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，以增加对生长因子/受体相互作用的结合和运输方面如何影响哺乳动物细胞增殖的定量理解。并利用这种增加的理解来建议旁分泌和自分泌细胞培养系统的设计原则。成纤维细胞系上的表皮生长因子（EGF）及其受体（EGFR）将作为焦点系统。数学模型描述了EGF/EGFR运输及其外源性EGF在配体/受体参数方面对细胞生长调节的影响，这些模型最近得到了发展和实验验证。这些模型现在将用于研究细胞培养系统的扰动策略，并将扩展到旁分泌和自分泌EGF来源。扩展模型将通过使用转染了编码EGF和/或EGFR基因的细胞构建人工旁分泌和自分泌系统进行严格测试。通过定点诱变对这些基因进行修饰，可以直接改变系统的关键速率参数。这些研究将确定可以有效操纵的参数，以调节生物技术目的的细胞生长。

This project will combine approaches of biochemical engineering, cell biology, and molecular biology to increase quantitative understanding of how binding and trafficking aspects of growth factor/receptor interactions influence mammalian cell proliferation, and to exploit this increased understanding to suggest design principles for paracrine and autocrine cell culture systems. Epidermal Growth Factor (EGF) and its receptor (EGFR) on fibroblastic cell lines will serve as the system of focus. Mathematical models which describe EGF/EGFR trafficking and its effect on cell growth regulation by exogenous EGF in terms of mechanistic ligand/receptor parameters have been recently developed and experimentally validated. These models will now be used to investigate strategies for perturbation of cell culture systems, and will be extended to paracrine and autocrine EGF sources. The extended models will be critically tested by constructing artificial paracrine and autocrine systems using cells transfected with genes encoding EGF and/or EGFR. Modification of these genes by site-directed mutagenesis will allow key rate parameters of the system to be directly varied. These studies will identify parameters that can be usefully manipulated to regulate cell growth for biotechnological purposes.