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.

项目摘要 尽管对心脏发展的理解越来越复杂，但其基础的机制 先天性心脏缺陷（CHD）的原因，外观和表型多样性，最常见的先天缺陷， 不太了解。这代表了风险分层的障碍，改善治疗方法和预防 冠心。我们研究了异质综合征的遗传和发育基础，多系统疾病 至少部分归因于异常心脏循环形态发生的CHD谱。 尽管已证明早期胚胎期间左右（LR）轴形成异常 发育导致杂种中的CHD杂质，遇到的心室形态学缺陷更多 与LR轴的简单破坏所预期的要多。我们假设这是严重的杂种 CHD由心脏祖细胞异常细胞命运产生 从后来的LR图案介导的CHD。来自X-小鼠模型的单细胞RNA测序（SCRNA-SEQ） 链接的异际，ZIC3无效小鼠，支持了这一假设。数据证明了异常中胚层与 心脏病发生之前的神经外胚层分配，导致心肌细胞命运异常。与 我们关于ZIC3 NULL小鼠异常原始条纹形成的数据，这些研究揭示了必需品的必要性 研究ZIC3基因调节网络（GRN）在从细胞多能到LR的过渡期间 构图了解多样化的CHD的机械基础。我们将配对这个调查 凭借我们在良好的型号中的基因组分析和整个基因组测序（WGS）数据方面的专业知识 杂质冠心病患者队列。我们的初步数据表明，冠心病的稀有变体负担增加 候选基因，表明复杂的CHD可能是由多个的组合相互作用引起的 敏感性等位基因。这种方法将用于测试候选基因的单基因和寡疾病 协会。这项研究的目的是：1）检验以下假设：将ZIC3 DNA占用数据整合到 ZIC3功能丧失引起的转录和表观遗传变化将确定心脏的新型GRN 形态发生； 2）检验ZIC3 GRN内遗传变异与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 形态发生缺陷会导致冠心病。通过识别胃肠道过渡状态期间的ZIC3途径 对于心脏病，我们将发现心脏形成所需的不同ZIC3 GRN。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将提供至关重要的 翻译信息。总的来说，我们将确定新颖的ZIC3 GRNS对腔室很重要 形态发生，并提供了有关途径的单基因和寡源性贡献的基本信息 调节心脏细胞的命运，以发展人类异质股的发展。