13 Functional Proteomics Reverse Phase Protein Array Core

项目摘要：功能蛋白质组学反相蛋白质阵列(RPPA)核心 功能蛋白质组反相蛋白质阵列(RPPA)核心提供MD Anderson癌症 中心成员和癌症研究社区拥有强大的、高通量、定量、成本- 有效的功能蛋白质组平台。RPPA测量蛋白质表达水平和翻译后水平 对大量肿瘤样本或细胞系进行系统生物学分析所需的修改。RPPA CORE于2008年开始运作，陆以玲博士担任主任，最近又增加了雷汉·阿克巴尼博士担任 联席导演。目前的服务包括蛋白质提取、细胞/肿瘤裂解物的机器人阵列、探查 经过验证的抗体，并向客户报告蛋白质表达和修饰。使用以下工具分析数据 定制软件(ArrayPro和超级曲线拟合)，以提供信号强度、曲线构造和 相对蛋白质浓度。重要的是，该设施不断扩大抗体库并拥有 对其流水线进行了广泛的修订，重点是改进交付时间、数据质量和准确性。至 确保RPPA资源的效用，Lu博士和Akbani博士在MD Anderson有多个积极的合作， 在国内和国际上为他们的科学活动和职业发展提供支持。卢博士是一位 NCI研究专家(R50)拨款，Akbani博士获得了2个U24拨款和NCI RPPA数据 代工合同。自开始运作以来，该设施已经处理了来自605名调查人员的155,855个样本 (183名癌症中心成员和422名外部调查人员)，抗体在150到302之间。作为授权书 年，所有样本中的72%来自同行审查资助的用户，CCSG乳腺癌和妇科 这两个癌症项目都占到了样本的25%。许多外部用户来自其他NCI 指定的癌症中心，包括77个美国学术机构。在过去的6年里，RPPA核心 支持285种出版物，其中230种(81%)在使用IF&>5的期刊上，101种(35%)在使用IF&>10的期刊上，包括 备受瞩目的期刊如《N Engl J Med》、《自然》和《细胞》。此外，它还提供了功能蛋白质组 来自代表32种疾病类型的癌症基因组图谱和RPPA核心的7991个样本的分析 目前是2个获得批准的NCI基因组表征中心之一。估计的业务预算总额 目前的43日元是1,194,149美元，其中15%(183,777美元)来自CCSG。在过去的6年资助期内，MD Anderson 提供了1221,041美元的资本设备支助。在下一个赠款期间，请拨187,727美元/年(14%) 来自CCSG的。RPPA的具体目标是：目的1)为患者提供功能性蛋白质组评估， 动物模型，并将细胞系样本交给MD安德森中心成员，采用稳健、经济、公正的方式 基于抗体的RPPA方法分析主要信号通路网络的功能状态；目标2) 扩大用于RPPA的有效抗体库；以及目标3)改进RPPA的质量控制 RPPA数据；以及目标4)改进RPPA技术，将其扩展到包括空间定向和 多元分析的定量方法。

PROJECT SUMMARY: FUNCTIONAL PROTEOMICS REVERSE PHASE PROTEIN ARRAY (RPPA) CORE The Functional Proteomics Reverse Phase Protein Array (RPPA) Core provides the MD Anderson Cancer Center members and the cancer research community with a powerful, high-throughput, quantitative, cost- effective functional proteomic platform. RPPA measures levels of protein expression and post-translational modification needed for systems biology analysis across large numbers of tumor samples or cell lines. The RPPA Core commenced operations in 2008 with Dr. Yiling Lu as director and recently added Dr. Rehan Akbani as co-director. Current services include protein extraction, robotic arraying of cell/tumor lysates, probing with validated antibodies, and reporting protein expression and modifications to clients. Data are analyzed using customized software (ArrayPro and Supercurve Fitting) to provide signal intensity, curve construction, and relative protein concentration. Importantly, the facility continuously expands the antibody repertoire and has extensively revised its pipeline with emphasis on improved delivery times, quality and accuracy of data. To ensure the utility of the RPPA resource, Drs. Lu and Akbani have multiple active collaborations at MD Anderson, nationally and internationally for their scientific activities and career development. Dr. Lu is the recipient of an NCI Research Specialist (R50) grant, and Dr. Akbani has received 2 U24 grants and an NCI RPPA data generation contract. Since initiating operation, the facility has processed 155,855 samples from 605 investigators (183 cancer center members and 422 external investigators) with between 150 and 302 antibodies. In grant Yr42, 72% of all samples were from peer review-funded users, with the CCSG Breast Cancer and Gynecological Cancers programs both responsible for 25% of the samples. Many of the external users are from other NCI designated cancer centers, including 77 U.S. academic institutions. Over the past 6 years, the RPPA Core has supported 285 publications, with 230 (81%) in journals with IF >5 and 101 (35%) in those with IF >10, including high-profile journals such as N Engl J Med, Nature, and Cell. Furthermore, it provided functional proteomic analysis for 7991 samples from The Cancer Genome Atlas, representing 32 disease types, and the RPPA Core is currently 1 of 2 approved NCI Genomic Characterization Centers. The estimated total operating budget for the current Yr43 is $1,194,149, with 15% ($183,777) from the CCSG. For the past 6-year grant period, MD Anderson provided $1,221,041 of capital equipment support. In the next grant period, $187,727/year (14%) is requested from the CCSG. The RPPA specific aims are: Aim 1) To provide functional proteomic assessment of patient, animal model, and cell line samples to MD Anderson center members using a robust, cost-effective, unbiased antibody-based RPPA approach to analyze the functional status of major signaling pathway networks; Aim 2) To expand the repertoire of validated antibodies for use with RPPA; and Aim 3) To improve quality control of the RPPA data; and Aim 4) To improve the RPPA technology by expanding it to include spatial orientation and quantitative approaches for multiplex analysis.