Molecular probes for a vOTU from CCHFV using a fluorogenic peptide

使用荧光肽对来自 CCHFV 的 vOTU 进行分子探针

基本信息

批准号：
8547834
负责人：
Scott Dusan Pegan
金额：
$ 0.43万
依托单位：
UNIVERSITY OF DENVER (COLORADO SEMINARY)
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-20 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8547834
关键词：
Address Affect Afghanistan Africa American Americas Animal Diseases Area Arterivirus Asia Biological Assay Bunyaviridae Cell Line Central Asia Cessation of life Chemicals Congo Crimean Hemorrhagic Fever Crimean-Congo Hemorrhagic Fever Virus Cysteine Protease DNA-Directed RNA Polymerase Deubiquitination Disease Down-Regulation Eastern Europe Effectiveness Enzymes Equine arteritis virus Europe Evaluation Excision FDA approved Family Family suidae Fever Fluorescence Fluorescence Resonance Energy Transfer Fluorogenic Substrate Gel Genes Hemorrhage Homologous Gene Human Immune Immune response Immune system In Vitro Inhibitory Concentration 50 Interferons Kinetics Lead Left Libraries Link Mammalian Cell Measures Middle East Military Personnel Modeling Modification Molecular Bank Molecular Probes Nairovirus Natural Immunity Papain Peptide Hydrolases Peptides Phylogenetic Analysis Plant Diseases Plant Viruses Play Polyubiquitin Porcine Reproductive and Respiratory Syndrome Porcine respiratory and reproductive syndrome virus Process Protease Domain Proteins RNA Regulation Relative (related person)Reporting Reproduction Research Personnel Rice Rice stripe virus Risk Role Route Russia SARS Coronavirus Protease Pathway Signal Transduction Simulate Soldier Specificity Structure Structure-Activity Relationship Syndrome Techniques Testing Therapeutic Ticks Toxic effect Ubiquitin United States Vaccines Variant Viral Viral Proteins Virulence Factors Virus Virus Diseases base cost effective drug discovery health economics high throughput screening human disease improved in vitro Assay in vivo information gathering inhibitor/antagonist insight member mortality ovarian neoplasm pathogen preference prophylactic prostration public health relevance repository respiratory response scaffold screening small molecule trafficking transmission process vector viral RNA

项目摘要

DESCRIPTION (provided by applicant): Crimean-Congo hemorrhagic fever (CCHF) virus is a ssRNA (-) Nairovirus that produces fever, prostration, and severe hemorrhages in humans. Fatality rates for CCHF range from 5-70% based on phylogenetic variation of the virus, transmission route, and different treatment facilities. Originally identified in Russia and the Congo, CCHF has rapidly spread across large sections of Europe, Asia, and Africa. Recently, U.S. citizen and military traffic has increased substantially to the regions affected by CCHF, specifically South Central Asia. As a result, there is a substantial risk for transmission of CCHF and/or its tick vector to the United States of America. This risk has been recently highlighted by a CCHF death of an American Soldier based in Afghanistan in 2009. Currently, there is no FDA approved vaccine or therapeutic for treatment of CCHF. Recent reports have identified a viral homologue of the ovarian tumor protease (vOTU) and implicated its involvement down-regulation of the Interferon type 1 immune response through cleavage of post- translational modifying proteins ubiquitin (Ub) and Ub-like interferon-simulated gene 15 (ISG15). Additionally, homologues of CCHFV's vOTU have been suggested to perform similar roles in the economically devastating ssRNA (+) Arteriviruses, Porcine Respiratory and Reproduction Syndrome and Equine Arteritis viruses. Even plant viruses such as the damaging rice stripe virus have been shown to possess a viral ovarian tumor domain protease homologue. This proposal will implement a cost effective fluorescent-based enzymatic assay to identify molecule probes for a vOTU from CCHFV as well as assess whether the molecular probe scaffold can serve as a basis to selectively inhibit other viral and eukaryotic ovarian tumor domain proteases. Specifically a high-throughput screening campaign using a fluorogenic peptide and the MLPCN library will occur at a MLPCN facility. Molecular probes initially identified for the vOTU from CCHFV will be subsequently optimized using secondary, orthogonal, and tertiary in vitro assays as well as in vivo ones. The resulting information provided by molecular probes inhibiting the function of vOTU from CCHFV will allow the necessary insight into not only the role of CCHF's vOTU, but vOTUs in general, in viral evasion of the innate immune system. Additionally, information gathered from vOTU molecular probes would also assist in determining the practicality of developing prophylactics targeting vOTUs and their eukaryotic superfamily relatives that negatively regulate the human innate response.

描述（由申请人提供）：Crimean-Congo出血热（CCHF）病毒是一种ssRNA（ - ）奈罗内病毒，可在人类中产生发烧，俯卧和严重的出血。基于病毒，传播途径和不同治疗设施的系统发育变化，CCHF的死亡率范围为5-70％。 CCHF最初在俄罗斯和刚果中被确定，已迅速遍及欧洲，亚洲和非洲的大部分地区。最近，美国公民和军事交通已大大增加了受CCHF影响的地区，特别是中亚南亚的影响。结果，CCHF和/或其tick矢量传播到美利坚合众国存在很大的风险。最近，2009年，一名美国士兵在阿富汗的一名美国士兵死亡的CCHF死亡最近强调了这种风险。目前，尚无FDA批准的疫苗或治疗CCHF治疗的疫苗。最近的报告已经确定了卵巢肿瘤蛋白酶（FOTU）的病毒同源物，并暗示了其参与其对1型干扰素1型免疫反应的下调，这是通过裂解后翻译后修饰的蛋白质泛素（UB）和Ub样干扰素模拟的基因15（ISG15）。此外，已经建议使用CCHFV的投票的同源物在经济毁灭性的ssRNA（+）动脉病毒，猪呼吸和繁殖综合征和马动脉炎病毒中发挥相似的作用。甚至诸如破坏水稻条纹病毒之类的植物病毒也已证明具有病毒卵巢肿瘤结构蛋白酶同源物。该提案将实施基于荧光的酶促测定法，以鉴定来自CCHFV的投票的分子探针，并评估分子探针支架是否可以作为选择性抑制其他病毒和真核卵巢肿瘤结构蛋白的基础。特别是使用荧光肽和MLPCN库的高通量筛查活动将在MLPCN设施中发生。最初从CCHFV鉴定出的投票的分子探针将随后使用次级，正交和第三级的体外测定以及体内进行优化。由CCHFV抑制投票功能的分子探针提供的最终信息将使必要的了解不仅对CCHF的votu的作用，而且对总体投票的作用，在逃避先天免疫系统的病毒式逃避中。此外，从投票分子探针中收集的信息还将有助于确定针对投票的预防学及其真核生物超级家族亲戚的实用性，从而对人类先天的反应产生负面的调节。