Dissecting the establishment and regulation of human pluripotency

剖析人类多能性的建立和调节

• 批准号: 8535278
• 负责人: Alexander Meissner
• 金额: $ 210.8万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8535278
• 关键词: Cell Culture Techniques; Cell Line; Cells; DNA-Protein Interaction; Disease model; Ensure; Epigenetic Process; Functional RNA; Gene Expression; Genetic; Genomics; Goals; Human; Human Development; Inheritance Patterns; Maintenance; Molecular; Preclinical Drug Evaluation; Property; Proteins; Proteomics; RNA; Recruitment Activity; Regenerative Medicine; Regulation; Repression; Role; Technology; Variant; X Chromosome; data integration; human disease; induced pluripotent stem cell; innovation; insight; pluripotency; programs; tool; transcription factor

DESCRIPTION (provided by applicant): Induced pluripotent stem (iPS) cells have a tremendous potential for aidvancing our understanding of human development and disease. To help unlock this potential we have organized a program to (A) comprehensively identify the genetic and epigenetic components of the regulatory network that maintains cells in a pluripotent state; (B) characterize culture-induced variation in the activites of these components in pluripotent cells; and (C) characterize temporal variation in their activities during induction of pluripotency with defined factors. To achieve these goals, we have formulated four interdependent projects: Project I (Meissner) will (1) characterize transcriptional coregulators and small non-coding RNAs that modulate the activity of the core pluripotency transcription factors, and (2) define and isolate subpopulations from pluripotent cell cultures to characterize their transcriptional and epigenetic states. Project II (Rinn) will characterize long non-coding RNAs expressed in pluripotent cells and elucidate their role in remodeling the epigenetic landscape during reprogramming. Project III (Mikkelsen) will characterize the cis-regulatory modules that direct activation, maintenace and repression of gene expression in pluripotent cells by recruiting transcription factors and their coregulators to key genomic loci. Project IV (Eggan) will characterize the inheritance patterns and maintenance of inactivated X chromosomes during reprogramming and in pluripotent cell cultures. The four projects rely on complementary use of innovative high-throughput genomic and proteomic technologies to profile high-quality iPS cell lines. The integration of data and insights from each of the projects will generate a comprehensive view of protein-protein, protein-RNA and protein-DNA interactions essential to the maintenace of pluripotency (goal A). This intergrated view will then guide studies of culture- induced and temporal variation in the network (goals B and C).

描述(申请人提供)：诱导多能干细胞(IPS)在帮助我们了解人类发育和疾病方面具有巨大的潜力。为了帮助释放这种潜力，我们组织了一个计划，以(A)全面识别维持细胞处于多能性状态的调控网络的遗传和表观遗传成分；(B)表征培养诱导的多能性细胞中这些成分活性的变化；以及(C)表征在诱导多能性的过程中，它们活动的时间变化。为了实现这些目标，我们制定了四个相互依存的项目： 项目I(Meissner)将(1)表征转录辅助调节因子和调节核心多能性转录因子活性的非编码小RNA，以及(2)定义和分离多潜能细胞培养的亚群，以表征它们的转录和表观遗传状态。 项目II(RINN)将描述在多能细胞中表达的长非编码RNA，并阐明它们在重新编程过程中重塑表观遗传格局的作用。 项目III(Mikkelsen)将描述顺式调控模块的特征，这些模块通过招募转录因子及其辅助调节因子到关键基因组座位来指导多能细胞中基因表达的激活、维持和抑制。 项目四(Eggan)将描述在重新编程和多能细胞培养过程中失活X染色体的遗传模式和维持。 这四个项目依赖于互补使用创新的高通量基因组和蛋白质组技术来描述高质量的iPS细胞系。综合每个项目的数据和见解，将全面了解对维持多能性至关重要的蛋白质-蛋白质、蛋白质-RNA和蛋白质-DNA相互作用(目标A)。然后，这种综合的观点将指导对网络中文化诱导和时间变化的研究(目标B和C)。