CAREER: Discovering Upstream Effectors to Cell Fate Determination

点击翻译按钮获取中文摘要

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.