Genome-Wide Profiling of REST/NRSF Targets in Adult Neural Stem Cells

成体神经干细胞 REST/NRSF 靶标的全基因组分析

• 批准号: 8274637
• 负责人: Jenny Hsieh
• 金额: $ 19.86万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-06 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8274637
• 关键词: Activities of Daily Living; Address; Adult; Applications Grants; Binding; Bioinformatics; Biological Assay; Brain; Brain Diseases; Brain Injuries; Cell Culture Techniques; Cells; ChIP-seq; Chromatin; Clinical; Cytoplasmic Granules; Data; Development; Disease; Epigenetic Process; Exploratory/Developmental Grant; Funding; Gene Expression; Gene Expression Profile; Gene Targeting; Genome; Genomics; Goals; Grant; Hippocampus (Brain); Homeostasis; In Vitro; Individual; Knockout Mice; Knowledge; Laboratories; Lead; Learning; Life; Link; Memory; Mental disorders; Mission; Modification; Molecular; Natural regeneration; Nerve Regeneration; Nerve Tissue; Nervous System Trauma; Neuronal Differentiation; Neurons; Neurosciences; Pharmaceutical Preparations; Physiological; Production; Proliferating; Public Health; RNA; Reporter; Research; Research Proposals; Role; Stem cells; Structure of germinal center of lymph node; System; Techniques; Testing; Therapeutic; Time; Transcription Repressor/Corepressor; United States; Work; adult neurogenesis; cell type; cognitive change; cognitive function; epigenomics; exhaust; genome wide association study; genome-wide; high risk; improved; innovation; insight; nerve stem cell; nervous system disorder; neurodevelopment; neurogenesis; neuron loss; newborn neuron; prevent; programs; repaired; research study; response; stem cell biology

DESCRIPTION (provided by applicant): An essential guardian of the neuronal genome is the transcriptional repressor REST/NRSF. The long-term goal is to better understand the transcriptional and epigenetic circuitry important for neuronal cell fate, in both physiological and pathological contexts. The objective of this grant proposal is to generate a genome-wide view of REST/NRSF target genes by performing ChIP-Seq and RNA-seq in adult neural stem cells. The central hypothesis is REST/NRSF is essential for stem cell quiescence and is a master negative regulator of adult hippocampal neurogenesis. The rationale for the proposed research is that understanding developmental mechanisms controlling the neuronal lineage program have the potential to further therapeutic approaches to an increasing number of mental disorders linked to neuronal loss or aberrant neuronal function. Thus, the proposed study is relevant to that part of NIH's mission that relates to gaining fundamental knowledge that will potentially help treat neurological disorders. Guided by strong preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Genome-wide identification of REST/NRSF targets in adult NSC chromatin and 2) Transcriptome analysis of adult NSCs during quiescence, proliferation, and neuronal differentiation. Aim 1 will focus on performing ChIP-Seq of REST/NRSF in adult hippocampal neural stem cells. Aim 2 will focus on performing RNA-seq from adult NSCs under conditions of quiescence, proliferation, and neuronal differentiation. Aim 2 will also identify which targets are dependent on REST/NRSF by comparing ChIP-Seq targets to gene expression analysis from REST/NRSF conditional knockout mice generated in our laboratory. The conceptual framework and approach is innovative, because it is the first study to apply state-of-the-art deep sequencing techniques to a physiologically relevant adult neural stem cell system in order to identify a genome-wide view of REST/NRSF target genes. The proposed R21 grant is significant, because it is expected to advance and expand our basic understanding of transcriptional networks regulating stem cell quiescence, proliferation, and neuronal differentiation which may be of particular importance for restoring cognitive function after brain damage or disease.

描述（由申请人提供）：神经元基因组的一个重要监护人是转录抑制因子REST/NRSF。长期目标是更好地了解在生理和病理背景下对神经元细胞命运重要的转录和表观遗传电路。这项资助提案的目的是通过在成人神经干细胞中进行ChIP-Seq和RNA-seq来产生REST/NRSF靶基因的全基因组视图。中心假设是REST/NRSF是干细胞静止所必需的，并且是成年海马神经发生的主要负调节因子。拟议研究的基本原理是，了解控制神经元谱系程序的发育机制有可能进一步治疗越来越多的与神经元丢失或异常神经元功能相关的精神障碍。因此，拟议的研究与NIH的使命的一部分有关，即获得可能有助于治疗神经系统疾病的基础知识。 在强有力的初步数据的指导下，该假设将通过追求两个特定目标进行测试：1）成人NSC染色质中REST/NRSF靶标的全基因组鉴定和2）静止、增殖和神经元分化期间成人NSC的转录组分析。目的1将致力于在成年海马神经干细胞中进行REST/NRSF的ChIP-Seq。目标2将专注于在静止、增殖和神经元分化的条件下从成年NSC执行RNA-seq。目标2还将通过比较ChIP-Seq靶标与我们实验室产生的REST/NRSF条件性敲除小鼠的基因表达分析来确定哪些靶标依赖于REST/NRSF。概念框架和方法是创新的，因为它是第一个将最先进的深度测序技术应用于生理相关的成人神经干细胞系统的研究，以确定REST/NRSF靶基因的全基因组视图。拟议的R21赠款意义重大，因为它有望推进和扩大我们对调节干细胞静止、增殖和神经元分化的转录网络的基本理解，这可能对脑损伤或疾病后恢复认知功能特别重要。