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：
8774293
负责人：
BRUCE D GELB
金额：
$ 4.35万
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/8774293
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 Health 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 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）：心血管生物学研究的长期障碍是无法在长期细胞培养中维持人类心肌细胞。自2007年以来，当证明了将终末分化的人类细胞（如皮肤成纤维细胞）重编程为多能干细胞的可能性，就可以在体外产生人类心肌细胞，从而可以研究人类原发性心肌疾病。对于这个项目，我们打算研究遗传的儿童心肌疾病：与RAS信号传导异常和与HRAS信号增加有关的RAS信号传导异常和与心房相关的心动过速相关的肥厚心肌病（HCM）。 “ rasopathies”是由编码RAS/MAPK途径蛋白的基因中的错义突变引起的常染色体显性疾病家族。 HCM在RASOPARIS中很常见，多灶性心脏心动过速（MAT）是在一种疾病Costello综合征中特别观察到的，这是由功能获得的HRAS突变引起的。对于特定的目标1，PIS假设相关的HCM通过信号传导途径激活而产生，这在特定疾病之间有所不同。为了测试这一点，他们将使用现有的人类IPSC线进行两种表现出心肌细胞肥大的rasopathies。携带豹子的分离性室心肌细胞和心脏反性综合征引起的突变将在信号转导，细胞内钙处理和细胞之间的收缩力方面进行表征。对于特定的目标2，PIS假设Costello综合征中的MAT是由HRAS的信号改变引起的细胞内钙处理引起的。为此，将评估与Costello综合征IPSC系不同的心房心肌细胞，以CA2+瞬态，电生理学和对相关抗心律失常药物的反应进行评估。对于特定的目标3，PIS假设与rasopathy相关的HCM将被解决突变特异性信号扰动的抑制剂逆转。为了研究这一点，IPSC衍生的豹子和心faciociociathorme综合征心肌细胞将用信号通路抑制剂治疗以逆转肥大。将建立剂量反应曲线。组合疗法将在较低剂量下测试功效。将确定疗法对豹综合征心肌细胞自噬缺陷的影响。从广义上讲，这些研究将利用新的IPSC技术的力量来阐明与rasopathies相关的心肌疾病的发病机理：HCM和心房心律不齐。这些研究可能会对这些心肌疾病的新型治疗策略的发展产生重要影响，为此，我们目前的方法无法治愈。