HGFならびにLIMキナーゼによる高次細胞機能の調節とがん特性の研究

1999 Fiscal Year Annual Research Report

• Project/Area Number: 06283216
• 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 / 肝硬変 / 慢性腎不全 / 器官再生 / 浸潤・転移阻止 / HGFアンタゴニスト / 血管新生 / LIMキナーゼ

Research Abstract

平成6-11年の6年間に私達はHGFが主に上皮細胞や血管内皮細胞などを標的細胞とし、機能的な細胞社会の構築(組織化)に必須な生物活性であるmitogen,motogen,morphogen活性に加え、anti-apoptosis作用による組織破壊の防止やbFGFに匹敵する強いangiogenesis作用を有することを明らかにした。そして私達はHGFが肝再生のみならず腎再生や肺再生など多くの組織器官の再生修復因子であり、一方、発生過程にあっては、肝、腎、肺などの多くの器官形成に必須の因子であることも明らかにした。また組織・器官の形成や再生因子であるHGFは広範囲な臓器傷害や組織破壊によって起こるさまざまな難治性疾患に対して極めて顕著な治療効果を発揮することを多くの疾患モデル動物を用いて実証した。たとえばHGFは劇症肝炎をはじめ、急性腎不全や急性心筋梗塞の発症防止と共に治癒を促し、さらに慢性疾患の肝線維症/肝硬変、腎硬化症、肺線維症などの線維化疾患に対し顕著な発症防止と治癒・機能改善作用を示す。
他方、HGFは癌と宿主の相互作用を介して多くの癌細胞の悪性化に関与すること、この機構が癌細胞が産生分泌するHGF誘導因子(IL-1,bFGF,TGF-α,PGsなど)とそれに応答して宿主間質により産生されるHGFとの間に形成されるHGFインデューサー/HGFループにあることを明らかにした。すなわち癌の浸潤・転移などの癌悪性化をブロックするにはこのループを断ち切ることにあると考えHGF・アンタゴニスト(NK4)を作製した。NK4はヌードマウスを用いたin vivo実験によって多くのヒト悪性癌(胆のう癌、肺癌、乳癌、膵癌なと)の浸潤・転移を抑制した。またNK4には腫瘍血管新生阻害作用もあり、新しい制癌剤になりうると考えられる。
LIMキナーゼ(LIMK)は私達の研究室によりc-seaのチロシンキナーゼドメインをプローブに8年前にクローニングした分子内にLIMドメインとPDZドメインを有するユニークなプロテインキナーゼである。LIMKはアクチン結合タンパク質のコフィリンをリン酸化し、アクチン細胞骨格を制御することが明らかになった。LIMK/はRacのLIMK2はRho/Cdc42の下流でそれぞれlamliopodiaとstress fiber形成/filopodia形成の制御を行っていることを明らかにした。

Research Products

• [Publications] T. Ueki, et al.: "Hepatocyle growth factor gene therapy of liver cirrhosis in rats"Nature Medicine. 5. 226-230 (1999)
• [Publications] Y. Liu, et al.: "Upregulation of hepatocyte growth factor receptor : an amplification and targeting mechanism for hepatocyte growth factor action in acute renal failure"Kidney Intern. 55. 442-453 (1999)
• [Publications] M. Tahara, K. Matsumoto, T. Nukiwa, and T. Nakamura: "Hepatocyte growth factor leads to recovery from alcohol-induced fatty liver in rats"J. Clin. Invest.. 103. 313-320 (1999)
• [Publications] J. Wojta, et al.: "Hepatocyte growth factor increases expression of vascular endothelial growth factor and plasminogen activator inhibitor-1 in human keratinocytes and the vascular endothelial growth factor receptor f1k-1 in human endothelial cells Evidence for a paracrine loop in wound healing"Lab. Invest. 79. 427-438 (1999)
• [Publications] Z. Jiao, et al.: "Effect of D-galactosamine hydrochloride and partial hepatectomy on spontaneous hepatic injury and hepatocarcinogenesis in Long Evans Cinamon rats"Jpn. J. Cancer Res.. 90. 496-504 (1999)
• [Publications] H. Azuma, et al.: "Effect of hepatocyte growth factor on cluonic rejection in rat renal allografts"Transplat. Proc.. 31. 854-855 (1999)
• [Publications] S. Takada, et al.: "Effect of hepatocyte growth factor on tacrolimus-induced nephrotoxicity in spontaneously hypertensive rats"Transpl. Int.. 12. 27-32 (1999)
• [Publications] H. Ueda, et al.: "Gene transfection of hepatocyte growth factor attenuates reperfusion injury in the heart"Annals. Thoracic Surg.. 67. 1726-1731 (1999)
• [Publications] K. Kosai, K. Matsumoto, and T. Nakamura: "Hepatocyte growth factor prevents endotoxin-induced lethal hepatic failure in mice"Hepatology. 30. 151-159 (1999)
• [Publications] R. Hasina, et al.: "Autocrine and paracrine motility factors and their involvement in invasiveness in a human oral carcinoma cell line"Brit. J. Cancer. 80. 1708-1717 (1999)
• [Publications] K. Kitajima, U. Koshimizu and T. Nakamura: "Expression of a novel type of classic cadherin, PB-cadherin in developing brain and limb buds"Develop. Dyn.. 215. 206-214 (1999)
• [Publications] N. Taniura-Matsumoto, K. Matsumoto and T. Nakamura: "Prostaglandin production in mouse mammary carcinoma cells confers invasive growth potential by inducing hepatocyte growth factor in fibroblasts"Brit. J. Cancer. 81. 194-202 (1999)
• [Publications] M. Kato, Y. Kato, T. Nakamura and Y. Sugiyama: "Efficient extraction by the liver governs the overall elimination of hepatocyte growth factor in rats"J. Pharmacol. Exp. Therap.. 290. 373-379 (1999)
• [Publications] Morishita, et al.: "Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as a cytokine supplement therapy"Hypertention. 33. 1379-1384 (1999)
• [Publications] W. G. Jiang, et al.: "Hepatocyte growth factor/scatter factor decreases the expression of occuludin and transendothelial resistance (TER) and increased paracellular permeability inhuman vascular endothelial cells"J. Cell. Pliysiol.. 181. 319-329 (1999)
• [Publications] C. C. Fehlner-Gardiner, et al.: "Characterization of a functional relationship between hepatocyte growth factor and mouse bone marrow-derived mast cells"Differentiation. 65. 27-42 (1999)
• [Publications] S. Deleu, et al.: "Immediate early gene expression in dog thyrocytes in response to growth proliferation and differentiation stimuli"J. Cell. Physiol.. 181. 342-354 (1999)
• [Publications] S. Deleu, et al.: "IFG-1 or insulin, and the TSH cyclic AMP cascade separately control dog and human thyroid cell growth and DNA synthesis, and complement each other in inducing mitogenesis"Mole. Cell. Endocrin.. 149. 41-51 (1999)
• [Publications] S. Hayashi, et al.: "Potential role of hepatocyte growth factor, a novel angiogenic growth factor in peripheral arterial disease : Downregulation of HGF in response to hypoxia in vascular cells"Circulation. 100. II-301-308 (1999)
• [Publications] T. Nagahori, et al.: "Interferon-gamma upregulates the c-MEt/HGF receptor expression in alvedar epithelial cells"Am. J. Respir. Cell Mol. Biol.. 21. 490-497 (1999)
• [Publications] O. Amano, U. Koshimizu, T. Nakamura and S. Iseki: "Enhancement by hepatocyte growth factor (HGF) of bone and cartilage formation during embryonic mouse mandibular development in vitro"Arch. Oral Biol.. 44. 935-946 (1999)
• [Publications] K. Nishimura, et al.: "Prostate stromal cell-derived hepatocyte growth factor induces invasion of prostate cancer cells DU145 through Tumor-stromal interaction"Prostate. 41. 145-153 (1999)
• [Publications] W. Sun, H. Funakoshi and T. Nakamura: "Differential expression of hepatocyte growth factor and its receptor, c-Met in the rat retina during development"Brain Res.. 851. 46-53 (1999)
• [Publications] A. Uchiyama, et al.: "Hepatocyte growth factor and invasion-stimulatory activity are induced in pleural fluid by surgery in lung cancer patients"Br. J. Cancer. 81. 721-726 (1999)
• [Publications] K. Matsumoto, et al.: "Prevention of renal damage by Angiotensin II blockade, accompanied by increased renal hepatocyte growth factor in experimental hypertensive rats"Hypertension. 34. 279-284 (1999)
• [Publications] Y. Okura, et al.: "Analysis of neurotrophic factor of hepatocyte growth factor (HGF) in the adult hypoglossal nerve axotomy model"Eur. J. Neurosci.. 11. 4139-4144 (1999)
• [Publications] T. Sumi, K. Matsumoto, Y. Takai and T. Nakamura: "Confilin phosphorylation and actin cytoskeletal dynamics regulated by Rho-and Cdc42-activated LIM-kinase 2"J. Cell Biol.. 147. 1519-1532 (1999)
• [Publications] M. K. Jones, et al.: "HGF triggers activation of Cox-2 gene in rat gastric epithelical cells Action mediated through the ERK2 signalling pathway"FASEB J.. 13. 2186-2194 (1999)
• [Publications] K. Kawano, et al.: "Phosphorylation of myosin-binding subunit (MBS) of myosin phosphatase by Rho-kinase in vivo"J. Cell Biol.. 147. 1023-1037 (1999)
• [Publications] F. Tamura, et al.: "FK506 promotes liver regeneration by suppressing natural killer cell activity"J. Gastroenterol. and Hepatol.. (in press). (2000)
• [Publications] A. Uchiyama, et al.: "Significance of locally-secreted hepatocyte growth factor after thoracic cancer surgery as a stimulator for postoperative cancer invasion"Jpn. J. Chin. Oncol.. (in press). (2000)
• [Publications] W. G. Jiang, et al.: "Antagonistic effect of NK4, a novel HGF variant, on in vitro angiogenesis of human vascular endothelial cells"Clin. Cancer Res.. (in press). (2000)
• [Publications] S. Hiscox, et al.: "Inhibition of HGF/SF-induced breast cancer cell matility and invasion by the HGF/SF variant, NK4"Breast Cancer Res. & Treat.. (in press). (2000)
• [Publications] M. Nakano, et al.: "Hepatocyte growth factor is essential for amelioration of hyperglycemia in strepto zotocin-induced diabetic mice receiving a marginal mass of intrahepatic islet grafts"Transplantation. (in press). (2000)
• [Publications] T. Yamada, et al.: "Enhanced expression of hepatocyte growth factor by pulmonary ischemia reperfusion injury in the rat"Am. J. Resp. Crit Care Med.. (in press). (2000)
• [Publications] S. Mizuno, et al.: "Reciprocal balance between HGF and TFG-β a key determinant for pathological phenotypes in chronic renal disease"Kidney Intern.. (in press). (2000)
• [Publications] C. Parr, et al.: "NK4, a new HGF/SF variant, is an antagonist to the influence of HGF/SF on the matility and invasion of colon cancer cells"Intern. J. Cancer. (in press). (2000)
• [Publications] 中村敏一: "新しい腫瘍血管新生制御物質-malignostatin(NK4)"細胞工学. 18. 223-225 (1999)
• [Publications] 久場敬司、松本邦夫、中村敏一: "HGFによる血管新生とその分子内クリングル断片(NK4)による腫瘍血管新生阻害"実験医学. 17. 734-740 (1999)
• [Publications] 水野信哉、中村敏一: "HGFによる急性および慢性腎不全の治療"腎と透析. 46. 494-498 (1999)
• [Publications] 久場敬司、中村敏一: "HGFと癌の浸潤・転移"Surgery Frontier 6 (No2) : 27-38. 6. 27-38 (1999)
• [Publications] 小清水右一、中村敏一: "HGF-肝細胞癌に対するHGFの抗腫瘍作用"血液・免疫・腫瘍. 4. 40-45 (1999)
• [Publications] 中村隆弘、中村敏一: "肝再生の分子機序"別冊:医学のあゆみ、消化器疾患. 2. 68-73 (1999)
• [Publications] 久場敬司、中村敏一: "医学研究その15:血管新生(Angiogenesis)"The Lung. 7. 94-99 (1999)
• [Publications] 水野信哉、中村敏一: "肝細胞増殖因子による腎再生"最新医学. 54. 59-69 (1999)
• [Publications] 角智行、中村敏一: "細胞遊走における増殖因子の役割"医学のあゆみ. 191. 137-142 (1999)
• [Publications] 水野信哉、中村敏一: "肝再生とHGF"現代医療. 31. 69-78 (1999)
• [Publications] 松本邦夫、中村敏一: "HGFの臨床応用に向けて"Medical Academy News. 720/721. 12-12 (1999)
• [Publications] 船越洋、中村敏一: "肝細胞増殖因子(HGF)の検査値と各種疾患"日本臨牀. 37. 821-826 (1999)
• [Publications] 小清水右一、中村敏一: "形質転換成長因子(TGF)"日本臨牀. 37. 802-804 (1999)
• [Publications] 中村敏一、大熊勝治: "肝細胞の培養法"細胞生物学実験法. 1. 27-47 (1999)
• [Publications] 中村敏一、小池克郎、関谷剛男、近藤寿人: "タンパク質分析法"分子生物学プロトコール 改訂2版. 11章. 269-284 (1999)
• [Publications] 中村敏一、長澤俊彦: "HGFによる腎疾患の治療"Annual Review 腎臓2000. XII. 233-240 (2000)