Proteomics Shared Resource

蛋白质组学共享资源

• 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
• 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

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）是一个全方位服务的设施， 在肽合成，蛋白质分析，蛋白质和肽分离，质量方面提供基本服务 在项目计划，样本准备，数据获取方面的光谱，蛋白质组学应用和专业知识 和分析。该专业人士在医学科学大楼的三楼总共占据了4920平方英尺的 位于罗切斯特（MCR）地点的梅奥诊所。 8位职业员工有广泛的知识， 提供的服务专业知识。职业主管Daniel J. McCormick博士提供了有远见的方向 关于所有运营，人员配备和服务的计划，招聘和执行决定。与之合作 机构委员会和中心已有33年以上，麦考密克博士能够提供科学 高端核心仪器的方向和支持，以及蛋白质化学的新技术，蛋白质组学 和质谱（MS）支持MCCC成员在癌症研究中的计划。在上一个 资助时期，有6个新的服务线（例如，无标签的差异蛋白质组学，通过 SCX分馏和TIO2富集，FFPE和冷冻组织的蛋白质组学分析，高/低pH LC 样品分馏，从头/MS/MS蛋白测序和复杂聚糖的MS分析） 开发以支持MCCC成员的蛋白质分析需求。科学成就包括： 鉴定与髓样癌中Tat2蛋白相互作用的关键蛋白质；开发代理商 抑制TGFß作用（通过SMAD信号传导），并防止PDX细胞系中的胶质母细胞瘤迁移；识别 在Alisertib治疗的人乳腺癌中，关键的磷蛋白（例如CXCR4）下调； Sox2中磷酸化位点的映射位点，Sox2是肺中“茎”表型的转录激活剂 鳞状细胞癌;癌蛋白抗原的合成，用于自适应中幼稚T细胞的体外激活 治疗癌症的T细胞疗法（例如胰腺）；以及对Spink1激活的蛋白质的差异分析 卵巢癌的增殖。 MCCC成员的利用率很强，总平均使用率为 在当前资助期（2013-2017）中，有99名成员（72名同行评审）为51％。 Pro由MCCC使用 来自所有3个站点的成员，包括来自佛罗里达州梅奥诊所的12名成员和梅奥诊所的5名成员 在亚利桑那州。 Pro向MCCC成员提供免费咨询，项目设计和数据解释， 并以付费服务提供服务；项目和服务请求的标准化表格已打开 资源的内部网站。比较Pro的许多服务的利率与外部核心的比较 供应商于2017年完成，并表明核心利率具有很高的竞争力。未来的方向 Pro包括：用高级MS仪器和数据无关的深层测序蛋白质组学 获取（DIA）方法；自上而下的MS蛋白测序；离子迁移率分离（IMS）质谱； 基因组/蛋白质组学数据集的蛋白质组学整合以及糖蛋白的MS分析。