2002 Fiscal Year Annual Research Report

NK4(HGFアンタゴニスト/血管新生抑制因子)による悪性腫瘍の治療研究

Research Project

Project/Area Number	14030049
Research Institution	Osaka University
Principal Investigator	松本邦夫大阪大学, 医学系研究科, 助教授 (90201780)
Keywords	NK4 / HGF / 腫瘍血管新生 / 転移 / 浸潤 / 遺伝子治療 / がん / 血管内皮細胞
Research Abstract	悪性癌の本態というべき浸潤・転移を高効率に阻止することが癌の治癒率向上をもたらすと考えられる。私達はHGFが癌細胞の浸潤・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を調製した。その後、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するとともに、NK4が実験動物に移植した癌に対して、浸潤・転移を抑制するとともに、腫瘍血管新生阻害を介して癌の成長を抑制す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環として、1)NK4遺伝子治療の制癌作用の解析、ならびに2)NK4による血管内皮細胞の増殖抑制に関与する細胞内情報伝達の作用点を明らかにする目的で研究を行った。 NK4の生体内発現にはCMVプロモーターの下流にヒトNK4 cDNAを連結した発現カセットを有する組換えアデノウイルス(AdCMV-NK4)を使用した。Lewis肺癌細胞ならびにB16F10メラノーマ細胞を尾静脈から移植する肺転移モデルを用いてAdCMV-NK4投与の効果を調べたところ、AdCMV-NK4投与により肺転移の個数はコントロールに比べ1/10以下に強く抑制された。さらに、マウス皮下に移植されたLewis肺癌ならびにB16F10メラノーマに対しても、AdCMV-NK4を尾静脈から1回投与することにより強い成長阻害作用が認められた。一方、NK4による血管内皮細胞の増殖抑制機構を調べるため、ヒト血管内皮細胞の培養系にNK4を添加したところ、NK4はbFGFによって誘導されるERK1/2の活性化には無効であった。一方、bFGFはヒト血管内皮細胞においてGSK3-βならびにAktの活性化(リン酸化)を引き起こしたが、NK4はbFGFによって誘導されるGSK3-βならびにAktの活性化を容量依存的に抑制した。また、NK4はbFGFによって誘導されるcyclin D1の発現上昇を阻害した。以上の結果から、1)NK4遺伝子治療は転移阻害、血管新生阻害を介した癌の成長阻害をもたらす制癌法になる可能性、2)NK4の血管内皮細胞の増殖抑制に関与するシグナル伝達系はAktならびにGSK3-βを介してもたらされるcyclin D1ならびにRBリン酸化経路にあることが明らかになった。

Research Products

(26 results)

All Other

All Publications (26 results)

[Publications] Morishita R et al.: "Impairment of collateral formation in lipoprotein(a) transgenic mice : therapeutic angiogenesis induced by human hepatocyte growth factor gene"Circulation. 105. 1491-1496 (2002)
[Publications] Taniyama Y et al.: "Development of safe and efficient novel nonviral gene transfer using ultrasound : enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle"Gene Ther. 9. 372-380 (2002)
[Publications] Sakamaki Y et al.: "Hepatocyte growth factor stimulates proliferation of respiratory epithelial cells during postpneumonectomy compensatory lung growth in mice"Am J Respir Cell Mol Biol. 26. 525-533 (2002)
[Publications] Yoshimura S et al.: "Gene transfer of hepatocyte growth factor to subarachnoid space in cerebral hypoperfusion model"Hypertension. 39. 1028-1034 (2002)
[Publications] Kishi Y.A.et al.: "Molecular cloning, expression and partial characterization of Xksy, Xenopus member of the Sky family of receptor tyrosine kinases"Gene. 288. 29-40 (2002)
[Publications] Sumi T et al.: "LIM-kinases in actin cytoskeletal regulation and their physiological role"Seikagaku. 74. 402-407 (2002)
[Publications] Taniyama Y et al.: "Angiogenesis and antifibrotic action by hepatocyte growth factor in cardiomyopathy. Hypertension"Hypertension. 40. 47-53 (2002)
[Publications] Hirao S et al.: "Tumor suppression effect using NK4, a molecule acting as an antagonist of HGF, on human gastric carcinomas"Cancer Gene Ther.. 9. 700-707 (2002)
[Publications] Nakagami H et al.: "Hepatocyte growth factor prevents endothelial cell death through inhibition of bax translocation from cytosol to mitochondrial membrane"Diabetes. 51. 2604-2011 (2002)
[Publications] To Y et al.: "A two-way interaction between hepatocyte growth factor and interleukin-6 in tissue invasion of lung cancer cell line"Am J Respir Cell Mol Biol. 27. 220-226 (2002)
[Publications] Maehara N et al.: "Gene transduction of NK4, HGF antagonist, inhibits in vitro invasion and in vivo growth of human pancreatic cancer"Clin Exp Metastasis. 19. 417-426 (2002)
[Publications] Saimura M et al.: "Intraperitoneal injection of adenovirus-mediated NK4 gene suppresses peritoneal dissemination of pancreatic cancer cell line AsPC-1 in nude mice"Cancer Gene Ther.. 9. 799-806 (2002)
[Publications] Ono M et al.: "In vivo gene transfection with hepatocyte growth factor via the pulmonary artery induces angiogenesis in the rat lung"Circulation. 106. 1264-1269 (2002)
[Publications] Saimura M et al.: "Tumor suppression through angiogenesis inhibition by SUIT-2 pancreatic cancer cells genetically engineered to secrete NK4"Clin Cancer Res.. 8. 3243-3249 (2002)
[Publications] Matsumoto K et al.: "Renotropic role and therapeutic potential of HGF in the kidney"Nephrol Dial Transplant. 17. 59-61 (2002)
[Publications] Kikuchi T et al.: "Tumor suppression induced by intratumor administration of adenovirus vector expressing NK4, a 4-kringle antagonist of hepatocyte growth factor, and naive dendritic cells"Blood. 100. 3950-3959 (2002)
[Publications] Nakanishi K et al.: "Gene transfer of human hepatocyte growth factor into rat skin wounds mediated by liposomes coated with the sendai virus(hemagglutinating virus of Japan)"Am J Pathol.. 161. 1761-1772 (2002)
[Publications] Qian LW et al.: "Co-cultivation of panceatic cancer cells with orthotopic tumor-derived fibroblasts : fibroblasts stimulate tumor cell invasion via HGF secretion whereas cancer cells exert a minor regulative effect on fibroblasts HGF production"Cancer Lett. 190. 105-112 (2003)
[Publications] Yoshida S et al.: "Neutralization of hepatocyte growth factor leads to retarded cutaneous wound healing associated with decreased neovascularization and granulation tissue formation"J Invest Dermatol. 120. 335-343 (2003)
[Publications] Koike H et al.: "Enhanced angiogenesis and improvement of neuropathy by cotransfection of human hepatocyte growth factor and prostacyclin synthase gene"FASEB J. (印刷中). (2003)
[Publications] Qian L.W.et al.: "Radiation stimulates HGF receptor/c-Met expression that leads to amplifying cellular response to HGF stimulation via upregulated receptor tyrosine phosphorylation and MAP kinase activity in pancreatic cancer cells"Int J Cancer. 104. 542-549 (2003)
[Publications] Bessho K et al.: "Counteractive Effects of HGF on PDGF-induced Mesangial Cell Proliferation in a Rat Model of Glomerulonephritis"Am J Physiol Renal Physiol. (印刷中). (2003)
[Publications] 町出充, 他: "HGFによる肝再生医療の展開"再生医療. 1-2. 47-55 (2002)
[Publications] 松本邦夫: "肝臓の初期発生と血管内皮細胞"医学のあゆみ. 200. 647-649 (2002)
[Publications] 田村隆彦, 他: "肝癌、膵癌の発生ならびに悪性化におけるオートクリン・パラクリン増殖因子の意義"Biotherapy. (印刷中). (2003)
[Publications] 松本邦夫, 中村敏一(分担執筆): ""肝再生と肝臓病の克服-21世紀の肝臓病治療"バイオサイエンスの新世紀第14刊、生命工学:新しい生命へのアプローチ"共立出版. 80-94 (2002)

2002 Fiscal Year Annual Research Report

NK4(HGFアンタゴニスト/血管新生抑制因子)による悪性腫瘍の治療研究

Principal Investigator

松本 邦夫 大阪大学, 医学系研究科, 助教授 (90201780)

Research Products

[Publications] Morishita R et al.: "Impairment of collateral formation in lipoprotein(a) transgenic mice : therapeutic angiogenesis induced by human hepatocyte growth factor gene"Circulation. 105. 1491-1496 (2002)

[Publications] Taniyama Y et al.: "Development of safe and efficient novel nonviral gene transfer using ultrasound : enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle"Gene Ther. 9. 372-380 (2002)

[Publications] Sakamaki Y et al.: "Hepatocyte growth factor stimulates proliferation of respiratory epithelial cells during postpneumonectomy compensatory lung growth in mice"Am J Respir Cell Mol Biol. 26. 525-533 (2002)

[Publications] Yoshimura S et al.: "Gene transfer of hepatocyte growth factor to subarachnoid space in cerebral hypoperfusion model"Hypertension. 39. 1028-1034 (2002)

[Publications] Kishi Y.A.et al.: "Molecular cloning, expression and partial characterization of Xksy, Xenopus member of the Sky family of receptor tyrosine kinases"Gene. 288. 29-40 (2002)

[Publications] Sumi T et al.: "LIM-kinases in actin cytoskeletal regulation and their physiological role"Seikagaku. 74. 402-407 (2002)

[Publications] Taniyama Y et al.: "Angiogenesis and antifibrotic action by hepatocyte growth factor in cardiomyopathy. Hypertension"Hypertension. 40. 47-53 (2002)

[Publications] Hirao S et al.: "Tumor suppression effect using NK4, a molecule acting as an antagonist of HGF, on human gastric carcinomas"Cancer Gene Ther.. 9. 700-707 (2002)

[Publications] Nakagami H et al.: "Hepatocyte growth factor prevents endothelial cell death through inhibition of bax translocation from cytosol to mitochondrial membrane"Diabetes. 51. 2604-2011 (2002)

[Publications] To Y et al.: "A two-way interaction between hepatocyte growth factor and interleukin-6 in tissue invasion of lung cancer cell line"Am J Respir Cell Mol Biol. 27. 220-226 (2002)

[Publications] Maehara N et al.: "Gene transduction of NK4, HGF antagonist, inhibits in vitro invasion and in vivo growth of human pancreatic cancer"Clin Exp Metastasis. 19. 417-426 (2002)

[Publications] Saimura M et al.: "Intraperitoneal injection of adenovirus-mediated NK4 gene suppresses peritoneal dissemination of pancreatic cancer cell line AsPC-1 in nude mice"Cancer Gene Ther.. 9. 799-806 (2002)

[Publications] Ono M et al.: "In vivo gene transfection with hepatocyte growth factor via the pulmonary artery induces angiogenesis in the rat lung"Circulation. 106. 1264-1269 (2002)

[Publications] Saimura M et al.: "Tumor suppression through angiogenesis inhibition by SUIT-2 pancreatic cancer cells genetically engineered to secrete NK4"Clin Cancer Res.. 8. 3243-3249 (2002)

[Publications] Matsumoto K et al.: "Renotropic role and therapeutic potential of HGF in the kidney"Nephrol Dial Transplant. 17. 59-61 (2002)

[Publications] Kikuchi T et al.: "Tumor suppression induced by intratumor administration of adenovirus vector expressing NK4, a 4-kringle antagonist of hepatocyte growth factor, and naive dendritic cells"Blood. 100. 3950-3959 (2002)

[Publications] Nakanishi K et al.: "Gene transfer of human hepatocyte growth factor into rat skin wounds mediated by liposomes coated with the sendai virus(hemagglutinating virus of Japan)"Am J Pathol.. 161. 1761-1772 (2002)

[Publications] Qian LW et al.: "Co-cultivation of panceatic cancer cells with orthotopic tumor-derived fibroblasts : fibroblasts stimulate tumor cell invasion via HGF secretion whereas cancer cells exert a minor regulative effect on fibroblasts HGF production"Cancer Lett. 190. 105-112 (2003)

[Publications] Yoshida S et al.: "Neutralization of hepatocyte growth factor leads to retarded cutaneous wound healing associated with decreased neovascularization and granulation tissue formation"J Invest Dermatol. 120. 335-343 (2003)

[Publications] Koike H et al.: "Enhanced angiogenesis and improvement of neuropathy by cotransfection of human hepatocyte growth factor and prostacyclin synthase gene"FASEB J. (印刷中). (2003)

[Publications] Qian L.W.et al.: "Radiation stimulates HGF receptor/c-Met expression that leads to amplifying cellular response to HGF stimulation via upregulated receptor tyrosine phosphorylation and MAP kinase activity in pancreatic cancer cells"Int J Cancer. 104. 542-549 (2003)

[Publications] Bessho K et al.: "Counteractive Effects of HGF on PDGF-induced Mesangial Cell Proliferation in a Rat Model of Glomerulonephritis"Am J Physiol Renal Physiol. (印刷中). (2003)

[Publications] 町出充, 他: "HGFによる肝再生医療の展開"再生医療. 1-2. 47-55 (2002)

[Publications] 松本邦夫: "肝臓の初期発生と血管内皮細胞"医学のあゆみ. 200. 647-649 (2002)

[Publications] 田村隆彦, 他: "肝癌、膵癌の発生ならびに悪性化におけるオートクリン・パラクリン増殖因子の意義"Biotherapy. (印刷中). (2003)

[Publications] 松本邦夫, 中村敏一(分担執筆): ""肝再生と肝臓病の克服-21世紀の肝臓病治療"バイオサイエンスの新世紀第14刊、生命工学:新しい生命へのアプローチ"共立出版. 80-94 (2002)

松本邦夫大阪大学, 医学系研究科, 助教授 (90201780)