肺がんの薬剤耐性と転移因子を分子標的とした免疫遺伝子治療

• 研究課題/領域番号: 08266242
• 研究種目: 特定領域研究(A)
• 配分区分: 補助金
• 研究機関: 徳島大学
• 研究代表者: 曽根 三郎 徳島大学, 医学部, 教授 (40145024)
• 研究分担者: 篠原 勉 徳島大学, 医学部・附属病院, 助手 (70284289) ; 西岡 安彦 徳島大学, 医学部, 講師 (70274199) ; 清水 英治 徳島大学, 医学部, 講師 (50187449) ; 楊河 宏章 徳島大学, 医学部附属病院, 助手 (50263827)
• 研究期間 (年度): 1996 – 1999
• 研究課題ステータス: 完了 (1999年度)
• 配分額: 39,000千円 (直接経費: 39,000千円); 1999年度: 10,000千円 (直接経費: 10,000千円); 1998年度: 10,000千円 (直接経費: 10,000千円); 1997年度: 10,000千円 (直接経費: 10,000千円); 1996年度: 9,000千円 (直接経費: 9,000千円)
• キーワード: 肺がん / 転移 / 多剤耐性 / P糖蛋白質 / 遺伝子治療 / 免疫療法 / SCIDマウス / 肺癌 / 転移モデル / ガンクリオシドGM2 / モノクローナル抗体 / ADCC / NK細胞 / サイトカイン / 薬剤耐性 / 免疫遺伝子治療 / 抗体治療

研究概要

多剤耐性(MDR)癌細胞が発現するP糖蛋白質(P-gp)に対する抗体MRK16はマクロファージ(MΦ)の抗体依存性細胞障害反応(ADCC)を誘導し、MDR癌細胞を障害する。本年度は、肺癌の遠隔転移とMDRの克服を目的に、MDR肺癌細胞の多臓器転移に対する抗P-gp抗体の治療効果を検討し、更にMΦのADCCを強化すべくMDR肺癌細胞へMΦ活性化因子M-CSFまたはMΦ遊走因子MCP-1の遺伝子導入を行い、抗P-gp抗体との併用効果を検討し、以下の成果を得た。1、ヒト小細胞肺癌株H69及びP-gp陽性MDR株H69/VPはNK細胞除去SCIDマウスに静注した場合、多臓器(肝、腎、リンパ節)転移を形成した。ビンデシン治療はH69の転移形成を抑制したがH69/VPのそれを全く抑制せず、H69/VPの転移巣での抗癌剤耐性を確認した。2、抗P-gp抗体(マウス抗体MRK16とキメラ抗体MH162)は用量依存性にH69/VPの多臓器転移形成を抑制した。3、H69/VPにM-CSFまたはMCP-1遺伝子を導入し生物活性を有するM-CSFまたはMCP-1産生株を得た。これらの株を用い上記転移モデルにてMRK16治療の転移抑制効果を検討した。(1)H69/VPに対する遺伝子導入はin vitroでの増殖能及びP-gp発現に影響しなかった。(2)M-CSF遺伝子導入はMΦ活性化により肝及びリンパ節転移を抑制したが腎転移には影響しなかった。しかし、MRK16併用にて各臓器の転移形成が完全に抑制された。(3)MCP-1遺伝子導入は転移形成に影響しなかったが、MRK16併用において転移抑制効果を有意に増強した。その機序として、転移巣で産生されたMCP-1がMΦ集積を促進しADCCを増強させたと考えられた。以上より、MDR肺癌に対する抗転移療法を開発する上で、免疫エフェクター細胞の転移局所への動員と活性化を試みるアプローチの重要性が示唆された。

研究成果

• [文献書誌] Sone.S.,: "Heterogeneity of multiorgan metastases of human lung cancer cells genetically engineered to produce cytokines and reversal using chimeric monoclonal antibodies in natural killer cell-depleted severe combined immunodeficient mice"Cancer Chemother.Pharmacol.,. 43. S26-31 (1999)
• [文献書誌] Nokihara,H.,: "Monocyte chemoattractant protein-1 gene modification of multidrug-resistant human lung cancer enhances antimetastatic effect of therapy with anti-P-glycoprotein antibody in SCID mice"Int.J.Cancer,. 80. 773-780 (1999)
• [文献書誌] Yano,S.,: "Combined therapy with anti-P-glycoprotein antibody and macrophage colony-stimulating factor gene transduction for multiorgan metastases of multidrug-resistant human small cell lung cancer in NK cell-depleted SCID mice"Int.J.Cancer,. 82. 105-111 (1999)
• [文献書誌] Nakamura,K.,: "Apoptosis induction of human lung cancer cell line in multicellular heterospheroids with humanized antiganglioside GM2 monoclonal antibody"Cancer Res.,. 59. 5323-2330 (1999)
• [文献書誌] Torisu,H.,: "Macrophage infiltration correlates with tumjor stage and angiogenesis in human malignant melanoma : possible involvement of TNFalpha and IL-1alpha"Int.J.Cancer,. 85. 182-188 (2000)
• [文献書誌] Yanagawa,H.,: "Vascular endothelial growth factor in malignant pleural effusion associated with lung cancer"Cancer Immunol.Immunother.,. 48. 396-400 (1999)
• [文献書誌] Parajuli,P.,: "Cytolysis of human dendritic cells by autologous lymphokine-activated killer cells : participation of both T cells and NK cells in the killing"J.Leukoc.Biol.,. 65. 764-770 (1999)
• [文献書誌] Kawano,T.: "Comparison of suppressive effects of a new anti-inflammatory compound, FR167653, on production of PGE2 and inflammatory cytokines, human monocytes, and alveolar macrophages in response to endotoxin"J.Leukoc.Biol.,. 65. 80-86 (1999)
• [文献書誌] Yamamoto,A.,: "Infrequent presence of anti-c-Myc antibodies and absence of c-Myc oncoprotein in sera from lung cancer patients"Oncology,. 56. 129-133 (1999)
• [文献書誌] Sumitomo,K.,: "Activation of RB tumor suppressor protein and growth suppression of small cell lung carcinoma cells by reintroduction of p16INK4A gene"Int.J.Oncol.,. 14. 1075-1080 (1999)
• [文献書誌] Yanagawa,H.,: "Presence and potent immunosuppressive role of interleukin-10 in malignant pleural effusion due to lung cancer"Cancer Lett.,. 136. 27-32 (1999)
• [文献書誌] Parajuli,P.,: "Therapeutic efficacy of a new topoisomerase I and II inhibitor TAS-103, against both P-glycoprotein-expressing and -nonexpressing drug-resistant human small-cell lung cancer"Oncol.Res.,. 11. 219-224 (1999)
• [文献書誌] Namikawa,O.,: "Analysis of antibodies to p16INK4A tumor suppressor gene products in lung cancer patients"Int.J.Oncol.,. 14. 681-685 (1999)
• [文献書誌] Ichikawa,W.,: "Characterization of immunoglobulin binding factor in sputum from patients with chronic airway diseases"Respirology,. 4. 375-381 (1999)
• [文献書誌] Yanagawa,H.,: "Production of interleukin-10 by alveolar macrophages from lung cancer patients"Respir.Med.,. 93. 666-671 (1999)
• [文献書誌] Yanagawa,H.,: "A case of resectable lung adenocarcinoma associated with sarcoidosis"Med.Oncol.,. 16. 216-220 (1999)
• [文献書誌] Ogushi,F.,: "Decreased prostaglandin E2 synthesis by lung fibroblasts isolated from rats with bleomycin-induced lung fibrosis"Int.J.Exp.Pathol.,. 80. 41-49 (1999)
• [文献書誌] Nishimura,N.,: "Down-regulation by a new anti-inflammatory compound, FR167653, of differentiation and maturation of human monocytes and bone marrow CD34+ cells to dentritic cells"Int.J.Immunopharmacol.. (in press).
• [文献書誌] Sumitomo,K.,: "Expression of a TGF-b1 inducible gene, TSC-36, causes growth inhibition in human lung cancer cell lines"Cancer Letters. (in press).
• [文献書誌] Hiramatsu,K. et al.: "Generation of killer activity by interleukin-12 of mononuclear cells in malignant pleural effusions due to lung cancer" Cancer Immunol Immunother. 46. 1-6 (1998)
• [文献書誌] Takeuchi,E. et al.: "Comparative analysis of interleukin 15 and interleukin 2 for induction of killer activity and of type 2 cytokine production by mononuclear cells from lung cancer patients" Br.J.Cancer. 78. 616-620 (1998)
• [文献書誌] Hanibuchi,M. et al.: "Therapeutic efficacy of mouse-human chimeric anti-ganglioside GM2 monoclonal antibody against multiple organ micrometastases of human lung cancer in NK cell-depleted SCID mice" Int.J.Cancer. 78. 480-485 (1998)
• [文献書誌] Shrivastava,P.et al.: "Circumvention of multidrug resistance by a quinoline derivative,MS-209,in multidrug-resistant human small-cell lung cancer cells and its synergistic interaction with cyclospirin A or verapamil" Cancer Chemother Pharmacol. 42. 483-490 (1998)
• [文献書誌] Pai,K.et al.: "Different effect of Interleukin 10 on Interleukin 12-and Interleukin 2-Mcdiated Killer Induction From blood Mononuclear Cells" Oncol.Res.10. 295-300 (1998)
• [文献書誌] Nokihara,H.et al.: "Monocyte chemoattractant protein-1 gene modification of multidrug-resistant human lung cancer enhances antimetastatic effect of therapy with anti-P-glycoprotein antibody in SCID mice" Int.J.Cancer. 80. 773-780 (1999)
• [文献書誌] Yano,S.et al.: "Model of malignant pleural effusion of human lung adenocarcinoma in SCID mice" Oncol.Res.(in press). (1997)
• [文献書誌] Nishioka,Y.et al.: "Combined therapy of multidrug-resistnt human lung cancer with anti-P-glycoprotein antibody and monocyte chemoattractant protein-1 gene transduction : The possibility of immunological overcoming of multidrug resistance." Int.J.Cancer. 71. 170-177 (1997)
• [文献書誌] Yanagawa,H.et al.: "Intrapleural instillation of IFN-gamma in patients with malignant pleurisy due to lung cancer" Cancer Immunol.Immunother.45. 93-99 (1997)
• [文献書誌] Shinohara,T.et al.: "Combination therapy with antibody and interleukin-2 gene transfer against multidrug-resistant cancer cells" Jpn.J.Cancer Res.88. 1100-1107 (1997)
• [文献書誌] Yano,S.et al.: "Macrophage colony-stimulating factor-gene transduction into human lung cancer cells differentially regulates metastasis formations in various organ micro-environments of NK-cell depleted SCID mice." Cancer Res.57. 784-790 (1997)
• [文献書誌] Fujiki,F.et al.: "Prevention of adenocarcinoma colon 26-induced cachexia by interleukin-10 gene transfer" Cancer Res.57. 94-99 (1997)
• [文献書誌] Kawanishi,M.et al.: "Allelotype and replication error phenotype of small cell lung carcinoma" Carcinogenesis. 18. 2057-2062 (1997)
• [文献書誌] Yano,S.: "Macrophage colony-stimulating factor-gene transduction into human lung cancer cells differentially regulates metastasis formations in various organ micro-environments of NK-cell depleted SCID mice." Cancer Res.57. 784-790 (1997)
• [文献書誌] Fujiki,F.: "Prevention of adenocarcinoma colon 26-induced cachexia by interleukin-10 gene transfer." Cancer Res.57. 94-99 (1997)
• [文献書誌] Nishioka,Y.: "Monocyte chemoattractant protein-1 gene-transduction into multidrug-resistant human lung cacer cells enhances the therapeutic efficacy of anti-P-glycoprotein antibody." Int.J.Cancer.
• [文献書誌] Takeuchi,E.: "Induction by interleukin-15 of human killer cell activity against lung cancer cell lines and its regulatory mechanism." Jpn.J.Cancer Res.87. 1251-1258 (1996)
• [文献書誌] Yano,S.: "T helper 2 cytokines differently regulate monocyte chemoattractant protein-1(MCP-1)production by human peripheral blood monocytes and alveolalr macrophages." J.Immunol.157. 2660-2665 (1996)
• [文献書誌] Yano.S.: "Novel metastasis model of human lung cancer in SCID mice depleted of NK cells." Int.J.Cancer. 67. 211-217 (1996)