Mapping the changing epigenetic landscape in muscle stem cells

绘制肌肉干细胞不断变化的表观遗传图谱

• 批准号: 10251662
• 负责人: Payel Sen
• 金额: $ 45.95万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10251662
• 关键词: 3-Dimensional; Acetylation; Aging; Cell Aging; Cell Separation; Cell physiology; Cells; Chromatin; Collaborations; Epigenetic Process; Functional disorder; Future; Gene Expression; Histone Acetylation; Human; In Vitro; Injury; Investigation; Lead; Literature; Maps; Methods; Modification; Molecular Conformation; Mus; Muscle; Muscle satellite cell; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Population; Proteins; Protocols documentation; Reporting; Skeletal Muscle; System; Techniques; Technology; age related; aged; base; epigenome; experimental study; functional decline; genome-wide; histone methylation; histone modification; improved; in vivo; muscle aging; regenerative; senescence; stem cells; tissue repair; transcription factor

Muscle stem cells (MuCS) are a population of muscle-resident stem cells that are essential for efficient tissue repair post-injury. During aging, MuSC dysfunction is a major contributor to the decline in regenerative potential of muscle tissue. Emerging evidence indicates epigenetic changes accumulate (erosion and drift) and may lead to a functional decline in MuSCs during aging. In this project, a low-input chromatin mapping technology will be used to acquire genome-wide maps of histone modification, especially acetylation, and transcription factor profiles in MuSCs from young and aged mice and bulk human skeletal muscle. In the past year, we have optimized a FACS based MuSC isolation protocol in collaboration with Dr. Vittorio Sartorelli (NIAMS). Using this method, we now routinely isolate >300,000 cells/young mouse. From old mice, the average yield is slightly lower, as reported in literature. We have also optimized a low-input chromatin mapping technique (CUT&RUN) in the lab and will now apply this method to our isolated cells. In the future, our comprehensive chromatin maps will be validated in MuSCs isolated from human skeletal muscle tissue. Furthermore, we will generate 3D map chromatin conformation maps in mice and humans. Finally, we will use this information to screen for epigenetic regulators of muscle aging.

肌肉干细胞 (MuCS) 是一组肌肉驻留干细胞，对于损伤后有效的组织修复至关重要。在衰老过程中，MuSC 功能障碍是肌肉组织再生潜力下降的主要原因。新的证据表明表观遗传变化不断累积（侵蚀和漂移），并可能导致衰老过程中 MuSC 的功能下降。在该项目中，低输入染色质作图技术将用于获取年轻和老年小鼠和大量人类骨骼肌 MuSC 中组蛋白修饰（尤其是乙酰化）的全基因组图谱和转录因子谱。 在过去的一年中，我们与 Vittorio Sartorelli 博士 (NIAMS) 合作优化了基于 FACS 的 MuSC 分离方案。使用这种方法，我们现在通常分离每只幼鼠> 300,000 个细胞。据文献报道，年老小鼠的平均产量略低。我们还在实验室中优化了低输入染色质定位技术 (CUT&RUN)，现在将该方法应用于我们的分离细胞。 将来，我们的综合染色质图谱将在从人类骨骼肌组织中分离的 MuSC 中得到验证。此外，我们将生成小鼠和人类的 3D 图谱染色质构象图。最后，我们将利用这些信息来筛选肌肉衰老的表观遗传调节因子。