The role of Tet1 in myofibroblast differentiation

Tet1在肌成纤维细胞分化中的作用

• 批准号: 10201053
• 负责人: SEM H PHAN
• 金额: $ 60.64万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10201053
• 关键词: Achievement; Address; Alveolar; Appearance; Area; Base Excision Repairs; Binding; Binding Proteins; Biological; Cell Differentiation process; Cell division; Cells; Complex; CpG Islands; DNA; DNA Methylation; DNA Modification Methylases; DNA Sequence; Data; Derivation procedure; Development; Developmental Biology; Differentiation Antigens; Dioxygenases; Disease; Elements; Epigenetic Process; Epithelial Cells; Extracellular Matrix; Fibroblasts; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Hour; Impaired healing; Impairment; In Vitro; Light; Lung; Malignant Neoplasms; Mediating; Mesenchymal Differentiation; Molecular; Myofibroblast; Neoplasm Metastasis; Process; Protein Isoforms; Proteins; Proteomics; Regulation; Role; Signal Pathway; Smooth Muscle Actin Staining Method; Specificity; Stimulus; Testing; Tetanus Helper Peptide; Thymine DNA Glycosylase; Time; Transgenic Mice; angiogenesis; base; carcinogenesis; cell type; cytokine; demethylation; epigenetic regulation; epigenomics; healing; in vivo; organ growth; overexpression; oxidation; programs; promoter; recruit; repaired; response; stem cells; tissue injury; tissue repair; transcription factor; tumor growth; wound

Project Summary: Myofibroblasts are differentiated mesenchymal cells with identified roles in development, cancer, tissue repair/remodeling. Regulation of myofibroblast differentiation involves multiple signaling pathways and respective downstream transcription factors as well as significant modulation by epigenetic factors such as DNA methylation. Abundant studies focused on identification of transcription factors and their role in regulation of the α-smooth muscle actin (Acta2) gene as a marker of differentiation, but the mechanism of epigenetic regulation is not as well understood. Select CpG islands in the Acta2 gene are differentially methylated in myofibroblasts vs. fibroblasts and epithelial cells. Impairment of DNA methylation enhances differentiation while the converse inhibits differentiation. Active demethylation by the Tet (Ten-eleven translocation) proteins, which are methylcytosine dioxygenases, is implicated in cell differentiation. Their importance in cell differentiation is suggested in studies of embryonal and other stem cells, but their significance in regulation of myofibroblast differentiation is unknown. Preliminary data showed selective induction of Tet1 and not the other 2 isoforms (Tet2 and Tet3) in myofibroblast differentiation, while Tet1 deficiency caused impaired differentiation both in vivo and in vitro. Thus the selective importance of the Tet1 isoform is suggested in differentiation. While Tet1 does not globally regulate all genes by demethylation, the identity of at least one Tet1 regulated target gene, Acta2 could be inferred from the preliminary data. Based on the previous findings and preliminary data we hypothesized that Tet1 importantly regulates myofibroblast differentiation by demethylation of regulatory DNA sequences in select target genes essential for the differentiation process. Select recruitment to the relevant target DNA sequences is mediated by Tet1 binding proteins/transcription factors with binding specificity for these DNA regions. To test this hypothesis the Specific Aims are as follows, 1) to screen for and identify Tet1 target genes important in myofibroblast differentiation in fibroblasts, 2) to study the mechanism of Tet1 regulation of key target genes essential for myofibroblast differentiation, and 3) to assess the in vivo significance of Tet1 regulation of myofibroblast differentiation. The studies will use epigenomic approaches to assess differentially methylated genes and transgenic mice to evaluate the biological importance of Tet1 and select differentiation relevant genes in specific cell types in vivo. Achievement of the goals will shed new light on the epigenetic regulation of myofibroblast differentiation of relevance to development, cancer and tissue repair/remodeling.

项目摘要： 肌纤维细胞是分化的间充质细胞，在发育，癌症，组织中鉴定出作用 维修/改建。肌纤维细胞分化的调节涉及多个信号通路， 特定的下游转录因子以及表观遗传因素（例如 DNA甲基化。大量研究重点是鉴定转录因子及其在调节中的作用 α-平滑肌肌动蛋白（ACTA2）基因作为分化的标记，但表观遗传学的机理 监管没有那么充分理解。 ACTA2基因中的精选CpG岛在不同的甲基化中被不同 肌纤维细胞与成纤维细胞和上皮细胞。 DNA甲基化的损害增强了分化 匡威抑制分化。 TET（十个易位）蛋白的活性去甲基化， 在细胞分化中实现了甲基二糖苷的二加氧酶。它们在细胞中的重要性 在胚胎和其他干细胞的研究中提出了分化，但它们在调节中的重要性 肌纤维细胞的分化未知。初步数据显示了TET1的选择性诱导，而不是其他数据 肌纤维细胞分化中的2种同工型（TET2和TET3），而TET1缺乏症会受损 体内和体外。在分化中提出了TET1同工型的选择性重要性。尽管 TET1不通过脱甲基化来调节所有基因，至少一个TET1调控靶标的身份 基因，可以从初步数据中推断出ACTA2。根据先前的发现和初步数据 我们假设TET1重要地通过脱甲基来调节肌纤维细胞分化 在分化过程中必不可少的选择靶基因中的DNA序列。选择招聘 相关的靶DNA序列是由具有结合的TET1结合蛋白/转录因子介导的 这些DNA区域的特异性。为了检验该假设，具体目的如下：1）筛选和 确定在成纤维细胞中肌纤维细胞分化重要的TET1靶基因，2）研究 TET1调节关键靶基因对肌纤维细胞分化必不可少的，3）评估体内 TET1调节肌纤维细胞分化的重要性。研究将使用表观基因质学方法 评估甲基化基因和转基因小鼠的不同，以评估TET1和 在体内特定细胞类型中选择分化相关的基因。实现目标将使新的光明 关于与发育，癌症和组织相关性的肌纤维细胞分化的表观遗传调节 维修/改建。