The role of ZIC3 within cardiomyocyte precursors in cardiac morphogenesis

ZIC3 在心肌细胞前体细胞中在心脏形态发生中的作用

• 批准号: 10495949
• 负责人: Stephanie M Ware
• 金额: $ 48.8万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-15 至 2028-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10495949
• 关键词: Abnormal Cell; Alleles; Animals; Binding; Biological Assay; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cells; Child; Chromatin; Complex; Congenital Abnormality; Congenital Heart Defects; DNA; Data; Data Set; Defect; Development; Developmental Biology; Disease; Embryo; Embryonic Development; Epiblast; Epigenetic Process; Epitopes; Exhibits; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Variation; Genetic study; Genomics; Goals; Growth; Heart; Heart Abnormalities; Human; Human Genetics; Indiana; Investigation; Knockout Mice; Lead; Left; Link; Mediating; Mesoderm; Molecular; Morphogenesis; Multiomic Data; Mus; Neuroectoderm; Normal tissue morphology; Outcome; Pathway interactions; Patients; Pattern; Pediatric Cardiac Genomics Consortium; Penetrance; Phenotype; Play; Predisposition; Prevention; Primitive Streaks; Research; Resources; Risk Factors; Role; Severities; Situs Inversus; Specific qualifier value; Susceptibility Gene; Syndrome; System; Testing; Therapeutic; Tissues; Transposase; United States National Institutes of Health; Variant; Ventricular; Zinc Fingers; biobank; cardiogenesis; cohort; combinatorial; embryo tissue; embryonic stem cell; epigenome; exome; exome sequencing; gastrulation; gene regulatory network; genetic variant; genome sequencing; human embryonic stem cell; improved; in vivo; in vivo Model; insight; loss of function; mouse model; multiple omics; mutation screening; novel; pluripotency; progenitor; programs; rare variant; risk stratification; single-cell RNA sequencing; somitogenesis; stem cells; transcription factor; transcriptome; whole genome

PROJECT SUMMARY Despite an increasingly sophisticated understanding of cardiac development, the mechanisms underlying the causes, penetrance, and phenotypic diversity of congenital heart defects (CHDs), the most common birth defect, are not well understood. This represents a barrier to risk stratification, improved therapeutics, and prevention of CHD. We have studied the genetic and developmental basis of heterotaxy syndrome, a multisystem disorder with a spectrum of CHDs that are attributable, at least in part, to abnormal cardiac looping morphogenesis. Although it has been proven that abnormalities in left-right (LR) axis formation during early embryonic development lead to CHDs in heterotaxy, the ventricular chamber morphogenic defects encountered are more diverse than would be expected from simple disruption of the LR axis. We hypothesize that severe heterotaxy CHDs result from abnormal cell fate of cardiac progenitors and that this is a distinct CHD causing mechanism from later LR patterning-mediated CHDs. Single cell RNA sequencing (scRNA-seq) from a mouse model of X- linked heterotaxy, Zic3 null mice, supports this hypothesis. The data demonstrate an abnormal mesoderm versus neuroectoderm allocation prior to cardiogenesis that results in abnormal cardiomyocyte cell fate. Combined with our data on abnormal primitive streak formation in Zic3 null mice, these studies reveal an essential need to investigate the ZIC3 gene regulatory networks (GRNs) during the transition from cell pluripotency through LR patterning to understand the mechanistic underpinnings of a diverse set CHDs. We will pair this investigation with our expertise in genomic analyses and whole genome sequencing (WGS) data in our well-phenotyped cohort of heterotaxy CHD patients. Our preliminary data demonstrate increased rare variant burden in CHD candidate genes, suggesting that complex CHD can result from combinatorial interactions of multiple susceptibility alleles. This approach will be used to test candidate genes for monogenic and oligogenic disease association. The aims of this study are to: 1) test the hypothesis that integrating ZIC3 DNA-occupancy data with transcriptional and epigenetic changes caused by ZIC3 loss-of-function will identify novel GRNs for cardiac morphogenesis; and 2) test the hypothesis that genetic variation within the ZIC3 GRN is associated with CHD. The overarching hypothesis of this proposal is that ZIC3 regulatory network genes are risk factors for ventricular morphogenesis defects that result in CHD. By identifying ZIC3 pathways during transition states from gastrulation to cardiogenesis, we will discover distinct ZIC3 GRNs required for heart formation. The integration of this multiomics data will provide novel insight into cardiac progenitor cell specification. Investigating the importance of these ZIC3 GRNs and developmental-stage specific pathways to human CHD susceptibility will provide critical translational information. Collectively, we will have identified novel ZIC3 GRNs important for chamber morphogenesis and provided essential information on monogenic and oligogenic contributions of pathways regulating cardiac cell fate to the development of human heterotaxy CHDs.

项目概要 尽管对心脏发育的了解日益深入，但心脏发育背后的机制 先天性心脏病 (CHD)（最常见的出生缺陷）的原因、外显率和表型多样性， 没有被很好地理解。这对风险分层、改进治疗和预防造成了障碍 冠心病。我们研究了异位综合征（一种多系统疾病）的遗传和发育基础 一系列冠心病至少部分归因于异常的心脏循环形态发生。 尽管已证明早期胚胎时期左右（LR）轴形成异常 发育导致冠心病的异位性，遇到的心室形态缺陷较多 与简单破坏 LR 轴所预期的不同。我们假设严重的异质性 CHD 是由心脏祖细胞异常细胞命运引起的，这是一种独特的 CHD 引起机制 来自后来的 LR 模式介导的先心病。 X-小鼠模型的单细胞 RNA 测序 (scRNA-seq) 连锁异位，Zic3 缺失小鼠，支持了这一假设。数据表明中胚层与 心脏发生前的神经外胚层分配导致心肌细胞命运异常。结合 我们关于 Zic3 缺失小鼠中异常原始条纹形成的数据，这些研究揭示了必要的 研究从细胞多能性到 LR 转变过程中的 ZIC3 基因调控网络 (GRN) 模式来理解不同类型先心病的机制基础。我们将配对这项调查 凭借我们在基因组分析和全基因组测序 (WGS) 数据方面的专业知识，我们的表型良好 异位冠心病患者队列。我们的初步数据表明先心病的罕见变异负担增加 候选基因，表明复杂的 CHD 可能是由多个基因的组合相互作用引起的 易感性等位基因。该方法将用于测试单基因和寡基因疾病的候选基因 协会。本研究的目的是：1）检验将 ZIC3 DNA 占用数据与 ZIC3 功能丧失引起的转录和表观遗传变化将识别心脏疾病的新 GRN 形态发生； 2) 检验 ZIC3 GRN 内的遗传变异与 CHD 相关的假设。 该提案的总体假设是 ZIC3 调节网络基因是心室的危险因素 导致 CHD 的形态发生缺陷。通过识别原肠胚形成过渡状态期间的 ZIC3 途径 对于心脏发生，我们将发现心脏形成所需的独特 ZIC3 GRN。这个的整合 多组学数据将为心​​脏祖细胞规范提供新的见解。调查重要性 这些 ZIC3 GRN 和人类 CHD 易感性的发育阶段特定途径的研究将提供关键的 翻译信息。总的来说，我们将确定对腔室重要的新型 ZIC3 GRN 形态发生并提供有关途径的单基因和寡基因贡献的重要信息 调节心脏细胞命运与人类异位先心病的发展。