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.

项目摘要 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 基因表达在分化为专业细胞谱系的基因表达模式以及这些遗传如何 与细胞分化受损相关的病理皮肤状况，程序会改变。 对基因组和染色质生物学的研究表明，除了信号传导/转录因子 - 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，通过调节共价DNA/组蛋白修饰和高阶染色质重塑。 DNA甲基化并随后将5-甲基胞嘧啶氧化成5-羟基甲基胞嘧啶（5HMC）为 关键的表观遗传事件调节哺乳动物的发育和干细胞分化。 5-的氧化 TET1/2/3家族酶催化甲基胞嘧啶，并作为DNA的重要步骤 脱甲基化。最近的数据表明，TET蛋白在许多生物学过程中都起着重要作用 发育，癌症和细胞重编程，而所有三个TET基因的遗传消融都是致命的。 然而，TET介导的DNA羟基化在控制基因表达中的作用 表皮末端分化过程中的角质形成细胞和毛囊中的角膜细胞仍然被遮盖。我们的初步 数据表明，毛囊凸起以及分化表皮中5HMC修饰的DNA丰富 和头发基质角质形成细胞。此外，遗传TET3消融导致表皮屏障的改变 胚胎发育过程中的形成，而SHH-CRE介导了头发中所有三个TET基因的消融 基质角质形成细胞会导致发轴结构和毛制蛋白基因表达的改变。 在此提案中，我们将检验以下假设：TET蛋白是调节基因的关键决定因素 通过DNA脱甲基化在区分表皮和毛囊角质形成细胞中的表达程序 谱系特异性基因的启动子和增强子，以及控制表皮屏障的关键基因 形成和毛囊循环。该假设将通过两个具体目的解决： 1）定义TET1，TET2和TET3在控制表皮发育，终端中的作用 角质形成细胞分化和表皮屏障维护。 2）定义不同TET基因对自我更新和的共同特征和差异影响 毛囊上皮干细胞的分化潜力及其在生理过程中的后代 依赖头发周期的皮肤再生。 该项目将对我们当前对调节表观遗传机制的了解产生根本性的影响 基因组在干细胞分化过程中重组，并将促进朝向 通过靶向TET酶和表观基因组来开发新型范例来治疗皮肤疾病。