TRPC6 inhibition therapy to rescue cardiac muscle dysfunction in muscular dystrophy

TRPC6 抑制疗法可挽救肌营养不良症患者的心肌功能障碍

• 批准号: 10541224
• 负责人: Brian Leei Lin
• 金额: $ 13.03万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10541224
• 关键词: Acute; Address; Adult; Arrhythmia; Attenuated; Biological Assay; Biological Availability; Biomedical Engineering; Blinded; Blood Vessels; Calcium; Cardiac; Cardiac Myocytes; Cations; Cell Death; Cell Line; Cell Separation; Cells; Chronic; Clinical; Clinical Trials; Complementary therapies; Custom; Data; Defect; Deformity; Disease; Duchenne muscular dystrophy; Dystrophin; Environment; Fibroblasts; Fibrosis; Functional disorder; Future; Gene Deletion; Genetic; Goals; Heart Diseases; Heart failure; Histologic; Histopathology; Human; Human Engineering; Hyperactivity; Immobilization; Inflammatory; Ion Channel; Kidney Diseases; Length; Longevity; Lung diseases; Measures; Mechanical Stress; Mechanics; Mediating; Membrane; Mentors; Methods; Modeling; Molecular; Mus; Muscle; Muscle Cells; Muscle Weakness; Muscle function; Muscular Dystrophies; Myocardial dysfunction; Myocardium; Myopathy; Pathologic; Pathology; Pathway interactions; Patients; Pharmaceutical Preparations; Profibrotic signal; Receptor, Angiotensin, Type 1; Reporting; Research; Residual state; Role; Sampling; Sarcolemma; Signal Pathway; Signal Transduction; Skeletal Muscle; Spinal; Stress; Stress Tests; Stretching; Striated Muscles; System; Testing; Time; Tissue Model; Training; Treatment Efficacy; Utrophin; Vertebral column; biological adaptation to stress; cardiac tissue engineering; career; cell injury; cell type; clinically relevant; coronary fibrosis; dystrophic cardiomyopathy; efficacy testing; experience; gene therapy; heart function; improved; in vivo; induced pluripotent stem cell; inhibitor; mdx mouse; mechanical load; mechanical stimulus; micro-dystrophin; mortality; mouse model; muscle physiology; muscular dystrophy mouse model; novel; novel therapeutics; nuclear factors of activated T-cells; pharmacologic; post-doctoral training; prevent; randomized placebo controlled trial; receptor; response; sealant; single-cell RNA sequencing; skeletal; skills; small molecule inhibitor; stem cells; success; transcriptomics

Project Summary This proposal tests the efficacy and mechanisms by which a recently developed, bioavailable, selective small- molecule inhibitor of Transient Receptor Potential Canonical 6 (TRPC6) ameliorates cardiac and skeletal myopathy in Duchenne muscular dystrophy (DMD). This research fulfills the candidate’s long-term goals of advancing novel therapies for dystrophic cardiomyopathy and applying mechanosensitive signaling assays in mice and engineered heart tissues (EHT) to study disease. DMD results from a loss of dystrophin, inducing profound progressive muscle weakness, spinal deformities, fibrosis, heart failure, and early mortality. TRPC6 is a mechanosensitive, non-voltage gated cation channel expressed in muscle cells that is hyper-activated in DMD, mediating excessive mechanical stress-induced force/Ca2+ responses, arrhythmias, cardiac dysfunction, and muscle fibrosis. During the candidate’s postdoctoral training, he led projects assessing the impact of blocking TRPC6 genetically and pharmacologically in models of cardiac fibrosis. Preliminary data in DMD show genetic or pharmacological TRPC6 inhibition prolongs lifespan in severe DMD models by 2-3 fold, ameliorating fibrosis and associated pathology, and improving heart and skeletal muscle function. The candidate first reported on the TRPC6 drug inhibitor (BI 749327) in 2019, and its clinical derivatives are now in human trials for lung and renal disease. In this proposal, the candidate addresses key questions whose answers will importantly inform future DMD translational efforts, and is organized into three aims. Aim 1 tests the efficacy of chronic TRPC6 inhibition by BI 749327 to prevent and reverse DMD skeletal and cardiac muscle dysfunction, histopathology, and TRPC6- NFAT, pro-fibrotic, and inflammatory signaling. Cell-type expression is analyzed by single-cell RNAseq to identify how subpopulations of cardiac cells that express TRPC6 (fibroblasts, vascular, and myocytes) are impacted by the treatment. Aim 2 tests the capacity of chronic TRPC6 suppression to restore mechanical activation-induced defects in force and calcium in isolated DMD mouse cardiomyocytes, and to obviate effects of membrane sealants and other mechanosensitive-activated pathways. I will further test the role of TRPC6 pathobiology in a novel human DMD EHT model of mechanosensitive activation using the same mechanical stimuli as in mouse cardiomyocytes. Aim 3 tests the efficacy of micro-dystrophin (μDys) gene therapy to treat TRPC6 pathobiology in the recently-developed D2.mdx DMD mouse model or the combination of μDys with BI 749327 provides additive benefits. The proposal uniquely combines the candidate’s prior training with expertise from his mentors. The candidate has the unique skills needed to conduct these studies, combining biomedical engineering, muscle physiology, and molecular signaling experience. He will expand into single-cell transcriptomics, stem cell derived EHT and their mechanical analysis, and DMD gene therapy. The environment at Johns Hopkins and expert mentoring team provides every opportunity for success for the candidate and project.

项目概要 该提案测试了最近开发的、生物可利用的、选择性的小分子药物的功效和机制。 瞬时受体电位 Canonical 6 (TRPC6) 的分子抑制剂可改善心脏和骨骼 杜氏肌营养不良症 (DMD) 中的肌病。这项研究实现了候选人的长期目标 推进营养不良性心肌病的新疗法并应用机械敏感信号分析 小鼠和工程心脏组织（EHT）来研究疾病。 DMD 是由肌营养不良蛋白缺失引起的， 严重的进行性肌肉无力、脊柱畸形、纤维化、心力衰竭和早期死亡。 TRPC6 是 在 DMD 中过度激活的肌肉细胞中表达的机械敏感、非电压门控阳离子通道， 介导过度机械应力引起的力/Ca2+反应、心律失常、心脏功能障碍和 肌肉纤维化。在该候选人的博士后培训期间，他领导了评估阻塞影响的项目 TRPC6 在心脏纤维化模型中的遗传和药理学作用。 DMD 的初步数据显示遗传因素 或药理 TRPC6 抑制可将严重 DMD 模型的寿命延长 2-3 倍，改善纤维化 和相关的病理学，并改善心脏和骨骼肌功能。候选人首先报告了 2019年推出TRPC6药物抑制剂（BI 749327），其临床衍生物目前正在进行肺和肾的人体试验 疾病。在这份提案中，候选人提出了一些关键问题，这些问题的答案将对未来产生重要影响。 DMD 转化工作分为三个目标。目标 1 测试慢性 TRPC6 抑制的功效 通过 BI 749327 预防和逆转 DMD 骨骼和心肌功能障碍、组织病理学和 TRPC6- NFAT、促纤维化和炎症信号传导。通过单细胞 RNAseq 分析细胞类型表达来识别 表达 TRPC6 的心肌细胞亚群（成纤维细胞、血管细胞和心肌细胞）如何受到影响 治疗。目标 2 测试慢性 TRPC6 抑制恢复机械激活诱导的能力 分离的 DMD 小鼠心肌细胞中的力和钙缺陷，并消除膜的影响 密封剂和其他机械敏感激活途径。我将进一步测试 TRPC6 病理学在 使用与小鼠相同的机械刺激的新型人类 DMD EHT 机械敏感激活模型 心肌细胞。目标 3 测试微肌营养不良蛋白 (μDys) 基因疗法治疗 TRPC6 病理学的功效 在最近开发的 D2.mdx DMD 小鼠模型中或 μDys 与 BI 749327 的组合提供了 附加效益。该提案独特地将候选人之前的培训与其导师的专业知识结合起来。 候选人拥有进行这些研究所需的独特技能，结合生物医学工程、肌肉 生理学和分子信号传导经验。他将扩展到单细胞转录组学、干细胞衍生 EHT 及其机械分析，以及 DMD 基因治疗。约翰霍普金斯大学的环境和专家 指导团队为候选人和项目提供一切成功的机会。