Biodefense Proteomics Collaboratory

生物防御蛋白质组学实验室

• 批准号: 6604414
• 负责人: David G Gorenstein
• 金额: $ 121.5万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-15 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6604414
• 关键词: Arenaviridae; Lassa virus; New World Arenavirus; biotechnology; bioterrorism /chemical warfare; combinatorial chemistry; communicable disease diagnosis; cooperative study; cytokine; diagnosis design /evaluation; flow cytometry; gene expression; guinea pigs; inflammation; laboratory mouse; mass spectrometry; molecular biology information system; oligonucleotides; proteomics; shock; transcription factor; virus diseases

DESCRIPTION (provided by applicant): In vitro enzymatic combinatorial selection and split-synthesis chemical combinatorial methods will be used to develop a "ThioAptamer Chip" (TACh) for proteomics - a diagnostic tool to identify and quantify the differential expression of key proteins in response to pathogens of concern for bioterrorism threat (BT). This new proteomics technology will utilize our proprietary thioselection and phosphorothioate-modified oligonucleotide "thioaptamers," combined with the surface enhanced laser desorption/ionization (SELDI) mass spectroscopy technology of our collaborating partner, Ciphergen, to target both rodent and human proteomes. In particular we will study the inflammatory response of cytokines and key transcription factors (e.g., NF-kappaB) challenged with BT agents. The five NF-?B/Rel family proteins can combine to form 15 homo- and heterodimers, each performing a specific signaling function upon translocation across the cell nuclear membrane and binding to a gene's promoter region. In partnership with Ciphergen, we will also develop new, massively parallel, thioaptamer bead-based screening of the proteome with SELDI mass-spectrometric methods to identify uncharacterized proteins involved in the immune response to BT viruses. Our results from the TACh/SELDI approaches will be validated by 2D gel mass spectrometric proteomic methods. We will also apply bioinformatic analyses to correlate changes in protein expression with available genomic data on changes in gene expression as a result of inflammation after viral infection or shock. Elucidating these protein expression changes will allow early diagnosis and enhanced prognosis of viral disease, and subsequent development of effective pharmacological and immunological interventions. Specific initial viral targets include arenaviruses, Pichinde and Lassa (the latter on both the NIH and CDC class A lists).

描述（由申请人提供）：将使用体外酶促组合选择和分裂合成化学组合方法来开发用于蛋白质组学的“硫代患者芯片”（TACH） - 一种诊断工具，用于识别和量化对生物侵蚀性的病原体响应的钥匙蛋白的差异表达（BT）。这项新的蛋白质组学技术将利用我们的专有硫代和磷酸盐剂修饰的寡核苷酸“ ThioAptamers”，结合了我们协作伙伴的表面增强激光解吸/离子化（SELDI）质谱技术（SELDI）质谱技术。特别是，我们将研究细胞因子和关键转录因子（例如NF-kappab）的炎症反应。五个NF-？B/REL家族蛋白可以组合形成15种同二聚体，每个二聚体都在跨细胞核膜易位并与基因的启动子区域结合时，都会执行特定的信号传导功能。与Ciphergen合作，我们还将使用SELDI质谱法对基于蛋白质组的基于蛋白质组的新型，大规模平行的，硫代蛋白珠的筛选，以鉴定参与BT病毒免疫反应的未表征的蛋白质。我们来自TACH/SELDI方法的结果将通过2D凝胶质谱蛋白质组学方法验证。我们还将应用生物信息学分析，以将蛋白质表达的变化与病毒感染或休克后炎症导致基因表达变化的可用基因组数据相关联。阐明这些蛋白质表达的变化将允许早期诊断和增强病毒疾病的预后，并随后发展有效的药理和免疫学干预措施。特定的初始病毒靶标包括体育症病毒，Pichinde和Lassa（后者是NIH和CDC A类列表）。