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.

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。这种情况是完全穿透的，但表现力不同，为鉴定基因提供了机会。 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 采用人和动物模型遗传学方法对22q11.2DS患者进行遗传分析。约60%的 22q11.2DS患者合并冠心病。在受影响的患者中，大多数都有心脏流出道(OFT)缺陷， 从而在胚胎发育过程中扰乱了主动脉和肺动脉干的形成。地球上的畸形 患者从轻到重不等，大约一半的患者需要手术才能在出生后存活。典型调查结果 包括法洛四联症、永存动脉干或右主动脉弓合并室间隔缺损。 OFT来源于从胚胎咽部器官迁移到心脏的细胞。TBX1基因， 编码T-box转录因子在咽部器官中表达。我们的假设是 Tbx1的单倍性不足，映射到22q11.2上的关键缺失区域，以及 基因组对患者的整体表型有贡献。为了确定冠心病的遗传修饰因素，我们获得了 1182例22q11.2DS受试者的全基因组序列(WGS)。最近我们获得了一份合同 来自NIH CIDR的895名手持DNA的额外受试者的WGS。我们进一步发起了与 G2MH网络将以另外500美元获得WGS，所有费用都不会对此计划产生任何影响，WG总数为2577 22q11.2DS受试者。我们目标1的目标是研究罕见的单核苷酸编码、非编码和 用于识别冠心病遗传修饰因子的结构变异。我们建议采用基于基因集的方法来聚焦 关于从我们最近的单细胞转录实验中确定的TBX1网络中的基因 TBX1功能丧失的小鼠胚胎。我们还将调查用于散发性遗传病遗传学研究的基因组。 以确定22q11.2DS的修饰符是否也可作为冠心病的危险因素 普通冠心病。我们正在利用一种新的贝叶斯优先排序方法对基于基因集的基因进行加权 它们的功能重要性。作为初步数据，我们确定了染色质调节基因 22q11.2ds受试者患冠心病和散发性冠心病的风险增加。在目标2中，我们将执行函数和 利用小鼠模型对这个项目中发现的基因进行机制研究。我们将进一步演绎生物 在斑马鱼模型系统中使用基因编辑对该计划中确定的DNA变体进行验证研究。 本计划将阐明22q11.2ds的CHD的分子发病机制，这些因素也可能是 普通人群中的散发性冠心病。了解这些应该会改进诊断，并在未来有所帮助， 以了解受影响患者不同结局的基础。