13 Functional Proteomics Reverse Phase Protein Array Core

项目总结：功能蛋白质组学反相蛋白质阵列（RPPA）核心 功能蛋白质组学反相蛋白质阵列（RPPA）核心提供MD安德森癌症 中心成员和癌症研究界拥有强大的，高通量的，定量的，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 在大量肿瘤样品或细胞系中进行系统生物学分析所需的修饰。RPPA Core于2008年开始运营，由Yiling Lu博士担任董事，最近又增加了Rehan Akbani博士 联合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， 验证抗体，并向客户报告蛋白质表达和修饰。数据分析使用 定制软件（ArrayPro和Supercurve Fitting）提供信号强度、曲线构建和 相对蛋白质浓度重要的是，该设施不断扩大抗体库， 广泛修订了其管道，重点是改进数据的交付时间、质量和准确性。到 确保RPPA资源的效用，Lu博士和Akbani博士在MD安德森有多个积极的合作， 在国内和国际上为他们的科学活动和职业发展。卢博士是一位 NCI研究专家（R50）资助，Akbani博士获得了2项U24赠款和一项NCI RPPA数据 发电合同。自开始运作以来，该设施已处理了来自605名调查员的155 855份样本 (183癌症中心成员和422名外部研究人员），抗体在150到302之间。在格兰特 42年，72%的样本来自同行评审资助的用户，CCSG乳腺癌和妇科 癌症计划都负责25%的样本。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核心已经 支持了285篇出版物，其中230篇（81%）发表在IF >5的期刊上，101篇（35%）发表在IF >10的期刊上，包括 高知名度的期刊，如N Engl J Med，Nature和Cell。此外，它还提供了功能蛋白质组学 分析来自癌症基因组图谱的7991个样本，代表32种疾病类型，以及RPPA核心 目前是2个批准的NCI基因组表征中心之一。2000年的估计总业务预算 目前的43年为1，194，149美元，其中15%（183，777美元）来自CCSG。在过去的6年赠款期间，MD安德森 提供了1 221 041美元的资本设备支助。在下一个赠款期，申请187，727美元/年（14%） 从CCSG。RPPA的具体目标是：目标1）提供患者的功能蛋白质组学评估， MD安德森中心成员使用一个强大的，具有成本效益的，公正的， 基于抗体的RPPA方法分析主要信号通路网络的功能状态;目的2） 为了扩大与RPPA一起使用的经验证的抗体的库;以及目的3）为了改进RPPA的质量控制， 目标4）通过扩大RPPA技术以包括空间定向和 多重分析的定量方法。

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.