The regulatory role of chromatin interaction in pluripotency and differentiation

染色质相互作用在多能性和分化中的调节作用

• 批准号: 8918305
• 负责人: Justin Brumbaugh
• 金额: $ 5.8万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8918305
• 关键词: Adult; Architecture; Area; Basic Science; Binding; Biochemistry; Biological; Biology; Cell Nucleus; Cells; Cellular biology; ChIP-seq; Chromatin; Chromatin Structure; Chromosomes; Complex; Coupled; DNA; Data; Data Analyses; Development; Disease model; Distant; Epigenetic Process; Gene Expression; Gene Expression Regulation; Gene Structure; Genes; Genetic Transcription; Genomic Segment; Genomics; Goals; Health; Human; Immunoprecipitation; Institution; Knock-out; Knowledge; Laboratories; Learning; Maps; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Mentors; Methods; Molecular Conformation; Monitor; Multipotent Stem Cells; Play; Pluripotent Stem Cells; Process; Proteins; RNA; Regenerative Medicine; Regulation; Regulatory Element; Relative (related person); Research; Research Design; Research Project Grants; Resources; Role; Sampling; Site; Source; Staging; Stem cells; Students; System; Technology; Testing; Therapeutic Human Experimentation; Tissue Engineering; Training; Training Programs; Western Blotting; Work; Writing; adult stem cell; career development; cell type; chromatin modification; cohesin; deep sequencing; drug testing; embryonic stem cell; epigenetic regulation; genome-wide; improved; induced pluripotent stem cell; instrumentation; interest; large-scale database; meetings; multipotent cell; nerve stem cell; novel; overexpression; pluripotency; public health relevance; research study; self-renewal; stem cell biology; tool; trait

DESCRIPTION (provided by applicant): Pluripotent stem cells are a renewable cell source with the capacity to differentiate into any specialized cell in the adult body. These traits make pluripotent cells a tractable tool for studying normal and dysfunctional biological networks in the context of development. Therefore, pluripotent cells have the potential to greatly impact human health through drug testing, disease modeling, tissue engineering, and regenerative medicine. Harnessing this potential requires a detailed understanding of the biology that governs pluripotency and directs differentiation. The mechanisms that guide these processes have been studied extensively at the protein and RNA level; however, the role of epigenetic regulation has only recently come into focus. A largely unexplored aspect of epigenetic regulation is the spatial organization of genes relative to distant genomic regions and regulatory elements. It is our central hypothesis that genomic regions important for pluripotency associate in the nucleus and change upon induction of differentiation and reprogramming. Charting DNA interaction and its dynamics will resolve this regulatory system in pluripotent cells and may provide a means to manipulate these cells for research and therapeutic purposes. Specific Aims: The first aim in this proposal will map long-range DNA interaction for key pluripotency genes in pluripotent cells. The second aim tracks chromatin interactions at various stages of development and also explores the dynamics of DNA interaction during reprogramming. The third aim identifies novel protein components that mediate chromatin interaction and applies biochemistry to characterize these proteins. Study Design: We propose research that combines expertise in stem cell biology with emerging genomic technology. Specifically, we will use modified circular chromosome conformation capture (m4C) to examine DNA interaction for key pluripotency genes Oct4 and Sox2 in pluripotent and multipotent stem cells. We will also track chromatin organization following the induction of pluripotency by sampling cells at intermediate stages of reprogramming. By characterizing proteins that mediate chromatin structure, we will determine key epigenetic regulators. Through targeted knockdown of these regulators, we will establish their functional role in pluripotency. These data, coupled to existing gene expression, chromatin modification, and ChIP results will reveal in unparalleled detail the epigenetic regulatory mechanisms at play during pluripotency, differentiation, and reprogramming. The sponsoring laboratory has numerous resources available to accomplish these goals, including instrumentation and technical training for m4C and deep sequencing. Resources are also available through the host institution for computational support and large-scale data analysis. The training program includes numerous opportunities for career development through cutting edge research, grant writing, and student mentoring.

描述（由申请人提供）：多能干细胞是一种可再生的细胞来源，具有分化成成人体内任何特化细胞的能力。这些特性使多能性细胞成为研究人体正常和不正常生物网络的一种易于操作的工具