Developmental mechanisms of human congenital heart disease

摘要 本P01更新申请是对NIH NICHD研究所RFA HD-16-009的回应， 确定结构性出生缺陷的发育机制。本次更新将继续在目前的 主题是识别导致先天性心脏病的基因和机制。先天性类 作为P01焦点的心脏病被称为圆锥动脉干和相关主动脉弓缺损， 作为CTRD。动物模型的研究表明， 在胚胎发育期间引起圆锥干心脏缺陷和/或主动脉弓异常。特别是 心脏流出道和主动脉弓来源于胚胎内共有的前体细胞群 咽器CTRD的分子基础是极其复杂的，其基于以下基因研究： 非综合征性CTRD（NS-CTRD;项目2），其中可能存在全谱遗传变异， 责任为了解决这一复杂的出生缺陷，该计划的更新包括三个 相互依存的项目。在项目1中，我们将鉴定22q11.2缺失中CTRD的遗传修饰物 综合征（22q11.2DS;又称迪乔治综合征，腭心面综合征）。这个想法是，删除 它本身可以使基因组对进一步的遗传损伤敏感，这可能揭示了 更多异质性NS-CTRD中的疾病。我们对这一目标有积极的发现， 22q11.2DS队列中SLC 2A 3重复和组蛋白修饰基因的表达。但由于 22 q11-CTRD和NS-CTRD队列遗传学的复杂性，以及有限的样本量，我们 转向专注于发育中的咽器的生物学（包括额外的新兴的 途径）来检查基因组，而不是采取不可知论的方法。生物分析核心将 提供生物统计和生物信息学监督。在项目2中，我们将检查常见和罕见的DNA 严重CTRD的基因组变异，以及了解严重CTRD与 轻度CTRD，特别是孤立的主动脉弓异常。在项目3中，我们将使用 22q11.2DS，其中我们关注缺失区域中的两个特定基因，Tbx 1，编码T-box 转录因子和Crkl，编码用于细胞内信号传导的衔接子，在形成主动脉弓中起作用。 其次，我们将从这些显微解剖的咽器中鉴定转录组， 模型和其他模型（Lgdel/+），以生成一个交互式基因网络，称为PA-INet，用于 项目1和2，并在生物分析核心组织。最后，我们将对top进行功能分析。 基因和基因座，首先是生物信息学，然后是细胞培养，最后是小鼠模型。我们相信我们的 战略，重点是22q11.2DS和发展机制，将有可能获得独特的 这一重要的人类先天性结构缺陷

ABSTRACT This P01 renewal application is in response to RFA, HD-16-009 from the NICHD institute of NIH to determine developmental mechanisms of structural birth defects. This renewal will continue on the current theme to identify genes and mechanisms responsible for congenital heart disease. The class of congenital heart disease that is the focus of the P01 is termed, conotruncal and related aortic arch defects, referred to as CTRDs. Studies of animal models indicate that there are similar developmental mechanisms disrupted during embryogenesis causing conotruncal heart defects and/or aortic arch anomalies. In particular, the cardiac outflow tract and aortic arch derive from shared precursor cell populations from within the embryonic pharyngeal apparatus. The molecular basis of CTRDs is extremely complex, based upon genetic studies of non-syndromic CTRDs (NS-CTRDs; Project 2), in which the full spectrum of genetic variation is likely responsible. In order to tackle this complex birth defect, this program renewal consists of three interdependent projects. In Project 1, we will identify genetic modifiers of CTRDs in 22q11.2 deletion syndrome (22q11.2DS; aka DiGeorge syndrome velo-cardio-facial syndrome). The idea is that the deletion itself serves to sensitize the genome for further genetic insults that may shed light onto the mechanisms of disease in more heterogeneous NS-CTRDs. We have positive findings towards this goal in the discoveries of the SLC2A3 duplication and histone modifier genes in the 22q11.2DS cohort. However, due to the complexity of the genetics in 22q11-CTRD and NS-CTRD cohorts, as well as limited sample sizes, we have turned to a focus on the biology of the developing pharyngeal apparatus (including additional emerging pathways) to examine gene-sets rather than taking agnostic approaches. The Bio-analytics Core will provide the biostatistics and bioinformatics oversight. In Project 2, we will examine common and rare DNA variants in gene-sets for severe CTRDs, as well as understand the genetic architecture of severe versus mild CTRDs, specifically, isolated aortic arch anomalies. In Project 3, we will use mouse models of 22q11.2DS, where we focus on two particular genes in the deleted region, Tbx1, encoding a T-box transcription factor and Crkl, encoding an adaptor for intracellular signaling, in forming the aortic arch. Secondly, we will identify the transcriptomes from the microdissected pharyngeal apparatus from these models and others (Lgdel/+) to generate an interactive gene network, termed the PA-INet to be used in Projects 1 and 2 and organized in the Bio-analytic Core. Finally, we will perform functional analysis of top genes and loci, first bioinformatically, but then in cell culture and finally in mouse models. We believe our strategy, focusing on 22q11.2DS and developmental mechanisms, will make it possible to gain unique inroads into this important human structural birth defect.