Epigenetic regulation of PDE signaling in dilated cardiomyopathy

扩张型心肌病 PDE 信号的表观遗传调控

• 批准号: 10321971
• 负责人: Haodi Wu
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321971
• 关键词: Adrenergic Agents; Advisory Committees; Affect; Alleles; American; Binding; Biological Assay; CRISPR interference; CRISPR/Cas technology; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular system; Career Mobility; Cell Nucleus; Cells; ChIP-seq; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Cues; Cyclic Nucleotides; DNA; Data; Development; Dilated Cardiomyopathy; Disease; Disease model; Dominant-Negative Mutation; Elements; Enzymes; Epigenetic Process; Family; Gene Expression; Generations; Genes; Genetic; Goals; Health Care Costs; Heart failure; High-Throughput Nucleotide Sequencing; Histones; Human; Impairment; Induced Mutation; Institutes; Knowledge; Lead; Light; Mammalian Cell; Mentors; Mentorship; Modeling; Modification; Molecular; Molecular Biology Techniques; Molecular Target; Mutation; Nuclear; Nucleic Acid Regulatory Sequences; Pathogenesis; Pathologic; Pathology; Patients; Pattern; Phase; Phenotype; Phosphodiesterase Inhibitors; Positioning Attribute; Program Development; Promoter Regions; Proteins; Publishing; RNA Interference; Regulation; Regulatory Element; Research; Research Personnel; Research Project Grants; Resolution; Resources; Role; Signal Transduction; Standardization; Structure; Technology; Testing; Therapeutic; Training; Transcriptional Regulation; Work; base; career; career development; design; disease phenotype; epigenetic regulation; genome editing; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; innovation; meetings; molecular subtypes; novel; novel therapeutic intervention; personalized medicine; phosphoric diester hydrolase; professor; programs; promoter; recruit; reduce symptoms; screening; small hairpin RNA; small molecule; stem cell biology; stem cells; tool; transcriptome sequencing; whole genome

PROJECT SUMMARY This proposal describes a five-year career development program to prepare Dr. Haodi Wu for a career as an independent investigator. This program will build on Dr. Haodi Wu’s background as a cell biologist and biophysicist by providing him expertise in molecular biology techniques such as sequencing technology and genome-editing to advance our understanding of the pathogenesis and treatment of dilated cardiomyopathy (DCM). Dr. Haodi Wu will be mentored by Dr. Joseph Wu, professor and Director of Stanford Cardiovascular Institute and a pioneer of stem cell biology research and cardiovascular disease modeling with iPSC derived cardiomyocytes (iPSC-CMs), and co-mentored by Dr. Michael Snyder, professor and Chair of Stanford Genetics Department and a world-renowned expert in omics and personalized medicine. The K99 phase of Dr. Haodi Wu’s training will consist of structured mentorship by the primary mentor and co-mentor, close interactions with collaborators, complementary meeting with advisory committee, formal coursework, a provocative research project, and a program of career transition. Patient specific iPSC-CMs present a unique opportunity to define the cellular phenotype, elucidate pathology mechanism and develop potential treatment of DCM. In Dr. Haodi Wu’s previous works, he generated iPSC-CM models from a DCM family, and showed patient specific iPSC-CMs recapitulate key phenotype and molecular cues of DCM. He also identified the subtype-specific epigenetic activation of phosphodiesterase (PDEs) as a key molecular regulation that contributes to DCM pathogenesis. Moreover, he established multiple signaling and functional assays on iPSC-CMs, which allow for examination of the molecular mechanism in DCM iPSC-CM models with a level of depth and resolution never before achieved. With the current advancement in high-throughput sequencing and the cutting-edge CRISPR/dCas9 molecular tools, Dr. Haodi Wu is in a unique position to uncover the unknown mechanisms that underlie the remodeled PDE expression pattern in human DCM cardiomyocytes, and to develop novel molecular approaches and compounds that target molecular basis of DCM in a gene and regulatory element specific way. In the K99 phase, Dr. Haodi Wu will generate and characterize genome edited DCM iPSC-CM models via introduction of dominant negative mutations (Aim1). With the platform, Dr. Haodi Wu will integrate both RNA-seq and ChIP-seq data to identify the key epigenetic regulatory mechanism of PDE expression in DCM cardiomyocytes with TNNT2 mutation (Aim2). Basing on the knowledge, he will develop novel molecular approaches and small molecules that restore PDE expression and signaling in R00 phase (Aim3). Collectively, Dr. Haodi Wu’s proposed work will create a valuable platform to study the detailed role of single DCM mutation in human cardiomyocytes, and reveal mutation-specific molecular mechanism of PDE regulations underlying DCM pathogenesis. Additionally, this work will cast new light on the application of novel molecular tools and PDE inhibitors in targeting on the molecular basis of DCM disease, which will be carried out by Dr. Haodi Wu as an independent investigator.

项目摘要 该建议描述了一项为期五年的职业发展计划，以准备Haodi Wu博士作为职业 独立研究者。该计划将以Haodi Wu博士为基础，作为细胞生物学家和 生物物理学家通过为他提供分子生物学技术的专业知识，例如测序技术和 基因组编辑以提高我们对扩张心肌病的发病机理和治疗的理解 （DCM）。 Haodi Wu博士将由Stanford心血管教授兼主任Joseph Wu博士考虑 IPSC的研究所和干细胞生物学研究和心血管疾病建模的先驱 斯坦福大学遗传学教授兼主席迈克尔·斯奈德（Michael Snyder）博士，心肌细胞（IPSC-CMS），并由 部门和世界知名的OMICS和个性化医学专家。 Haodi Wu博士的K99阶段 培训将包括主要的心理和同事的结构化心态，与 合作者，与咨询委员会的完整会议，正式课程，挑衅性研究 项目和职业过渡计划。患者特定的IPSC-CM为定义的独特机会 细胞表型，阐明病理机制和发展DCM的潜在处理。在Haodi Wu博士 他以前的工作，从DCM家族产生了IPSC-CM型号，并显示了患者特定的IPSC-CMS 概括DCM的关键表型和分子提示。他还确定了亚型特异性表观遗传学 磷酸二酯酶（PDES）的激活是有助于DCM发病机理的关键分子调节。 此外，他在IPSC-CM上建立了多个信号传导和功能测定，这允许检查 DCM IPSC-CM模型中具有深度和分辨率水平的分子机制从未实现。 随着高通量测序和尖端CRIS/DCAS9分子的当前进步 工具，Haodi Wu博士处于独特的位置，可以揭示删除的未知机制 人DCM心肌细胞中的PDE表达模式，并开发新的分子方法和 以基因和调节元件的特定方式靶向DCM的分子基础的化合物。在K99阶段， Haodi Wu博士将通过引入主导地位生成和表征基因组编辑的DCM IPSC-CM模型 负突变（AIM1）。使用该平台，Haodi Wu博士将将RNA-Seq和Chip-Seq数据集成到 用TNNT2确定DCM心肌细胞中PDE表达的关键表观遗传调节机制 突变（AIM2）。基于知识，他将开发新的分子方法和小分子 该恢复R00相中的PDE表达和信号传导（AIM3）。 Haodi Wu博士的拟议工作总的来说 创建一个有价值的平台来研究人类心肌细胞中单个DCM突变的详细作用，并 揭示了DCM发病机理的PDE法规的突变特异性分子机制。此外， 这项工作将对新型分子工具和PDE抑制剂在靶向靶向时的应用宣传 DCM疾病的分子基础，该疾病将由Haodi Wu博士作为独立研究者进行。