Epigenomic drivers of human muscle progenitor cells in development and disease

人类肌肉祖细胞发育和疾病的表观基因组驱动因素

• 批准号: 8814714
• 负责人: AMY JO WAGERS
• 金额: $ 33.8万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-08 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8814714
• 关键词: Affect; Automobile Driving; Biological Markers; Biology; Birth; Cancer cell line; Cell Count; Cell Line; Cell Maintenance; Cell Therapy; Cells; Clinical; Data; Data Set; Degenerative Disorder; Derivation procedure; Development; Diagnostic; Disease; Disease Progression; Disease model; Distal; Drug Targeting; Duchenne muscular dystrophy; Dystrophin; Enhancers; Environment; Epigenetic Process; Evaluation; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Genetic Transcription; Goals; Government; Growth; Human; Injury; Investigation; Laboratories; Maintenance; Maps; Modeling; Molecular; Muscle; Muscle Cells; Muscle Fibers; Muscle function; Muscle satellite cell; Mutation; Myoblasts; Myopathy; Natural regeneration; Nucleic Acid Regulatory Sequences; Pathology; Pathway Analysis; Pathway interactions; Patients; Persons; Play; Pluripotent Stem Cells; Population; Process; Proliferating; RNA Sequences; Regulation; Regulatory Element; Research; Resolution; Resources; Role; Severities; Signal Transduction; Skeletal Muscle; Specific qualifier value; Staging; Stem cells; Stretching; Technology; Testing; Therapeutic; Tissues; Translating; Transplantation; United States National Institutes of Health; Universities; Work; advanced disease; base; chromatin immunoprecipitation; epigenomics; gene correction; human disease; improved; induced pluripotent stem cell; innovation; interest; male; malignant state; muscle form; muscle regeneration; new therapeutic target; novel; novel strategies; precursor cell; progenitor; programs; public health relevance; regenerative; repaired; satellite cell; scaffold; small molecule; stem; stem cell biology; success; transcription factor; transcriptome sequencing

DESCRIPTION (provided by applicant): There is great need for new therapies for Duchenne Muscular Dystrophy (DMD), a currently incurable degenerative muscle disorder that affects 1 in every 3500 male births. Promising options include the transplantation of normal or "gene-corrected" muscle progenitors, which could engraft affected muscle fibers to correct the underlying genetic defects, as well as small molecule based therapies that could correct disrupted myogenic processes to support improved muscle function. Unfortunately, the rarity of regenerative muscle precursor cells and lack of relevant models that strongly mimic the progression and severity of human DMD has created many obstacles to understanding disease mechanisms and hindered the development of curative therapies. However, we believe that new innovations in epigenomic analysis and muscle stem cell biology, developed in part in our laboratories, have created new opportunities to discover and advance novel therapies in DMD. This project will exploit the combined expertise of the Wagers and Rinn research groups, which share common laboratory space in the Department of Stem Cell and Regenerative Biology at Harvard University. Studies in the Wagers lab have established novel strategies to isolate highly enriched populations of skeletal muscle progenitors (satellite cells) from human muscle and to derive engraftable myogenic precursor cells from human induced pluripotent stem cells. Studies in the Rinn lab have defined novel computational pipelines for the annotation and analysis of gene regulatory elements that specify cell fate and function. This project will combine these innovative advances with the unique resources provided by the NIH Epigenomics Roadmap Program to identify epigenetic regulators that drive human muscle progenitor cell identity and pinpoint those that may play a role in initiating or promoting DMD pathology. Our analyses will concentrate particularly on regulatory enhancers (e.g., "Stretch" or "Super" Enhancers, SEs), which represent regulatory "hubs" that frequently interact with the epigenetic and transcriptional machinery to control gene expression. We will use ChIP-Seq and transcriptional datasets to define muscle progenitor-specific SEs and their target genes, including candidate "master" transcriptional regulators of the muscle progenitor fate. Regulatory SEs and their targets will also be defined in skeletal muscle progenitors and terminal muscle cells derived from healthy or DMD patient induced pluripotent stem cells (iPSCs), using isogenic iPSC lines and novel culture-based strategies for generating and identifying myogenic precursors and their progeny at distinct stages of muscle differentiation. Together, this work will define the core regulatory and transcriptional landscapes that uniquely enforce the skeletal muscle progenitor cell fate, identify DMD-related alterations in enhancer activity that may drive disease progression, and further the application of iPSC-based technologies for the discovery of novel therapies for DMD and other muscle disorders.

描述（由申请人提供）：对Duchenne肌肉营养不良（DMD）的新疗法非常需要，这是一种目前无法治愈的退行性肌肉障碍，每3500个男性出生中会影响1个。有希望的选择包括正常或“基因校正”的肌肉祖细胞的移植，这些肌肉可能会植入受影响的肌肉纤维以纠正潜在的遗传缺陷，以及基于小分子的小分子疗法，这些疗法可以纠正肌原性过程以支持改善肌肉功能。不幸的是，再生肌肉前体细胞的稀有性以及缺乏强烈模仿人DMD的进展和严重程度的相关模型，为理解疾病机制的发展带来了许多障碍，并阻碍了治疗疗法的发展。但是，我们认为表观基因组分析和肌肉干细胞生物学的新创新是在我们的实验室中开发的，为发现和推进DMD中的新型疗法创造了新的机会。该项目将利用赌注和Rinn研究小组的综合专业知识，这些研究小组在哈佛大学的干细胞和再生生物学系中共享共同的实验室空间。 Wagers实验室中的研究已经建立了新的策略，以将骨骼肌祖细胞（卫星细胞）的高度富集群体与人类肌肉分离，并从人类诱导的多能干细胞中得出植入的肌源性前体细胞。 Rinn实验室中的研究定义了新的计算管道，用于对指定细胞命运和功能的基因调节元件的注释和分析。该项目将将这些创新的进步与NIH表观基因组路线图计划提供的独特资源相结合，以确定驱动人类肌肉祖细胞细胞身份的表观遗传调节器，并确定可能在发起或促进DMD病理学中发挥作用的那些。我们的分析将特别集中在调节性增强子（例如“拉伸”或“超级”增强子，SES）上，该增强剂代表了经常与表观遗传学和转录机械相互作用以控制基因表达的调节“枢纽”。我们将使用CHIP-SEQ和转录数据集来定义肌肉祖细胞特异性SES及其目标基因，包括肌肉祖细胞命运的候选“主”转录调节剂。调节性SES及其靶标也将在骨骼肌祖细胞和末端肌肉细胞中定义，这些肌肉细胞来自健康或DMD患者诱导的多能干细胞（IPSC），使用异基因IPSC线条和基于新型培养的策略，用于产生和识别肌发型前体及其在肌肉分化的不同阶段的弟弟。总之，这项工作将定义核心的调节和转录景观，这些核心和转录景观独特地强制执行骨骼肌肉祖细胞命运，确定可能驱动疾病进展的增强剂活性的与DMD相关的变化，并进一步推动IPSC基于IPSC的技术在基于IPSC的技术中用于发现DMD和其他肌肉分散剂的新疗法。