Role of Energy Metabolism in Patterning the Vertebrate Musculo-Skeletal Axis

能量代谢在脊椎动物肌肉骨骼轴模式中的作用

• 批准号: 10211585
• 负责人: OLIVIER POURQUIE
• 金额: $ 67.01万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10211585
• 关键词: Acetylation; Adipose tissue; Cell Differentiation process; Cells; Characteristics; Chick Embryo; Chickens; Complex; Congenital Abnormality; Congenital Scoliosis; Connective Tissue; Defect; Development; Embryo; Energy Metabolism; Enzymes; Exhibits; Funding; Gene Expression Regulation; Global Change; Glycolysis; Goals; Human; Human Development; Image; In Vitro; Knowledge; Lead; Light; Link; Longevity; Malignant Neoplasms; Mammals; Mediating; Mesoderm; Mesoderm Cell; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Molecular; Mus; Musculoskeletal; Paraxial Mesoderm; Pattern; Physiological; Play; Population; Post-Translational Protein Processing; Primitive Streaks; Process; Production; Property; Protein Acetylation; Proteomics; Proxy; Regulation; Reporter; Respiration; Role; Segmentation Clock Pathway; Signal Transduction; Skeletal Muscle; Skeleton; Somites; Species Specificity; Spinal Dysraphism; Structure; System; Tail; Time; Tissues; Transcriptional Regulation; Vertebral column; WNT Signaling Pathway; Warburg Effect; Work; aerobic glycolysis; beta catenin; cancer cell; cell fate specification; differentiation protocol; embryo cell; embryonic stem cell; epigenomics; experimental study; extracellular; in vivo; induced pluripotent stem cell; live cell imaging; malformation; metabolomics; method development; mouse development; multiple omics; pH gradient; progenitor; protein function; single-cell RNA sequencing; spine bone structure; stem cells; transcriptomics

Project Summary/Abstract The proposed project is focused on the crosstalk between metabolism and cell fate during development of the paraxial mesoderm, the tissue which forms skeletal muscles and vertebrae. One striking characteristic of this tissue is its segmentation into repeated structures termed somites, a process driven by a molecular oscillator called segmentation clock [1]. Defects in paraxial mesoderm development can lead to severe malformations such as congenital scoliosis, spina bifida or caudal agenesis. The paraxial mesoderm arises from a population of progenitors located in the primitive streak and tail bud. Remarkably, these progenitor cells exhibit aerobic glycolysis and an inverted intra- vs. extracellular pH gradient, which are characteristic of the Warburg effect of cancer cells [2, 3]. In the tailbud, we demonstrated that glycolysis increases the intracellular pH to promote acetylation of -catenin and Wnt activation, which ultimately leads to paraxial mesoderm induction [3]. As these processes are difficult to study in vivo, we have developed in vitro systems in which embryonic stem (ES) cells or induced pluripotent stem (iPS) cells can be efficiently differentiated toward the paraxial mesoderm fate recapitulating the normal features of its metabolism, signaling and even oscillations of the segmentation clock [4-7]. We will now take advantage of these in vitro systems, as well as mouse and chicken embryos, to characterize in detail the role of aerobic glycolysis in paraxial mesoderm development to see how it relates to the Warburg effect. In this application, we propose to carry out large scale multi-omics experiments (metabolomics, transcriptomics, proteomics, epigenomics) and live imaging of cellular metabolic state to characterize the impact of metabolic transitions on the regulation of gene expression, protein function and cell fate. As the physiological significance of the Warburg effect is not well understood, carefully dissecting its role in the embryo might help shed light on its role in cancer. Finally, we propose to use in vivo, ex vivo and in vitro systems recapitulating the oscillations of the segmentation clock to study the role of metabolism in the control of the oscillatory period. We will analyze the differences in metabolism regulation between mouse and human paraxial mesoderm cells, with special focus on mitochondrial respiration. The period of the oscillations diverges significantly between the two species and can be used as a proxy for developmental timing to try to understand why human development proceeds more slowly than mouse development. We expect these experiments to shed light on the molecular basis of developmental timing, which is tightly linked to longevity in mammals.

项目概要/摘要 拟议的项目重点关注细胞发育过程中新陈代谢和细胞命运之间的串扰 轴旁中胚层，形成骨骼肌和椎骨的组织。这其中有一个显着的特点 组织被分割成称为体节的重复结构，这是一个由分子振荡器驱动的过程 称为分段时钟[1]。轴旁中胚层发育缺陷可导致严重畸形 例如先天性脊柱侧凸、脊柱裂或尾椎发育不全。近轴中胚层起源于群体 祖细胞位​​于原条和尾芽中。值得注意的是，这些祖细胞表现出有氧能力 糖酵解和细胞内与细胞外 pH 梯度相反，这是 Warburg 效应的特征 癌细胞 [2, 3]。在尾芽中，我们证明糖酵解会增加细胞内 pH 值，从而促进 -连环蛋白的乙酰化和 Wnt 激活，最终导致轴旁中胚层诱导 [3]。正如这些 体内过程很难研究，我们开发了体外系统，其中胚胎干（ES）细胞 或诱导多能干（iPS）细胞可以有效地分化为近轴中胚层命运 概括其新陈代谢、信号传导甚至分段时钟振荡的正常特征 [4-7]。我们现在将利用这些体外系统以及小鼠和鸡胚胎来 详细描述有氧糖酵解在轴旁中胚层发育中的作用，看看它与 瓦尔堡效应。在此应用中，我们建议进行大规模多组学实验 （代谢组学、转录组学、蛋白质组学、表观基因组学）和细胞代谢状态的实时成像 表征代谢转变对基因表达、蛋白质功能和细胞调节的影响 命运。由于瓦尔堡效应的生理意义尚不清楚，仔细剖析其作用 胚胎中的这种现象可能有助于阐明其在癌症中的作用。最后，我们建议使用体内、离体和体外 系统重现分段时钟的振荡，以研究新陈代谢在控制中的作用 的振荡周期。我们将分析小鼠和人类代谢调节的差异 近轴中胚层细胞，特别关注线粒体呼吸。振荡周期发散 两个物种之间存在显着差异，可以用作发育时间的代理来尝试理解 为什么人类发育比小鼠发育慢。我们期望这些实验能够 揭示了发育时间的分子基础，这与哺乳动物的寿命密切相关。