Molecular and Functional Evaluation of NOTCH1 Deficient iPSC Derived Cardiomyocytes in Hypoplastic Left Heart Syndrome

项目概要/摘要： 该提案概述了作为医学博士Anita Saraf的物理学家科学家的5年职业发展计划， 博士作为主要研究者。萨拉夫博士是匹兹堡大学的终身助理教授 医学院。她通过医学科学家培训计划完成了她的研究生培训， Baylor College of Medicine，Houston，TX，博士在生物工程通过威廉马什赖斯 大学随后，她通过ABIM研究途径完成了她的居住和奖学金， 埃默里大学，亚特兰大，佐治亚州，并完成了成人先天性心脏病的专科培训。博士 Saraf的研究生研究主要集中在涉及生物标志物的转化和基础科学研究 单心室Fontan患者的特征，并了解这些生物标志物在心肌细胞中的作用 来源于诱导多能干细胞（iPSC）。在Bernhard Kühn博士的共同指导下， 和托伦·芬克尔（共同导师），萨拉夫博士建议进一步提高她在基因编辑方面的研究技能， 心肌细胞生理学，长期目标是建立自己作为一个领导者的翻译和基础 先天性心脏病的科学研究 在这项提案中，Saraf博士研究了NOTCH 1亚型突变在诱导心律失常中的作用。 和心力衰竭。使用来自NOTCH 1的iPSC 左心室发育不全综合征（HLHS）和工程类似的左心室发育不全患者 通过CRISPR基因编辑控制iPSC中的突变，Saraf博士提出了以下目标：（1） 确定炎性细胞因子（发现在单心室患者中慢性升高）对 （2）确定系统压力对亚型NOTCH 1心肌细胞的影响。 NOTCH 1心肌细胞，相对于异常的钙处理和收缩性。 由于缺乏可行的先天性心脏病动物模型，iPSC衍生的心肌细胞一起 基因编辑技术为了解心肌细胞异常提供了一个强大的平台 生理学，发现新的治疗靶点，并创造可行的动物模型，具有类似的亚形态 未来的突变。除了研究培训外，Kühn、Finkel和Saraf博士还制定了明确的 职业发展的时间轴，包括出版物、在会议上发言和领导能力课程 这将使萨拉夫博士成为这一领域的领导者。

Project Summary/Abstract: This proposal outlines a 5-year mentored career development plan as a physician-scientist for Anita Saraf, MD, PhD as the principal investigator. Dr. Saraf is a tenure-track Assistant Professor at University of Pittsburgh School of Medicine. She completed her graduate training through the Medical Scientist Training Program at Baylor College of Medicine, Houston, TX with a Ph.D. in Bioengineering through the William Marsh Rice University. Subsequently, she completed her residency and fellowship through the ABIM research pathway at Emory University, Atlanta, GA and completed subspeciality training in Adult Congenital Heart Disease. Dr. Saraf’s post-graduate research has focused on translational and basic science studies involving biomarker profiles in single ventricle Fontan patients and understanding the effect of these biomarkers in cardiomyocytes derived from induced pluripotent stem cells (iPSCs). Under the combined mentorship of Drs. Bernhard Kühn and Toren Finkel (co-mentor), Dr. Saraf proposes to further her research skill-set in gene-editing and cardiomyocyte physiology, with a long-term goal of establishing herself as a leader in translational and basic science research in congenital heart disease. In this proposal, Dr. Saraf investigates the role of NOTCH1 hypomorphic mutations in inducing arrhythmias and heart failure in patient-derived and engineered cardiomyocytes from iPSCs. Using iPSCs from NOTCH1 hypomorphic patients with hypoplastic left heart syndrome (HLHS) and engineering similar hypomorphic mutations in control iPSCs with CRISPR gene-editing, Dr. Saraf proposes the following objectives: (1) Determine the influence of inflammatory cytokines (found to chronically elevated in univentricular patients) on hypomorphic NOTCH1 cardiomyocytes and (2) Determine the influence of systemic pressures on hypomorphic NOTCH1 cardiomyocytes, with respect to abnormal calcium-handling and contractility. Given the lack of viable animal models for congenital heart disease, iPSC-derived cardiomyocytes together with gene-editing technology provides a powerful platform to understand abnormalities in cardiomyocyte physiology, discover novel targets for therapy, and create viable animal models with similar hypomorphic mutations in the future. In addition to research training, Drs. Kühn, Finkel and Saraf have formulated a clear timeline for career development involving publications, presentations at meetings and courses in leadership that will transition Dr. Saraf to independence as a leader in this field.