Epigenetic regulation of lineage specification in colon epithelial cells

结肠上皮细胞谱系规范的表观遗传调控

• 批准号: 10676682
• 负责人: Anne R. Meyer
• 金额: $ 6.95万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10676682
• 关键词: Address; Biological Models; Blood; Cancer Patient; Candidate Disease Gene; Cell Differentiation process; Cell Fate Control; Cell Lineage; Cell Maintenance; Cell physiology; Cells; Chromatin; Chromatin Structure; Colitis; Colon; Complementary RNA; DNA; DNA Sequence; DNA-Directed RNA Polymerase; Data; Dedications; Disease; Dominant-Negative Mutation; Epigenetic Process; Epithelial Cells; Epithelium; Fluorescence-Activated Cell Sorting; Gene Expression; Genes; Genetic Transcription; Goblet Cells; Health; Histone H3; Histones; Homeostasis; Human; Knockout Mice; Lysine; Maintenance; Malignant Neoplasms; Messenger RNA; Methionine; Methylation; Methyltransferase; Modeling; Mucous body substance; Mus; Mutation; N-terminal; Organoids; Physiology; Play; Polymerase; Population; Post-Translational Protein Processing; Process; Production; Proteins; Protocols documentation; RNA Polymerase II; Role; Site; Sorting; Specific qualifier value; System; Tail; Testing; Testis; Tissues; Water; Work; absorption; candidate identification; cell type; comparative; epigenetic regulation; epigenomic profiling; experimental study; genome-wide; histone methylation; histone modification; inhibitor; innovation; knock-down; mutant; mutant mouse model; novel; programs; recruit; scaffold; self-renewal; single-cell RNA sequencing; small hairpin RNA; stem cells; tool; transcriptome sequencing

PROJECT SUMMARY The colon is a responsive tissue, and the epithelium replaces itself every 3-4 days. The colon has dedicated stem cells that divide for self-renewal and differentiation into specialized cell types. Differentiated colon cells perform distinct functions including water absorption and mucus secretion. In many tissues, differentiation is driven by epigenetic mechanisms including histone modifications. Methylation of histone H3 at lysine 36 (H3K36) is associated with gene bodies at sites of transcriptional activity. H3K36 methylation contributes to differentiation and stem cell maintenance in rapidly renewing tissues including blood and testes. Based on preliminary data, I hypothesize that H3K36 methylation governs cell fate decisions and reinforces cell identity in colon epithelial cells, specifically colonocyte and goblet cell lineages. To address this hypothesis, I will pursue two specific aims. First, I plan to characterize the H3K36 methylation signature in colon epithelial cells. Second, I will determine the functional role of H3K36 methylation in colon cell identity. Although, establishing a direct connection between specific histone modifications and cellular processes is challenging due to redundancies in the system, I can overcome this obstacle by taking advantage of a lysine (K)-to-methionine (M) mutation first discovered in cancer patients to precisely suppress H3K36 methylation. I will combine multiple cutting-edge tools to specifically trace and manipulate H3K36 methylation in mice and organoid model systems to advance the understanding of chromatin and lineage specification in the colon. I will also utilize novel sorting protocols to purify distinct colon cell populations for comparative epigenetic and transcriptional analysis. Collectively, this work is significant because it will establish the role of H3K36 methylation in controlling cell-type specific gene expression and lineage decisions in the colon. Revealing the role histone modifications play in colon homeostasis is the first step to understanding how these regulatory mechanisms break down in diseases such as colitis/cancer and could impact human health.

项目摘要 结肠是一种反应性组织，上皮每3-4天就自身替代。结肠献身于 分裂自我更新和分化为专用细胞类型的干细胞。分化的结肠细胞 执行不同的功能，包括吸水和粘液分泌。在许多组织中，分化是 由包括组蛋白修饰的表观遗传机制驱动。在赖氨酸36（H3K36）处组蛋白H3的甲基化 与转录活性部位的基因体有关。 H3K36甲基化有助于分化 以及包括血液和睾丸在内的快速更新组织中的干细胞维持。基于初步数据，我 假设H3K36甲基化控制细胞命运决策并增强结肠上皮的细胞身份 细胞，特别是结肠细胞和杯状细胞谱系。为了解决这一假设，我将追求两个具体的目标。 首先，我计划表征结肠上皮细胞中H3K36甲基化特征。第二，我将确定 H3K36甲基化在结肠细胞身份中的功能作用。虽然，建立直接联系 由于系统中的冗余，特定的组蛋白修饰和蜂窝过程很具有挑战性，我可以 通过利用赖氨酸（K） - 甲硫代（M）突变来克服这一障碍 患者精确抑制H3K36甲基化。我将结合多个尖端工具以专门跟踪 并操纵小鼠和器官模型系统中的H3K36甲基化，以提高对 结肠中的染色质和谱系规范。我还将利用新颖的排序方案净化不同的结肠 比较表观遗传和转录分析的细胞群。总的来说，这项工作很重要 因为它将确定H3K36甲基化在控制细胞类型特异性基因表达和 结肠中的血统决定。揭示组蛋白修饰在结肠体内稳态中的作用是第一步 了解这些调节机制如何在结肠炎/癌症等疾病中分解 影响人类健康。