PRDM16 regulation of metabolism in the intestinal stem cell niche

项目摘要 长期目标和培训目标：凭借该奖项，Rachel Stine博士将获得支持、指导 她的最终目标是成为一名独立调查员， 代谢控制的干细胞在肠道生态位。这项研究非常适合美国宇航局的使命。 NIDDK，因为它涉及消化和代谢紊乱。宾夕法尼亚大学提供所有 完成这项提案所需的科学资源，以及两个示范性的研究计划，重点是 代谢研究和肠道生物学。Stine博士召集了一群著名的 科学家担任她的导师，咨询委员会和合作者。她制定了一个培训计划， 提高她的出版记录，以项目赠款的形式获得独立资助，并申请 并通过完成该奖项获得独立地位。斯坦博士独特的研究项目 旨在回答有关肠道干细胞生物学的基本问题，并最终将提供深入了解 干细胞及其分化子细胞的代谢控制的改变如何导致疾病状态。 博士Stine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，导师制和 在更广泛的科学界的互动;并建立技能，成功地确保一个独立的 建立新的实验室并开始实验。背景和研究目的：肠道干细胞具有 每隔几天就迅速分裂和补充肠壁。Stine博士完成的初步研究 显示在成年小鼠中转录因子PRDM 16缺失导致严重的肠消耗 五天内死亡Prdm 16缺失后的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， FAO的药理学抑制阻断肠类肠的出芽和生长，特别是在近端 小肠中PRDM 16高度表达。PRDM 16缺陷和对细胞增殖的药理学抑制 FAO可以通过补充乙酸来拯救，乙酸可以补充乙酰辅酶A的库。目标1将 确定哪些肠道祖细胞群体需要高水平的PRDM 16和FAO， 更有针对性地分析这些途径如何调节肠道分化。这一目标还将探讨 在小鼠中发现的机制是否适用于人类系统。目标2：为什么粮农组织 特别是乙酰辅酶A生产所需的，即使在其他营养素的存在下。因为乙酰辅酶A 促进组蛋白的乙酰化、组蛋白谱分析以及乙酰辅酶A途径中的遗传扰动 将被用来探索这些机制。这项建议将确定如何代谢变化，在肠道 干细胞和祖细胞转化为细胞行为的变化， 在这个系统中，

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.