al.: "Both simultaneous and delayed administration of hepatocyte growth factor(HGF)repress the fibrotic changes in murine lung injury induced by bleomycin." Am.J.Resp.&Clin.Care Med.156. 1937-1944 (1997)
• [Publications] R.Morishita et al.: "Potential role of a novel vascular modulator,hepatocyte growth factor(HGF),in cardiovascular disease:characterization and regulation of local HGF system." J.Atherosclerosis and Thrombosis. 4. 12-19 (1997)
• [Publications] U.Koshimizu et al.: "Hepatotrophic activities of HGF in liver regeneration after injury and the clinical potentiality for liver diseases." "Liver and Environmental Xenobiotis"Narosa-Elsevier Verlag,. 220-229 (1997)
• [Publications] K.Matsumoto et al.: "HGF:its organotrophic role and therapeutic potential." "Plasminogen-Related Growth Factors". 212. 198-214 (1997)
• [Publications] K.Matsumoto et al.: "Hepatocyte growth factor(HGF)as a rissue organizer for organogenesis and tegeneration." Biochem.Biophys.Res.Commun.(Mini-Review). 239. 639-644 (1997)
• [Publications] K.Matsumoto et al.: "Pathophysiological aspects and therapeutic potential of hepatocyte growth factor in liver disease." Internal Medicine and Endocrinology:Pathophysiological and clinical interactions(eds by A.Fabbrini and V.Santiemma.). pp.95-106 (1997)
• [Publications] D.Schuppan et al.: "Collagens in the liver extracellular matrix hepatocyte growth factor." Gastroenterology.114. 139-152 (1998)
• [Publications] M.Lyon, et al.: "Hepatocyte growth factor/scatter factor binds with high affinity to dematan sulphate." J.Biol.Chem.273. 271-278 (1998)
• [Publications] Y.Yo et al.: "Actions of hepatocyte growth factor as a local modulator in the kidney:Potential role in pathogenesis of renal disease." Kindney Intern. 53. 50-58 (1998)
• [Publications] K.Liu et al.: "Protamine enhances the proliferative activity of hepatocyte growth factor in rats." Am.J.Physiol.(in press.). (1997)
• [Publications] R.Gal-Levi et al.: "Hepatocyte growth factor as a potent regulator of skeletal muscle satellite cell proliferation and differentiation." Biochem.Biophys.Acta.(in press.). (1997)
• [Publications] K.Liu et al.: "Change in hepatic handling of hepatocyte growth factor during liver regeneration in rats." Am.J.Physiol.269(in press.). (1998)
• [Publications] S.Mizuno et al.: "Hepatocyte growth factor prevents renal fibrosis and dysfunction in mouse model of chronio renal disease." J.Clin.Invest.(in press.). (1998)
• [Publications] H.Ohmichi et al.: "Hepatocyte growth factor(HGF)acts as a mesenchymal-derived morphogenic factor during fetal lung develpment." Development.(in press.). (1998)
• [Publications] T.Miyazawa et al.: "Protection of hippocampal neurons from ischemia-induced delayed neuronal death by hepatocyte growth factor:A novel neurotrophic factor." J.Celebral Blood Flow Metabolism.(in press.). (1998)
• [Publications] M.T.Donato et al.: "Human hepatocyte growth factor down regulated the expression of cytochrome P450 isozymes in human hepatocytes in primaoy culture." J.Pharmacl Exp.Therap.(in press.). (1998)
• [Publications] K.Kosai et al.: "Hepatocyte growth factor abrogates Fas-induced lethal liver apoptosis in mice." Biochem.Biophys.Res.Commun.(in press.). (1998)
• [Publications] F.Tamura et al.: "FK506 promotes liver regeneration by suppressing natural killer cell activity.natural killer cell activity." J.Gastroenterol.and Hepatol.(in press.). (1998)
• [Publications] W.G.Jiang et al.: "Hepatocyte growth factor/scatter factor,its molecular,cellular and clinical impacts in cancer." Crit.Review Onclo.Hematol.(in press.). (1998)
• [Publications] 中村敏一: "標準分子医化学" 医学書院, 843-852 (1997)
• [Publications] 中村敏一: "分子生物学プロトコール改訂版 タンパク質の生化学的分析" 南江堂(印刷中), (1997)
• [Publications] 中村敏一: "肝臓病-今日の診断と最新の治療" 国際医書出版, pp.299-310 (1997)
• [Publications] 中村敏一: "生命体システムの修復" 講談社サイエンティフィック, 124 (1997)
• [Publications] 松本邦夫、中村敏一: "病理学キーワード" 文光社, 40 (1997)