Development of gene therapy for the vascular proliferative diseases by ribozyme targetting TGF-β mRNA

核酶靶向TGF-β mRNA开发血管增殖性疾病基因治疗

• 批准号: 12670699
• 负责人: FUKUDA Noboru
• 金额: $ 2.11万
• 依托单位: Nihon University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670699/
• 关键词: ribozyme; transforming growth factor-β; gene therapy; vascular smooth muscle cell; proliferation; gromerulosclerosis; RNA; キメラ; 増殖; RT-PCR; ウエスタンブロット

We designed hammerhead ribozyme targetting TGF-β_1 mRNA which cleaved the synthetic substrate RNA. We developed DNA-RNA chimeric ribozyme, which significantly inhibited growth of vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR). DNA-RNA chimeric ribozyme inhibited TGF-β_1 mRNA and protein in VSMC from SHR. DNA-RNA chimeric ribozyme significantly suppressed expression of TGF-β_1, fibronectin, collagen type I mRNAs in mesangial cells from SHR-SP. In vivo experiments, FITC-labeled ribozyme was efficiently distributed in aorta and kidney. DNA-RNA chimeric ribozyme considerably inhibited hypertrophy of renal capirally artery and gromerulosclerosis in SHR-SP in vivo. We are appling DNA-RNA chimeric ribozyme on coronary stenosis after angioplasty in pig, by which efficiency, dosage, and period of treatment will be determined to develop the DNA-RNA chimeric ribozyme to TGF-β_1 as a gene therapy for arterial proliferative diseases and gromerulosclerosis.

我们设计了针对TGF-β_1 mRNA的锤头核酶，用于切割合成的底物RNA。我们研制的DNA-RNA嵌合核酶能显著抑制自发性高血压大鼠血管平滑肌细胞（VSMC）的生长。DNA-RNA嵌合核酶抑制SHR VSMC TGF-β_1 mRNA和蛋白表达。DNA-RNA嵌合核酶显著抑制shrr - sp系膜细胞中TGF-β_1、纤维连接蛋白、胶原I型mrna的表达。在体内实验中，fitc标记的核酶在主动脉和肾脏中有效分布。DNA-RNA嵌合核酶在体内显著抑制SHR-SP患者肾动脉肥大和肾小球硬化。我们将DNA-RNA嵌合核酶应用于猪血管成形术后冠状动脉狭窄，通过确定疗效、剂量和治疗周期，开发DNA-RNA嵌合核酶TGF-β_1作为动脉增殖性疾病和肾血管性硬化的基因治疗方法。

项目成果

Fukuda N, Hu W-Y et al.: "Angiotensin II upregulates transforming growth factor-β type I receptors on rat vascular smooth muscle cells"American Journal of Hypertension. 13. 191-198 (2000)

Fukuda N、Hu W-Y 等：“血管紧张素 II 上调大鼠血管平滑肌细胞上的转化生长因子-β I 型受体”美国高血压杂志 13. 191-198 (2000)。

Hu W-Y, Fukuda N, Kanmatsuse K: "Phenotypic modulation by fibronectin enhances the angiotensin II-generating system in cultured vascular smooth muscle cells"Arteriosclerosis Thrombosis and Vascular Biology. 20. 1500-1505 (2000)

Hu W-Y、Fukuda N、Kanmatsuse K：“纤连蛋白的表型调节增强了培养的血管平滑肌细胞中的血管紧张素 II 生成系统”动脉硬化血栓形成和血管生物学。

Hu W-Y, Fukuda N et al.: "Hammerhead ribozyme targeting human PDGF A-chain mRMA inhibited the proliferation of human vascular smooth muscle cells"Atherosclerosis. 158. 321-329 (2001)

Hu W-Y、Fukuda N等人：“靶向人PDGF A链mRMA的锤头核酶抑制人血管平滑肌细胞增殖”动脉粥样硬化。

Satoh C, Fukuda N et al.: "Role of endogenous angiotensin II in the increased expression of growth factors in vascular smooth muscle cells from spontaneously hypertensive rats"Journal of Cardiovascular Pharmacology. 37. 108-118 (2001)

Satoh C、Fukuda N 等人：“内源性血管紧张素 II 在自发性高血压大鼠血管平滑肌细胞中生长因子表达增加中的作用”心血管药理学杂志。

Kishioka H, Fukuda N et al.: "Effect of methylene methylimino-linkaged antisense oliigonucleotide to PDGF A-chain on growth of vascular smooth muscle cells from spontaneously hypertensive rats"European Journal of Pharmacology. 392. 129-132 (2000)

Kishioka H、Fukuda N 等人：“亚甲基甲基亚氨基连接的反义寡核苷酸对 PDGF A 链对自发性高血压大鼠血管平滑肌细胞生长的影响”欧洲药理学杂志。