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Regulation of hematopoiesis by epigenetic timing control

表观遗传时序控制对造血的调节

基本信息

批准号：
10065914
负责人：
Nicholas Pease
金额：
$ 3.76万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-16 至 2021-06-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10065914
关键词：
Address Alleles Biological Biological Models Biological Process Blood Blood Cells CRISPR/Cas technology Cell Differentiation process Cell Lineage Cell Therapy Cell division Cells Characteristics Collaborations DNA DNA-Protein Interaction Development Developmental Gene Developmental Process Disease Educational workshop Elements Environment Epigenetic Process Exhibits Fellowship Gene Activation Gene Expression Regulation Generations Genes Genomic approach Genomics Goals Hematological Disease Hematopoiesis Hematopoietic Hematopoietic stem cells Homeostasis Human Human Development Hybrids Individual Measures Mechanics Mentors Modeling Molecular Monitor Morphology Mouse Strains Multipotent Stem Cells Mutation Outcome Pathogenesis Play Population Prevalence Process Public Health Recurrence Regulation Regulatory Element Reporter Reporter Genes Research Research Personnel Research Training Role Single Nucleotide Polymorphism System T-Lymphocyte Testing Time Tissues Training Untranslated RNA Variant Work X Inactivation career development cell type chromatin remodeling engineered stem cells epigenetic regulation epigenome editing genome editing genome-wide genomic locus hematopoietic gene hematopoietic stem cell differentiation human disease insight mouse model novel strategies reconstitution self-renewal skills training stem cell differentiation stem cell fate stem cells training opportunity transcription factor

项目摘要

Project Summary: The timing of stem cell fate decisions is critical for multicellular tissue size and function. In many developmental processes, stem cells differentiate only after a long time-delay spanning multiple days and cell divisions, allowing rare populations of multipotent progenitors to expand exponentially. Variation in differentiation timing generates dramatic changes in tissue size and morphology and likely contributes to human development and disease. However, the mechanism underlying timing control during differentiation remains unclear. This proposal will use hematopoiesis a model system to understand how epigenetic regulation contributes to the timing of cell fate decisions. Recent evidence suggests that epigenetic mechanisms acting at individual genomic loci in cis can change slowly over the course of multiple days and cell divisions, implementing rate-limiting steps to developmental gene activation. However, it remains unclear if this is a paradigm for timing control broadly utilized during cell differentiation. This proposal aims to determine the prevalence of epigenetic timing mechanisms (Aim 1) and uncover how they can be regulated by non-coding DNA elements (Aim 2). Epigenetic regulation, unlike transcription factor regulation in trans, functions at each gene copy independently in the same cell (e.g. X-chromosome inactivation). Therefore, to investigate epigenetic timing control prevalence and mechanisms, this proposal will use mouse models in which the activity of individual gene copies can be monitored separately. In Aim 1, a F1 hybrid mouse strain that harbors frequent single nucleotide polymorphisms will be used to analyze individual alleles genome-wide by CUT&Tag. This single-cell genomics approach can distinguish epigenetic states by quantifying of protein-DNA interactions and will reveal the generality of epigenetic timing mechanisms during hematopoiesis. In Aim 2, a two-copy gene reporter system will be used to provide a highly sensitive readout for epigenetic regulation in hematopoietic progenitors undergoing differentiation. CRISPR/Cas9 approaches will be used to efficiently identify functional non-coding DNA elements and elucidate how they contribute to epigenetic timing control. This fellowship will provide an opportunity for training in computational genomics and CRISPR/Cas9 genome and epigenome editing. Experts in these fields will serve as mentors for skills training and career development. This training plan incorporates tailored courses, seminars and workshops that will enhance the training environment and facilitate the transition to a postdoctoral research fellowship. The expected outcome of this proposal will establish a new model for timing control during tissue development and homeostasis. It will also provide new insights into blood disease pathogenesis and inspire new approaches for epigenetic reprogramming in cell-based therapies.

项目概要：干细胞命运决定的时机对于多细胞组织的大小和功能至关重要。在许多在发育过程中，干细胞只有在跨越多天的长时间延迟后才分化，分裂，使罕见的多能祖细胞群体呈指数级增长。分化变异定时产生组织大小和形态的巨大变化，可能有助于人类发育和疾病然而，分化过程中的时序控制机制仍不清楚。本建议将利用造血系统的一个模型来了解表观遗传是如何调控的有助于决定细胞命运的时间。最近的证据表明，表观遗传机制的作用， cis中的单个基因组位点可以在多天和细胞分裂的过程中缓慢变化，发育基因激活的限速步骤。然而，目前还不清楚这是否是一个时机选择的范例在细胞分化过程中广泛使用的控制。该提案旨在确定表观遗传时间机制（Aim 1）的普遍性，并揭示它们如何被非编码DNA元件调控（目的2）。表观遗传调控，不同于转录反式因子调节在同一细胞中的每个基因拷贝独立地起作用（例如X染色体失活）。因此，为了研究表观遗传时间控制的流行和机制，本建议将使用小鼠模型，其中单个基因拷贝的活性可以单独监测。在Aim 1中，F1 携带频繁单核苷酸多态性杂交小鼠品系将用于分析个体等位基因全基因组的切割和标签。这种单细胞基因组学方法可以区分表观遗传状态，定量的蛋白质-DNA相互作用，并将揭示普遍性的表观遗传定时机制，造血在目标2中，将使用双拷贝基因报告系统来提供高灵敏度的读出，造血祖细胞分化过程中的表观遗传调控。CRISPR/Cas9方法将在用于有效地识别功能性非编码DNA元件，并阐明它们如何有助于表观遗传定时控制该奖学金将提供计算基因组学和CRISPR/Cas9培训的机会基因组和表观基因组编辑。这些领域的专家将担任技能培训和职业生涯的导师发展该培训计划包括量身定制的课程、研讨会和讲习班，培训环境和促进过渡到博士后研究奖学金。的预期成果这一建议将建立一个新的模型，在组织发育和稳态的时间控制。它将还为血液病的发病机制提供了新的见解，并启发了表观遗传学的新方法。细胞治疗中的重编程。