Genome-wide Analysis of Cardiac Development in Zebrafish

描述(由申请人提供)： 我们建议与斑马鱼发育生物学家、斑马鱼心脏生理学家以及染色质结构、全基因组基因网络图谱、生物工程和生物信息学专家合作建立一个多学科的斑马鱼心脏发育研究中心。我们将研究发育过程中不同阶段的基因调控网络，这些网络在多个分子水平上控制正常的心脏发育。心脏发育的几种不同的扰动将作为评估心脏发育基因调控网络变化的“输入”，包括遗传突变和形态突变、胚胎细胞谱系操作和药物治疗。“输出”将是对调控转录程序的染色质结构的分析，包括DNA甲基化和有助于转录激活(H3K4me3)或转录抑制(H3K27me3)的染色质标记的表观遗传修饰的分析，全基因组基因表达谱(包括mRNAs、microRNAs、Sine RNAs和其他小RNA)以及参与心肌细胞发育过程中的图案化、形态发生和生理学的转录调控因子。对斑马鱼体内正常和大量异常特征的生物信息学比较将揭示心脏发育缺陷的分子特征。这种心脏发育的多层分子图谱还没有在任何生物体中进行过。当心脏发育联盟和儿科心脏基因组学联盟出现新的候选基因或其他干扰(包括药理学或环境)时，它们将被纳入这一多层分子图谱计划，以快速获得基因组范围的分子签名。我们的基础设施将使这些数据集随时可供财团进行跨物种比较，以揭示与人类发育性心脏缺陷相关的保守分子特征。相关性(见说明)：获得心脏发育缺陷的多层分子图谱将揭示心脏发育缺陷的根本原因，长期目标是将这些见解应用于心脏缺陷儿童的治疗。

DESCRIPTION (provided by applicant): We propose to build a multidisciplinary Zebrafish Cardiac Development Research Center with a collaborative group of zebrafish developmental biologists, zebrafish cardiac physiologists, and experts in chromatin structure, genome-wide gene network profiling, bioengineering and bioinformatics. We will investigate gene regulatory networks at distinct steps in development that control normal cardiac development at multiple molecular levels. Several diverse perturbations of cardiac development will serve as "inputs" to assess changes in cardiac development gene regulatory networks, including genetic mutants and morphants, embryological cell lineage manipulations and pharmacological treatments. The "outputs" will be analyses of chromatin structure that regulates transcriptional programs, epigenetic modifications including DNA methylation and chromatin marks that contribute to transcriptional activation (H3K4me3) or transcriptional repression (H3K9me3, and H3K27me3), genome-wide gene expression profiles (including mRNAs, microRNAs, SINE RNAs, other small RNAs) and transcriptional regulators involved in the patterning, morphogenesis and physiology of cardiomyocytes during development. Bioinformatics comparisons of normal and a large number of aberrant profiles within zebrafish will uncover molecular signatures of cardiac developmental defects. This multi-layered molecular profiling of cardiac development has not been performed in any organism. As new candidate genes or other perturbations (including pharmacological or environmental) arise within the Cardiac Development Consortium and Pediatric Cardiac Genomics Consortium, they will be incorporated into this multi-layered molecular profiling program to rapidly obtain genome-wide molecular signatures. Our infrastructure will make these datasets readily accessible to the consortia for trans-species comparisons to uncover conserved molecular signatures relevant to human developmental heart defects. RELEVANCE (See instructions): Obtaining multi-layered molecular profiles of cardiac developmental defects will reveal the underlying causes of cardiac developmental defects, with the long term goal of applying these insights to treatment of children with heart defects.