Developing novel RPPA for the detection of metastatic prostate cancer

项目摘要 反相蛋白质芯片（RPPA）已成为一种很有前途的基于抗体的高度定量分析技术 蛋白质组学技术适用于分析数百至数千例患者样本中的蛋白质。的 RPPA的吞吐量、灵敏度和成本效益，以及其处理微小 样本量，推动了该技术在基础，临床前和临床中的应用 研究领域。这项技术在很大程度上依赖于缺乏高质量的单特异性 然而，由于限制，抗体仅集中于检测少数关键信号分子。 高质量磷酸特异性抗体的可用性。在这项NIH STTR I期研究中，我们将 开发基于金属离子官能化可溶性纳米聚合物的新型RPPA平台， 用于信号分子的灵敏、高通量分析的商业产品， 抗体的局限性。新的RPPA平台将被应用于区分侵略性和 惰性人前列腺肿瘤异种移植小鼠模型。我们假设前列腺癌可以 通过测量关键癌基因上的磷酸化变化进行分类，从而建立RPPA平台 可用作发现和临床前工具，以区分侵袭性肿瘤和惰性肿瘤。的 将完成以下目标。目的#1：用于捕获和纯化的官能化RPPA的优化 磷丙汀的检测。目的#2：惰性和攻击性的通路激活分析 前列腺癌异种移植小鼠模型。到第一阶段研究完成时，我们预计 可建立灵敏度高、动态范围宽、性能优良的分析平台 可重复性和可承受的成本。

PROJECT SUMMARY Reverse-phase protein array (RPPA) has emerged as a promising antibody-based highly quantitative proteomic technology suitable for profiling proteins in hundreds to thousands of patient samples. The throughput, sensitivity, and cost effectiveness of RPPA, together with its ability to deal with minuscule sample amounts, have propelled applications of the technology in basic, preclinical and clinical research fields. The technology, which relies heavily on the paucity of high-quality monospecific antibodies, however, is only centered on detecting a few key signaling molecules due to limited availability of high quality phosphospecific antibodies. In this NIH STTR Phase I study, we will develop a novel RPPA platform based on metal ion-functionalized soluble nanopolymers into commercial products for sensitive, high throughput profiling of signaling molecules without the limitation of antibodies. The novel RPPA platform will be applied to distinguish aggressive from indolent human prostate tumors in xenograft mouse models. We hypothesis that prostate cancer can be classified by measuring phosphorylation changes on key oncogenes and thus a RPPA platform can be used as a discovery and preclinical tool to distinguish aggressive from indolent tumors. The following aims will be completed. Aim #1: Optimization of functionalized RPPA for capture and detection of phosphopropteins. Aim #2: Pathway-activation profiling in indolent and aggressive prostate cancer xenograft mouse models. By the completion of Phase I study, we expect that an analytical platform can be established with high sensitivity, wide dynamic range, excellent reproducibility, and affordable cost.