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

项目摘要 丝状病毒感染人类、非人类灵长类动物、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，但持续不断的 缺乏有效和广泛可用的疫苗或抗病毒药物方案，因为难以确保 即使在现代化的医院环境下，患者的康复也受到持续风险的出现和迅速 新疾病的国际传播。该项目将揭示靶向糖蛋白(GP)的新机会。 通过识别和鉴定识别和抑制丝状病毒GP的核酸适配子。 丝状病毒在其表面膜上显示一个三聚体GP，这种GP是它们在感染细胞过程中获得的 集合。GPS是中和抗体和非中和抗体的结合靶标，而 中和的分子机制开始出现。然而，非保守的和严重的- 糖基化粘蛋白样域(MLD)阻断了抗体对GP表面的大部分访问，包括NCP1 在EBOV GP(“GPEBOV”)的情况下的结合位点。较小的配体，如适配子，可能会到达 并阻断抗体无法接触到的中和表面，它们可以帮助识别 新的中和点。 我们的长期目标是开发适配子作为工具，用于识别新的中和表位和 剖析病毒发病机制中的分子和细胞事件。目前的提案将确定可行性 并确定机械学研究的初步线索。它利用互补性 参与实验室在病毒学和RNA生物化学方面的专业知识；关于我们在高级 适配子选择和信息学方法；关于我们最近对机制和多功能性的批判性见解 病毒组装过程中糖蛋白的掺入；以及EBOV的新合作者的加入 专业知识。第一个目标是通过以下方式识别中和GP假型和传染性病毒的适体 识别在GPEBOV和GPMARV之间共享(或唯一)的表位。第二个目标是确定 并评估针对已知中和表位的适配子。

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.