Resubmission of Diversity Supplement

重新提交多样性补充材料

• 批准号: 10579764
• 负责人: James Bayrer
• 金额: $ 12.16万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-14 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579764
• 关键词: Address; Adult; Autoimmune Diseases; Automobile Driving; Binding; Biochemical; Biological Models; Biology; CRISPR-mediated transcriptional activation; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cell Proliferation; Cell Survival; Cells; Chromatin; Complement; Complex; Coupled; Disease; Drug Receptors; Enteroendocrine Cell; Environment; Epithelial; Epithelial Cells; Epithelial Physiology; Epitopes; Equilibrium; Evaluation; Funding; Genes; Genetic; Genetic Transcription; Goals; Health; Hormones; Infection; Inflammatory; Injury; Intestines; Investigation; Knock-out; Knowledge; Lead; Liquid substance; Maps; Mediating; Modeling; Molecular; Mucous Membrane; NR5A2 gene; Nuclear Receptors; Organoids; Outcome; Pharmacology; Pharmacotherapy; Physiology; Population; Positioning Attribute; Post-Translational Protein Processing; Process; Production; Property; Regulation; Regulator Genes; Research; Role; Secretory Cell; Shapes; Signal Transduction; Sumoylation Pathway; Supporting Cell; Surface; Surveys; Technology; Therapeutic; Tissues; Work; base; cell type; crypt cell; design; directed differentiation; drug development; enteritis; epithelial repair; epithelial stem cell; experimental study; follow-up; gastrointestinal epithelium; healing; in vivo; injury recovery; insight; intercellular communication; intestinal crypt; intestinal epithelium; intestinal injury; mouse model; mutant; novel; overexpression; pluripotency factor; progenitor; programs; receptor function; repaired; response to injury; restoration; stem; stem cell division; stem cell population; stem cell survival; stem cells; stemness; tool; transcription factor

Project Summary/Abstract Background: The primary epithelial stem cell in the intestine has been known for over a decade, leading to important observations in epithelial renewal and injury recovery. These fundamental studies have demonstrated that the intestinal epithelium is a balanced continuum between stemness in the crypt base and differentiation towards the mucosal surface, with complex signaling networks influencing stemness and fate of the differentiating progenitor cells. Several groups have now shown that destruction of intestinal stem cells and/or extensive epithelial damage can be repaired by the dedifferentiation of progenitor cells. Thus, the balance between stemness and differentiation can be tipped by as yet undefined molecular mechanisms. We discovered that the nuclear receptor Liver Receptor Homolog-1 (LRH-1; NR5A2) is essential for both stem cell maintenance and for directing differentiation of the highly specialized enteroendocrine cell lineage. Therefore, this unusual transcription factor has dual roles promoting stemness and cell differentiation, making it an attractive target for elucidating the mechanisms responsible for gut epithelial renewal and healing. Approach: In this application, we will 1) determine the contribution of LRH-1 to epithelial repair and stem cell restitution in enteritis models, 2) determine how SUMOylation of LRH-1 modulates receptor function in stem and progenitor cells, and 3) determine and validate the cell-specific gut LRH-1 “targetome.” We will accomplish this through the use of lineage-traced mouse models and novel implementations of the intestinal organoid platform. Following successful completion of our aims, we will have mechanistically evaluated the contribution of a druggable nuclear receptor to 1) balancing stem cell renewal, differentiation, and restitution of damaged epithelium 2) provided the first ever detailed evaluation of SUMOylated LRH-1 in the gut, and 3) generated a bona fide LRH-1 gut targetome that will be invaluable for receptor drug development. Goals: This proposal is designed to expand upon a key finding uncovered during my funded K08 investigation and advance our mechanistic understanding of how the gut balances stemness vs differentiation during normal and diseased states. Underlying this work is our hypothesis that LRH-1 is differentially modulated within unique cellular contexts in stem and progenitor cells to drive diverse gene programs simultaneously supporting cell renewal and specialized cell differentiation. Our experiments are designed to maximally exploit the extraordinary confluence of intestinal epithelial physiology and nuclear receptor biology framed by LRH-1.

项目概要/摘要 背景： 肠道中的初级上皮干细胞已为人所知已有十多年了，这导致了重要的研究 上皮更新和损伤恢复的观察结果这些基础研究表明 肠上皮是隐窝基底干性和分化之间的平衡连续体 粘膜表面，具有影响分化的干性和命运的复杂信号网络 一些研究小组现已表明，肠道干细胞和/或广泛的破坏。 上皮损伤可以通过祖细胞的去分化来修复，从而达到平衡。 干性和分化可能由尚未明确的分子机制决定。 我们发现核受体肝受体同源物-1 (LRH-1; NR5A2) 对于两者都是必需的 干细胞维持和指导高度专业化的肠内分泌细胞谱系的分化。 因此，这种不寻常的转录因子具有促进干性和细胞分化的双重作用，使其成为 阐明肠道上皮更新和愈合机制的一个有吸引力的目标。 方法： 在此应用中，我们将 1) 确定 LRH-1 对上皮修复和干细胞恢复的贡献 肠炎模型，2) 确定 LRH-1 的 SUMO 化如何调节干细胞和祖细胞中的受体功能 细胞，3) 确定并验证细胞特异性肠道 LRH-1“目标组”。 使用谱系追踪小鼠模型和肠道类器官平台的新颖实现。 成功完成我们的目标后，我们将机械地评估 可药物核受体 1) 平衡干细胞更新、分化和受损恢复 上皮细胞 2) 首次对肠道中的 SUMOylated LRH-1 进行了详细评估，并且 3) 产生了 真正的 LRH-1 肠道靶组对于受体药物开发具有无价的价值。 目标： 该提案旨在扩展我资助的 K08 调查期间发现的一项重要发现 增进我们对肠道在正常和不同阶段如何平衡干性与分化的机械理解 这项工作的基础是我们的假设，即 LRH-1 在独特的范围内受到差异调节。 干细胞和祖细胞中的细胞环境可同时驱动多种基因程序支持细胞 我们的实验旨在最大限度地利用细胞的更新和专门的分化。 LRH-1 构建了肠上皮生理学和核受体生物学的非凡融合。