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

基本信息

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
项目类别：
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

项目摘要

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；迪乔治综合征）是一种罕见的出生缺陷疾病，影响 1/4000 活产数。这种情况是完全渗透的，但表达性各不相同，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通过采用人类和动物模型遗传方法，在 22q11.2DS 患者中（CHD）。大约 60% 22q11.2DS 患者患有 CHD。在受影响的患者中，大多数有心脏流出道（OFT）缺陷，从而破坏胚胎发生过程中主动脉和肺干的形成。畸形现象在患者的病情从轻度到重度不等，大约一半需要手术才能在出生后生存。典型发现包括法洛四联症、持续性动脉干或伴有室间隔缺损的右侧主动脉弓。 OFT 源自从胚胎咽器迁移到心脏的细胞。 TBX1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 TBX1 的单倍体不足，映射到 22q11.2 上的关键删除区域，以及其他位置的修饰符基因组对患者的整体表型有贡献。为了确定 CHD 的遗传修饰因素，我们获得了来自 1,182 名 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 名额外受试者的全基因组测序（WGS），其中手头有 DNA。我们进一步启动了与 G2MH 网络将另外 500 名患者获得 WGS，所有这些都无需该计划承担任何费用，总共 2,577 名患者获得 WGS 具有 22q11.2DS 的受试者。我们的目标 1 是研究罕见的单核苷酸编码、非编码和结构变异来识别冠心病的遗传修饰物。我们建议采取基于基因集的方法，重点关注 TBX1 网络中的基因是从我们最近的单细胞转录组实验中鉴定出来的 Tbx1 小鼠胚胎功能丧失。我们还将研究用于散发性遗传研究的基因集一般人群中的 CHD 以确定 22q11.2DS 的修饰因子是否也可以作为以下疾病的危险因素一般冠心病。我们正在利用一种新颖的贝叶斯优先级方法来对基于基因组中的基因进行加权根据它们的功能重要性。作为初步数据，我们确定了染色质调控基因可能增加受试者 22q11.2DS 和散发性 CHD 的风险。在目标 2 中，我们将执行功能性和使用小鼠模型对该程序中发现的基因进行机制研究。我们将进一步开展生物使用斑马鱼模型系统中的基因编辑对该程序中识别的 DNA 变异进行验证研究。该计划将阐明 22q11.2DS 中 CHD 的分子发病机制，这也可能是以下疾病的危险因素：普通人群中的散发性冠心病。了解这些应该可以改善诊断并在未来有所帮助，了解受影响患者不同结果的基础。