2001 年度実績報告書

NK4(HGFアンタゴニスト/抗血管新生因子)による悪性腫瘍の治療研究

研究課題

研究課題/領域番号	13218075
研究機関	大阪大学
研究代表者	松本邦夫大阪大学, 医学系研究科, 助教授 (90201780)
キーワード	HGF / c-Met / NK4 / 浸潤 / 転移 / 血管新生 / クリングルドメイン / 腫瘍血管新生
研究概要	[目的]私達はHGFが多くの癌細胞の浸潤・転移を促す宿主因子であること基づき、HGFに対するアンタゴニストとして作用するNK4を調製するとともに、NK4にはHGFアンタゴニスト活性に加え、血管新生阻止能をもつ2機能性分子であることを明らかにした。本年度はNK4による制癌研究を行う一端としてNK4の血管新生阻害機構を明らかにする目的で研究を行った。 [結果]ヒト血管内皮細胞の倍養系にNK4を添加したところ、NK4はbFGFによって誘導されるDNA合成ならびにS期への移行を阻害した。また、NK4はbFGFによって誘導されるERK1/2の活性化を阻害しなかったが、DNA合成に先立つRB蛋白質のリン酸化を強く抑制した。したがって、NK4による血管内皮細胞の増殖阻害の作用点は、少なくともERK1/2の下流でRB蛋白質のリン酸化制御に関与する分子であることが明らかになった。一方、NK4による血管新生阻害作用が当初のHGFアンタゴニスト作用と独立のものであることを明らかにするため、NK4分子の欠失変異体を調製しその生物活性を調べた。NK4分子内のN末端ヘアピンドメインならびに第1クリングルドメインはc-Met/HGFレセプターへの結合に必須の領域である。そこで、NK4からN末端ヘアピンドメインを欠く変異体(K1-4)を発現、精製した。K1-4はNK4と異なりHGFに対するアンタゴニスト作用を失っていたが、血管新生阻害作用を有していた。したがって、血管新生阻害に関与するNK4の分子内領域は4個のクリングルドメイン内にあることが明らかになった。 [考察]血管新生阻害は主にNK4分子内の4個のクリングルドメインを介して発揮されることから、NK4はクリングルドメインを介して血管内皮細胞上のc-Metレセプターとは異なる結合分子に作用し、この結合分子からのシグナルがRB蛋白質のリン酸化阻害に関与するものと考えられる。本NK4結合分子の同定はNK4による血管新生阻害の分子機構の解明や血管新生阻害作用をもつ新たな分子を見いだすための標的分子となるものと考えられる。

研究成果

(32件)

すべてその他

すべて文献書誌 (32件)

[文献書誌] Matsumoto, K., et al.: "Liver organogenesis promoted by endothelial cells prior to vascular function"Science. 294. 559-563 (2001)
[文献書誌] Tomioka, D., et al.: "Inhibition of growth, invasion, and metastasis of human pancreatic carcinoma cells by NK4 in an orthotopic mouse model"Cancer Res.. 61. 7518-7524 (2001)
[文献書誌] Sumi, T., et al.: "Specific activation of LIM-kinase 2 via phosphorylation of the threonine-505 by ROCK, a Rho-dependent protein kinase"J. Biol. Chem.. 276. 670-676 (2001)
[文献書誌] Matsumoto, K., et al.: "Hepatocyte growth factor : renotropic role and potential therapeutics for renal diseases"Kid. Intern.. 59. 2023-2038 (2001)
[文献書誌] Sumi, T., et al.: "Activation of LIM kinases by myotonic dystrophy kinase-related Cdc42-binding kinase alpha"J Biol Chem.. 276. 23092-23096 (2001)
[文献書誌] Kuroiwa, T., et al.: "Hepatocyte growth factor ameliorates acute graft-versus-host disease and promotes hematopoietic function"J Clin Invest.. 107. 1365-1373 (2001)
[文献書誌] Maehara, N., et al.: "NK4, a four-kringle antagonist of HGF, inhibits spreading and invasion of human pancreatic cancer cells"Br. J. Cancer. 84. 864-873 (2001)
[文献書誌] Mizuno, S., et al.: "Hepatocyte growth factor suppresses interstitial fibrosis in a mouse model of obstructive nephropathy"Kidney Intern.. 59. 1304-1314 (2001)
[文献書誌] Nakamura, T., et al.: "Molecular cloning and characterization of Kremen, a novel kringle-containing transmembrane protein"Biochim. Biophys. Acta. 1518. 63-72 (2001)
[文献書誌] Taniyama, Y., et al.: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat and rabbit hindlimb ischemia models : preclinical study for treatment of peripheral arterial disease"Gene Therapy. 8. 181-189 (2001)
[文献書誌] Azuma, H., et al.: "Hepatocyte growth factor prevents development of chronic allograft nephropathy in an experimental rat transplant model"J. Am. Soc. Nephrol.. 12. 1280-1292 (2001)
[文献書誌] Matsumoto, K., et al.: "Inhibition of neointima by angiotensin-conveitng enzyme inhibitor in porcine coronary artery balloon-injury model"Hypertension. 37. 270-264 (2001)
[文献書誌] Nakagami, H., et al.: "Mitogenic and antiapoptotic actions of hepatocyte growth factor through ERK, STAT3, and AKT in endothelial cells"Hypertension. 37. 581-586 (2001)
[文献書誌] Yamamoto, K., et al.: "Contribution of bcl-2, but not bcl-xl and bax, to antiapoptotic actions of hepatocyte growth factor in hypoxia-conditioned human endothelial cells"Hypertension. 37. 1341-1348 (2001)
[文献書誌] Ueda, H., et al.: "A potential cardioprotective role of hepatocyte growth factor in myocardial infarction in rats"Cardiovascular Res.. 51. 41-50 (2001)
[文献書誌] Tsuzuki, N., et al.: "Hepatocyte growth factor reduces the infarct volume after transient focal cerebral ischemia in rats"Neurol Res.. 23. 417-424 (2001)
[文献書誌] Parr, C., et al.: "The HGF/SF-induced phosphorylation of paxillin, matrix adhesion, and invasion of prostate cancer cells were suppressed by NK4,an HGF/SF variant"Biochem Biophys Res Commun.. 285. 1330-1337 (2001)
[文献書誌] Beppu, K., et al.: "Hepatocyte growth factor production by peripheral blood mononuclear cells of recurrent cancer patients"Anticancer Res.. 21. 2195-2200 (2001)
[文献書誌] Aoki, M., et al.: "Inhibition of neointimal formation after balloon injury by cilostazol, accompanied by improvement of endothelial dysfunction and induction of hepatocyte growth factor in rat diabetes model"Diabetologia. 44. 1034-1042 (2001)
[文献書誌] Taniyama, Y., et al.: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat diabetic hind limb ischemia model : molecular mechanisms of delayed angiogenesis in diabetes"Circulation. 104. 2344-2350 (2001)
[文献書誌] Hayashi, K., et al.: "Gene therapy for preventing neuronal death using hepatocyte growth factor : in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CA1 neurons"Gene Therpy. 8. 1167-1173 (2001)
[文献書誌] Shibuki, H., et al.: "Expression and neuroprotective effect of hepatocyte growth factor in retinal ischemia-reperfusion injury"Invest. Ophthalmol. Vis. Sci.. 43. 528-536 (2002)
[文献書誌] Maemondo, M, et al.: "Targeting angiogenesis and HGF function using an adenovirul vector expressing the HGF-antagonist NK4 for cancer therapy"Mol. Therapy. 5. 177-185 (2002)
[文献書誌] Sumi, T., et al.: "Mitosis-dependent phosphorylation and activation of LIM-kinase-1"Biochem. Biophys. Res. Commun.. (印刷中). (2002)
[文献書誌] Sakamaki, Y., et al.: "Hepatocyte growth factor stimulates proliferation of respiratory epithelial cells during postneumonectomy compensatory lung growth in mice"Am. J. Respir. Cell Mol. Biol.. (印刷中). (2002)
[文献書誌] Funakoshi, H., et al.: "Identification of Gas6,a putative ligand for Sky and Axl receptor tyrosine kinases, as a novel neurotrophic factor for hippocampal neurons"J. Neurosci. Res.. (印刷中). (2002)
[文献書誌] Kishi, A.Y., et al.: "Molecular cloning, expression and partial characterization of Kksy, a novel Xenopus member of Sky receptor tyrosine kinase"Gene. (印刷中). (2002)
[文献書誌] 松本邦夫, 中村敏一: "増殖因子による再生医学の展開-HGFを中心にして-"Biotherapy. 15. 95-104 (2001)
[文献書誌] 松本邦夫, 他: "HGFによる肺の再生と線維化の阻止"呼吸. 20. 935-939 (2001)
[文献書誌] 富岡大策, 他: "NK4/malignostatinによる腫瘍血管新生阻止作用とその制がん効果"最新医学. 56. 1799-1807 (2001)
[文献書誌] 松本邦夫: "肝臓の初期発生を支える血管内皮細胞"実験医学. 20. 468-471 (2002)
[文献書誌] Matsumoto, K, et al.: "分担:HGF-c-Met receptor pathway in tumor invasion-metastasis and potential cancer treatment with NK4書名:Growth Factors and Their Receptors in Cancer Metastasis"Kluwer Academic Publisher(分担部分). 35 (2001)

2001 年度 実績報告書

NK4(HGFアンタゴニスト/抗血管新生因子)による悪性腫瘍の治療研究

研究代表者

松本 邦夫 大阪大学, 医学系研究科, 助教授 (90201780)

研究成果

[文献書誌] Matsumoto, K., et al.: "Liver organogenesis promoted by endothelial cells prior to vascular function"Science. 294. 559-563 (2001)

[文献書誌] Tomioka, D., et al.: "Inhibition of growth, invasion, and metastasis of human pancreatic carcinoma cells by NK4 in an orthotopic mouse model"Cancer Res.. 61. 7518-7524 (2001)

[文献書誌] Sumi, T., et al.: "Specific activation of LIM-kinase 2 via phosphorylation of the threonine-505 by ROCK, a Rho-dependent protein kinase"J. Biol. Chem.. 276. 670-676 (2001)

[文献書誌] Matsumoto, K., et al.: "Hepatocyte growth factor : renotropic role and potential therapeutics for renal diseases"Kid. Intern.. 59. 2023-2038 (2001)

[文献書誌] Sumi, T., et al.: "Activation of LIM kinases by myotonic dystrophy kinase-related Cdc42-binding kinase alpha"J Biol Chem.. 276. 23092-23096 (2001)

[文献書誌] Kuroiwa, T., et al.: "Hepatocyte growth factor ameliorates acute graft-versus-host disease and promotes hematopoietic function"J Clin Invest.. 107. 1365-1373 (2001)

[文献書誌] Maehara, N., et al.: "NK4, a four-kringle antagonist of HGF, inhibits spreading and invasion of human pancreatic cancer cells"Br. J. Cancer. 84. 864-873 (2001)

[文献書誌] Mizuno, S., et al.: "Hepatocyte growth factor suppresses interstitial fibrosis in a mouse model of obstructive nephropathy"Kidney Intern.. 59. 1304-1314 (2001)

[文献書誌] Nakamura, T., et al.: "Molecular cloning and characterization of Kremen, a novel kringle-containing transmembrane protein"Biochim. Biophys. Acta. 1518. 63-72 (2001)

[文献書誌] Taniyama, Y., et al.: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat and rabbit hindlimb ischemia models : preclinical study for treatment of peripheral arterial disease"Gene Therapy. 8. 181-189 (2001)

[文献書誌] Azuma, H., et al.: "Hepatocyte growth factor prevents development of chronic allograft nephropathy in an experimental rat transplant model"J. Am. Soc. Nephrol.. 12. 1280-1292 (2001)

[文献書誌] Matsumoto, K., et al.: "Inhibition of neointima by angiotensin-conveitng enzyme inhibitor in porcine coronary artery balloon-injury model"Hypertension. 37. 270-264 (2001)

[文献書誌] Nakagami, H., et al.: "Mitogenic and antiapoptotic actions of hepatocyte growth factor through ERK, STAT3, and AKT in endothelial cells"Hypertension. 37. 581-586 (2001)

[文献書誌] Yamamoto, K., et al.: "Contribution of bcl-2, but not bcl-xl and bax, to antiapoptotic actions of hepatocyte growth factor in hypoxia-conditioned human endothelial cells"Hypertension. 37. 1341-1348 (2001)

[文献書誌] Ueda, H., et al.: "A potential cardioprotective role of hepatocyte growth factor in myocardial infarction in rats"Cardiovascular Res.. 51. 41-50 (2001)

[文献書誌] Tsuzuki, N., et al.: "Hepatocyte growth factor reduces the infarct volume after transient focal cerebral ischemia in rats"Neurol Res.. 23. 417-424 (2001)

[文献書誌] Parr, C., et al.: "The HGF/SF-induced phosphorylation of paxillin, matrix adhesion, and invasion of prostate cancer cells were suppressed by NK4,an HGF/SF variant"Biochem Biophys Res Commun.. 285. 1330-1337 (2001)

[文献書誌] Beppu, K., et al.: "Hepatocyte growth factor production by peripheral blood mononuclear cells of recurrent cancer patients"Anticancer Res.. 21. 2195-2200 (2001)

[文献書誌] Aoki, M., et al.: "Inhibition of neointimal formation after balloon injury by cilostazol, accompanied by improvement of endothelial dysfunction and induction of hepatocyte growth factor in rat diabetes model"Diabetologia. 44. 1034-1042 (2001)

[文献書誌] Taniyama, Y., et al.: "Therapeutic angiogenesis induced by human hepatocyte growth factor gene in rat diabetic hind limb ischemia model : molecular mechanisms of delayed angiogenesis in diabetes"Circulation. 104. 2344-2350 (2001)

[文献書誌] Hayashi, K., et al.: "Gene therapy for preventing neuronal death using hepatocyte growth factor : in vivo gene transfer of HGF to subarachnoid space prevents delayed neuronal death in gerbil hippocampal CA1 neurons"Gene Therpy. 8. 1167-1173 (2001)

[文献書誌] Shibuki, H., et al.: "Expression and neuroprotective effect of hepatocyte growth factor in retinal ischemia-reperfusion injury"Invest. Ophthalmol. Vis. Sci.. 43. 528-536 (2002)

[文献書誌] Maemondo, M, et al.: "Targeting angiogenesis and HGF function using an adenovirul vector expressing the HGF-antagonist NK4 for cancer therapy"Mol. Therapy. 5. 177-185 (2002)

[文献書誌] Sumi, T., et al.: "Mitosis-dependent phosphorylation and activation of LIM-kinase-1"Biochem. Biophys. Res. Commun.. (印刷中). (2002)

[文献書誌] Sakamaki, Y., et al.: "Hepatocyte growth factor stimulates proliferation of respiratory epithelial cells during postneumonectomy compensatory lung growth in mice"Am. J. Respir. Cell Mol. Biol.. (印刷中). (2002)

[文献書誌] Funakoshi, H., et al.: "Identification of Gas6,a putative ligand for Sky and Axl receptor tyrosine kinases, as a novel neurotrophic factor for hippocampal neurons"J. Neurosci. Res.. (印刷中). (2002)

[文献書誌] Kishi, A.Y., et al.: "Molecular cloning, expression and partial characterization of Kksy, a novel Xenopus member of Sky receptor tyrosine kinase"Gene. (印刷中). (2002)

[文献書誌] 松本邦夫, 中村敏一: "増殖因子による再生医学の展開-HGFを中心にして-"Biotherapy. 15. 95-104 (2001)

[文献書誌] 松本邦夫, 他: "HGFによる肺の再生と線維化の阻止"呼吸. 20. 935-939 (2001)

[文献書誌] 富岡大策, 他: "NK4/malignostatinによる腫瘍血管新生阻止作用とその制がん効果"最新医学. 56. 1799-1807 (2001)

[文献書誌] 松本邦夫: "肝臓の初期発生を支える血管内皮細胞"実験医学. 20. 468-471 (2002)

[文献書誌] Matsumoto, K, et al.: "分担:HGF-c-Met receptor pathway in tumor invasion-metastasis and potential cancer treatment with NK4書名:Growth Factors and Their Receptors in Cancer Metastasis"Kluwer Academic Publisher(分担部分). 35 (2001)

2001 年度実績報告書

松本邦夫大阪大学, 医学系研究科, 助教授 (90201780)