Discovery of Novel Genetic Variants Underlying Unexplained Hypertrophic Cardiomyopathy by Whole Genome Sequencing, RNA-seq and iPSC modeling

通过全基因组测序、RNA-seq 和 iPSC 建模发现不明原因肥厚性心肌病的新遗传变异

• 批准号: 10335270
• 负责人: Alireza Haghighi
• 金额: $ 16.96万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10335270
• 关键词: Advisory Committees; Age; Alleles; Animal Model; Award; Bioinformatics; Blood; CRISPR/Cas technology; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cardiology; Cardiomyopathies; Cardiovascular Diseases; Cardiovascular system; Caring; Childhood; Clinical; Code; Collaborations; Computational Biology; Consumption; DNA; Data; Disease; Disease model; Early Diagnosis; Environment; Etiology; Exhibits; Extramural Activities; Five-Year Plans; Foundations; Funding; Future; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Genomics; Goals; Grant; Health; Heart; Heart Transplantation; Hospitals; Human; Human Genetics; Hypertrophic Cardiomyopathy; Inherited; Institutes; Investigation; Laboratory Organism; Lead; Left ventricular structure; Medical Care Costs; Medicine; Mentors; Mentorship; Methods; Modeling; Molecular; Morbidity - disease rate; Mutation; National Heart, Lung, and Blood Institute; Null Lymphocytes; Online Mendelian Inheritance In Man; Onset of illness; Pathogenesis; Pathogenicity; Pathway interactions; Patient Care; Patients; Persons; Phenotype; Physiology; Play; Prevalence; Proteins; RNA Splicing; Relaxation; Research; Research Personnel; Role; Sarcomeres; Scientist; Solid; Spliced Genes; Sudden Death; Tissue Sample; Tissues; Training; Trans-Omics for Precision Medicine; Translating; Untranslated RNA; Variant; Woman; base; career; career development; differential expression; effective therapy; epigenomics; exome sequencing; experimental study; gene function; genetic variant; genome sequencing; improved; in silico; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; insight; instructor; loss of function; medical schools; mortality; mutant; mutation carrier; novel; novel diagnostics; novel therapeutic intervention; proband; protein function; rare variant; research and development; skills; statistics; stem cell model; sudden cardiac death; symposium; transcriptome sequencing; variant of unknown significance; whole genome

PROJECT SUMMARY This proposal describes a five-year plan for Dr. Alireza Haghighi's career development towards becoming an independent cardiovascular investigator. Currently an Instructor in Medicine and Clinical Molecular Geneticist in Brigham and Women's Hospital (BWH), and Harvard Medical School (HMS), Dr. Haghighi's long-term career goals are to translate genomic research into clinical advances for cardiovascular patients. His career development and research plan leverages his training in human genetics and the unique training environment of BWH, HMS, and the Broad Institute that will provide him with new skills in statistics, computational biology, and induced pluripotent stem cell (iPSC) disease modeling, in order to shed novel insight into the mechanisms underlying hypertrophic cardiomyopathy (HCM). His career development goals will be achieved through scientific investigation and collaborations, and supported by mentorship, didactic coursework and conferences. Dr. Haghighi has assembled a Mentoring and Scientific Advisory Committee that includes world experts in cardiology, and human genetics: Drs. Christine Seidman (primary mentor), Jon Seidman (co-mentor), Shamil Sunyaev (advisor) and Mark Daly (advisor), who have mentored countless young investigators to successful independent investigators. HCM is the most common inherited cardiovascular diseases and the leading cause of sudden death in young people. Dominant mutations in sarcomere proteins cause HCM in 30-60% of patients- data that has enabled mechanistic studies that propel new therapeutic strategies to treat HCM. These opportunities are profoundly limited in sarcomere-negative (unexplained) HCM patients in whom there is no understanding of how or why disease emerges. Dr. Haghighi's whole exome sequencing (WES) analyses in genetically unexplained HCM patients concluded that non-coding variants may play a role in pathogenesis of this disease. This proposal seeks to define genetic etiologies (with a focus on non-coding variants) for HCM in patients that remain unsolved after WES sequencing by whole genome sequencing (WGS). To determine the functional impact of newly identified non-coding variants, Dr. Haghighi will model and characterize prioritized candidate variants in human isogenic cardiomyocytes derived from induced pluripotent stem cells (iPSC-CMs). Using these approaches he will explore three specific aims that (1) Identify rare variants in non-coding regions of known HCM genes and prioritized genes in blood and heart tissue samples of unexplained HCM patients by WGS, (2) Define dysregulated gene expressions and splicing effects of identified variants by RNAseq, and (3) Assess the effects of prioritized non-coding variants in human iPSCs. By providing fundamental new insights into the functional role of non-coding variants that may uncover new causes of HCM (thereby improving patient care), these studies will support NHLBI goals to improve our understanding of the molecular basis of health and disease. In addition, successful completion of the proposed studies advance Dr. Haghighi's transition to an independent clinical- scientist and provide him with a solid foundation from which he will apply for extramural funding.

项目摘要 该提案描述了Alireza Haghighi博士职业发展的五年计划， 独立心血管研究者。目前是医学讲师和临床分子遗传学家， 布里格姆妇女医院（BWH）和哈佛医学院（HMS）， 博士Haghighi的长期职业目标 将基因组研究转化为心血管患者的临床进展。他的职业发展和 研究计划利用他在人类遗传学方面的培训以及BWH，HMS和 布罗德研究所将为他提供统计学，计算生物学和诱导多能干细胞方面的新技能 细胞（iPSC）疾病建模，以揭示肥大细胞的潜在机制， 心肌病（HCM）。他的职业发展目标将通过科学考察实现， 合作，并得到导师，教学课程和会议的支持。哈格海迪博士已经召集了一个 指导和科学咨询委员会，包括世界心脏病学专家，人类遗传学：博士。 克莉丝汀塞德曼（主要导师），乔恩塞德曼（共同导师），沙米尔Sunyaev（顾问）和马克戴利（顾问）， 他们指导了无数年轻的调查员成为成功的独立调查员。 肥厚型心肌病是最常见的遗传性心血管疾病，也是老年人猝死的主要原因。 年轻人肌节蛋白的显性突变导致30-60%的患者HCM-数据显示， 使机制研究能够推动新的治疗策略来治疗HCM。这些机会 在肌节阴性（原因不明）的HCM患者中， 或者为什么疾病会出现Haghighi博士的全外显子组测序（WES）分析在遗传上无法解释 HCM患者的结论是，非编码变异可能在这种疾病的发病机制中发挥作用。这项建议 旨在确定HCM患者的遗传病因（重点是非编码变异）， 在通过全基因组测序（WGS）进行WES测序之后。要确定新的功能影响， Haghighi博士将对人类中优先的候选变异进行建模和表征， 来源于诱导多能干细胞（iPSC-CM）的同基因心肌细胞。使用这些方法，他 将探索三个具体目标，（1）确定已知HCM基因非编码区的罕见变异， 通过WGS在不明原因的HCM患者的血液和心脏组织样品中优先考虑基因，（2）定义 通过RNAseq鉴定的变体的失调基因表达和剪接效应，以及（3）评估效应 在人类iPSC中优先的非编码变体。通过提供对职能作用的基本新见解， 非编码变异可能揭示HCM的新原因（从而改善患者护理），这些研究将 支持NHLBI的目标，以提高我们对健康和疾病的分子基础的理解。此外，本发明还提供了一种方法， 成功完成拟议的研究 博士哈格海吉向独立临床医生的转变- 科学家，并为他提供了一个坚实的基础，他将申请校外资助。