DNA hydroxymethylation and Tet-enzymes in the control of the skin development and hair growth

DNA 羟甲基化和 Tet 酶在控制皮肤发育和毛发生长中的作用

• 批准号: 10433970
• 负责人: VLADIMIR A BOTCHKAREV
• 金额: $ 35.94万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10433970
• 关键词: Ablation; Acetylation; Address; Biological Process; Biology; C-terminal; CDKN2A gene; CDKN2B gene; Catalytic Domain; Cell Cycle; Cell Differentiation process; Cell Lineage; Cell Proliferation; Chromatin; Chronic; Complex; CpG Islands; Cysteine; DNA; DNA Binding; DNA Methylation; Data; Defect; Development; Differentiated Gene; Embryo; Embryonic Development; Enhancers; Enzymes; Epidermis; Epigenetic Process; Epithelial Cells; Event; Exhibits; Family; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Genetically Engineered Mouse; Genome; Goals; Growth; Hair; Hair follicle structure; Hair shaft structure; Health; Histone H3; Homeostasis; Human; Impairment; In Vitro; Keratin; Knockout Mice; Knowledge; Litter Size; Maintenance; Malignant Neoplasms; Mammals; Mediating; Mesoderm; Methylation; Modeling; Multipotent Stem Cells; Mus; N-terminal; Nucleic Acid Regulatory Sequences; Paraxial Mesoderm; Pathologic; Pattern; Physiological; Play; Population; Primitive Streaks; Process; Proteins; Psoriasis; Quality of life; Regulation; Regulator Genes; Research; Role; Signal Transduction; Skin; Skin Cancer; Testing; Tetanus Helper Peptide; Therapeutic Intervention; bone morphogenetic protein receptors; chromatin remodeling; demethylation; epigenome; epithelial stem cell; epithelial wound; gastrulation; histone modification; imprint; inhibitor; keratinocyte; keratinocyte differentiation; novel; novel strategies; oxidation; premature; programs; promoter; self-renewal; skin barrier; skin disorder; skin organogenesis; skin regeneration; stem cell differentiation; stem cells; tissue regeneration; transcription factor; transcriptome

PROJECT SUMMARY The long-term goal of this project is to understand how epithelial stem cells in the skin establish distinct patterns of gene expression during their differentiation into specialized cell lineages and how these genetic programs are altered in pathological skin conditions associated with impaired cell differentiation. Research into genome and chromatin biology has revealed that in addition to signaling/transcription factor- dependent regulatory mechanisms, lineage-specific gene expression programs are also regulated epigenetically, i.e., via regulation of covalent DNA/histone modifications and higher-order chromatin remodeling. DNA methylation and subsequent oxidation of 5-methylcytosine into 5-hydroxymethylcytosine (5hmC) are key epigenetic events regulating development and stem cell differentiation in mammals. Oxidation of 5- methylcytosine is catalyzed by the TET1/2/3 family enzymes and serve as an important step in DNA demethylation. Recent data reveal that Tet proteins plays essential roles in many biological processes including development, cancer and cellular reprogramming, while genetic ablation of all three Tet genes is lethal. However, the role of Tet-mediated DNA hydroxymethylation in the control of gene expression in keratinocytes during terminal differentiation in the epidermis and hair follicle remain obscured. Our preliminary data reveal that 5hmC-modified DNA is abundant in the hair follicle bulge, as well as in differentiating epidermal and hair matrix keratinocytes. Furthermore, genetic Tet3 ablation results in alterations of epidermal barrier formation during embryonic development, while Shh-Cre mediated ablation of all three Tet genes in the hair matrix keratinocytes results in alterations of the hair shaft structure and hair keratin gene expression. In this proposal, we will test the hypothesis that Tet proteins serve as critical determinants that regulate gene expression programmes in differentiating epidermal and hair follicle keratinocytes via DNA demethylation at the promoters and enhancers of lineage-specific genes, as well as of key genes that control epidermal barrier formation and hair follicle cycling. This hypothesis will be addressed via two Specific Aims: 1) Define the roles of Tet1, Tet2 and Tet3 in the control of epidermal development, terminal keratinocyte differentiation and epidermal barrier maintenance. 2) Define the common features and differential impact of the distinct Tet genes on self-renewing and differentiation potentials of the hair follicle epithelial stem cells and their progenies during physiological hair cycle-dependent skin regeneration. This project will have a fundamental impact on our current knowledge of epigenetic mechanisms that regulate genome reorganization in stem cells during their differentiation and will promote the progress towards the development of novel paradigms for treatment of skin disorders via targeting Tet enzymes and epigenome.

项目概要 该项目的长期目标是了解皮肤中的上皮干细胞如何建立独特的 细胞分化为特化细胞谱系期间的基因表达模式以及这些基因如何 程序在与细胞分化受损相关的病理性皮肤状况下发生改变。 对基因组和染色质生物学的研究表明，除了信号/转录因子之外， 依赖的调节机制，谱系特异性基因表达程序也受到调节 表观遗传学，即通过共价 DNA/组蛋白修饰和高阶染色质重塑的调节。 DNA 甲基化和随后 5-甲基胞嘧啶氧化成 5-羟甲基胞嘧啶 (5hmC) 是 调节哺乳动物发育和干细胞分化的关键表观遗传事件。 5-的氧化 甲基胞嘧啶由 TET1/2/3 家族酶催化，是 DNA 合成的重要步骤 去甲基化。最近的数据表明 Tet 蛋白在许多生物过程中发挥着重要作用，包括 发育、癌症和细胞重编程，而所有三个 Tet 基因的基因消除都是致命的。 然而，Tet 介导的 DNA 羟甲基化在基因表达控制中的作用 表皮和毛囊终末分化期间的角质形成细胞仍然模糊。我们的初步 数据显示，5hmC 修饰的 DNA 在毛囊隆起以及分化表皮中大量存在。 和毛发基质角质形成细胞。此外，基因 Tet3 消融会导致表皮屏障的改变 胚胎发育期间的形成，而 Shh-Cre 介导头发中所有三个 Tet 基因的消融 基质角质形成细胞导致毛干结构和毛发角蛋白基因表达的改变。 在本提案中，我们将检验 Tet 蛋白作为调节基因的关键决定因素的假设 通过 DNA 去甲基化区分表皮和毛囊角质形成细胞的表达程序 谱系特异性基因以及控制表皮屏障的关键基因的启动子和增强子 形成和毛囊循环。这一假设将通过两个具体目标来解决： 1) 定义Tet1、Tet2和Tet3在控制表皮发育、终末期的作用 角质形成细胞分化和表皮屏障维持。 2) 定义不同 Tet 基因的共同特征和对自我更新和影响的不同影响 毛囊上皮干细胞及其后代在生理过程中的分化潜能 毛发周期依赖性皮肤再生。 该项目将对我们目前对调节表观遗传机制的了解产生根本性影响。 干细胞在分化过程中的基因组重组将促进干细胞的进展 通过针对 Tet 酶和表观基因组开发治疗皮肤病的新范例。