Human Induced Pluripotent Cell Models of Pediatric Cardiac Disorders

人类诱导的小儿心脏病多能细胞模型

基本信息

批准号：
8583749
负责人：
BRUCE D GELB
金额：
$ 40.34万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8583749
关键词：
Address Affect Animal Model Anti-Arrhythmia Agents Arrhythmia Atrial Tachycardia Autophagocytosis Biology Calcium Cardiac Cardiac Myocytes Cardiomyopathies Cardiovascular Abnormalities Cardiovascular system Cell Culture Techniques Cell Line Cell model Cells Child Childhood Combined Modality Therapy Complex Congenital Heart Defects Costello syndrome Defect Development Disease Dose Drug usage Electrophysiology (science)Exhibits Family Fibroblasts Genes HRAS gene Heart Atrium Heart Diseases Human Hypertrophic Cardiomyopathy Hypertrophy In Vitro Inborn Genetic Diseases Infant Inherited LEOPARD Syndrome Live Birth Lung diseases MAP Kinase Gene MAPK Signaling Pathway Pathway Medical Missense Mutation Modeling Morbidity - disease rate Mus Muscle Mutation Myocardium Myopathy Pathogenesis Pathway interactions Patients Pharmaceutical Preparations Phenotype Pluripotent Stem Cells Primary Myocardial Diseases Proteins Research Role Ryanodine Receptor Calcium Release Channel Signal Pathway Signal Transduction Skin Tachycardia Testing Ventricular cardiofaciocutaneous syndrome clinical care efficacy testing gain of function gain of function mutation in vivo Model induced pluripotent stem cell inhibitor/antagonist insight mortality mouse model novel therapeutic intervention novel therapeutics older patient public health relevance response small molecule stem cell technology

项目摘要

DESCRIPTION (provided by applicant): A long-standing barrier in cardiovascular biology research has been the inability to maintain human cardiomyocytes in long-term cell culture. Since 2007 when the possibility of reprogramming terminally differentiated human cells like skin fibroblasts into pluripotent stem cells was demonstrated, it became possible to generate human cardiomyocytes in vitro, enabling the study of human primary myocardial diseases. For this project, we intend to study inherited forms of childhood myocardial disease: hypertrophic cardiomyopathy (HCM) associated with RAS signaling abnormalities and atrial muscle-related tachycardia associated with increased HRAS signaling. The "RASopathies" are a family of autosomal dominant disorders caused by missense mutations in genes encoding RAS/MAPK pathway proteins. HCM is common in the RASopathies and multifocal atrial tachycardia (MAT) is specifically observed in one disorder, Costello syndrome, which is caused by gain-of-function HRAS mutations. For SPECIFIC AIM 1, the PIs hypothesize that RASopathy- associated HCM arises through signaling pathway activation that differs among the specific disorders. To test this, they will use existing human iPSC lines for two RASopathies that exhibit cardiomyocyte hypertrophy. Isolated ventricular cardiomyocytes harboring LEOPARD and cardiofaciocutaneous syndrome-causing mutations will be characterized with respect to signal transduction, intracellular calcium handling and contractility among cells. For SPECIFIC AIM 2, the PIs hypothesize that MAT in Costello syndrome is caused by perturbations in intracellular calcium handling induced by altered signaling from HRAS. To test this, atrial cardiomyocytes, differentiated from Costello syndrome iPSC lines, will be assessed for Ca2+ transients, electrophysiology and response to relevant anti-arrhythmic drugs. For SPECIFIC AIM 3, the PIs hypothesize that RASopathy-associated HCM will be reversed by inhibitors that address mutation-specific signaling perturbations. To study this, iPSC-derived LEOPARD and cardiofaciocutaneous syndrome cardiomyocytes will be treated with signaling pathway inhibitors to reverse hypertrophy. Dose response curves will be established. Combination therapies will be tested for efficacy at lower doses. Effects of therapies on the autophagy defect in LEOPARD syndrome cardiomyocytes will be determined. Broadly, these studies will harness the power of the new iPSC technology to elucidate the pathogenesis of myocardial disease associated with RASopathies: HCM and atrial arrhythmias. These studies could have important impact on the development of novel therapeutic strategies for these myocardial diseases, for which our current approaches are not curative.

描述（由申请人提供）：心血管生物学研究中长期存在的障碍是无法在长期细胞培养中维持人类心肌细胞。自2007年证明将皮肤成纤维细胞等终末分化的人类细胞重编程为多能干细胞的可能性以来，体外产生人类心肌细胞成为可能，从而使人类原发性心肌疾病的研究成为可能。在这个项目中，我们打算研究儿童心肌疾病的遗传形式：与 RAS 信号异常相关的肥厚性心肌病 (HCM) 和与 HRAS 信号增强相关的心房肌相关心动过速。 “RASopathies”是由编码 RAS/MAPK 途径蛋白的基因错义突变引起的常染色体显性遗传疾病家族。 HCM 在 RAS 病中很常见，而多灶性房性心动过速 (MAT) 在一种疾病 Costello 综合征中特别观察到，该疾病是由功能获得性 HRAS 突变引起的。对于 SPECIFIC AIM 1，PI 假设 RAS 病相关的 HCM 是通过特定疾病之间不同的信号通路激活而产生的。为了测试这一点，他们将使用现有的人类 iPSC 系来治疗两种表现出心肌细胞肥大的 RASopathies。含有 LEOPARD 和心面皮肤综合征引起突变的离体心室心肌细胞将在信号转导、细胞内钙处理和细胞间收缩性方面进行表征。对于 SPECIFIC AIM 2，PI 假设 Costello 综合征中的 MAT 是由 HRAS 信号改变引起的细胞内钙处理扰动引起的。为了测试这一点，将评估从 Costello 综合征 iPSC 系分化而来的心房心肌细胞的 Ca2+ 瞬变、电生理学和对相关抗心律失常药物的反应。对于 SPECIFIC AIM 3，PI 假设 RAS 病相关的 HCM 将通过解决突变特异性信号传导扰动的抑制剂来逆转。为了研究这一点，将用信号通路抑制剂治疗 iPSC 衍生的 LEOPARD 和心面皮肤综合征心肌细胞以逆转肥大。将建立剂量响应曲线。将测试较低剂量联合疗法的疗效。将确定治疗对 LEOPARD 综合征心肌细胞自噬缺陷的影响。总的来说，这些研究将利用新 iPSC 技术的力量来阐明与 RASopathies 相关的心肌疾病的发病机制：HCM 和房性心律失常。这些研究可能会对这些心肌疾病的新治疗策略的开发产生重要影响，而我们目前的方法无法治愈这些疾病。