Proteomic Predictors of Clinical Outcome of Targeted Therapies in Prostate Cancer

前列腺癌靶向治疗临床结果的蛋白质组预测因子

• 批准号: 7067896
• 负责人: DAVID B AGUS
• 金额: $ 65.89万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-12-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7067896
• 关键词: antineoplastics; bioimaging /biomedical imaging; biological signal transduction; biomarker; cell surface receptors; clinical research; cooperative study; epidermal growth factor; flow cytometry; gene expression profiling; glycoproteins; growth factor receptors; intravital microscopy; microarray technology; microprocessor /microchip; nanotechnology; neoplasm /cancer diagnosis; neoplasm /cancer therapy; neoplastic process; phosphotransferases; prognosis; protein quantitation /detection; proteomics; secretion; technology /technique development

The lack of biomarkers for assessment of which patients may respond to pathway-targeted therapy creates a profound need for the application of integrated technologies for the discovery and translational validation of such biomarkers. Recent advances in proteomic methods and computational bioinformatics processing have enabled application of integrated proteomic technologies to the discovery of biomarkers. If used by ex vivo nano-sensor devices and in vivo nanoparticle imaging methods such biomarkers may provide effective new tools to cancer therapy development and use. We propose to discover candidate markers by integrating two directed and two comprehensive proteomic technologies: a) intracellular-signaling protein chips consisting of a directed analysis method which quantitates known, intracellular signaling proteins; b) living-cell capture sensor arrays which represent a nanosensor approach for directed analysis of the cell-surface and secretory proteomes; c) biotin-capture-based cell-surface profiling methods consisting of a comprehensive analysis method that identifies and quantitates the abundance of cell-surface proteins; d) solid-phase extraction of glycoprotein (SPEG) profiling, which is a comprehensive analysis method for the study of secreted proteins and blood. Biomarker discovery will initially be disease- (prostate cancer) and pathway- (human epidermal growth factor receptor (Her)-kinase axis) focused and will provide a foundation our CCNE-TR Center will use to produce, evaluate and validate nanosensors and nanoparticle-based imaging. We will first define the cell-surface and secretory proteomes of androgen-independent prostate cancer and identify proteins within these and the intracellular proteome that are indicative of the perturbations to the Her-kinase axis. We will analyze these sub-proteomes in primary culture models of androgen-independent prostate cancer by treatment with the Her-kinase targeted therapeutics, 2C4 (a humanized monoclonal antibody that binds epitopes on Her-2 that prevent ligand-mediated Her-2 heterodimerization) and geftinib (a small molecule inhibitor that competes for the ATP binding site on epidermal growth factor receptor). We will additionally stimulate with ligands targeting each of the receptor-dimer partners of this axis. Axis-response-informative proteins will be evaluated for their Her-kinase and prostate cancer specificity, by comparison with a database of gene and protein expression in other cell lines of different tissue origin available to the investigator group. Next we will integrate our in vitro results with information of the protein expression patterns of human xenograft models to identify a panel of markers with utility for predicting and/or for monitoring response. The bank of blood and viable tissue samples developed by the UCLA Prostate SPORE for use in this project represent different pre and post-treatment time points and a diverse collection of androgen-independent xenograft models characterized for sensitivity to the Her-kinase-targeted therapeutics. Lastly we will validate the utility of the biomarkers discovered in Specific Aims 1 and 2 for use with ex vivo nanosensor devices and in vivo nanoparticle imaging by generating affinity reagents for protein candidates; We will also validate the panel's ability to guiding human therapeutic intervention by using SPORE tissue and serum samples of prostate cancer patients treated with Her-kinase-directed therapies.

缺乏用于评估哪些患者可能对路径靶向治疗有反应的生物标记物造成了 迫切需要应用集成技术来发现和翻译验证 这样的生物标志物。蛋白质组学方法和计算生物信息学处理的最新进展 使综合蛋白质组学技术能够应用于发现生物标记物。如果在体外使用 纳米传感器件和体内纳米粒子成像方法等生物标志物可能提供有效的新的 癌症治疗工具的开发和使用。我们建议通过整合两个候选标记来发现候选标记 定向和两种全面的蛋白质组学技术：a)细胞内信号蛋白质芯片，包括 对已知的细胞内信号蛋白进行量化的直接分析方法；b)活细胞捕获 传感器阵列代表了一种直接分析细胞表面和分泌物的纳米传感器方法 蛋白质组；c)基于生物素捕获的细胞表面图谱方法，包括综合分析 鉴定和定量细胞表面蛋白质丰度的方法；d)固相萃取 糖蛋白(SpeG)谱，这是一种研究分泌蛋白的综合分析方法 还有鲜血。生物标记物的发现最初将是疾病(前列腺癌)和途径(人类表皮) 生长因子受体(HER)-激动轴)，并将为我们的CCNE-TR中心提供一个基础 生产、评估和验证纳米传感器和基于纳米颗粒的成像。我们将首先定义细胞表面 和雄激素非依赖性前列腺癌的分泌蛋白质组，并鉴定其中的蛋白质 以及细胞内的蛋白质组，这些蛋白质组表明了对HER-激酶轴的扰动。我们将分析 雄激素非依赖性前列腺癌原代培养模型中的这些亚蛋白质组 HER-激酶靶向治疗药物2C4(一种人源化的单抗，结合HER-2上的表位 阻止配体介导HER-2异二聚化)和吉夫替尼(一种竞争的小分子抑制剂 用于表皮生长因子受体上的ATP结合位点)。我们还会用配体来刺激 靶向这个轴的每一个受体-二聚体伙伴。轴向反应信息蛋白质将是 通过与基因和前列腺癌数据库的比较，评估他们的HER-激酶和前列腺癌的特异性 不同组织来源的其他细胞系的蛋白质表达可供研究小组使用。接下来我们将 将我们的体外结果与人类异种移植模型的蛋白质表达模式信息相结合，以 确定一组具有用于预测和/或监控响应的标记。血库和 加州大学洛杉矶分校前列腺分生孢子为该项目开发的活组织样本代表了不同的 以及治疗后时间点和各种雄激素非依赖性异种移植模型的收集 特点是对Her-Kinase靶向治疗药物敏感。最后，我们将验证 用于体外纳米传感器装置和体内的特定AIMS 1和2中发现的生物标记物 通过为候选蛋白质生成亲和试剂进行纳米颗粒成像；我们还将验证该小组的 利用前列腺孢子组织和血清标本指导人类治疗干预的能力 接受HER-激酶导向治疗的癌症患者。