Molecular Mechanisms of Cell Fate Decisions in Hair Follicle Stem Cells

毛囊干细胞细胞命运决定的分子机制

• 批准号: 8324927
• 负责人: Tudorita Tumbar
• 金额: $ 36.47万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-16 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8324927
• 关键词: Ablation; Acute; Address; Adopted; Adult; Behavior; Biological Assay; Biological Models; CCL4 gene; Cell Culture Techniques; Cell Fate Control; Cell Lineage; Cell Maintenance; Cell Separation; Cell Transplantation; Cell surface; Cells; Characteristics; Chromatin; Clinic; Co-Immunoprecipitations; Cultured Cells; DNA; Data; Developmental Gene; Disease; Enzymes; Epigenetic Process; Frequencies; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genome; Genomics; Germ; Goals; Hair; Hair follicle structure; Heterogeneity; Histone H2B; Histones; Homeostasis; Immunofluorescence Immunologic; Label; Light; Link; Location; Maps; Memory; Messenger RNA; Microarray Analysis; Modeling; Molecular; Morphology; Mus; Normal tissue morphology; Nuclear; Pathway interactions; Pattern; Pharmaceutical Preparations; Phase; Physiologic pulse; Population; Precipitation; Prevalence; Proliferating; Regulation; Relative (related person); Skin; Sorting - Cell Movement; Staging; Stem cells; Structure; System; Testing; Tetanus Helper Peptide; Time; Tissues; Western Blotting; Work; adult stem cell; base; cell behavior; cell type; chromatin immunoprecipitation; gene repression; genome-wide; histone modification; insight; knockout gene; mRNA Expression; novel; progenitor; programs; promoter; protein complex; protein protein interaction; public health relevance; regenerative therapy; research study; self-renewal; stem cell fate; stem cell niche; stem cell population; therapeutic target; tissue regeneration; tool; transcription factor

DESCRIPTION (provided by applicant): Deciphering the molecular mechanisms of adult stem cell (SC) fate decisions is essential for understanding disease and for tissue regenerative therapy. The overall goal of this proposal is to utilize hair follicles and skin as model systems to address at the molecular level the interplay of transcriptional factors and epigenetic regulation in fate choice decisions of a vertebrate adult stem cell population in its native tissue. Because hair follicle stem cells synchronously proliferate and return to quiescence in the mouse skin, this system allows mechanistic insight that is otherwise more difficult to achieve. In Aim (1) we plan to investigate in more depth the relationship between more dormant versus more active cell subpopulations in the hair follicle stem cell niche, by characterizing their morphogenetic and self-renewal ability in cell culture and cell transplantation assays. The existence of these two kinds of cell subpopulations in stem cell niches appears to be a relatively common feature for at least several vertebrate tissues. Their interplay in maintaining tissue homeostasis is important for accurate description of stem cell behavior and fate decisions. In Aim (2) we will examine the distribution of histone marks across the genome at two stages of stem cell activity by chromatin immuno-precipitation and sequencing, and attempt to examine a potential link between two known adult tissue stem cells features: quiescence and plasticity. In Aim (3) we will employ two transcription factors Runx1 and Gata6 as fate acquisition regulators. We will test their potential implication in remodeling the stem cell histone epigenome via interactions with a battery of histone modifying enzymes we found expressed in the hair follicle in a relevant pattern. We will induce acute loss of Runx1 and Gata6 in mouse skin and examine the mRNA expression level of specific histone modifying enzymes in the relevant hair follicle cell populations. In addition we will perform co-immunoprecipitation or pull-down experiments to examine a potential direct protein-protein interaction between Runx1 or Gata6 and specific histone modifying enzymes. This will begin to address the mechanisms of fate acquisition in a vertebrate adult stem cell population and shed light on the interplay between transcription factors that work as master regulator in orchestrating cell fate and global remodeling of histone epigenetic marks. PUBLIC HEALTH RELEVANCE: A large number of diseases are due to miss-regulation of cell fate acquisition of adult tissue stem cells during their activity to maintain normal tissue homeostasis. There has been recently great promise for developing novel drug therapeutics that target specific histone modifying enzymes, some already present in clinics. This proposal seeks to utilize a versatile vertebrate model system, mouse skin and hair follicles, to gain better understanding of the interplay between histone epigenetic marks and transcription factors that control cell fate decisions in adult vertebrate stem cells.

描述（由申请人提供）：破译成体干细胞（SC）命运决定的分子机制对于理解疾病和组织再生治疗至关重要。该提案的总体目标是利用毛囊和皮肤作为模型系统，在分子水平上解决转录因子和表观遗传调控在命运选择中的相互作用 脊椎动物成体干细胞群在其天然组织中的决定。因为头发 毛囊干细胞在小鼠皮肤中同步增殖并恢复静止状态， 该系统可以提供机械洞察力，而这在其他情况下是很难实现的。瞄准 (1) 我们计划更深入地研究更多休眠与更多休眠之间的关系 通过表征毛囊干细胞生态位中更活跃的细胞亚群 细胞培养和细胞移植测定中的形态发生和自我更新能力。 干细胞生态位中这两种细胞亚群的存在似乎是 至少对于几种脊椎动物组织而言，这是一个相对常见的特征。他们的相互作用 维持组织稳态对于准确描述干细胞很重要 行为和命运的决定。在目标 (2) 中，我们将检查组蛋白标记的分布 通过染色质免疫沉淀在干细胞活性的两个阶段跨越基因组 和测序，并尝试检查两个已知成人组织之间的潜在联系 干细胞的特点：静止性和可塑性。在目标 (3) 中，我们将雇用两个 转录因子 Runx1 和 Gata6 作为命运获取调节剂。我们将测试他们的 通过相互作用重塑干细胞组蛋白表观基因组的潜在意义 我们发现在毛囊中表达的一组组蛋白修饰酶 相关模式。我们将诱导小鼠皮肤中 Runx1 和 Gata6 的急性丢失， 检查特定组蛋白修饰酶的 mRNA 表达水平 相关毛囊细胞群。此外，我们将进行免疫共沉淀 或下拉实验来检查潜在的直接蛋白质-蛋白质相互作用 Runx1 或 Gata6 与特定组蛋白修饰酶之间。这将开始 解决脊椎动物成体干细胞群的命运获得机制 并揭示了作为主控的转录因子之间的相互作用 协调细胞命运和组蛋白表观遗传标记全局重塑的调节因子。 公共健康相关性：许多疾病是由于成体组织干细胞在维持正常组织稳态的活动过程中细胞命运获得的错误调节所致。最近，开发针对特定组蛋白修饰酶的新型药物疗法前景广阔，其中一些已经应用于临床。该提案旨在利用多功能脊椎动物模型系统、小鼠皮肤和毛囊，更好地了解组蛋白表观遗传标记和控制成体脊椎动物干细胞细胞命运决定的转录因子之间的相互作用。