Biodefense Proteomics Collaboratory

描述（由申请人提供）：体外酶组合选择和分裂合成化学组合方法将用于开发用于蛋白质组学的“thiioaptamer Chip”(TACh) -一种诊断工具，用于识别和量化应对生物恐怖主义威胁（BT）的关键蛋白质的差异表达。这项新的蛋白质组学技术将利用我们专有的硫代选择和磷硫代修饰的寡核苷酸“硫代核酸”，结合我们合作伙伴Ciphergen的表面增强激光解吸/电离（SELDI）质谱技术，针对啮齿动物和人类的蛋白质组学。特别是，我们将研究细胞因子和关键转录因子（如nf - κ b）在BT制剂挑战下的炎症反应。5个NF-？B/Rel家族蛋白可以结合形成15个同源二聚体和异源二聚体，每个二聚体在跨核膜易位和与基因启动子区域结合时发挥特定的信号功能。通过与Ciphergen的合作，我们还将开发新的、大规模平行的、基于硫帕坦珠的蛋白质组筛选和SELDI质谱方法，以鉴定与BT病毒免疫反应有关的未表征的蛋白质。我们的TACh/SELDI方法的结果将通过2D凝胶质谱蛋白质组学方法进行验证。我们还将应用生物信息学分析，将蛋白质表达的变化与病毒感染或休克后炎症导致的基因表达变化的现有基因组数据联系起来。阐明这些蛋白表达变化将有助于病毒性疾病的早期诊断和改善预后，以及随后开发有效的药理学和免疫学干预措施。具体的初始病毒目标包括沙粒病毒、皮钦德病毒和拉沙病毒（后者在美国国立卫生研究院和美国疾病控制与预防中心的A类名单上）。

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).