Investigating the Time-Dependent Reversibility of DCM-Induced Epigenetic, Matrix, and Functional Remodeling

研究 DCM 诱导的表观遗传、基质和功能重塑的时间依赖性可逆性

• 批准号: 10812314
• 负责人: Isabella Reichardt
• 金额: $ 4.26万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2025-09-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10812314
• 关键词: ATAC-seq; Aftercare; Age; Age Months; Cardiac; Cardiology; Cardiovascular Diseases; Cardiovascular system; Cells; Chromatin; Collagen; Congestive Heart Failure; Coupled; Cytoskeleton; DNA Methylation; Data; Deposition; Dilated Cardiomyopathy; Disease; Disease Progression; Dose; Doxycycline; EFRAC; Engineering; Ensure; Epigenetic Process; Etiology; Extracellular Matrix; Family suidae; Fellowship; Fibroblasts; Fibrosis; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic Transcription; Genome; Genomics; Goals; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Human; Hypertrophy; In Vitro; Induced Mutation; Inherited; Injury; Learning; Left ventricular structure; Link; Mechanics; Medicine; Memory; Mentors; Mentorship; Mesenchymal Stem Cells; Microfilaments; Modeling; Mus; Muscle Cells; Mutate; Mutation; Myocardial dysfunction; Myocardium; Myosin ATPase; Nature; Organ; Outcome; Patients; Periodicity; Pharmacological Treatment; Phenotype; Physicians; Point Mutation; Progressive Disease; Proteomics; Repression; Research; Resolution; Running; Scientist; Signal Transduction; Structure; Symptoms; Technology; Testing; Therapeutic; Time; Training; Transgenic Mice; Treatment Failure; Troponin C; United States National Institutes of Health; career; career development; chromatin remodeling; clinically relevant; effective therapy; epigenetic memory; epigenome; experimental study; familial dilated cardiomyopathy; healing; heart function; improved; inhibitor; mechanical stimulus; mouse model; multiple omics; mutant; palliative; patient variability; pharmacologic; pre-doctoral; pressure; professor; response; screening; skills; therapeutic target; tool; transcriptome sequencing; transgene expression; translational therapeutics

Project Summary Dilated cardiomyopathy (DCM) is a highly prevalent inherited cardiac disease, characterized by systolic dysfunction, eccentric hypertrophy, and left ventricle dilation. While pharmacologic and mechanical treatments have been shown to partially improve cardiac function, these results are often short lived and highly variable from patient to patient. Moreover, recent studies have demonstrated that epigenetic and matrix dysregulation can persist, even in patients with improved systolic function after treatment. Given that aberrant chromatin remodeling and extracellular matrix (ECM) deposition have been identified as drivers of dilated remodeling—and that chromatin and ECM remodeling can become irreversible over time—it is crucial to understand the time- dependent effects of DCM mutations on reversing maladaptive remodeling at the genome, myocyte, and matrix levels. The central hypothesis of this proposal is that the reversibility of the DCM phenotype is time-dependent due to the accumulation of permanent ECM and chromatin remodeling as the disease progresses. To complete this proposal, I will utilize a DCM mouse model created by the Davis lab which contains a mutation (I61Q) in cardiac troponin C (cTnC) that directly decreases the myofilament’s Ca2+ sensitivity, leading to eccentric hypertrophy, systolic dysfunction, and left ventricle dilation. Importantly, expression of this mutant can be temporally controlled with doxycycline to specifically test our central hypothesis. In this proposal I will 1) Determine the time-dependent effects of I61Q cTnC expression on myocyte, matrix, and chromatin accessibility, 2) Examine the reversibility of DCM remodeling in myocyte, matrix, and chromatin accessibility at different stages of the disease, and 3) Determine if myocytes retain epigenetic memories of the mechanical disequilibrium caused by DCM. Improving our understanding of the time-dependent reversibility of DCM remodeling will better inform therapeutic targets and treatment windows for patients with DCM. Moreover, completion of this project will enhance Bella’s training as an independent scientist and prepare her to one day become a professor in cardiovascular engineering. Receiving the NIH F31 predoctoral fellowship will facilitate important experiments and training that will aid in her pursuit of this career goal. In this project, Bella will gain expertise in multi-omic approaches—such as proteomics, RNAseq, and ATACseq—which are growing in popularity due to their unbiased screening capabilities. The UW Genomics Core will assist Bella in learning how to effectively use these tools, which will not only benefit this project but will also be an incredibly useful skillset for Bella’s future career. Given the clinical relevance of this project, we have engaged Farid Moussavi-Harami, MD, an acting physician- scientist who practices cardiology within the UW Department of Medicine. Dr. Moussavi-Harami’s input as a clinician will be crucial for ensuring our research questions and aims remain relevant to patients, and his mentorship throughout this project will enhance Bella’s training and career development as she aims to eventually run a lab with an emphasis in translational therapeutics and technologies for cardiovascular diseases.

项目摘要 扩张心肌病（DCM）是一种高度普遍的遗传性心脏病，其特征是收缩 功能障碍，偏心肥大和左心室词典。而药物和机械处理 已显示已部分改善心脏功能，这些结果通常是短暂的，并且高度可变 从病人到病人。此外，最近的研究表明表观遗传和基质失调 即使在治疗后收缩功能改善的患者中，也可以持续存在。鉴于异常染色质 重塑和细胞外基质（ECM）沉积物已被确定为扩张的驱动因素，并 随着时间的流逝，染色质和ECM的重塑可能变得不可逆转 - 了解时间至关重要。 DCM突变对基因组，心肌和基质逆转适应不良重塑的依赖性影响 水平。该提议的中心假设是DCM表型的可逆性是时间依赖性的 由于疾病进展，永久性ECM和染色质重塑的积累。完成 该提案，我将利用戴维斯实验室创建的DCM鼠标模型，该模型包含突变（i61q） 心脏肌钙蛋白C（CTNC）直接降低肌丝的Ca2+敏感性，导致偏心 肥大，收缩功能障碍和左心室词典。重要的是，这个突变体的表达可能是 用强力霉素暂时控制，以专门检验我们的中心假设。在这个建议中，我将1） 确定i61q CTNC表达对肌细胞，基质和染色质可及性的时间依赖性影响， 2）检查在不同阶段，在心肌细胞，基质和染色质可及性中重塑DCM的可逆性 疾病，3）确定肌细胞是否保留了引起的机械障碍的表观遗传记忆 由DCM。提高我们对DCM重塑时间相关的可逆性的理解将更好地告知 DCM患者的治疗靶标和治疗窗口。而且，该项目的完成将 增强贝拉作为独立科学家的培训，并准备她成为一名教授 心血管工程。接受NIH F31奖学金奖学金将有助于重要实验 和培训将有助于她追求这一职业目标。在这个项目中，贝拉将获得多运动的专业知识 方法（例如蛋白质组学，RNASEQ和ATACSEQ），由于它们 公正的筛选功能。 UW基因组学核心将有助于贝拉学习如何有效使用这些 工具不仅将使该项目受益，而且还将成为贝拉未来职业的一个非常有用的技能。 鉴于该项目的临床相关性，我们聘请 科学家在大学医学系中练习心脏病学。 Moussavi-Harami博士的意见 临床医生对于确保我们的研究问题和目标至关重要 整个项目中的遗产将增强贝拉的培训和职业发展，她的目标是 最终运行一个实验室，重点是翻译疗法和心血管疾病的技术。