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

NOTCH1 缺陷 iPSC 来源的心肌细胞在左心发育不全综合征中的分子和功能评估

• 批准号: 10590735
• 负责人: Anita Saraf
• 金额: $ 16.11万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-10 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590735
• 关键词: Adult; Animal Model; Architecture; Arrhythmia; Award; Basic Science; Biological Markers; Biomedical Engineering; Calcium; Cardiac; Cardiac Myocytes; Cardiomyopathies; Cellular Stress; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Common Ventricle; Complex; Congenital Heart Defects; Data; Development; Development Plans; Doctor of Philosophy; Engineering; Etiology; Evaluation; Exposure to; Face; Fellowship; Future; Genes; Goals; Heart; Heart failure; Hypoplastic Left Heart Syndrome; Incidence; Inflammation; Inflammatory; Interleukin-6; Leadership; Left; Life; Medical; Medicine; Mentors; Mentorship; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Mutation; Myocardial; Myocardial dysfunction; NOTCH1 gene; Pathologic; Pathway interactions; Patients; Pattern; Physicians; Physiologic intraventricular pressure; Physiology; Population; Positioning Attribute; Principal Investigator; Proteins; Publications; Reactive Oxygen Species; Research; Research Training; Residencies; Rice; Role; Scientist; Signal Transduction; Stress; TNF gene; Technology; Therapeutic Intervention; Training; Training Programs; Translational Research; Universities; Ventricular; cardiogenesis; cardioprotection; career development; college; congenital heart disorder; cytokine; experience; in vivo; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; medical schools; meetings; mortality; new therapeutic target; preclinical study; pressure; professor; skills; systemic inflammatory response; tenure track; timeline; tool

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.

项目摘要/摘要： 这份提案概述了作为医学博士安妮塔·萨拉夫的一名内科科学家的5年指导职业发展计划。 博士生担任首席研究员。萨拉夫博士是匹兹堡大学终身教职助理教授 医学院。她通过医学科学家培训计划完成了研究生培训， 贝勒医学院，德克萨斯州休斯敦，通过威廉·马什·赖斯获得生物工程博士学位 大学。随后，她通过ABIM研究途径完成了她的住院医生和研究员资格 埃默里大学，亚特兰大，佐治亚州，并完成成人先天性心脏病专业培训。Dr。 萨拉夫的研究生研究侧重于涉及生物标记物的翻译和基础科学研究 单心室Fontan患者的特征及其对心肌细胞生物标志物的影响 来源于诱导多能干细胞(IPSCs)。在伯恩哈德·库恩博士的共同指导下 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和 心肌细胞生理学，长期目标是将自己确立为翻译和基础领域的领导者 先天性心脏病的科学研究。 在这项提案中，萨拉夫博士研究了NOTCH1亚型突变在诱发心律失常中的作用 患者来源的心肌细胞和来自ipscs的工程心肌细胞中的心力衰竭。使用NOTCH1中的IPSC 左心发育不全综合征(HLHS)的低形态患者和工程相似的低形态患者 在CRISPR基因编辑的对照IPSCs中的突变，Saraf博士提出了以下目标：(1) 确定炎性细胞因子(发现在单室患者中慢性升高)对 亚型NOTCH1心肌细胞和(2)决定全身压力对亚型的影响 NOTCH1心肌细胞，与异常的钙处理和收缩能力有关。 鉴于缺乏可行的先天性心脏病动物模型，IPSC来源的心肌细胞共同 基因编辑技术为了解心肌细胞的异常提供了一个强大的平台 生理学，发现新的治疗靶点，并创建具有类似亚形态的可行动物模型 未来的突变。除了研究培训，库恩、芬克尔和萨拉夫博士还制定了明确的 职业发展时间表，包括出版物、在会议上的演讲和领导力课程 这将使萨拉夫博士成为这一领域的领导者，走向独立。