Promoting Diversity via Single-cell Metabolomics and Proteomics: The Missing Link to Understanding Vertebrate Embryonic Patterning

通过单细胞代谢组学和蛋白质组学促进多样性：理解脊椎动物胚胎模式缺失的环节

• 批准号: 10170538
• 负责人: Peter Nemes
• 金额: $ 7.22万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10170538
• 关键词: Barker Hypothesis; Behavioral Assay; Biochemical; Biological; Biology; Biomedical Research; Capillary Electrophoresis; Cell Differentiation process; Cell Lineage; Cells; Chemistry; Cognitive; Data; Development; Doctor of Philosophy; Embryo; Embryonic Development; Generations; Genes; Germ Layers; Goals; Human; Immersion; Impairment; Individual; Knowledge; Laboratory Research; Learning; Link; Molecular; Outcome; Pattern; Process; Production; Proteins; Proteomics; Research; Research Training; Role; Spectrometry, Mass, Electrospray Ionization; Structural Congenital Anomalies; Students; Technology; Testing; Time; Tissues; Training; Transcript; Underrepresented Students; Vertebrates; Work; Xenopus; Xenopus laevis; blastomere structure; career; career development; empowered; experience; frontier; gene function; graduate student; innovation; interdisciplinary approach; metabolome; metabolomics; next generation sequencing; precursor cell; programs; research and development; skills; small molecule; stem cells; vertebrate embryology; vertebrate embryos; zygote

Abstract The goal of this project is to enhance the diversity of the biomedical research workforce by training a PhD Graduate Student from underrepresented backgrounds to enable their original research and career development at the frontiers of chemistry and biology. The student will learn advanced bioanalytical chemistry and vertebrate embryology while elucidating the mechanism of action underlying cell fate changes by small molecules called metabolites, which the Nemes Research Laboratory has recently discovered. Understanding embryogenesis requires knowledge of all the molecules produced as the zygote differentiates into the three primary germ layers of the embryo. Four decades of innovative embryological manipulations, testing of gene functions one gene at a time, and recently, Next-Generation Sequencing have identified multiple transcripts and abundant proteins that are essential to the patterning of the vertebrate embryo. However, very little is known about the contribution of small molecules called metabolites to the formation of the germ layers and the long-term development and functioning of the embryo. The proposed training–research program fills this knowledge gap in technology and biology by empowering the PhD Graduate Student to conduct original research at the chemistry-biology interface. The student will develop skills in bioanalytical chemistry, specifically quantitative metabolomics by capillary electrophoresis and ultrasensitive electrospray ionization mass spectrometry to enable the characterization of the metabolomic states of cells and tissues. Further, the student will also develop the required biomedical–biological skills to study the developmental and cognitive implications of cell fate decisions, including classical embryological manipulations, cell fate tracking, Xenopus laevis biology, and behavioral assays. The outcomes of this interdisciplinary approach will help illuminate the role of the metabolome for the establishment of these important precursor cells and tissues. Because these molecular processes are highly conserved across vertebrates, the data collected from Xenopus are likely to have high relevance to human structural birth defects. The new biochemical information that will be obtained in individual embryonic cells and their progeny (cell lineage) at several critical developmental time points will also advance other research fields that involve cell differentiation (e.g., of stem cells) and the developmental origins of adult disease. This project will provide immersive cross-disciplinary training–research experience to enable the student to pursue an independent career while diversifying the biomedical research workforce. Nemes-Abstract-1|1

摘要 该项目的目标是通过培养一名博士生来提高生物医学研究人员的多样性 来自代表性不足背景的研究生，使他们的原创研究和职业生涯 在化学和生物学的前沿发展。学生将学习高级生物分析化学 和脊椎动物胚胎学，同时阐明小细胞凋亡引起细胞命运变化的作用机制。 尼弥斯研究实验室最近发现了一种叫做代谢物的分子。理解 胚胎发生需要了解受精卵分化为三种细胞时产生的所有分子。 胚胎的初级胚层。四十年来，创新的胚胎学操作，基因测试 一次只对一个基因起作用，最近，下一代测序技术已经鉴定出多个转录本 以及丰富的蛋白质，这些蛋白质是脊椎动物胚胎形成所必需的。然而，很少有 已知称为代谢物的小分子对胚层形成的贡献， 胚胎的长期发育和功能。拟议的培训研究计划填补了这一点 技术和生物学的知识差距，授权博士研究生进行原创 化学-生物学界面的研究。学生将发展生物分析化学的技能， 毛细管电泳-超灵敏电喷雾离子化特异性定量代谢组学 质谱法，以能够表征细胞和组织的代谢组学状态。此夕h 学生还将培养研究发育和认知所需的生物医学生物技能 细胞命运决定的影响，包括经典的胚胎学操作，细胞命运跟踪，非洲爪蟾 lievis生物学和行为分析。这种跨学科方法的结果将有助于阐明 代谢组在建立这些重要的前体细胞和组织中的作用。因为这些 分子过程在脊椎动物中是高度保守的，从非洲爪蟾收集的数据很可能 与人类结构性出生缺陷有很高的相关性。新的生化信息将在 在几个关键的发育时间点的单个胚胎细胞和它们的后代（细胞谱系）也将 推进涉及细胞分化的其他研究领域（例如，干细胞的发育起源 成人疾病。该项目将提供身临其境的跨学科培训研究经验， 学生追求独立的职业生涯，同时多样化的生物医学研究队伍。 尼弥斯-摘要-1| 1