Developing novel RPPA for the detection of metastatic prostate cancer

项目总结 反相蛋白质阵列(RPPA)是一种基于抗体的高效定量检测技术 适用于分析数百到数千个患者样本中的蛋白质的蛋白质组学技术。这个 RPPA的吞吐量、敏感度和成本效益，以及处理细微问题的能力 样本量，推动了该技术在基础、临床前和临床上的应用 研究领域。这项技术严重依赖于高质量的单一品种的匮乏 然而，由于有限的限制，抗体仅集中在检测少数关键的信号分子上。 高质量的磷酸特异性抗体的可用性。在这项NIH STTR第一阶段研究中，我们将 开发一种基于金属离子功能化可溶纳米聚合物的新型RPPA平台 用于敏感、高通量分析信号分子的商业产品，无需 抗体的限制。新的RPPA平台将被应用于区分攻击性和 小鼠异种移植模型中的惰性人前列腺癌。我们假设前列腺癌可以 通过测量关键癌基因的磷酸化变化进行分类，从而建立RPPA平台 可作为一种发现和临床前工具来区分侵袭性和惰性肿瘤。这个 将完成以下目标。目标1：优化功能化的RPPA，用于捕获和 磷蛋白的检测。目标2：惰性和攻击性的通路激活特征 前列腺癌异种移植小鼠模型。在完成第一阶段研究后，我们预计会有 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.