Quantitation of protein interactions in cancer cells

描述（由申请人提供）：蛋白质相互作用是生物系统中蛋白质功能的关键决定因素。尽管定量蛋白质相互作用信息对癌症研究的所有领域都有潜力，但对原生生物系统中蛋白质相互作用的无偏见或大规模定量是当今技术无法满足的挑战。在原生细胞、患者样本或组织中大规模鉴定和定量蛋白质相互作用的能力目前尚不存在。量化蛋白质相互作用的能力的提高将对了解癌症、转移和抗癌耐药性的发展产生重大影响。本项目旨在利用先进的蛋白相互作用报告（PIR）技术开发和应用与癌细胞的定量交联。细胞培养氨基酸稳定同位素标记（SILAC）将与PIR技术相结合，以定量细胞中的蛋白质水平和蛋白质相互作用。这些功能将应用于顺铂、紫杉醇和SN-38耐药的癌细胞，以便定量与药物敏感癌细胞的相互作用。该项目将提供癌细胞中蛋白质相互作用的第一个相对定量数据，以及第一个在蛋白质相互作用水平上与耐药性相关的功能调节的无偏测量。

DESCRIPTION (provided by applicant): Protein interactions are key determinants of protein function in biological systems. Despite the potential that quantitative protein interaction information could have for all areas of cancer research, unbiased or large-scale quantitation of protein interactions within native living systems is a challenge that is unmet by today's technology. The capacity to identify and quantitate protein interactions on a large-scale within native cells, patient samples, or tissues does not currently exist. Improved capabilities to quantitate protein interactions will have a major impact on the understanding of cancer, metastasis and the development of anti-cancer drug resistance. This project aims to develop and apply quantitative cross-linking with cancer cells with advanced Protein Interaction Reporter (PIR) technology. Stable Isotope Label of Amino acids in Cell culture (SILAC) will be combined with PIR technology to allow quantitation of protein levels and protein interactions in cells. These capabilities will be applied to cisplatin-, taxol-, and SN-38 resistant cancer cells to allow quantitation of interactions relative to drug sensitive cancer cells. This project will provide the first relative quantitation data on protein interactions in cancer cells and the first unbiased measurements of functional regulation at the protein interaction level relevant to drug resistance. PUBLIC HEALTH RELEVANCE: Drug resistance in cancer treatment is the primary reason for therapy failure and will likely remain a primary factor leading to cancer patient death until functional regulation that supports drug resistance can be better understood. Functional regulation in all cells is achieved by changes in protein abundance, localization, interactions and topological features. This project will develop and apply advanced technology to help visualize changes in functional regulation in cancer cells that have acquired drug resistance.