Using Mechanical Loading to Develop Human iPSC-Derived Models of Cardiomyopathy

使用机械负载开发人类 iPSC 衍生的心肌病模型

• 批准号: 8909611
• 负责人: Venkatesh Hariharan
• 金额: $ 2.93万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-16 至 2015-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8909611
• 关键词: Accounting; Actins; Action Potentials; Acute; Adrenergic Agents; Affect; Anti-Arrhythmia Agents; Apoptosis; Arrhythmia; Arrhythmogenic Right Ventricular Dysplasia; Biochemical; Biological Assay; Biomechanics; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cardiovascular Diseases; Cells; Chronic; Complex; Connexin 43; Counseling; Cues; Custom; Defect; Deposition; Development; Devices; Disease; Disease Progression; Electrophysiology (science); Engineering; Environment; Exercise; Extracellular Matrix; Fatty acid glycerol esters; Frequencies; Gap Junctions; Generations; Genes; Heart; Heart Diseases; Histology; Human; Hybrids; Individual; Intercellular Junctions; Isoproterenol; Keratoderma; Link; Maps; Measurement; Measures; Mechanical Stimulation; Mechanical Stress; Mechanics; Modeling; Molecular; Muscle Cells; Mutation; Optics; Pathogenesis; Patients; Penetrance; Phenotype; Play; Process; Property; Proteins; Rattus; Regulation; Risk; Role; Slice; Staging; Strenuous Exercise; Structure; Syndrome; Techniques; Technology; Testing; Tissue Model; Tissue Viability; Tissues; Troponin T; Variant; Work; Zebrafish; adrenergic; cell growth; cohesion; design; disease phenotype; disease-causing mutation; experience; human tissue; in vivo; induced pluripotent stem cell; insight; lipid biosynthesis; mimetics; monolayer; mortality; mutation carrier; plakoglobin; prevent; protein expression; prototype; public health relevance; response; sudden cardiac death; trafficking

 DESCRIPTION (provided by applicant): Arrhythmogenic right ventricular cardiomyopathy (ARVC) is the most arrhythmogenic form of human heart disease, and one of the leading causes of sudden cardiac death in the young. Numerous studies have established the link between endurance exercise and disease penetrance in desmosomal mutation carriers, but the exact mechanism by which exercise alters disease pathogenesis and arrhythmic risk is unclear. There is increasing evidence that mechanical loading can play a defining role in cardiac tissue remodeling, including the regulation of key intercellular proteins such as plakoglobin and connexin-43. For cardiomyopathies affecting anchoring cell-cell junctions, such as ARVC, this lends to the hypothesis that mechanical stimulation may play a role in disease pathogenesis by augmenting electrophysiological or biomechanical defects. Thus far, much of the work characterizing the effects of mechanical loading on ARVC pathogenesis has not been verified in humans - a deficit that limits mechanistic insights, treatment options and patient counseling. In this study, I propose to investigate the influence of mechanical stimulation on cellular and tissue aspects of the disease (apoptosis, fatty deposits, gap junction remodeling, arrhythmia) in 3D tissue models of ARVC. In particular, we will apply strain to engineered heart slices seeded with induced pluripotent stem cells from clinically verified ARVC patients (a hybrid of acellular tissue and organotypic heart slice technology that recapitulates a 3D microenvironment for cell growth and enables mechanical interactions). Following mechanical stimulation, electrophysiological measurements will be used to measure changes in the cardiac action potential, and to determine if mechanical stimulation can cause ectopic rhythms in iPSC-derived model of ARVC. Next, we test the hypothesis that mechanical loading can recapitulate the overt stage phenotype of ARVC. Specifically, we determine if chronic mechanical loading can increase fat deposition, and alter the expression or localization of key proteins (i.e. connexin-43, plakoglobin, troponin-T a-sarcomeric actin, etc.). Finally, we utilize our platform for pharmacological interrogation by determining if treatment with anti-arrhythmic agents or SB216763, a compound shown to have restorative effects in a zebrafish model of ARVC, can be used to prevent the pro-arrhythmic effects of ß-adrenergic stimulation with isoproterenol, and the development of an overt-stage disease phenotype. Collectively, these studies will provide insight into the role of physio- mimetic mechanical stimulation in the progression of ARVC.

 描述（通过应用提供）：心律不齐的右心室心肌病（ARVC）是人类心脏病的最心律失常形式，也是年轻人心脏突然死亡的主要原因之一。大量研究已经建立了耐力运动与脱骨突变携带者中疾病的渗透之间的联系，但是锻炼改变疾病发病机理和心律失常风险的确切机制尚不清楚。越来越多的证据表明，机械负荷可以在心脏组织重塑中起定义作用，包括调节关键细胞间蛋白，例如斑块蛋白和连接蛋白43。对于影响锚定细胞 - 细胞连接（例如ARVC）的心肌病，这证明了机械刺激可能通过增加电生理或生物力学缺陷来在疾病发病机理中起作用。在人类中，尚未验证那些表征机械负荷对ARVC发病机理的影响的许多工作 - 这种防御限制了机械见解，治疗选择和患者咨询。在这项研究中，我建议研究机械刺激对细胞和组织的影响 ARVC 3D组织模型中疾病的各个方面（凋亡，脂肪沉积，间隙连接重塑，心律不齐）。特别是，我们将在临床验证的ARVC患者的诱导多能干细胞（细胞组织组织的杂种）中施加应变 和有机心脏切片技术，概括了3D微环境的细胞生长并实现机械相互作用）。机械刺激后，将使用电生理测量来测量心脏作用电位的变化，并确定机械刺激是否会导致IPSC衍生的ARVC模型中的生态节奏。接下来，我们测试了机械负荷可以概括ARVC的明显阶段表型的假设。具体而言，我们确定慢性机械负荷是否可以增加脂肪沉积，并改变关键蛋白的表达或定位（即连接蛋白43，plakoglobin，troponin-t a-sarmomeric肌动蛋白等）。 Finally, we utilize our platform for pharmaceutical interrogation by determining if treatment with anti-arrhythmic agents or SB216763, a compound shown to have restorative effects in a zebrafish model of ARVC, can be used to prevent the Pro-arrhythmic effects of ß-adrenegic simulation with isoproterenol, and the development of an overt-stage disease phenotype.总的来说，这些研究将洞悉生理机械模拟在ARVC进展中的作用。