Strain-specific and pan-filoviral aptamer recognition of Ebola virus glycoproteins

项目摘要 丝状病毒感染人类、非人灵长类动物、蝙蝠和其他哺乳动物。几种丝状病毒引起 人类的出血热疾病，包括最近在西非爆发的埃博拉病毒（EBOV）。 有希望的干预措施即将出现，但持续的危机突出表明需要新的战略。 缺乏有效和广泛可用的疫苗或抗病毒药物方案，难以保证 即使在现代医院环境中，患者的康复也受到持续风险的影响， 新疾病的国际传播。该项目将揭示靶向糖蛋白（GP）的新机会 通过鉴定和表征识别和抑制丝状病毒GP的核酸适体。 丝状病毒在其表面膜上显示三聚体GP，它们在感染过程中从感染的细胞获得。 组装件. GP是中和抗体和非中和抗体（Ab）的结合靶标，并且GP是中和抗体和非中和抗体（Ab）的结合靶标。 中和的分子机制开始出现。然而，非保守和严重- 糖基化粘蛋白样结构域（MLD）阻断Ab接近大部分GP表面，包括NCP 1 在EBOV GP（“GPEBOV”）的情况下，结合位点。较小的配体，如适体， 并阻止抗体无法到达的中和表面，它们可以帮助识别 新的中和点 我们的长期目标是开发适体作为鉴定新中和表位的工具， 剖析病毒发病机制中的分子和细胞事件。目前的建议将建立可行性 并为机制研究确定初步线索。它利用了互补性 参与实验室在病毒学和RNA生物化学方面的专业知识;我们最近在先进的 适体选择和信息学方法;关于我们最近对机制和多功能性的重要见解 在病毒装配过程中糖蛋白掺入的研究;以及增加与EBOV的新合作者 专业知识第一个目标将通过以下方式鉴定中和GP假型和感染性病毒的适体： 识别GPEBOV和GPMARV之间共有（或独特）的表位。第二个目标将确定 并评估靶向已知中和表位的适体。

Project Summary Filoviruses infect humans, non-human primates, bats and other mammals. Several filoviruses cause hemorrhagic fever diseases in humans, including the recent Ebola virus (EBOV) outbreak in western Africa. Promising interventions are on the horizon, but the need for new strategies is highlighted by the continued absence of an effective and widely-available vaccine or antiviral drug regimen, by the difficulty of assuring patient recovery even in modern hospital settings, and by the continuing risk of the emergence and rapid international spread of new diseases. This project will reveal new opportunities to target glycoproteins (GPs) by identifying and characterizing nucleic acid aptamers that recognize and inhibit filoviral GPs. Filoviruses display a trimeric GP on their surface membrane, which they acquire from infected cells during assembly. GPs are the binding targets for both neutralizing and non-neutralizing antibodies (Ab), and the molecular mechanisms of neutralization are beginning to emerge. However, the non-conserved and heavily- glycosylated mucin-like domain (MLD) blocks Ab access to much of the GP surface, including the NCP1 binding site in the case of EBOV GP (“GPEBOV”). Smaller ligands such as aptamers could potentially reach and block neutralization surfaces that are inaccessible to antibodies, and they could aid identification of new neutralization sites. Our long-term objective is to develop aptamers as tools for identifying new neutralizing epitopes and for dissecting molecular and cellular events in viral pathogenesis. The current proposal will establish feasibility of that approach and identify initial leads for mechanistic studies. It capitalizes on complementary expertise in virology and RNA biochemistry in the participating labs; on our recent achievements in advanced aptamer selection and informatics methods; on our recent critical insights into the mechanisms and versatility of glycoprotein incorporation during viral assembly; and on the addition of new collaborators with EBOV expertise. The first Aim will identify aptamers that neutralize GP-pseudotyped and infectious virus by recognizing epitopes that are shared between (or unique to) GPEBOV and GPMARV. The second Aim will identify and evaluate aptamers that target known neutralizing epitopes.