Metabolic control of exit from naïve pluripotency

退出幼稚多能性的代谢控制

• 批准号: 10387708
• 负责人: Benjamin Tonnu Jackson
• 金额: $ 5.1万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2026-04-14
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10387708
• 关键词: ATP Citrate (pro-S)-Lyase; Acetates; Acetyl Coenzyme A; Acetylation; Affect; Binding; Biochemical Reaction; Biological Assay; C-terminal binding protein; Cell Fate Control; Cells; ChIP-seq; Chemicals; Chromatin; Citrates; Clinical; Competence; Complex; Cytoplasm; Data; Deposition; Development; Disease; Embryonic Development; Enzymes; Equilibrium; Excision; Exhibits; Gene Expression; Gene Expression Regulation; Genetic; Genetic Transcription; Genomics; Glucose; Goals; Histone Acetylation; LIF gene; Laboratories; Link; MEKs; Malates; Mass Spectrum Analysis; Measures; Mediating; Memorial Sloan-Kettering Cancer Center; Metabolic; Metabolic Control; Metabolic Pathway; Metabolism; Mitochondria; Modification; Mus; NADH; Output; Oxidation-Reduction; Pharmacology; Physicians; Process; Pyruvate; Regulation; Reporter; Reporting; Research Proposals; Scientist; Shapes; Signal Transduction; Supplementation; Testing; Training; Transcription Repressor; Western Blotting; Work; antiporter; base; career; cell fate specification; cell type; chromatin remodeling; citrate carrier; cofactor; defined contribution; developmental disease; embryonic stem cell; experimental study; genetic approach; genomic locus; histone modification; insight; mutant; pluripotency; pluripotency factor; programs; self-renewal; sensor; stem cell differentiation; stem cells; tool

PROJECT SUMMARY Cellular metabolic pathways exhibit remarkable plasticity across different cell types in both development and disease. In addition to accompanying changes in cell state, metabolic rewiring has been shown to drive cell fate decisions programs by altering the chromatin landscape. The deposition of chemical modifications that decorate chromatin requires the intermediates of metabolic pathways, and several enzymes that remove these marks use metabolites as part of their enzymatic reaction. Therefore, fluctuations in metabolite levels have the capacity to shape chromatin to effect cell fate-specific gene expression, but the metabolic changes that drive chromatin reorganization and the enzymes that mediate metabolic control of cell fate during early development remain largely unknown. We have previously identified specific metabolites that control self-renewal of mouse embryonic stem cells (ESCs). Whether metabolism is altered as ESCs exit the self-renewing pluripotent state, and whether these metabolic changes are required for multi-lineage differentiation remains an open question. The goal of this research proposal is to characterize the metabolic rewiring that occurs during exit from naïve pluripotency and to determine the mechanisms by which this rewiring controls mouse ESC differentiation. Our preliminary data indicate that exit from naïve pluripotency is accompanied by an increase in the mitochondrial export of citrate. In Aim 1, we will use genetic and pharmacologic approaches to target the mitochondrial citrate transporter SLC25A1 or the downstream citrate-catabolizing enzyme ATP-citrate lyase to test the hypothesis that mitochondrially-derived citrate is required for early differentiation. We will investigate whether this metabolic change regulates cell fate through the deposition of citrate-derived histone acetylation marks. Preliminary data also shows changes in cellular redox state marked by an increase in the cytosolic NAD+/NADH ratio during early differentiation. In Aim 2, we will determine if this metabolic change is required for exit from naïve pluripotency by modulating the NAD+/NADH ratio using pharmacological or genetic tools. Further experiments will identify the mechanism by which cellular redox state signals to the chromatin landscape to dictate cell fate. These studies will reveal the mechanisms of metabolic control during exit from naïve pluripotency and will provide critical insight into how metabolic regulation contributes to changes in cell identity during embryonic development. The work and training plan outlined in this proposal will be completed in the laboratory of Dr. Lydia Finley with the co-advisement of Dr. Kristian Helin at Memorial Sloan Kettering Cancer Center and will ideally prepare the applicant for further clinical training and a career as an independent physician-scientist.

项目摘要 在发育和 疾病。除了参与细胞状态的参与变化外，代谢重新布线已被证明驱动细胞脂肪 通过更改染色质景观来决定计划。装饰化学修饰的沉积 染色质需要代谢途径的中间体，并消除这些标记的几种酶 代谢物作为其酶促反应的一部分。因此，代谢物水平的波动具有 形成染色质以影响细胞命运特异性基因表达，但代谢变化驱动染色质 早期发育过程中介导细胞脂肪代谢控制的重组和酶 在很大程度上未知。我们先前已经确定了控制小鼠胚胎自我更新的特定代谢产物 干细胞（ESC）。当ESC退出自我更新多能状态以及是否会改变新陈代谢 这些代谢变化是多部位分化所必需的，这仍然是一个悬而未决的问题。 该研究建议的目的是表征退出期间发生的代谢重新布线 幼稚的多能性，并确定这种重新向导控制小鼠ESC的机制 分化。我们的初步数据表明，从幼稚的多能退出是通过增加完成的 在AIM 1中，我们将使用遗传和药物方法来针对 线粒体柠檬酸转运蛋白转运蛋白SLC25A1或下游柠檬酸盐促成酶ATP-CITRATE裂解酶至 测试以下假设：线粒体衍生的柠檬酸盐是早期分化所必需的。我们将调查 这种代谢变化是否通过沉积柠檬酸盐衍生的组蛋白乙酰化来调节细胞脂肪 标记。初步数据还显示了细胞氧化还原状态的变化，该状态标志着胞质的增加 NAD+/NADH在早期分化过程中。在AIM 2中，我们将确定是否需要这种代谢变化 通过使用药物或遗传工具调节NAD+/NADH的比率，从幼稚的多能力中退出。更远 实验将确定细胞氧化还原态信号到染色质景观的机制 决定细胞命运。这些研究将揭示从幼稚多质齿退出期间代谢控制的机制 并将提供有关代谢调节如何促进细胞身份变化的关键见解 胚胎发展。该提案中概述的工作和培训计划将在实验室完成 Lydia Finley博士与Kristian Helin博士在纪念Sloan Kettering Cancer Center的共同审议和 理想情况下，将为申请人做好准备进行进一步的临床培训和作为独立身体科学家的职业。