Proteomics Shared Resource

蛋白质组学共享资源

基本信息

批准号：
10113579
负责人：
AKHILESH PANDEY
金额：
$ 16.09万
依托单位：
MAYO CLINIC ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10113579
关键词：
Achievement Antigens Area Arizona Basic Science Breast Adenocarcinoma CXCR4 gene Cell Line Clinic Complex Comprehensive Cancer Center Computers and Advanced Instrumentation Consultations Data Data Analyses Data Collection Data Set Development Disease Electronic Mail Experimental Designs Fee-for-Service Plans Floor Florida Fractionation Freezing Funding Future Genomics Glioblastoma Glycoproteins Grant Human In Vitro Investigation Knowledge Label Malignant Neoplasms Malignant neoplasm of ovary Mass Spectrum Analysis Mayo Clinic Cancer Center Medical Methodology Methods Molecular Myelogenous Oncoproteins Pancreas Peer Review Peptide Sequence Determination Peptide Synthesis Peptides Phenotype Phosphoproteins Phosphorylation Site Polysaccharides Post Translational Modification Analysis Post-Translational Protein Processing Preparation Protein Analysis Protein Chemistry Proteins Proteome Proteomics Proteomics Shared Resource Resource Sharing Resources SPINK1 gene Sampling Science Services Signal Transduction Site Software Tools Squamous Cell Lung Carcinoma Standardization T cell therapy T-Lymphocyte Tissues Transcription Coactivator Transforming Growth Factor beta Vendor Work anticancer research cost data acquisition data management deep sequencing design experience improved informatics tool insight instrument instrumentation ion mobility member migration new technology operation phosphoproteomics prevent programs proteogenomics recruit scaffold service providers square foot stemness tandem mass spectrometry titanium dioxide tool web portal web site

项目摘要

PROTEOMICS SHARED RESOURCE PROJECT SUMMARY The Mayo Clinic Cancer Center (MCCC) Proteomics Shared Resource (PRO) is a full-service facility that provides essential services in peptide synthesis, protein analysis, protein and peptide separation, mass spectrometry, proteomics applications, and expertise in project planning, sample preparation, data acquisition and analysis. The PRO occupies a total of 4920 sq. ft. on the 3rd floor of the Medical Sciences Building, and is located at the Mayo Clinic in Rochester (MCR) site. The 8 PRO staff members have broad knowledge and expertise in the services provided. The PRO Director, Dr. Daniel J. McCormick, provides visionary direction, planning, recruitment, and executive decisions regarding all operations, staffing, and services. Working with institutional committees and centers for over 33 years, Dr. McCormick has been able to provide scientific direction and support for high-end core instrumentation, and new technologies in protein chemistry, proteomics and mass spectrometry (MS) to support Programs of MCCC members in cancer research. In the previous funded period, 6 new service-lines (e.g., label-free differential proteomics, expanded phosphoproteomics by SCX fractionation and TiO2 enrichment, proteomics analysis of FFPE and Frozen tissue, high/low pH LC sample fractionation, de novo MS/MS protein sequencing, and MS analysis of complex glycans) were developed to support the protein analysis needs of MCCC members. Scientific achievements include: identification of key proteins that interact with TAT2 protein in myeloid cancers; development of agents that inhibit TGFß action (via SMAD signaling) and prevent glioblastoma migration in PDX cell lines; identification of key phosphoproteins (e.g., CXCR4) downregulated in Alisertib-treated human breast adenocarcinoma; mapping sites of phosphorylation in SOX2, a transcriptional activator of “stemness” phenotype in lung squamous cell carcinoma; synthesis of oncoprotein antigens for in vitro activation of naïve T-cells in adoptive T-cell therapies to treat cancer (e.g., pancreatic); and differential analysis of proteins activated by SPINK1 in ovarian cancer proliferation. PRO utilization by MCCC members has been strong with a total average usage of 51% by 99 members (72 peer-reviewed) in the current funded period (2013-2017). The PRO is used by MCCC members from all 3 sites, including 12 members from Mayo Clinic in Florida and 5 members from Mayo Clinic in Arizona. The PRO provides free consultation, project design, and data interpretation to MCCC members, and provides its services on fee-for-service basis; standardized forms for project and service requests are on the resources' internal web site. Comparison of rates for many services of the PRO to external cores and vendors, was completed in 2017 and indicated that core rates were highly competitive. Future directions of the PRO include: deep sequencing proteomics with advanced MS instrumentation and data-independent acquisition (DIA) methods; top-down MS protein sequencing; ion mobility separation (IMS) mass spectrometry; proteogenomics integration of genomics/proteomics data sets, and MS analysis of glycoproteins.

蛋白质组学共享资源项目摘要梅奥诊所癌症中心 (MCCC) 蛋白质组学共享资源 (PRO) 是一家提供全方位服务的机构，提供肽合成、蛋白质分析、蛋白质和肽分离、质量分析等方面的基本服务光谱分析、蛋白质组学应用以及项目规划、样品制备、数据采集方面的专业知识 PRO 占地 4920 平方英尺，位于医学科学大楼三楼。位于罗彻斯特梅奥诊所 (MCR) 的 8 名 PRO 工作人员拥有广泛的知识和能力。 PRO 总监 Daniel J. McCormick 博士提供富有远见的指导，有关所有运营、人员配置和服务的规划、招聘和执行决策。麦考密克博士在机构委员会和中心工作超过 33 年，能够提供科学的高端核心仪器以及蛋白质化学、蛋白质组学新技术的指导和支持和质谱 (MS)，以支持 MCCC 成员的癌症研究计划。资助期间，6 个新的服务线（例如，无标记差异蛋白质组学、通过 SCX 分级分离和 TiO2 富集、FFPE 和冷冻组织的蛋白质组学分析、高/低 pH 液相色谱样品分级分离、从头 MS/MS 蛋白质测序和复杂聚糖的 MS 分析）为支持 MCCC 成员的蛋白质分析需求而开发的科学成就包括：鉴定与骨髓癌中 TAT2 蛋白相互作用的关键蛋白；抑制 TGFβ 作用（通过 SMAD 信号传导）并防止 PDX 细胞系中胶质母细胞瘤的迁移；在 Alisertib 治疗的人乳腺癌中关键磷蛋白（例如 CXCR4）下调；绘制肺中“干细胞”表型转录激活因子 SOX2 的磷酸化位点鳞状细胞癌；用于体外激活过继幼稚 T 细胞的癌蛋白抗原的合成用于治疗癌症（例如胰腺癌）的 T 细胞疗法；以及 SPINK1 激活蛋白的差异分析 MCCC 成员的卵巢癌增殖率一直很高，平均总使用量为 MCCC 在当前资助期间（2013-2017 年）使用了 99 名成员（72 名同行评审）的 51%。来自所有 3 个地点的会员，其中 12 名会员来自佛罗里达州 Mayo Clinic，5 名会员来自 Mayo Clinic PRO 为 MCCC 会员提供免费咨询、项目设计和数据解释，并按服务收费提供服务；已制定项目和服务请求的标准化表格 PRO 的许多服务与外部核心的费率比较和资源的内部网站。于 2017 年完成，表明核心费率具有高度竞争力。 PRO 包括：具有先进 MS 仪器和数据独立性的深度测序蛋白质组学采集 (DIA) 方法；自上而下的 MS 蛋白质测序；基因组学/蛋白质组学数据集的蛋白质基因组学整合，以及糖蛋白的 MS 分析。