Reactivating regulatory programs for regeneration

重新启动再生监管计划

• 批准号: 10687448
• 负责人: Megan Lee Martik
• 金额: $ 134.44万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687448
• 关键词: ATAC-seq; Ablation; Automobile Driving; Cardiac; Cardiac Myocytes; Cells; Chromatin; Cicatrix; Clustered Regularly Interspaced Short Palindromic Repeats; Development; Developmental Gene; Failure; Gene Combinations; Genes; Genetic; Goals; Heart; Heart Injuries; Human; Impairment; Information Networks; Injury; Intervention; Laboratories; Modality; Modeling; Mus; Myocardial Infarction; Natural regeneration; Neural Crest; Organism; Organoids; Proliferating; Regenerative Medicine; Regenerative response; Regulator Genes; Therapeutic; Tissues; Zebrafish; cardiac regeneration; cardiac repair; functional genomics; gene network; gene regulatory network; human pluripotent stem cell; in vivo; multiple omics; next generation; programs; regenerative; repaired; self assembly; synergism; translational approach

PROJECT SUMMARY/ABSTRACT Human cardiac injury, such as a heart attack, leads to irreparable damage and life-long heart complications. Developing translational strategies for inducing heart repair has been limited to laboratory accessible models such as the zebrafish and mouse. Using the zebrafish, which can regenerate their heart after substantial injury, we have previously shown that neural crest-derived cardiomyocytes promote injury-induced proliferation of surrounding cardiomyocytes by re-activating developmental gene networks after injury. Importantly, genetic ablation of neural crest-derived cardiomyocytes leads to a failure of regeneration and a large scar. Now knowing the importance of neural crest-derived cardiomyocytes and re-activating developmental networks, many questions remained unanswered on how these networks are re-deployed after injury and if these networks remain silenced in human hearts after injury. Our current hypothesis is that human neural crest-derived cardiomyocytes are unable to redeploy developmental gene networks after injury and are therefore unable to induce repair mechanisms. Until recently, assessing gene regulatory dynamics in human-derived cardiac tissues was not possible. Now, self-assembling cardiac organoids derived from human pluripotent stem cells have presented a new avenue for exploring cardiac repair in human-derived tissues; however, these cardioid models do not contain cardiomyocytes derived from neural crest. Here, we propose to (i) assess the dynamic chromatin landscapes of the regenerating zebrafish heart using single cell ATAC-seq to unravel critical components necessary for re-activating developmental programs that control cardiac regeneration in the zebrafish, (ii) interrogate the reactivation of developmental programs in a human-derived cardioid model after injury using a multiomics approach, and finally, (iii) use next-generation CRISPR-based functional genomics screens to identify gene circuits responsible for “repair impairment” of human neural crest-derived cardiomyocytes. Ultimately, our goal is to combine gene regulatory network information from zebrafish repair circuits and our human-derived screen to identify optimal targets for potential intervention using any relevant therapeutic modality for driving cardiac repair in vivo post-injury.

项目概要/摘要 人类心脏损伤，例如心脏病发作，会导致不可挽回的损害和终生心脏并发症。 开发诱导心脏修复的转化策略仅限于实验室可访问的模型 例如斑马鱼和老鼠，利用斑马鱼的心脏在严重受伤后可以再生， 我们之前已经表明，神经嵴衍生的心肌细胞促进损伤诱导的增殖 损伤后通过重新激活发育基因网络来调节周围心肌细胞，重要的是遗传。 现在知道，神经嵴来源的心肌细胞的消融会导致再生失败和大疤痕。 神经嵴来源的心肌细胞和重新激活发育网络的重要性，许多 关于这些网络在受伤后如何重新部署以及这些网络是否 我们目前的假设是，人类的神经嵴源自受伤后的心脏。 心肌细胞在损伤后无法重新部署发育基因网络，因此无法 直到最近，人们还在评估人源性心脏组织中的基因调控动态。 现在，来自人类多能干细胞的自组装心脏类器官已经成为可能。 然而，这些心形模型为探索人源组织的心脏修复提供了新途径； 不包含来自神经嵴的心肌细胞在这里，我们建议（i）评估动态染色质。 使用单细胞 ATAC-seq 揭示斑马鱼心脏再生的关键组成部分 重新激活控制斑马鱼心脏再生的发育程序所必需的，（ii） 使用人源心形模型询问受伤后发育程序的重新激活 多组学方法，最后，(iii) 使用下一代基于 CRISPR 的功能基因组学筛选来识别 负责人类神经嵴衍生心肌细胞“修复损伤”的基因回路。 目标是将来自斑马鱼修复电路和我们人类来源的基因调控网络信息结合起来 使用任何相关的驾驶治疗方式进行筛选，以确定潜在干预的最佳目标 心脏损伤后的体内修复。