Gene Modulation of Acetylation Modifiers to Reveal Regulatory Links to Human Cardiac Electromechanics

乙酰化修饰剂的基因调节揭示与人类心脏机电的调节联系

基本信息

批准号：
10677295
负责人：
MARIA POZO
金额：
$ 3.79万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677295
关键词：
Acetylation Action Potentials Acute Arrhythmia Autoimmune Diseases Automobile Driving Autophagocytosis Behavior Biological Assay CRISPR interference CRISPR-mediated transcriptional activation Calcium Cardiac Cardiac Electrophysiologic Techniques Cardiac health Cardiotoxicity Cardiovascular Diseases Cardiovascular system Cell Cycle Cells Complex Computer Models Contractile Proteins Cytoskeletal Proteins Data Development Disease model Electrophysiology (science)Emergency Situation Epigenetic Process Event Exhibits Feedback Fibrosis Frequencies Functional disorder Gene Expression Genes Genetic Transcription Guide RNA Health Heart Heart Injuries Histone Deacetylase Histone Deacetylase Inhibitor Homeostasis Human Hypertrophy Immune System Diseases Individual Inflammatory Response Intervention Ion Channel Ischemia Libraries Link Malignant Neoplasms Measurement Measures Mechanics Mitochondria Modeling Morbidity - disease rate Oncology Optics Oxidative Stress Pathology Phenotype Predisposition Process Proteins Public Health RNA Interference Regulation Regulator Genes Reperfusion Injury Repression Research Role Sampling Shapes Small Interfering RNA Specificity Stimulus Techniques Therapeutic Tissues United States Validation Variant Work cancer therapy cardioprotection chromatin remodeling cost estimate epigenetic regulation epigenomics experience experimental study gene regulatory network gene repression heart function improved induced pluripotent stem cell derived cardiomyocytes interest knock-down loss of function mortality mouse model novel patient prognosis pharmacologic response screening tool transcription factor transcriptome sequencing transcriptomic profiling transcriptomics tumor progression voltage

项目摘要

PROJECT ABSTRACT Epigenetic regulation is critical for cardiac electromechanics and pathology. Epigenetic modulators, such as histone deacetylases (HDACs), are known master regulators of gene expression and influence cardiac function through chromatin remodeling, direct action on transcription factors (TFs), and action on cytoskeletal and contractile proteins, among others. Recently, novel pharmacological agents, HDAC inhibitors, have been developed as treatments for cancer and immune diseases, driving an interest in robust characterization of HDAC control in cardiac function. Our preliminary experiments focused on computational modeling of RNAi-informed transcriptomic data in human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CM) but saw limitations in knockdown efficiency and loss-of-function-only modulation using siRNAs. To extend and improve this work, we propose an experimental approach based on bidirectional perturbation (repression/activation) of individual HDAC genes in hiPSC-CM by CRISPR interference and activation (CRISPRi/a). Transcriptomic analysis of these samples will inform computational gene regulatory network (GRN) inference to model relationships between HDACs, TFs, and cardiac ion channels. GRN-predicted relationships will be validated by all-optical electromechanical assays measuring voltage, calcium, and contraction traces in hiPSC-CM. An iterative approach will allow feedback from functional experiments to refine our computational models. Such studies will advance our understanding of how certain HDACs drive electrophysiological phenotypes in the heart, which is critical in the fields of cardiac injury, cardiac therapeutics, and cardio-oncology.

项目摘要表观遗传调控对于心脏机电和病理学至关重要。表观遗传调节剂，例如组蛋白脱乙酰酶 (HDAC) 是已知的基因表达的主要调节因子并影响心脏功能通过染色质重塑、对转录因子 (TF) 的直接作用以及对细胞骨架和细胞因子的作用收缩蛋白等。最近，新型药物 HDAC 抑制剂已被开发出来。开发用于治疗癌症和免疫疾病，引发了人们对 HDAC 稳健表征的兴趣心脏功能的控制。我们的初步实验侧重于 RNAi 信息的计算建模人类诱导多能干细胞衍生的心肌细胞（hiPSC-CM）的转录组数据，但看到使用 siRNA 的敲低效率和仅功能丧失调节的局限性。扩展和改进在这项工作中，我们提出了一种基于双向扰动（抑制/激活）的实验方法 hiPSC-CM 中的单个 HDAC 基因通过 CRISPR 干扰和激活 (CRISPRi/a)。转录组学对这些样本的分析将为计算基因调控网络 (GRN) 模型推断提供信息 HDAC、TF 和心脏离子通道之间的关系。 GRN 预测的关系将通过以下方式验证全光学机电测定测量 hiPSC-CM 中的电压、钙和收缩痕迹。一个迭代方法将允许功能实验的反馈来完善我们的计算模型。这样的研究将加深我们对某些 HDAC 如何驱动心脏电生理表型的理解，这在心脏损伤、心脏治疗和心脏肿瘤学领域至关重要。