Dynamic readout of TGFb signaling and modeling of cell fate specification in huma

描述（由申请人提供）：从卵子到胚胎再到成体的转变需要细胞之间相互沟通，以便在正确的时间和正确的位置获得专门的命运。癌症在一定程度上是异常调控的结果。我们对信号传导途径的大部分信息来自细胞群体的遗传学和生物化学，刺激通常被简化为二元开/关。该项目将采用当代微流体技术，在多个小室中及时准确地控制信号分子的浓度，同时通过荧光标记的响应信号移动的蛋白质连续成像单个细胞的反应。我们将使用人类胚胎干细胞（hESC），因为它们与再生医学相关。我们将用TGF¿途径的配体刺激细胞，因为它们可以诱导这些细胞自然做出的第一轮发育选择。这条通路有两个相互干扰的分支，使其成为研究通路相互作用的自然环境。我们的数据将被整理和发展成一种预测工具，用于从hESCs中合理地重新编程专门的命运。

DESCRIPTION (provided by applicant): The transitions from egg to embryo to adult require that cells communicate with each other to acquire specialized fates at the right times in the correct places. Cancer is in part the result of aberrant regulation. Most of our information on signaling pathways comes from genetics and biochemistry on populations of cells, with stimuli typically reduced to a binary on/off. This project will adapt contemporary microfluidic technology to accurately control the concentrations of signaling molecules in time in multiple small chambers while continuously imaging the response of single cells via fluorescently tagged proteins that move in response to signals. We will use human embryonic stem cells (hESC) for this purpose because of their relevance to regenerative medicine. We will stimulate the cells with ligands from the TGF¿ pathway since they can induce the first round of developmental choices these cells naturally make. This pathway has two sub-branches that interfere, making it a natural context in which to study pathway interactions. Our data will be organized and developed into a predictive tool with which to rationally reprogram specialized fates from hESCs. PUBLIC HEALTH RELEVANCE: To use human embryonic stem cells for regenerative medicine requires the ability to direct the differentiation of these cells to the required fates without causing genetic damage. We will follow the first steps of stem cell differentiation using new high-resolution techniques drawn from Physics and integrate the results using topological methods into predictive models.