Role of ROCK isoform-mediated actin cytoskeleton modification in the pathogenesis of heart disease

ROCK亚型介导的肌动蛋白细胞骨架修饰在心脏病发病机制中的作用

• 批准号: 10542786
• 负责人: Jianjian Shi
• 金额: $ 44.5万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-15 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10542786
• 关键词: Actins; Adult; Aging; Animal Model; Autophagocytosis; Cardiac; Cardiac Myocytes; Cardiac health; Cardiovascular Diseases; Cause of Death; Cell physiology; Cerebrovascular Spasm; Cessation of life; Clinical Research; Clinical Trials; Compensation; Complex; Cytoskeletal Modeling; Cytoskeleton; Disease; Etiology; Extracellular Matrix Proteins; F-Actin; FRAP1 gene; Family; Fibroblasts; Fibrosis; Functional disorder; Genes; Genetic Processes; Goals; Grant; Guanosine Triphosphate Phosphohydrolases; Heart; Heart Diseases; Heart Hypertrophy; Heart Injuries; Heart failure; Human; Hypertrophy; Japan; Knock-out; Knockout Mice; Knowledge; LIM Domain Kinase 1; Mediating; Modification; Molecular; Mus; Muscle Contraction; Myocardial Ischemia; Myocardium; Myofibroblast; Myosin Light Chains; Names; Nature; Nuclear Translocation; Organelles; Outcome Study; Oxidative Stress; Pathogenesis; Pathologic; Pathway interactions; Phosphotransferases; Prevention; Process; Production; Protein Isoforms; ROCK1 gene; Regulation; Research; Rho-associated kinase; Role; Serum Response Factor; Signal Pathway; Signal Transduction; Smooth Muscle Myocytes; Stress; Stress Fibers; Subarachnoid Hemorrhage; Testing; Therapeutic; Therapeutic Effect; Time; Ventricular; Work; age related; biological adaptation to stress; cell type; coronary fibrosis; experimental study; fasudil; heart function; human disease; human model; in vivo; inducible Cre; inhibition of autophagy; inhibitor; interstitial; ischemic injury; member; misfolded protein; myocardin; myosin phosphatase; new therapeutic target; novel therapeutic intervention; pressure; response; senescence; therapeutic target; transcription factor; treatment trial

PROJECT ABSTRACT The prevention and treatment of heart disease remain challenging. Rho kinase (also named ROCK) has recently emerged as a potential therapeutic target for various cardiovascular diseases. A long-term goal of our past twenty years of research on ROCK pathophysiology is to define the roles and underlying mechanisms of ROCK-mediated signal pathway in regulating cardiac remodeling. The two members of the ROCK family, ROCK1 and ROCK2, have both shared and distinct cellular functions and can compensate each other in numerous single isoform knockout conditions. The majority of our knowledge on the cellular and molecular function of ROCKs comes from research on proliferative cell types in which ROCKs modulate actin cytoskeleton organization through promoting actomyocin contraction and F-actin stabilization. Cardiomyocytes stand apart from other cell types because they contain both sarcomeric and non-sarcomeric cytoskeleton. There is a gap in our knowledge on how ROCKs regulate sarcomeric and non-sarcomeric F-actin in cardiomyocytes and how these processes contribute to overall heart function. Recently, for the first time to use inducible approach to delete both ROCK isoforms in cardiomyocytes, we have discovered that although ROCKs are not required for maintaining sarcomeric cytoskeleton in adult hearts, they do participate in the regulation of non-sarcomeric F-actin organization, inhibit autophagy by promoting mammalian target of rapamycin (mTOR) activity and contribute to age-related cardiac fibrosis. In contrast, the non-sarcomeric F- actin dynamics are able to be maintained with the presence of either isoform in the cardiomyocytes where the other isoform has been deleted; this might be attributed to compensatory over-activation of the remaining isoform in cardiomyocytes having single ROCK isoform deletion. The proposed research aims to further elucidate the pathophysiological roles and downstream pathways of ROCK-mediated actin cytoskeleton changes in cardiomyocytes and fibroblasts under pathological stress sceneries. Aim 1 will determine if deletion of both ROCK1 and ROCK2 from adult cardiomyocytes limits the progression of heart failure in pathological hypertrophy and myocardial ischemic injury through activating autophagy and facilitating autophagic flux by inhibiting mTOR signaling. Aim 2 will determine if deletion of both ROCK1 and ROCK2 from adult fibroblasts limits the activation of myofibroblasts and fibrotic response through inhibition of F-actin regulated transcription factor activation including the serum response factor (SRF) and myocardin-related transcription factors (MRTFs); the direct contribution of ROCKs/F-actin/MRTFs/SRF axis in fibroblasts to cardiac fibrosis has never been demonstrated in vivo, and our preliminary results indicate that the inducible approach is required for double ROCK knockout in fibroblasts. The biomedical significance of this work is to provide the cutting-edge concepts for understanding pathophysiological roles of ROCKs in heart failure. The ultimate goal is to develop new therapeutic intervention to ameliorate compromised cardiac function.

项目摘要 心脏病的预防和治疗仍然充满挑战。 Rho 激酶（也称为 ROCK） 最近已成为多种心血管疾病的潜在治疗靶点。长期目标是 我们过去二十年对 ROCK 病理生理学的研究是为了定义其作用和潜在机制 ROCK介导的信号通路在调节心脏重构中的作用。 ROCK家族的两位成员， ROCK1 和 ROCK2 具有共同和不同的细胞功能，并且可以在 许多单一亚型敲除条件。我们关于细胞和分子的大部分知识 ROCK 的功能来自对增殖细胞类型的研究，其中 ROCK 调节肌动蛋白 通过促进肌动蛋白收缩和 F-肌动蛋白稳定来形成细胞骨架。心肌细胞 与其他细胞类型不同，因为它们同时含有肌节和非肌节细胞骨架。 我们对 ROCK 如何调节肌节和非肌节 F-肌动蛋白的知识存在差距。 心肌细胞以及这些过程如何影响整体心脏功能。最近第一次使用 通过诱导方法删除心肌细胞中的两种 ROCK 亚型，我们发现虽然 ROCK 不是维持成人心脏肌节细胞骨架所必需的，它们确实参与 调节非肌节 F-肌动蛋白组织，通过促进哺乳动物靶标抑制自噬 雷帕霉素 (mTOR) 活性并导致与年龄相关的心脏纤维化。相反，非肌节 F- 肌动蛋白动力学能够在心肌细胞中存在任一亚型的情况下维持，其中 其他亚型已被删除；这可能归因于剩余部分的代偿性过度激活 具有单个 ROCK 同工型缺失的心肌细胞中的同工型。拟议的研究旨在进一步 阐明 ROCK 介导的肌动蛋白细胞骨架的病理生理学作用和下游途径 病理应激环境下心肌细胞和成纤维细胞的变化。目标1将确定是否删除 来自成人心肌细胞的 ROCK1 和 ROCK2 限制了病理性心力衰竭的进展 通过激活自噬和促进自噬通量来抑制肥厚和心肌缺血损伤 抑制 mTOR 信号传导。目标 2 将确定是否从成体成纤维细胞中同时删除 ROCK1 和 ROCK2 通过抑制 F-肌动蛋白调节的转录来限制肌成纤维细胞的激活和纤维化反应 因子激活，包括血清反应因子 (SRF) 和心肌素相关转录因子 （MRTF）；成纤维细胞中的 ROCKs/F-actin/MRTFs/SRF 轴对心脏纤维化的直接贡献从未 已在体内得到证实，我们的初步结果表明，诱导方法是 成纤维细胞中的双重 ROCK 敲除。这项工作的生物医学意义在于提供了前沿的 用于理解 ROCK 在心力衰竭中的病理生理学作用的概念。最终目标是发展 新的治疗干预措施可改善受损的心脏功能。