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

描述(申请人提供)：从卵子到胚胎再到成体的转变要求细胞之间相互沟通，以在正确的时间和正确的地点获得特殊的命运。癌症在一定程度上是调控异常的结果。我们关于信号通路的大部分信息来自于细胞群体的遗传学和生物化学，刺激通常被减少到二进制开/关。该项目将采用当代微流控技术，及时准确地控制多个小室中信号分子的浓度，同时通过荧光标记的蛋白质对信号做出反应，连续成像单细胞的反应。我们将使用人类胚胎干细胞(HESC)来达到这一目的，因为它们与再生医学相关。我们将用转化生长因子途径中的配体刺激细胞，因为它们可以诱导这些细胞自然做出的第一轮发育选择。这条途径有两个相互干扰的分支，使其成为研究途径相互作用的自然环境。我们的数据将被组织和开发成一个预测工具，用来合理地重新规划人类胚胎干细胞的特殊命运。

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.