PRDM16 regulation of metabolism in the intestinal stem cell niche

PRDM16对肠道干细胞生态位代谢的调节

• 批准号: 10553635
• 负责人: Rachel Raeburn Webster Stine
• 金额: $ 11.89万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553635
• 关键词: 3-Dimensional; Acetates; Acetyl Coenzyme A; Adult; Advisory Committees; Affect; Amino Acids; Apoptosis; Award; Back; Biology; Carbon; Cell Differentiation process; Cell Lineage; Cell Maintenance; Cell Respiration; Cell physiology; Cells; Cellular Metabolic Process; Cessation of life; Citrates; Citric Acid Cycle; Communities; Cytoplasm; Data Analyses; Daughter; Defect; Dependence; Digestive System Disorders; Disease; Distal; Down-Regulation; Duodenum; Environment; Enzymes; Epigenetic Process; Epithelial Cells; Exhibits; Exposure to; Fatty Acids; Foundations; Funding; Future; Gastrointestinal tract structure; Gene Expression; Genes; Genetic; Glucose; Goals; Grant; Growth; Histone Acetylation; Histones; Human; Intestinal Diseases; Intestines; Investigation; Laboratories; Mentors; Mentorship; Metabolic; Metabolic Control; Metabolic Diseases; Metabolic Pathway; Metabolism; Methods; Mission; Mitochondria; Modeling; Mus; Mutant Strains Mice; National Institute of Diabetes and Digestive and Kidney Diseases; Nuclear; Nutrient; Organoids; Pathway interactions; Pennsylvania; Periodicity; Phenotype; Population; Positioning Attribute; Process; Production; Publications; Regulation; Research; Research Personnel; Resources; Role; Scientist; Secretory Cell; Secure; Signal Pathway; Signal Transduction; Small Intestines; Source; Supplementation; System; Techniques; Therapeutic Intervention; Tissues; Training; Translating; United States National Institutes of Health; Universities; Villus; building materials; career development; cell behavior; cell type; epigenetic regulation; etomoxir; fatty acid oxidation; genome-wide; histone modification; human tissue; in vivo; inhibitor; insight; intestinal crypt; intestinal epithelium; member; metabolic abnormality assessment; metabolic profile; mouse model; mutant; pharmacologic; programs; single cell sequencing; single-cell RNA sequencing; skills; stem; stem cell biology; stem cell function; stem cell niche; stem cell population; stem cells; success; tenure track; transcription factor; transcriptome sequencing; wasting

PROJECT SUMMARY Long term objectives and training aims: With this award, Dr. Rachel Stine will receive the support, mentorship and training required to reach her ultimate goal of becoming an independent investigator focused on the metabolic control of stem cells within the intestinal niche. This research is an excellent fit for the mission of the NIDDK as it relates to both digestive and metabolic disorders. The University of Pennsylvania offers all of the scientific resources required to complete this proposal, as well as two exemplary research programs focused on metabolic studies and intestinal biology respectively. Dr. Stine has assembled a group of renowned scientists to serve as her mentors, advisory committee and collaborators. She has developed a training plan to enhance her publication record, to secure independent funding in the form of project grants and to apply for and secure an independent position by the completion of this award. Dr. Stine’s distinctive research program seeks to answer basic questions about stem cell biology in the intestine, and will ultimately provide insight into how alterations in metabolic control of stem cells and their differentiating daughters can lead to a disease state. Dr. Stine will master techniques essential to her success in this proposal and her future independent research; integrate and expand her expertise in metabolism and intestinal biology through classes, mentorship and interactions within the broader scientific community; and build skills to successfully secure an independent position and start a new laboratory. Background and research aims: Intestinal stem cells have the capacity to rapidly divide and replenish the intestinal lining every few days. Preliminary studies completed by Dr. Stine show that deletion of the transcription factor PRDM16 in an adult mouse causes severe intestinal wasting within five days and death shortly after. RNAseq following Prdm16 deletion shows downregulation of metabolic genes in the intestinal crypt, particularly members of the fatty acid oxidation (FAO) pathway. Intriguingly, pharmacological inhibition of FAO blocks budding and growth of intestinal enteroids, specifically in the proximal small intestine where PRDM16 is highly expressed. Both PRDM16-deficiency and pharmacological inhibition of FAO can be rescued by supplementation with acetate, which can replenish pools of acetyl-CoA. Aim 1 will determine which intestinal progenitor cell populations require high levels of PRDM16 and FAO, allowing for more targeted analysis into how these pathways regulate intestinal differentiation. This aim will also explore whether mechanisms identified in mice are applicable to a human system. Aim 2 focuses on why FAO specifically is required for acetyl-CoA production, even in the presence of other nutrients. Because acetyl-CoA facilitates acetylation of histones, histone profiling as well as genetic perturbations in the acetyl-CoA pathway will be used to explore these mechanisms. This proposal will determine how metabolic changes in intestinal stem and progenitor cells translate to changes in cell behavior and provide fundamental insights into the role metabolism in this system.

项目总结 长期目标和培训目标：有了这个奖项，雷切尔·斯汀博士将获得支持、指导 以及达到她最终目标所需的培训，即成为一名专注于 干细胞在肠道内的代谢控制。这项研究非常适合 NIDDK，因为它与消化和代谢障碍有关。宾夕法尼亚大学提供所有 完成这项提议所需的科学资源，以及重点关注的两个示范性研究计划 分别在代谢研究和肠道生物学方面。斯汀博士召集了一群著名的 科学家担任她的导师、顾问委员会和合作者。她制定了一项培训计划，以 改善她的出版记录，以项目赠款的形式获得独立资金，并申请 并在完成本奖项后获得一个独立的职位。斯汀博士独特的研究计划 旨在回答有关肠道干细胞生物学的基本问题，并最终将提供对 干细胞及其分化子细胞代谢控制的改变如何导致疾病状态。 Stine博士将掌握对她在这项提案中的成功和她未来的独立研究至关重要的技术； 通过课程、指导和培训，整合和扩展她在代谢和肠道生物学方面的专业知识 在更广泛的科学界内进行互动；并培养技能，以成功地确保独立的 安置并启动一个新的实验室。背景和研究目的：肠道干细胞具有 每隔几天迅速分割和补充肠壁。Stine博士完成的初步研究 研究表明，成年小鼠体内转录因子PRDM16的缺失会导致严重的肠道萎缩 五天内死亡，不久后死亡。Prdm16缺失后的RNAseq显示代谢下调 肠道隐窝中的基因，特别是脂肪酸氧化(FAO)途径的成员。有趣的是， 粮农组织的药理抑制作用阻止了肠小球的萌发和生长，特别是在近端 PRDM16高表达的小肠。PRDM16缺乏症与药物抑制 粮农组织可以通过补充醋酸盐来拯救，醋酸盐可以补充乙酰辅酶A池。目标1将 确定哪些肠道祖细胞群体需要高水平的PRDM16和粮农组织，允许 对这些途径如何调节肠道分化进行更有针对性的分析。这一目标还将探索 在老鼠身上发现的机制是否适用于人类系统。目标2侧重于为什么粮农组织 特别是乙酰-辅酶A的生产，即使在其他营养物质存在的情况下也是如此。因为乙酰辅酶A 促进组蛋白的乙酰化、组蛋白图谱以及乙酰辅酶A途径中的遗传扰动 将被用来探索这些机制。这一提议将决定肠道新陈代谢的变化 干细胞和祖细胞转化为细胞行为的变化，并提供了对其作用的基本见解 在这个系统中的新陈代谢。