Genetic modifiers of congenital heart disease in 22q11.2 deletion syndrome

22q11.2缺失综合征先天性心脏病的遗传修饰

• 批准号: 10373375
• 负责人: BERNICE E MORROW
• 金额: $ 79.71万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10373375
• 关键词: 22q11; 22q11.2; ATAC-seq; Affect; Animal Model; Aorta; Award; Biological; Biological Models; Biology; Birth; CRISPR/Cas technology; Cardiac; Cells; ChIP-seq; Chromatin; Chromosomes; Code; Collaborations; Congenital Abnormality; Congenital Heart Defects; Contracts; DNA; Data; Defect; Development; DiGeorge Syndrome; Diagnostic; Disease; Elements; Embryo; Embryonic Development; Etiology; Future; Gene Expression Profile; General Population; Genes; Genetic; Genetic study; Genome; Goals; Hand; Heart; Heart Abnormalities; Human; Human Genetics; Immunofluorescence Immunologic; In Situ Hybridization; Incidence; Individual; Investigation; Joints; Left; Light; Live Birth; Lung; Modeling; Molecular; Mus; Mutant Strains Mice; Neural Crest; Nucleotides; Operative Surgical Procedures; Outcome; Pathogenesis; Patients; Persistent Truncus Arteriosus; Pharyngeal Apparatus; Phenotype; Quality Control; Regulator Genes; Reverse Transcriptase Polymerase Chain Reaction; Risk; Risk Factors; Sampling; Side; Single Nucleotide Polymorphism; Study models; Syndrome; System; Testing; Tetralogy of Fallot; United States National Institutes of Health; Untranslated RNA; Validation; Variant; Ventricular Septal Defects; Weight; Zebrafish; aortic arch; base; congenital heart disorder; cost; developmental genetics; differential expression; experimental study; functional genomics; gene discovery; genetic approach; genetic architecture; genetic association; genomic data; human model; improved; insertion/deletion mutation; interest; loss of function; malformation; mouse model; mutant; novel; phenotypic data; programs; single-cell RNA sequencing; spatiotemporal; stem cells; transcription factor; transcriptomics; validation studies; whole genome

ABSTRACT The 22q11.2 deletion syndrome (22q11.2DS; DiGeorge syndrome) is a rare birth defect disorder affecting 1/4000 live births. This condition is fully penetrant, but expressivity varies, providing the opportunity to identify genetic modifiers. Our main interest is to understand the developmental and genetic basis of congenital heart disease (CHD) in 22q11.2DS patients by taking human and animal model genetic approaches. Approximately 60% of patients with 22q11.2DS have CHD. Among affected patients, most have cardiac outflow tract (OFT) defects, thereby disrupting the formation of the aorta and pulmonary trunk during embryogenesis. The malformations in patients vary from mild to severe and approximately half require surgery for survival after birth. Typical findings include tetralogy of Fallot, persistent truncus arteriosus or right sided aortic arch with a ventricular septal defect. The OFT derives from cells that migrate from the embryonic pharyngeal apparatus to the heart. The TBX1 gene, encoding a T-box transcription factor is expressed in the pharyngeal apparatus. Our hypothesis is that haploinsufficiency of TBX1, mapping to the critical deleted region on 22q11.2, along with modifiers elsewhere in the genome contribute to the overall phenotype in patients. To identify genetic modifiers of CHD, we obtained whole genome sequence (WGS) from 1,182 subjects with 22q11.2DS. Recently we were awarded a contract from NIH CIDR for WGS of 895 additional subjects with DNA in hand. We further initiated a collaboration with the G2MH Network to obtain WGS for another 500, all at no cost to this program, for a total of WGS from 2,577 subjects with 22q11.2DS. Our goal for Aim 1 is to investigate rare single nucleotide coding, non-coding and structural variants to identify genetic modifiers of CHD. We propose to take a gene-set based approach focusing on genes in the TBX1 network identified from our recent single cell transcriptomic experiments on control and Tbx1 loss of function mouse embryos. We will also investigate gene-sets used for genetic studies of sporadic CHD in the general population to determine whether modifiers for 22q11.2DS may also serve as risk factors for general CHD. We are utilizing a novel Bayesian prioritization approach to weight genes in gene-sets based upon their functional importance. As preliminary data we identified chromatin regulatory genes that might increase risk to CHD in subjects 22q11.2DS and for sporadic CHD. In Aim 2, we will perform functional and mechanistic studies of genes discovered in this program using mouse models. We will further perform biological validation studies of DNA variants identified in this program using gene editing in the zebrafish model system. This program will elucidate the molecular pathogenesis of CHD in 22q11.2DS that might also be risk factors for sporadic CHD in the general population. Understanding these should improve diagnostics and help in the future, for understanding the basis of varying outcomes in affected patients.

抽象的 22q11.2缺失综合征（22q11.2ds; digeorge综合征）是一种罕见的产生缺陷障碍，影响1/4000 活产。这种情况是完全渗透的，但表现力有所不同，提供了识别遗传的机会 修饰符。我们的主要兴趣是了解先天性心脏病的发育和遗传基础 （CHD）在22q11.2ds患者中，采用人类和动物模型遗传方法。大约60％ 22q11.2d的患者患有冠心病。在受影响的患者中，大多数患有心脏流出路（通常）缺陷， 从而破坏胚胎发生过程中主动脉和肺部躯干的形成。畸形 从轻度到重度和大约一半的患者需要手术才能出生后的生存。典型的发现 包括法洛的四部曲，持续的毛动脉桥或右侧主动脉弓具有心室间隔缺陷。 OFT源自从胚胎咽型迁移到心脏的细胞。 tbx1基因， 编码T-box转录因子在咽式设备中表达。我们的假设是 TBX1的单倍不足，映射到22q11.2的关键删除区域，以及其他位置的修饰符 基因组有助于患者的整体表型。为了鉴定冠心病的遗传修饰符，我们获得了 来自22q11.2ds的1,182名受试者的全基因组序列（WGS）。最近我们被授予合同 从NIH CIDR进行895名其他受试者的WG，手里拿着DNA。我们进一步与 G2MH网络将获得另外500个WG的G2MH网络，全部无需支付该计划，总共2,577个WGS 22q11.2ds的受试者。 AIM 1的目标是研究稀有的单核苷酸编码，非编码和 结构变体鉴定冠心病的遗传修饰剂。我们建议采用基于基因的方法聚焦 关于从我们最近的单细胞转录组实验在对照和对照和 TBX1功能小鼠胚胎的丧失。我们还将研究用于零星的遗传研究的基因组 一般人群中的CHD，以确定22q11.2ds的修饰符是否也可以作为风险因素 一般冠心。我们利用一种基于基因基因的新型贝叶斯优先级方法来体重基因 具有其功能重要性。作为初步数据，我们确定了可能的染色质调节基因 增加受试者22q11.2ds和零星冠心病的风险。在AIM 2中，我们将执行功能和 使用小鼠模型在该程序中发现的基因的机理研究。我们将进一步执行生物学 在该程序中使用斑马鱼模型系统中的基因编辑确定的DNA变体的验证研究。 该程序将阐明22q11.2ds中冠心病的分子发病机理，这也可能是危险因素 普通人群中零星的冠心病。了解这些应该改善诊断和将来的帮助， 了解受影响患者的结果不同的基础。