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

项目总结 唐氏综合征(DS)是最常见的染色体异常，每700名活产儿中就有一名受到影响。向上 到50%的先天DS患者，也被称为21三体(T21)，出生时就患有先天性心脏病 (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……里面 这是对父R01 HL130020-5的补充，我们建议利用我们建立的基于IPSC的 了解DS-CHD遗传学基础的实验模型。我们假设基因之间的相互作用 T21和非T21突变导致DS的CHD。病人。在目标1中，我们将执行全基因组- 对伴或不伴冠心病的DS患者进行测序以确定与SNV相关的单核苷酸变异(SNV) 患有DS-CHD。在目标2中，我们将验证SNV在存在CHD的情况下对CHD的致病性 T21。我们将使用CRISPR/Cas9校正SNV或将SNV插入到来自对照组和DS患者的IPSCs中，以建立 SNV和CHD之间的基因-表型关系。完成本附录的目标很可能 增加我们对DS相关CHD的了解，并扩大父母R01奖项的整体影响。 总而言之，这项包括行政补充建议将创造新的机会，以建立一个 功能和测序数据库以及IPSC生物资源库，将与多个 纪律调查员。深度临床表型、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 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.