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.

项目概要 细胞代谢途径在不同细胞类型的发育和发育过程中表现出显着的可塑性 除了伴随的细胞状态变化之外，代谢重新布线已被证明可以驱动细胞命运。 通过改变染色质景观来决定程序，装饰化学修饰的沉积。 染色质需要代谢途径的中间体，并且使用几种酶来去除这些标记 代谢物作为其酶反应的一部分，因此，代谢物水平的波动有能力。 塑造染色质以影响细胞命运特异性基因表达，但驱动染色质的代谢变化 重组和介导早期发育过程中细胞命运代谢控制的酶仍然存在 我们之前已经鉴定出控制小鼠胚胎自我更新的特定代谢物。 干细胞 (ESC) 的新陈代谢是否随着 ESC 退出自我更新的多能状态而改变，以及是否会改变。 多谱系分化所需的这些代谢变化仍然是一个悬而未决的问题。 本研究计划的目标是描述退出过程中发生的代谢重新布线的特征。 幼稚多能性并确定这种重新布线控制小鼠 ESC 的机制 我们的初步数据表明，幼稚多能性的退出伴随着增加。 在目标 1 中，我们将使用遗传和药理学方法来靶向柠檬酸的线粒体输出。 线粒体柠檬酸转运蛋白 SLC25A1 或下游柠檬酸分解酶 ATP-柠檬酸裂解酶 检验早期分化需要线粒体来源的柠檬酸的假设。 这种代谢变化是否通过柠檬酸盐衍生的组蛋白乙酰化的沉积来调节细胞命运 初步数据还显示细胞氧化还原状态的变化，其标志是胞质的增加。 在目标 2 中，我们将确定早期分化期间的 NAD+/NADH 比率是否需要这种代谢变化。 通过使用药理学或遗传工具调节 NAD+/NADH 比率进一步退出幼稚多能性。 实验将确定细胞氧化还原状态向染色质景观发出信号的机制 这些研究将揭示退出幼稚多能性期间的代谢控制机制。 并将提供关于代谢调节如何促进细胞身份变化的重要见解 本提案中概述的工作和培训计划将在实验室中完成。 纪念斯隆·凯特琳癌症中心的 Lydia Finley 博士和 Kristian Helin 博士的共同顾问 将为申请人提供进一步的临床培训和作为独立医师科学家的职业生涯的理想准备。