Determining the role of Tet1 in intestinal stem cell differentiation and self-renewal

确定 Tet1 在肠道干细胞分化和自我更新中的作用

• 批准号: 9806499
• 负责人: Adam David Gracz
• 金额: $ 11.66万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 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 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9806499
• 关键词: Ablation; Adult; Affect; Biological Assay; Biology; Cell Differentiation process; Cell Fate Control; Cell Proliferation; Cell physiology; Cells; Cellular Structures; Cellular biology; ChIP-seq; Chromatin; Chromatin Remodeling Factor; Complement; Computational Biology; Cytosine; DNA; DNA Methylation; Data; Dissection; Embryo; Ensure; Enzymes; Epigenetic Process; Epithelial; Equilibrium; Funding; Future; Genes; Genetic; Genetic Transcription; Genetic study; Genomics; Grant; Hematopoietic stem cells; Histone Deacetylase; Homeostasis; Hyperplasia; In Vitro; Inflammation; Intestines; Investigation; Knock-out; Knockout Mice; Life; Maps; Mediating; Methylation; Mitotic; Natural regeneration; Organoids; Pathway interactions; Phenotype; Population; Positioning Attribute; Regenerative Medicine; Regulation; Role; Stem cells; System; Testing; Tissues; Training; Traumatic injury; Wild Type Mouse; demethylation; design; embryonic stem cell; epigenetic regulation; epigenome; epigenomics; histological studies; in vivo; intestinal crypt; intestinal epithelium; new therapeutic target; progenitor; recruit; response; seal; self-renewal; skills; stem cell biology; stem cell differentiation; stem cell population; stemness; transcriptome sequencing; tumorigenesis

ABSTRACT Intestinal stem cells (ISCs) reside in intestinal crypts and undergo continual proliferation, self-renewal, and differentiation in order to maintain function of the intestinal epithelium, which is replaced approximately once a week throughout adult life. Precise regulation of cell fate decisions between self-renewal and differentiation is critical for ISC homeostasis and damage response, but the role of chromatin regulation in balancing these decisions remains poorly characterized. Our currently funded K01 focuses on dissecting the genomic regulatory network of Tet1, which catalyzes DNA demethylation and is specifically enriched in ISC populations. Preliminary data from these studies demonstrate that loss of Tet1 results in remarkably rapid crypt hyperplasia and increased ISC function, which occurs within 5 days of Tet1 ablation. The central hypothesis of the proposed study is that Tet1 negatively regulates ISC activity by promoting differentiation. To test this hypothesis and expand on our K01, we will: (1) characterize the phenotypic role of Tet1 in ISC proliferation and differentiation in vivo, at 5 days and 4 weeks post-ablation (Aim 1) and (2) map the Tet1-dependent genomic landscape of 5-hydroxymethylcytosine (5hmC), the primary product of TET1 catalytic activity, in ISCs, progenitors, and post-mitotic cells (Aim 2). Collectively, these Aims will expand on and integrate with our ongoing K01 studies to provide sufficient preliminary data for future, R01-funded investigation of Tet1-mediated mechanisms involved in ISC homeostasis and regeneration.

摘要 肠干细胞（ISCs）存在于肠隐窝中，并经历持续增殖、自我更新和分化。 为了维持肠上皮细胞的功能，肠上皮细胞大约一次被替换， 一周的成年生活。在自我更新和分化之间对细胞命运决定的精确调节是 对于ISC稳态和损伤反应至关重要，但染色质调节在平衡这些方面的作用 决定的特点仍然很差。我们目前资助的K 01专注于解剖基因组 Tet 1的调节网络，它催化DNA去甲基化并在ISC群体中特异性富集。 这些研究的初步数据表明，Tet 1的缺失导致隐窝显著快速增生 增加ISC功能，这发生在Tet 1消融后5天内。的中心假设 Tet 1通过促进分化负调节ISC活性。为了验证这一 假设并扩展我们的K 01，我们将：（1）表征Tet 1在ISC增殖中的表型作用， 体内分化，在消融后5天和4周（目标1）和（2）绘制Tet 1依赖性基因组图谱 ISCs中TET 1催化活性的主要产物5-羟甲基胞嘧啶（5 hmC）的分布， 祖细胞和有丝分裂后细胞（Aim 2）。总的来说，这些目标将扩展并整合我们的 正在进行的K 01研究，为未来R 01资助的Tet 1介导的研究提供足够的初步数据。 ISC的稳态和再生机制。