GST-π/アンチセンスbcr/abl遺伝子導入によるCML急性転化の抑制

• Project/Area Number: 11877171
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Hematology
• Research Institution: Sapporo Medical University
• Principal Investigator: 新津 洋司郎 札幌医科大学, 医学部, 教授 (10045502)
• Co-Investigator(Kenkyū-buntansha): 高山 哲治 札幌医科大学, 医学部, 助手 (10284994)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥2,200,000 (Direct Cost: ¥2,200,000); Fiscal Year 2000: ¥500,000 (Direct Cost: ¥500,000); Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
• Keywords: GST-π / CML / bcr / Abl / in vivo selection

Research Abstract

[目的]本研究は白血病細胞にantisense bcr/abl遺伝子を導入しbcr/abl融合蛋白発現を抑制し急性転化を回避しようという試みである。これまでのantisense療法と異なり、抗癌剤耐性因子であるGST-π遺伝子を共導入しin vivoで抗癌剤投与し遺伝子導入細胞を増幅する試みである。本年度は抗癌剤投与によるin vivo selectionを確認すると共に遺伝子導入細胞の検討を2nd transplantation(2次骨髄移植)を行い検討した。
[方法]GST-π遺伝子導入マウス骨髄細胞をTBIを行った同系マウスに移植した。移植3週間後(骨髄再構築後)cyclophosphamide 100mg/kgを経静脈的に投与した。同療法を3クール施行後骨髄有核細胞を回収し、他の同系マウスに2次移植した。骨髄再構築後CFU-GM,CFU-Meg,BFU-EコロニーアッセイをおこないGST-π遺伝子の有無をPCR法で確認した。
[結果・考察]治療1コース目ではGST-π遺伝子導入マウス骨髄細胞移植マウス群と対照マウス群の末梢血白血球数の推移に差は認めなかった。投与3コース終了後の末梢血白血球数はGST-π遺伝子導入マウス群で対照群に比較し有意に高値であった(p<0.05)。2次骨髄移植施行マウス骨髄単核球より作成したCFU-GM,CFU-Meg,BFU-Eコロニー形成細胞中にGST-π遺伝子の発現を認めた。以上の結果から本法ではrepopulating stem cellに遺伝子導入可能でしかも抗癌剤投与により導入細胞のin vivo増幅が可能と考えられた。

Research Products

• [Publications] Sakamaki S,Niitsu Y, et al.: "Auto-antibodies against the specific epitope of human tropomyosin (s) detected by a peptide-based enzyme immunoassay in sera of patients with ulcerative colitis show antibody dependent cell-mediated cytotoxicity against HLA-DPw9 transfected L-cells."Gut. 47. 236-241 (2000)
• [Publications] Matsunaga T,Niitsu Y. et al.: "GST-p gene transduced hematopoietic progenitor cell transplantation overcomes the bone maroow toxicity of cyclophosphamide in mice"Hum Gene Ther. 11(12). 1679-1689 (2000)
• [Publications] Sate Y,Niitsu Y, et al.: "In vivo gene delivery to tumor cells by transferrin-streptavidin-DNA conjugate."FASEB J. 14. 2108-2118 (2000)
• [Publications] Mezawa S,Niitsu Y, et al.: "A study of carboplatin coated tube for the unresectable cholangiocarcinoma."Hepatology,. 32(5). 916-923 (2000)
• [Publications] Kohda K,Niitsu Y, et al.: "Long-term survival and late-onset complications of cancer patients treated with high-dose chemotherapy followed by autologous peripheral blood stem cell transplantation."Int J Hematol. 73(2). 251-257 (2001)
• [Publications] Lu Y,Niitsu Y, et al.: "Prevention of lethal acute-graft-versus-host disease in mice by oral administration of T helper 1 inhibitor-TAK-603."Blood. 97. 1123-1130 (2001)
• [Publications] Niitsu Y, Plate KH.: "Regulation of machinery for cancer cell growth, immortality, apoptosis and invation-The eighteenth international symposium of Sapporo Cancer Seminar"Jpn J Cancer Res.. 90. 1-2 (1999)
• [Publications] Matsunaga T, Niitsu Y, et al.: "Use of PCR serum in diagnosing and monitoring cytomegalovirus reactivation in bone marrow transplant recipients"Int J Hematol.. 69. 105-111 (1999)
• [Publications] Kogawa K, Niitsu Y, et al.: "Enhanced inhibition of experimental metastasis by the combination chemotherapy of Cu-Zn SOD and adriamycin"Clin Exp Metastasis.. 17. 239-244 (1999)
• [Publications] Sakamaki S, Niitsu Y, et al.: "Transforming growth factor-β1(TGF-β1) induces thrombopoietin from bone marrow stromal cells, which stimulates the expression of TGF-β1 receptor on megakaryocytes and, in tum, renders them susceptible to suppression by TGF-β1 itself with high specificity"Blood.. 94. 1961-1970 (1999)
• [Publications] Nobuoka A, Niitsu Y, et al.: "A case of malignant lymphome producing autoantibody against platelet glycoprotein Ib"Int J Hematology.. 200-206 (1999)
• [Publications] Homma H, Niitsu Y,: "Bilateral arterial infusion chemotherapy for advanced pancreatic cancer after superselective hemodynamic change"Cancer.. (in press).