Elucidating Genotype-Phenotype Relationship of Polygenic Dilated Cardiomyopathies: Administrative Supplement (INCLUDE)

项目摘要 唐氏综合症（DS）是最常见的染色体异常，每700个活产婴儿中就有一个患有这种疾病。起来 50%的先天性心脏病患者，也称为21三体（T21），出生时患有先天性心脏病 （CHD）。尽管DS-CHD的发生率很高，但DS相关CHD的确切机制仍不清楚。在 在对母公司R 01 HL 130020 -5的补充中，我们建议利用我们已建立的基于iPSC的 实验模型，以了解DS-CHD的遗传基础。我们假设基因间的相互作用 T21和非T21突变之间的差异导致DS中的CHD。患者在目标1中，我们将进行全基因组- 在伴有或不伴有CHD的DS患者中进行单核苷酸测序（WGS），以鉴定与 关于DS-CHD在目的2中，我们将验证在存在以下物质的情况下SNV对CHD发展的致病性： T21我们将使用CRISPR/Cas9来校正或将SNV插入到来自对照和DS患者的iPSC中，以建立 SNV和CHD之间的基因型-表型关系。完成这一补充的目标可能会 增加我们对DS相关CHD的了解，并扩大R 01奖项的总体影响。 总而言之，这项INCLUDE行政补充提案将创造新的机会， 功能和测序数据库以及iPSC生物储存库，将与多个国家广泛共享， 纪律调查员。深入的临床表型分析、iPSC、CRISPR/Cas9和 多组学方法无疑将有助于推动这一领域的发展，并更好地了解DS相关的心脏疾病。 病理学

PROJECT SUMMARY Down syndrome (DS) is the most common chromosomal abnormality affecting one in every 700 live births. Up to 50% of individuals born with DS, also known as trisomy 21 (T21), are born with a congenital heart disease (CHD). Despite the high frequency of DS-CHD, the exact mechanisms of DS-related CHD remain unknown. In this Supplement to the Parent R01 HL130020-5, we propose to leverage our established iPSC-based experimental models to understand the genetic basis of DS-CHD. We hypothesize that an interaction of genes between T21 and non-T21 mutations causes CHD in DS. Patients. In Aim 1, we will perform whole genome- sequencing (WGS) in DS patients with or without CHD to identify single nucleotide variants (SNV) associated with DS-CHD. In Aim 2, we will validate the pathogenicity of SNVs on development of CHDs in the presence of T21. We will use CRISPR/Cas9 to correct or insert the SNVs to iPSCs from control and DS patients to establish a genotype-phenotype relationship between SNVs and CHDs. Completing the aims of this supplement will likely increase our understanding of DS-related CHD as well as broaden the overall impact of the parent R01 award. In summary, this INCLUDE Administrative Supplement proposal will create novel opportunities to build a massive functional and sequencing database as well as iPSC biorepository that will be broadly shared with multi- disciplinary investigators. The combination of deep clinical phenotyping, the use of iPSC, CRISPR/Cas9 and multi-omic approaches will undoubtedly help to move this field forward and better understand DS-related cardiac pathologies.