13 Functional Proteomics Reverse Phase Protein Array Core

13 功能蛋白质组反相蛋白质芯片核心

• 批准号: 10212261
• 负责人: YILING LU
• 金额: $ 28.94万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-28 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212261
• 关键词: Algorithms; Animal Model; Antibodies; Antibody Repertoire; Apoptosis; Autophagocytosis; Bar Codes; Biological Assay; Budgets; Cancer Center; Cancer Center Support Grant; Capital; Cell Line; Cell surface; Cells; Cleaved cell; Client; Collaborations; Communities; Complement; Computer software; Contracts; Cultured Cells; Custom; DNA; DNA Repair; Data; Data Analyses; Development; Disease; Dot Immunoblotting; Ensure; Environment; Epithelial; Equipment; FRAP1 gene; Faculty; Funding; Gene Expression; Generations; Genomics; Grant; Immunoblot Analysis; Immunooncology; Individual; Institution; Integrins; International; Journals; Label; Location; MAP Kinase Gene; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Measures; Mediating; Mesenchymal; Messenger RNA; Metabolism; NCI-Designated Cancer Center; Nature; Noise; Pathway interactions; Patients; Peer Review; Performance; Phase; Post-Translational Protein Processing; Principal Investigator; Procedures; Process; Protein Array; Protein Array Analysis; Proteins; Proteomics; Publications; Pyroxylin; Quality Control; RNA; RNA Probes; Receptor Protein-Tyrosine Kinases; Reporting; Reproducibility of Results; Research; Research Personnel; Resources; Robotics; Sampling; Services; Signal Pathway; Signal Transduction; Slide; Solid Neoplasm; Space Perception; Specialist; Spectrophotometry; Stains; System; Systems Biology; Technology; The Cancer Genome Atlas; Therapeutic; Time; Time trend; Transforming Growth Factor alpha; Tumor Tissue; Update; Variant; anticancer research; base; beta catenin; career development; computerized data processing; cost effective; data management; data quality; functional status; improved; innovation; malignant breast neoplasm; member; nano-string; neoplastic cell; novel strategies; operation; programs; protein expression; receptor; single molecule; transcriptome sequencing; tumor; ultraviolet

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.

项目摘要：功能蛋白质组学反相蛋白阵列 (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 年开始运营，由卢一岭博士担任董事，最近又任命 Rehan Akbani 博士为董事 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排列、探测 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，该设施不断扩展抗体库，并拥有 广泛修改了其管道，重点是提高交付时间、数据质量和准确性。到 确保 RPPA 资源的效用，博士。 Lu 和 Akbani 在 MD 安德森大学进行了多项积极的合作， 在国内和国际上表彰他们的科学活动和职业发展。卢博士是获得者 NCI 研究专家 (R50) 资助，Akbani 博士已获得 2 项 U24 资助和 NCI RPPA 数据 发电合同。自投入运营以来，该设施已处理了 605 名研究人员的 155,855 份样本 （183 名癌症中心成员和 422 名外部研究人员）拥有 150 至 302 种抗体。拨款中 42 年，所有样本中 72% 来自同行评审资助的用户，CCSG 乳腺癌和妇科 癌症项目均占样本的 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 核心已经 支持 285 篇出版物，其中 230 篇 (81%) 发表在 IF >5 的期刊中，101 篇 (35%) 发表在 IF >10 的期刊中，包括 N Engl J Med、Nature 和 Cell 等知名期刊。此外，它还提供了功能性蛋白质组学 对来自癌症基因组图谱（代表 32 种疾病类型）和 RPPA 核心的 7991 个样本进行分析 目前是 2 个获批准的 NCI 基因组表征中心之一。预计总运营预算 当前第 43 年为 1,194,149 美元，其中 15%（183,777 美元）来自 CCSG。在过去 6 年的资助期内，MD 安德森 提供了 1,221,041 美元的资本设备支持。在下一个资助期内，要求每年 187,727 美元 (14%) 来自 CCSG。 RPPA 的具体目标是： 目标 1) 为患者提供功能蛋白质组学评估， 使用稳健、经济高效、公正的方法向 MD 安德森中心成员提供动物模型和细胞系样本 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 的经过验证的抗体库；目标 3) 提高质量控制 RPPA 数据；目标 4) 通过扩展 RPPA 技术以包括空间定向和 多重分析的定量方法。