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

基本信息

批准号：
10853562
负责人：
Rachel Raeburn Webster Stine
金额：
$ 10.39万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
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/10853562
关键词：
3-Dimensional Acetates Acetyl Coenzyme A Affect Apoptosis Back Carbon Cell Differentiation process Cell Lineage Cell Maintenance Cell Respiration Cell physiology Cells Citrates Citric Acid Cycle Cytoplasm Data Analyses Defect Dependence Disease Distal Duodenum Enzymes Epigenetic Process Epithelial Cells Exhibits Fatty Acids Foundations Funding Future Gene Expression Genes Histone Acetylation Histones Human Intestinal Diseases Intestines Investigation Metabolic Metabolic Pathway Metabolism Methods Mitochondria Modeling Mutant Strains Mice Nuclear Organoids Pathway interactions Periodicity Phenotype Population Positioning Attribute Process Production Publications Regulation Research Role Secretory Cell Signal Pathway Signal Transduction Small Intestines Source Techniques Tissues Training United States National Institutes of Health Villus career development cell type epigenetic regulation etomoxir fatty acid oxidation genome-wide histone modification human tissue in vivo inhibitor intestinal crypt intestinal epithelium metabolic profile mouse model mutant programs single cell sequencing single-cell RNA sequencing stem stem cell function stem cell niche stem cell population stem cells tenure track transcription factor

项目摘要

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.

项目概要长期目标和培训目标：通过此奖项，Rachel Stine 博士将获得支持和指导以及实现成为一名独立调查员的最终目标所需的培训肠道微环境内干细胞的代谢控制。这项研究非常适合该组织的使命 NIDDK 与消化和代谢紊乱有关。宾夕法尼亚大学提供所有完成该提案所需的科学资源，以及两个重点关注的示范性研究计划分别从事代谢研究和肠道生物学。斯坦博士聚集了一批著名的科学家们担任她的导师、顾问委员会和合作者。她制定了一个培训计划提高她的出版记录，以项目补助金的形式获得独立资金并申请并通过完成该奖项确保独立地位。 Stine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和未来独立研究中取得成功至关重要的技术；通过课程、指导和培训，整合和扩展她在新陈代谢和肠道生物学方面的专业知识更广泛的科学界内的互动；并培养技能以成功获得独立定位并启动一个新的实验室。背景和研究目的：肠道干细胞有能力每隔几天迅速分裂并补充肠道内壁。 Stine 博士完成的初步研究表明成年小鼠转录因子 PRDM16 的缺失会导致严重的肠道浪费五天内，不久后死亡。 Prdm16 删除后的 RNAseq 显示代谢下调肠隐窝中的基因，特别是脂肪酸氧化（FAO）途径的成员。有趣的是，药理学抑制FAO可阻止肠内肠类的出芽和生长，特别是在近端 PRDM16 高表达的小肠。 PRDM16 缺乏和药物抑制补充醋酸盐可以挽救FAO，醋酸盐可以补充乙酰辅酶A。目标1将确定哪些肠道祖细胞群需要高水平的PRDM16和FAO，从而允许更有针对性地分析这些途径如何调节肠道分化。这一目标还将探索在小鼠中发现的机制是否适用于人类系统。目标 2 重点关注粮农组织为何即使存在其他营养素，乙酰辅酶 A 的生产也特别需要。因为乙酰辅酶A 促进组蛋白乙酰化、组蛋白分析以及乙酰辅酶A途径中的遗传扰动将用于探索这些机制。该提案将确定肠道代谢变化如何干细胞和祖细胞转化为细胞行为的变化，并提供对其作用的基本见解在这个系统中进行新陈代谢。