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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要结肠是一种反应性组织，上皮每 3-4 天更换一次。结肠有专门的干细胞能够分裂自我更新并分化成特殊的细胞类型。分化的结肠细胞执行不同的功能，包括吸水和分泌粘液。在许多组织中，分化是由包括组蛋白修饰在内的表观遗传机制驱动。组蛋白 H3 在赖氨酸 36 处的甲基化 (H3K36) 与转录活性位点的基因体相关。 H3K36 甲基化有助于分化以及快速更新组织（包括血液和睾丸）中的干细胞维持。根据初步数据，我假设 H3K36 甲基化控制细胞命运决定并增强结肠上皮中的细胞身份细胞，特别是结肠细胞和杯状细胞谱系。为了解决这个假设，我将追求两个具体目标。首先，我计划表征结肠上皮细胞中的 H3K36 甲基化特征。其次，我将确定 H3K36 甲基化在结肠细胞身份中的功能作用。虽然，在之间建立直接连接由于系统冗余，特定的组蛋白修饰和细胞过程具有挑战性，我可以利用首次在癌症中发现的赖氨酸 (K) 到蛋氨酸 (M) 突变克服了这一障碍患者精确抑制 H3K36 甲基化。我会结合多种前沿工具来具体追踪并操纵小鼠和类器官模型系统中的 H3K36 甲基化，以促进对结肠中的染色质和谱系规范。我还将利用新颖的分选方案来纯化不同的结肠用于比较表观遗传和转录分析的细胞群。总的来说，这项工作意义重大因为它将确立 H3K36 甲基化在控制细胞类型特异性基因表达中的作用，结肠中的谱系决定。揭示组蛋白修饰在结肠稳态中的作用是第一步了解这些调节机制如何在结肠炎/癌症等疾病中崩溃，并可能影响人类健康。