Gene transfer into cardiovaseluar system using AAV vectors

使用 AAV 载体将基因转移至心血管系统

• 批准号: 10670675
• 负责人: IKEDA Uichi
• 金额: $ 1.92万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670675/
• 关键词: nitric oxide; adeno-associated virus; pulmonary hypertension; cardiac hypertrophy; gene therapy; アデノウイルス

We tried to use adeno-associated virus (AAV) vectors for gene therapy of various cardiovascular diseases such as pulmonary hypertension and cardiac hypertrophy. With a Grant-in-Aid for Scientific Research c, we revealed following new findings.(I) Transfer of AA V vectors into cardiovascular cells We first prepared AAV-LacZ . This AAV-LacZ could be transduced into rat vascular smooth muscles and cardiac myocytes with high efficiency. Furthermore, the expression of transduced LacZ gene persisted more than 2 weeks.(ii) ecNOS gene transfer into 293 cells We transduced endothelial nitric oxide synthase (ecNOS) gene into human embryonic kidney cell line 293 cells. The ecNOS protein expression was observed in the transfected cells, and ecNOS gene transfer markedly inhibited endothelin-1-induced proliferation of 293 cells.(iii) ecNOS gene transfer into cardiac myocytes We transduced AAV-ecNOS into rat cardiac myocytes. The ecNOS gene transfer significantly inhibited phenylephrine-induced protein synthesis of cardiac myocytes, which was recovered in the presence of the NOS inhibitor L-NMMA.Above findings suggest that ecNOS gene transfer using AAV vectors is useful for treatment of pulmonary hypertension and cardiac hypertrophy.

我们尝试将腺相关病毒（AAV）载体用于肺动脉高压、心肌肥厚等多种心血管疾病的基因治疗。通过科学研究补助金，我们发现了以下新发现。(I)将AAV载体转移到心血管细胞中我们首先制备AAV-LacZ。该AAV-LacZ可高效转导至大鼠血管平滑肌和心肌细胞。此外，转导的LacZ基因的表达持续超过2周。(ii)将内皮型一氧化氮合酶（endothelial nitric oxide synthase，ecNOS）基因导入人胚肾细胞系293细胞。转染细胞中可见ecNOS蛋白表达，ecNOS基因转染可明显抑制内皮素1诱导的293细胞增殖。(iii)ecNOS基因转染大鼠心肌细胞转染ecNOS基因可明显抑制苯肾上腺素诱导的心肌细胞蛋白质合成，并在NOS抑制剂L-NMMA作用下恢复，提示用腺相关病毒载体转染ecNOS基因可用于治疗肺动脉高压和心肌肥厚。

项目成果

Ikeda, U., Ikeda, M. et al: "Homocytstein increases nitric oxide synthesis in cytokine-stimulated vascular smooth muscle cells"Circulation. 99. 1230-1235 (1999)

Ikeda, U.、Ikeda, M. 等人：“同型半胱氨酸增加细胞因子刺激的血管平滑肌细胞中的一氧化氮合成”循环。

Nakagami,H, Ikeda,U., et al.: "Coronary artery disease and endothelial nitric oxide synthase an angiotensin-converting enzyme gene polymorphism."J. Thromb. Thrombolsis. 8. 191-195 (1999)

Nakagami,H, Ikeda,U., et al.：“冠状动脉疾病与内皮一氧化氮合酶和血管紧张素转换酶基因多态性。”J.

Maeda,Y.,Ikeda,U., Shimada, K.: "Endogenously generated nitric oxide by nitric oxide synthase gene transfer inhibits ceillular prolifertion."J. Pharmacol. Exp. Ther. 292. 387-393 (2000)

Maeda,Y.、Ikeda,U.、Shimada, K.：“通过一氧化氮合酶基因转移产生的内源性一氧化氮抑制细胞增殖。”J.

前田喜一、池田宇一、他: "心筋へのvascular endothelial growth factor 遺伝子導入による血管新生療法の試み"心筋の構造と代謝. 21. 13-18 (1999)

Kiichi Maeda、Uichi Ikeda 等人：“通过将血管内皮生长因子基因引入心肌来进行血管生成治疗的尝试”心肌结构和代谢。 21. 13-18 (1999)

Ikeda,U., Ikeda,M., et al.: "Homocystein increases nitric oxide synthesis in cytokine-stimulated vascular smooth muscle cells."Circulation. 99. 1230-1235 (1999)

Ikeda,U.、Ikeda,M. 等人：“同型半胱氨酸可增加细胞因子刺激的血管平滑肌细胞中的一氧化氮合成。”循环。