CAREER: Discovering Upstream Effectors to Cell Fate Determination

职业：发现细胞命运决定的上游效应器

• 批准号: 1651388
• 负责人: Peter Nemes
• 金额: $ 65万
• 依托单位: George Washington University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1651388&HistoricalAwards=false
• 关键词: CAREER; Discovering; Upstream; Effectors; Cell

Non-Technical ParagraphDuring normal development of the vertebrate embryo, cells of the embryo must develop into all the different types of tissues of the organism, but the complete set of biomolecules contributing to tissue formation is unknown. This project utilizes advanced instruments that have only recently been developed in analytical chemistry to determine changes in small molecules as individual cells form different types of tissues in the early developing frog embryo, which is an important model of vertebrate embryo development. The resulting data will provide previously unavailable insights into basic biological processes important for the formation of cells, tissues, organs, and organisms. These research efforts serve as the foundation for interdisciplinary training of diverse participants at the host university, as well as conferences, seminars, and national training centers such as Cold Spring Harbor Laboratory, NY. This work will train a new generation of scientists in both biology and chemistry, including many traditionally underrepresented populations in science, to allow them to address current challenges but also to ask new questions in these fields to better understand normal vertebrate embryo development and diseases.Technical ParagraphDecades of research has uncovered many genes and gene products with critical roles during development of the vertebrate embryo, but how small molecules (called metabolites) participate in cell developmental processes is not fully known. The PI's laboratory recently discovered metabolites capable of altering the normal dorsal-ventral fate of select stem cells in the early frog (Xenopus laevis) embryo, demonstrating that these molecules, too, are active players during patterning of the vertebrate body. The overall goal of this work is to determine the mechanism of action underlying metabolite-induced cell fate decisions. This will be accomplished through a systems cell biology approach, in which the molecular state of metabolite-injected cells will be characterized using unique single-cell mass spectrometry technologies that were developed and validated in the PI's laboratory. The project will identify how cell-fate altering metabolites perturb close-proximity metabolic networks as well as key proteins of metabolism and known signaling pathways of dorsal-ventral specification. Two single-cell mass spectrometry instruments will be used to perform flux analysis for the injected metabolites and to measure the relative translation of targeted proteins in fluorescently tracked cell clones that form from the metabolite-injected cells in the living frog embryo. The resulting data will identify gene candidates for functional tests via gene knock-down experiments to validate the proposed mechanism of action for metabolite-induced cell fate changes. Understanding small-molecule effects on cell fate commitment raises broad implications in diverse areas of the life sciences. The work will also train underrepresented groups in techniques bridging biology and analytical chemistry.

脊椎动物胚胎的正常发育的非技术段落，胚胎的细胞必须发展成生物体的所有不同类型的组织，但是造成组织形成的完整生物分子集尚不清楚。该项目利用了最近才在分析化学中开发的高级仪器来确定小分子的变化，因为单个细胞在早期发育的青蛙胚胎中形成不同类型的组织，这是脊椎动物胚胎发育的重要模型。最终的数据将提供以前对基本生物学过程的见解，对于细胞，组织，器官和生物的形成很重要。这些研究工作为托管大学的不同参与者以及纽约州冷泉港实验室等国家培训中心的各种参与者以及纽约州冷泉港诸如会议，研讨会和国家培训中心的跨学科培训的基础。 这项工作将培训生物学和化学方面的新一代科学家，包括许多传统上代表性不足的科学人群，使他们能够应对当前的挑战，同时在这些领域中提出新问题，以更好地理解正常的脊椎动物胚胎发育和疾病。研究的研究段落的研究中，许多基因和基因在许多基因中都遇到了许多摩尔群的摩洛群，但是在较小的基因中，却是多个垂直的垂直果实（垂直的垂直角色）（尚有垂直的垂直角色）（代谢产物）参与细胞发育过程尚不清楚。 PI的实验室最近发现了能够改变早期青蛙（Xenopus laevis）胚胎中精选干细胞正常背腹命运的代谢产物，表明这些分子在脊椎动物体的构图过程中也是活性参与者。这项工作的总体目标是确定代谢物引起的细胞命运决定的作用机理。这将通过系统细胞生物学方法来完成，其中将使用独特的单细胞质谱技术来表征代谢物注射细胞的分子状态，这些技术在PI实验室中开发和验证。该项目将确定细胞命运改变代谢物如何扰动近距离代谢网络以及代谢的关键蛋白质和背侧腹侧规范的已知信号通路。两种单细胞质谱仪器将用于对注射的代谢产物进行通量分析，并测量荧光蛋白在荧光细胞克隆中的相对翻译，这些细胞克隆从活的青蛙胚胎中的代谢物注射细胞形成。最终的数据将通过基因敲除实验来鉴定用于功能测试的基因候选物，以验证代谢物诱导的细胞命运变化所提出的作用机理。了解小分子对细胞命运承诺的影响会在生命科学的不同领域产生广泛的影响。 这项工作还将在桥接生物学和分析化学的技术方面培训代表性不足的群体。