Basic Research for Gene Therapy of Heart Failure by Transcoronay Gene Transfer

跨冠状基因转移心力衰竭基因治疗的基础研究

• 批准号: 12670677
• 负责人: SATOH Shinji
• 金额: $ 2.05万
• 依托单位: KYUSHU 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-12670677/
• 关键词: Heart Failure; Contractile Proteins; Myofibrillar Calcium Sensitivity; Adenovirus Vector; GTP-binding Protein; 組み替えアデノウイルス; 組み換えアデノウィルス

We performed basic experiments to investigate changes in the regulatory mechanisms of cardiac contractility through the GTP binding protein-related pathways in heart failure. The main purpose of this project was to determine possible target genes to prevent the process by which normal hearts develop heart failure. We obtained the following findings.(A) In canine failing hearts, 1) GTP-binding protein Gq-RhoA-Rho kinase signaling was upregulated in association with an increase in the phosphorylation level of myosin light chain, which caused an increase in myofibrillar Ca^<2+> sensitivity, 2) β-adrenergicstimulation-induced decreasein the myofibrillar Ca^<2+> sensitivity was attenuated. These abnormal regulation of the myofibrillar Ca^<2+> sensitivity may be related to decreased contractile function in the failing hearts.(B) In cardiacmuscle, as in smooth muscle, the phosphorylation of myosin light chain was modulatedby calmodulin and cyclic GMP, both of which were able to regulate the cardiacmyofibrillar Ca^<2+> sensitivity.(C) Using a selective Rho kinase inhibitor Y-27632, we tested the hypothesis that chronic inhibition of Rho kinase might be able to prevent the process of myocardial hypertrophy resulting in heart failure. We found that chronic inhibition of Rho kinase prevented myocardial hypertrophy induced by hypertension and thereby preserved myocardial contractile function. Rho kinase may be an important target in treatment of heart failure.(D) We are performing another experiment to investigate the function of the transient receptor potential (TRP) proteins in cardiac cells. The TRP protein superfamily consists of a diverse group of receptor-operated Ca^<2+> channels activated by Gq-dependent mechanisms. The hypothesis is that cultured cardiomyocytes transfected with the TRP gene, incorporated into adenovirus vector, develop hypertrophy when stimulated by angiotensin II.

我们进行了基础实验，以研究心力衰竭中通过GTP结合蛋白相关通路调节心肌收缩能力的机制的变化。该项目的主要目的是确定可能的靶基因，以防止正常心脏发生心力衰竭的过程。结果表明：(1)在心力衰竭犬，1)GTP结合蛋白Gq-RhoA-Rho激酶信号转导上调与肌球蛋白轻链磷酸化水平升高有关，从而导致肌原纤维钙敏感性增加；2)β肾上腺素能刺激引起肌原纤维钙敏感性降低。这些对肌原纤维钙敏感性的异常调节可能与衰竭心脏的收缩功能降低有关。(B)在心肌中，肌球蛋白轻链的磷酸化受到钙调蛋白和环鸟苷酸的调节，两者都能够调节心肌原纤维钙敏感性。(C)使用选择性Rho激酶抑制剂Y-27632，我们验证了长期抑制Rho激酶可能能够阻止心肌肥大导致心力衰竭的假说。我们发现，长期抑制Rho激酶可防止高血压引起的心肌肥厚，从而保护心肌收缩功能。Rho激酶可能是治疗心力衰竭的一个重要靶点。(D)我们正在进行另一项实验，以研究心肌细胞中瞬时受体电位(Trp)蛋白的功能。Trp蛋白超家族由一组不同的受体操纵的钙通道组成，这些通道由GQ依赖的机制激活。假说是，将Trp基因导入腺病毒载体的培养心肌细胞在血管紧张素II的刺激下会发生肥大。

项目成果

Satoh S., Makino N.: "Intracellular mechanisims of cGMP-mediated regulation of myocaridial contraction"Basic Res Cardiol. 96. 652-658 (2001)

Satoh S.，Makino N.：“cGMP 介导的心肌收缩调节的细胞内机制”Basic Res Cardiol。

Kinugawa et al.: "Treatment with dimethylthiourea prevents left ventricular remodeling and failure after experimental myocardial infarction in mice."Circ Res. 87. 392-398 (2001)

Kinukawa 等人：“用二甲基硫脲治疗可预防小鼠实验性心肌梗死后的左心室重塑和衰竭。”Circ Res。

Suematsu N, Satoh S, Kinugawa S, Tsutsui H, Hayashidani S, Nakamura R, Egashira K, Makino N, Takeshita A: "α_1-Adrenoceptor-Gq-RhoA signaling is upregulated to increase myofibrillar Ca^<2+> sensiivity in failing heart"Am J Physiol. 281. H637-H646 (2001)

Suematsu N、Satoh S、Kinukawa S、Ttsutsui H、Hayashidani S、Nakamura R、Egashira K、Makino N、Takeshita A：“α_1-肾上腺素受体-Gq-RhoA 信号传导上调，增加失败时肌原纤维 Ca^<2+> 敏感性心”Am J Physiol.281.H637-H646 (2001)

Mukai et al.(5番目): "Involvement of Rho-kinase in hypertensive vascular disease. -A novel therapeutic target in hypertension"FASEB J. 15. 1062-1064 (2001)

Mukai 等人 (5th)：“Rho 激酶参与高血压血管疾病。-高血压的新治疗靶点”FASEB J. 15. 1062-1064 (2001)

Suematsu et al.(2番目): "Effects of calmodulin and okadaic acid on myofibrillar Ca^<2+> sensitivity in cardiac myocytes"Basic Research in Cardiology. 97. 137-144 (2002)

Suematsu等人(2nd)：“钙调蛋白和冈田酸对心肌细胞中肌原纤维Ca 2+ 敏感性的影响”心脏病学基础研究97。137-144(2002)。