Mechanisms of Abnormal Diaphragm and Cardiac Development

膈肌异常和心脏发育的机制

• 批准号: 10404417
• 负责人: Daryl Armstrong Scott
• 金额: $ 12.16万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10404417
• 关键词: Address; Affect; Algorithms; Animal Model; Association Learning; Binding; Bioinformatics; Biological Assay; Birth; Candidate Disease Gene; Cardiac; Cardiac development; Cardiovascular system; Child; Chromosomes; Clinical; Clinical Data; Clinical Research; Congenital Abnormality; Congenital diaphragmatic hernia; Cytogenetics; Data; Development; Diaphragmatic Hernia; Discipline; Disease; Embryo; Encyclopedias; Endocardium; Endothelium; Foundations; GATA4 gene; Genes; Genetic; Genetic Transcription; Genomic Segment; Genomics; Goals; Grant; Heart; Hernia; Human; In Situ Hybridization; Individual; Intervention; Knowledge; Laboratories; Lead; Learning; Life; Luciferases; Mesenchymal; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Mus; Newborn Infant; Ontology; Pathogenicity; Pathway Analysis; Pathway interactions; Phenotype; Process; Proteins; Publishing; Recurrence; Research; Research Personnel; Respiratory Diaphragm; Role; SOX7 gene; Source; Syndrome; Techniques; Therapeutic Intervention; Training; United States National Institutes of Health; Update; Validation; WNT4 gene; Work; biochip; bioinformatics tool; cardiogenesis; congenital anomaly; data modeling; dosage; epigenomics; experimental study; gene discovery; genome database; human disease; innovation; large scale data; machine learning algorithm; malformation; microdeletion; molecular sequence database; mortality; mouse genome; mouse model; next generation sequencing; novel; parent grant; prevent; preventive intervention; research study; septal defect; transcription factor; transcriptome sequencing; web based interface

Project Summary Congenital diaphragmatic hernia (CDH) is a life-threatening birth defect that accounts for 8% of all major congenital anomalies. In cases where CDH co-occurs with cardiovascular malformations (CVMs), mortality rates increase from 30 to 60%, and long-term morbidity is common. Our goal is to identify genes that cause CDH and CDH/CVM, and to discover the mechanism by which they control diaphragm and heart development. Correctly identifying genes that contribute to specific phenotypes from the large list of candidate genes data generated in research and clinical studies is a major obstacle to progress in this and other research fields. In Specific Aim #1, we will address this challenge by generating ranked CDH and CDH/CVM pathogenicity scores for all RefSeq genes using a machine-learning algorithm that integrates data from large-scale genomic knowledge sources. We will use these scores to identify novel CDH and CDH/CVM genes from cytogenetically defined critical regions and form large next-generation sequencing databases as part of our multifaceted approach to novel gene discovery. We will also accelerate the pace at which human disease genes are discovered by making these scores and our machine-learning algorithm freely available. 8p23.1 microdeletions that encompass GATA4 and SOX7 are among the most frequently identified causes of CDH/CVM. In RNA-seq studies, we identified several CDH-associated genes that are dysregulated in the E15.5 diaphragms of Gata4flox/flox;Prx1-Cre embryos. These embryos are an ideal model of the sac hernias that comprise 20% of human CDH cases. In Specific Aim #2, we will combine data from RNA-seq and BioChIP-seq analyses with the priority scores generated in Aim #1 to identify primary GATA4 target genes whose dysregulation contribute to the development of sac CDH. We will then determine if alterations in the expression of these target genes can cause CDH, or can modify the CDH phenotypes of GATA4-deficient mice. We have shown that SOX7 deficiency causes septal defects by decreasing endothelial-to-mesenchymal transition (EMT) in the developing heart. In RNA-seq and in situ hybridization studies we have shown that the expression of Wnt4—a key regulator of EMT in the endocardium—is severely decreased in E9.5 Sox7-/- hearts. In Specific Aim #3 we will determine if Wnt4 is a primary target of SOX7, and whether modulation of WNT4 or its downstream effectors can rescue SOX7-related cardiac phenotypes. Several lines of evidence suggest that WNT4 is a novel CDH/CVM gene in humans. To confirm this association, and learn more about the role of WNT4 in diaphragm and heart development, we will determine if WNT4 deficiency causes CDH/CVM in mice, if Sox7 and Wnt4 interact genetically in the development of CDH/CVM and if SOX7 regulates Wnt4 transcription in the developing diaphragm. Through these studies we will identify novel CDH and CDH/CVM genes and pathways. The bioinformatic tools we develop in this grant will enhance gene discovery efforts across multiple research fields.

项目摘要 先天性diaphragmantic疝（CDH）是一种威胁生命的先天缺陷，占所有主要的8％ 先天性异常。如果CDH与心血管畸形（CVM）共同出现，则死亡率 利率从30％增加到60％，长期发病率很常见。我们的目标是确定引起的基因 CDH和CDH/CVM，并发现它们控制隔膜和心脏发育的机制。 正确识别从大型候选基因列表中有助于特定表型的基因 在研究和临床研究中产生的数据是该研究和其他研究领域进步的主要障碍。 在特定的目标＃1中，我们将通过产生排名的CDH和CDH/CVM致病性来应对这一挑战 使用机器学习算法的所有REFSEQ基因的分数，该算法整合了大规模基因组的数据 知识来源。我们将使用这些分数来鉴定细胞遗传学的新型CDH和CDH/CVM基因 定义的关键区域并形成大型下一代测序数据库，作为我们多方面的一部分 新型基因发现方法。我们还将加速人类疾病基因的速度 通过使这些分数和我们的机器学习算法免费发现。 8p23.1包含gata4和sox7的微缺失是最常见的原因之一 CDH/CVM。在RNA-seq研究中，我们确定了几个与CDH相关的基因，这些基因在 E15.5 gata4flox/flox; prx1-cre胚胎的膜片。这些胚胎是囊疝的理想模型 20％的人类CDH病例。在特定的目标＃2中，我们将结合RNA-Seq和Biochip-Seq的数据 AIM＃1中生成的优先分数分析以识别主要GATA4目标基因 失调有助于SAC CDH的发展。然后，我们将确定表达式是否发生变化 这些靶基因可能会导致CDH，也可以改变GATA4缺陷小鼠的CDH表型。 我们已经表明，Sox7缺乏通过减少内皮到间质会导致间隔缺陷 发育中心的过渡（EMT）。在RNA-seq和原位杂交研究中，我们表明 Wnt4的表达（内膜中EMT的关键调节剂）在E9.5 Sox7 - / - 心中得到了严重改进。 在特定的目标＃3中，我们将确定Wnt4是否是Sox7的主要目标，以及Wnt4或 它的下游效应可以挽救与Sox7相关的心脏表型。 几条证据表明，Wnt4是人类中新型的CDH/CVM基因。确认这一点 关联，并了解有关Wnt4在隔膜和心脏发育中的作用的更多信息，我们将确定是否是否 Wnt4缺乏会导致小鼠的CDH/CVM，如果Sox7和Wnt4在开发中始终相互作用 CDH/CVM和IF SOX7调节发育中的隔膜中的Wnt4转录。 通过这些研究，我们将确定新颖的CDH和CDH/CVM基因和途径。生物信息学 我们在这笔赠款中开发的工具将增强多个研究领域的基因发现工作。