REGULATORY GENOMIC STUDIES IN A COHORT OF IPS CELL DERIVED CARDIOMYOCYTES

IPS 细胞来源的心肌细胞群体的调控基因组研究

• 批准号: 8300829
• 负责人: SYLVIA M EVANS
• 金额: $ 140.31万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8300829
• 关键词: Biology; Biometry; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cell model; Cells; Cellular Assay; Cellular biology; Clinical Medicine; DNA; Developed Countries; Disease; Disease susceptibility; Epigenetic Process; Fibroblasts; Generations; Genetic Variation; Genomics; Genotype; Goals; Human Genetics; Individual; Measures; Molecular Genetics; Molecular Profiling; Morbidity - disease rate; Phase; Positioning Attribute; Procedures; Protocols documentation; Quantitative Trait Loci; Reagent; Regulator Genes; Research; Research Personnel; Signal Transduction; Treatment Protocols; United States; Validation; Variant; abstracting; cohort; genome wide association study; high throughput technology; induced pluripotent stem cell; keratinocyte; mortality; research study; stem cell biology; trait

DESCRIPTION (provided by applicant): The overarching goal of our project is to use IPSC derived cardiomyocytes from genotyped individuals as cellular models to investigate how human genetic variation influences the gene regulatory networks Involved In cardiac biology and disease. Despite current treatment regimens, cardiovascular diseases remain the leading cause of morbidity and mortality in the United States and developed countries. Genome-wide association studies have identified a number of loci associated with cardiovascular disease susceptibility. Our Study will clarify the functional significance of these findings by combining cellular reprogramming Strategies with integrated molecular profiling and cellular assays. We have assembled a team of highly accomplished researchers in stem cell biology, cardiac cell biology, genomics, molecular genetics/epigenetics, biostatistics, and clinical medicine, and are well positioned to achieve the project goals within five years. After collecting fibroblasts and keratinocytes from individuals in the UCSD TSP cohort, the project will be carried out in three phases. In PHASE I, we will establish standardized reagents and procedures for the generation of iPSCs. Additionally, we will take advantage of our ongoing research efforts to increase the efficiency of cardiomyocyte differentiation to 80%, a substantial increase over current protocols (~20%). In PHASE II, we will develop cutting-edge technologies for high throughput generation of IPSCs, which will enable us to generate 600 iPSC lines (3 lines each from 200 individuals) in ~ 24 months. We will also scale our optimized protocols for deriving cardiomyocytes from the IPSC lines. In PHASE III we will initially perform validation experiments to measure the genomic profile variability between isogenic (derived from the same individual) cardiomyocytes. We will then use the derived cardiomyocytes to 1) Identify and characterize the causal DNA variants underlying strong GWAS signals with electrocardiographic traits; and 2) Identify expressed quantitative traits loci (eQTL) in the cohort of IPSC derived cardiomyocytes at baseline (untreated) and after stimulation. (End of Abstract)

描述（由申请人提供）： 我们项目的总体目标是使用来自基因型个体的 IPSC 衍生心肌细胞作为细胞模型来研究人类遗传变异如何影响涉及心脏生物学和疾病的基因调控网络。尽管有目前的治疗方案，心血管疾病仍然是美国和发达国家发病和死亡的主要原因。全基因组关联研究已经确定了许多与心血管疾病易感性相关的位点。我们的研究将通过将细胞重编程策略与综合分子分析和细胞测定相结合来阐明这些发现的功能意义。我们组建了一支由干细胞生物学、心脏细胞生物学、基因组学、分子遗传学/表观遗传学、生物统计学和临床​​医学领域卓有成效的研究人员组成的团队，并有能力在五年内实现项目目标。在从 UCSD TSP 队列中的个体中收集成纤维细胞和角质形成细胞后，该项目将分三个阶段进行。在第一阶段，我们将建立用于生成 iPSC 的标准化试剂和程序。此外，我们将利用我们正在进行的研究工作，将心肌细胞分化的效率提高到 80%，比当前方案 (~20%) 大幅提高。在第二阶段，我们将开发用于高通量生成 IPSC 的尖端技术，这将使我们能够在约 24 个月内生成 600 个 iPSC 系（每个系来自 200 个人 3 个系）。我们还将扩展我们的优化方案，用于从 IPSC 系中获取心肌细胞。在第三阶段，我们将首先进行验证实验，以测量同基因（源自同一个体）心肌细胞之间的基因组谱变异性。然后，我们将使用衍生的心肌细胞来 1) 识别和表征具有心电图特征的强 GWAS 信号背后的因果 DNA 变异； 2) 鉴定基线（未处理）和刺激后 IPSC 衍生心肌细胞群中表达的数量性状位点 (eQTL)。 （摘要完）