Engineering Growth Factor/Receptor Processes

该项目将结合生物化学工程、细胞生物学和分子生物学的方法，在细胞和组织水平上增加对生长因子/受体动力学的定量理解和操作。工作将集中在自分泌生长因子信号的动态，特别是关于配体释放和受体占用的空间调节。这个方向的一个主要目标将是确定自分泌配体是否向细胞提供有关其组织“群落”而不是其局部微环境的信息，以及物理化学参数如何影响该信息。工作还将继续建立在当前受体贩运模型的基础上，以包括有关分选室的新实验信息，并扩展这些模型，以纳入相关信号转导途径的关键成分。这将允许定量评估改变受体运输对信号通路的影响，这些通路是先前细胞增殖反应研究的基础。了解这些对生长因子信号传导的综合动态影响将继续为设计改进的药理生长因子模拟物或拮抗剂提供合理的基础。通过将人表皮生长因子/受体系统转染到确定的细胞系中，分子生物学工具，如单克隆抗体、位点定向受体突变体和重组位点定向配体变异体，可用于生化工程目的。新资助期的工作分为三个具体目标，旨在促进对生长因子调节细胞功能中的受体和配体动力学的理解。* * *

9710143 Lauffenburger This project will combine approaches of biochemical engineering, cell biology, and molecular biology to increase the quantitative understanding and manipulation of growth factor/receptor dynamics at both the cell and tissue level. Work will focus on the dynamics of autocrine growth factor signaling, particularly with respect to the spatial regulation of ligand release and receptor occupancy. A major goal in this direction will be to determine whether autocrine ligands provided information to cells regarding their tissue "community" as opposed to their local microenvironment, and how physico-chemical parameters influence this information. Work will also continue to build on current models of receptor trafficking to include new experimental information about the sorting compartment, and to extend these models to incorporate key components of related signal transduction pathways. This will allow a quantitative evaluation of the effects of altered receptor trafficking on signaling pathways underlying previous studies of cell proliferation responses. Understanding these sorts of integrated dynamical influences on growth factor signaling will continue to generate a rational basis for the design of improved pharmacological growth factor mimics or antagonists. By using the human epidermal growth factor/receptor system transfected into defined cell lines, tools for molecular biology such as monoclonal antibodies, site-directed receptor mutants, and recombinant site- directed ligand variants can be used for biochemical engineering purposes. The work for the new grant period has been organized into three specific aims which are designed to facilitate the understanding of receptor and ligand dynamics in growth factor regulation of cell function. ***