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

用于阐明串扰信号和分泌组的人类 iPSC 模型：唐氏综合症行政补充

• 批准号: 9897087
• 负责人: Joseph C. Wu
• 金额: $ 31.16万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-19 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9897087
• 关键词: Administrative Supplement; Adult; Affect; Apoptosis; Attention; Award; Bioinformatics; CRISPR interference; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Line; Cell Nucleus; Cell model; Cells; Chromatin; Chromosome abnormality; Chromosomes, Human, Pair 21; Clinical; Collaborations; Communication; Complex; Cytogenetics; Data; Development; Disease; Down Syndrome; Endothelial Cells; Engineering; Experimental Models; Fibroblasts; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Genotype; Goals; Grant; Heart; Heart Abnormalities; Human; Impairment; In Vitro; Intervention; Live Birth; Maps; Medical Genetics; Metabolism; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Multiomic Data; Muscle Cells; Organoids; Parents; Pathogenesis; Pathology; Pathway interactions; Patient Recruitments; Patients; Pediatrics; Phenotype; Population; Publications; Research; Research Personnel; Role; Signal Transduction; Structural defect; Structure; Tetralogy of Fallot; Time; Tissues; Ventricular Septal Defects; Work; atrioventricular septal defect; base; biobank; cardiogenesis; cell type; clinical phenotype; comparative; congenital heart disorder; disease phenotype; dosage; genome editing; heart cell; in vivo Model; induced pluripotent stem cell; inherited cardiomyopathy; intercellular communication; mortality; multiple omics; next generation sequencing; novel; overexpression; programs; recruit; repository; response; stem cell technology; transcriptomics; Administrative Supplement; Adult; Affect; Apoptosis; Attention; Award; Bioinformatics; CRISPR interference; Candidate Disease Gene; Cardiac; Cardiac Myocytes; Cardiovascular system; Cell Line; Cell Nucleus; Cell model; Cells; Chromatin; Chromosome abnormality; Chromosomes, Human, Pair 21; Clinical; Collaborations; Communication; Complex; Cytogenetics; Data; Development; Disease; Down Syndrome; Endothelial Cells; Engineering; Experimental Models; Fibroblasts; Gene Expression; Genes; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Genotype; Goals; Grant; Heart; Heart Abnormalities; Human; Impairment; In Vitro; Intervention; Live Birth; Maps; Medical Genetics; Metabolism; Mitochondria; Modeling; Molecular; Morbidity - disease rate; Multiomic Data; Muscle Cells; Organoids; Parents; Pathogenesis; Pathology; Pathway interactions; Patient Recruitments; Patients; Pediatrics; Phenotype; Population; Publications; Research; Research Personnel; Role; Signal Transduction; Structural defect; Structure; Tetralogy of Fallot; Time; Tissues; Ventricular Septal Defects; Work; atrioventricular septal defect; base; biobank; cardiogenesis; cell type; clinical phenotype; comparative; congenital heart disorder; disease phenotype; dosage; genome editing; heart cell; in vivo Model; induced pluripotent stem cell; inherited cardiomyopathy; intercellular communication; mortality; multiple omics; next generation sequencing; novel; overexpression; programs; recruit; repository; response; stem cell technology; transcriptomics

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.

项目摘要 该申请正在响应于NOT-OD-19-071。唐氏综合症（DS）是最大的 在美国和全球的活产中，通常发生染色体异常。多达50％的DS 患者患有先天性心脏病（CHD），例如心室间隔缺陷，心室间隔 缺陷和法洛的四部曲。这些心脏异常引起的并发症会导致主要病毒和 该人群的死亡人数。然而，仍然不知道为什么额外的染色体21导致 这些潜在致命的冠心病的发展部分是由于缺乏足够的体内模型克服 种间遗传组成和无法进入受DS影响的人类心肌细胞的差异。 在父母R01 HL141371的补充中，我们建议利用患者衍生的人类诱导的 多能干细胞（IPSC）平台，用于研究DS患者中冠心病的机制。我们假设 心脏特异性，对剂量敏感的三方基因的过表达在CHR21上导致心脏缺陷 心脏串扰和心肌成熟受损。 AIM 1将产生40 ds特异性的生物座 IPSC线。为了研究细胞间串扰在与DS相关CHD发病机理中的作用，我们将 工程师ipsc-cardiac类器官类似于心肌细胞的心脏组织组成，内皮 细胞和成纤维细胞，并确定源自源自的心脏器官的分子和功能表型 DS IPSC。在AIM 2中，我们将使用Pan-Omic方法研究与DS相关的CHD的机制。 将通过基因组编辑策略进一步研究已确定的基因候选物的机制。 完成这种补充的目的可能会增加我们对DS相关冠心病的理解以及 扩大父母R01奖的整体影响。在父母奖中，我们使用IPSC技术来 鉴定体外遗传性心肌病的机制，并剖析串扰的作用 发病机理中的心血管细胞类型。与DS相关的CHD的基础机制涉及复杂的间隔 细胞通信导致发育和结构异常。因此，我们相信 使用DS和非DS CHD IPSC衍生的比较体外和生物信息学分析 心肌细胞可能会扩展我们对冠心病的理解，并促进新颖的发现 可能在其发病机理中至关重要的基因和途径。 总而言之，这项行政补充提案将创造新的机会来建立大规模的 功能和测序数据以及IPSC存储库，将与研究人员大致共享 CHD和DS。深层临床表型的结合，基于IPSC的工程心脏的使用 器官组织，多摩尼克分析和跨对信号研究无疑将有助于移动这一点 野外前进，更好地了解与DS相关的心脏病理。