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: 10553279
• 负责人: BERNICE E MORROW
• 金额: $ 76.31万
• 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/10553279
• 关键词: 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; Maps; Modeling; Molecular; Mus; Mutant Strains Mice; Neural Crest; Nucleotides; Operative Surgical Procedures; Outcome; Pathogenesis; Patients; Persistent Truncus Arteriosus; Pharyngeal Apparatus; Phenotype; Quality Control; Quantitative Reverse Transcriptase PCR; Regulator Genes; 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; cell motility; congenital heart disorder; cost; 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; model organism; mouse model; mutant; novel; phenotypic data; programs; risk prediction; 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.

摘要 22q11.2缺失综合征（22q11.2DS; DiGeorge综合征）是一种罕见的出生缺陷疾病，影响1/4000 活产这种情况是完全渗透的，但表现力不同，提供了机会，以确定遗传 修饰语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学方法。大约60%的 22q11.2DS患者有CHD。在受影响的患者中，大多数有心脏流出道（OFT）缺陷， 从而在胚胎发生期间破坏主动脉和肺动脉干的形成。畸形在 患者从轻度到重度不等，大约一半的患者在出生后需要手术才能生存。典型改变 包括法洛四联症、永存动脉干或右侧主动脉弓伴室间隔缺损。 OFT来源于从胚胎咽器迁移到心脏的细胞。TBX 1基因， 编码T-box转录因子的基因在咽器中表达。我们的假设是 TBX 1的单倍不足，映射到22 q11.2上的关键缺失区域，沿着其他地方的修饰， 基因组对患者的总体表型有贡献。为了鉴定CHD的遗传修饰物，我们获得了 全基因组序列（WGS）来自1，182例22q11.2DS受试者。最近我们得到了一份合同 来自NIH CIDR的895名额外的DNA在手的受试者的WGS。我们进一步发起了与 G2 MH网络为另外500人获得WGS，所有这些都是免费的，总共从2，577人获得WGS 22q11.2DS患者。我们的目标1是研究罕见的单核苷酸编码，非编码和 结构变异以鉴定CHD的遗传修饰剂。我们建议采取基于基因组的方法， 在TBX 1网络中的基因，从我们最近的单细胞转录组实验中确定的控制和 TBX 1小鼠胚胎功能丧失。我们还将研究用于散发性肝癌遗传研究的基因组， 一般人群中的CHD，以确定22q11.2DS的修饰因子是否也可作为CHD的风险因素。 一般冠心病。我们正在利用一种新的贝叶斯优先级方法来加权基因组中的基因， 其功能的重要性。作为初步的数据，我们确定了染色质调控基因， 增加22q11.2DS受试者和散发性CHD的CHD风险。在目标2中，我们将执行功能和 使用小鼠模型对该计划中发现的基因进行的机制研究。我们将进一步进行生物 在斑马鱼模型系统中使用基因编辑对该程序中鉴定的DNA变体进行验证研究。 该项目将阐明22q11.2DS中CHD的分子发病机制，这些分子发病机制也可能是CHD的危险因素。 一般人群中的散发性CHD。了解这些应该会改善诊断并在未来有所帮助， 了解受影响患者不同结局的基础。