Human iPSC Model for Elucidating Crosstalk Signaling and Secretomes: Down Syndrome Administrative Supplement

项目总结 此申请是针对NOT-OD-19-071提交的。唐氏综合征(DS)是最常见的 在美国和世界范围内，活产婴儿中常见的染色体异常。高达50%的DS 患有先天性心脏病(CHD)的患者，如室间隔缺损、房室间隔 缺陷和法洛四联症。这些心脏畸形的并发症会导致重大疾病和 这一人口中的死亡率。然而，为什么额外的21号染色体的存在会导致 这些潜在致命的CHD的发展，部分原因是缺乏足够的体内模型来克服 基因组成的种间差异和不能接触DS影响的人类心肌细胞。 在亲本R01 HL141371的这一补充中，我们建议利用患者来源的人类诱导 多能干细胞(IPSC)平台用于研究DS患者的CHD机制。我们假设 心脏特异的、剂量敏感的三体基因在chr21上的过度表达通过以下途径导致心脏缺陷 心脏串扰和心肌细胞成熟受损。AIM 1将生成一个包含40个特定DS的生物信息库 IPSC系列。为了研究细胞间串扰在DS相关冠心病发病机制中的作用，我们将 工程师IPSC-心脏有机体类似于心肌细胞的心脏组织组成，内皮细胞 细胞和成纤维细胞，并测定来源于心脏器官的分子和功能表型 DS iPSCs。在目标2中，我们将用全基因组方法研究DS相关CHD的发病机制。 已确定的候选基因的机制将通过基因组编辑策略进行进一步研究。 完成本附录的目标可能会增加我们对DS相关CHD以及 扩大母公司R01奖项的整体影响。在家长奖中，我们使用IPSC技术来 体外鉴定遗传性心肌病的机制并剖析串扰在其中的作用 心血管细胞在发病机制中的类型。DS相关CHD的发病机制涉及复杂的相互作用 细胞通讯导致发育和结构异常。因此，我们有信心 DS和非DS CHD IPSC来源的体外比较和生物信息学分析 心肌细胞可能会扩大我们对CHD的理解，并有助于发现新的 可能在其发病机制中起关键作用的基因和途径。 总而言之，这项行政副刊提案将创造新的机会，建立一个 功能和测序数据以及IPSC存储库，将与调查人员广泛共享 CHD和DS。结合深临床表型，使用IPSC为基础的工程心脏 有机组织、多基因组图谱和串扰信号研究无疑将有助于推动这一进程 更好地理解DS相关的心脏病理。

PROJECT SUMMARY This application is being submitted in response to NOT-OD-19-071. Down syndrome (DS) is the most commonly occurring chromosomal abnormality in live births in the US and worldwide. Up to 50% of DS patients suffer from congenital heart disease (CHD) such as ventricular septal defect, atrioventricular septal defect, and tetralogy of Fallot. Complications from these cardiac anomalies cause major morbidities and mortalities in this population. Yet, it is still unknown why the presence of extra chromosome 21 leads to the development of these potentially fatal CHD, due in part to a lack of adequate in vivo model overcoming interspecies differences in genetic composition and inability to access DS-affected human cardiomyocytes. In this Supplement to the Parent R01 HL141371, we propose to leverage patient-derived human induced pluripotent stem cell (iPSC) platform towards studying mechanisms of CHD in DS patients. We hypothesize overexpression of cardiac-specific, dosage-sensitive trisomic genes on chr21 leads to heart defects through impaired cardiac crosstalk and myocyte maturation. Aim 1 will generate a biorepository of 40 DS-specific iPSC lines. To investigate the role of intercellular crosstalk in the pathogenesis of DS-related CHD, we will engineer iPSC-cardiac organoids resembling the heart tissue composition of cardiomyocytes, endothelial cells, and fibroblasts and determine molecular and functional phenotypes of cardiac organoids derived from the DS iPSCs. In Aim 2, we will investigate the mechanism of DS-related CHD using a pan-omic approach. The mechanisms of identified gene candidates will be further investigated through genome editing strategy. 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 the parent award, we are using iPSC technology to identify mechanisms of genetic cardiomyopathy in vitro and dissecting the role of crosstalk between cardiovascular cell types in pathogenesis. Mechanism underlying DS-related CHD involves complex inter- cellular communication leading to developmental and structural anomalies. Hence, we are confident that a comparative in vitro and bioinformatics analysis utilizing both DS and non-DS CHD iPSC-derived cardiomyocytes will likely extend our understanding of CHD as well as facilitate the discovery of novel genes and pathways that may be critical in its pathogenesis. In summary, this Administrative Supplement proposal will create novel opportunities to build a massive functional and sequencing data as well as iPSC repository that will be broadly shared with investigators of CHD and DS. The combination of deep clinical phenotyping, the use of iPSC-based engineered cardiac organoid tissues, multi-omic profiling, and cross-talk signaling studies will undoubtedly help to move this field forward and better understand DS-related cardiac pathologies.