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

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.

项目概要 本申请是为了响应 NOT-OD-19-071 而提交的。唐氏综合症（DS）是最常见的 在美国和世界各地的活产中常见的染色体异常。高达 DS 的 50% 患有室间隔缺损、房室间隔缺损等先天性心脏病（CHD）的患者 缺陷和法洛四联症。这些心脏异常引起的并发症会导致严重的发病率和 该人群的死亡率。然而，目前仍不清楚为什么额外的 21 号染色体的存在会导致 这些潜在致命性冠心病的发生，部分原因是缺乏足够的体内模型来克服 遗传组成存在种间差异，并且无法获取受 DS 影响的人类心肌细胞。 在此对母体 R01 HL141371 的补充中，我们建议利用源自患者的人类诱导的 多能干细胞 (iPSC) 平台，用于研究 DS 患者的 CHD 机制。我们假设 chr21 上心脏特异性、剂量敏感的三体基因的过度表达通过以下方式导致心脏缺陷 心脏串扰和心肌细胞成熟受损。目标 1 将生成 40 个 DS 特定的生物存储库 iPSC 线。为了研究细胞间串扰在 DS 相关 CHD 发病机制中的作用，我们将 工程iPSC-心脏类器官类似于心肌细胞、内皮细胞的心脏组织组成 细胞和成纤维细胞，并确定源自心脏类器官的分子和功能表型 DS iPSC。在目标 2 中，我们将使用泛组学方法研究 DS 相关 CHD 的机制。 将通过基因组编辑策略进一步研究已确定的候选基因的机制。 完成本补充的目标可能会增加我们对 DS 相关的先心病以及 扩大母公司 R01 奖项的整体影响力。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之间串扰的作用 发病机制中的心血管细胞类型。 DS 相关 CHD 的潜在机制涉及复杂的内部机制 细胞通讯导致发育和结构异常。因此，我们有信心， 利用 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 CHD 的理解，并促进新发现 对其发病机制可能至关重要的基因和途径。 总之，这项行政补充提案将为建立大规模的 功能和测序数据以及 iPSC 存储库将与研究人员广泛共享 CHD 和 DS。深度临床表型分析的结合，基于 iPSC 的工程心脏的使用 类器官组织、多组学分析和串扰信号研究无疑将有助于推动这一进程 领域向前发展并更好地了解 DS 相关的心脏病理学。