Viral coat protein arrays for rapid development and screening of anti-fusogenic antivirals against Ebolavirus

埃博拉病毒感染始于病毒进入体内的特定细胞，通过与细胞表面结合的过程，然后融合，在此期间病毒将其遗传物质释放到细胞内。如果结合或融合中断，病毒将不再能够引起感染。这项工作的主要目标是开发阻止病毒融合到细胞的疗法。为了实现这一目标，PI将使用一种新型的体外平台，使她能够筛选防止病毒融合的化合物。在这项工作结束时，对社会的好处将是：1)一种筛查工具，可以很容易地发现化合物，以对抗未来的病毒爆发；以及2)候选抗体，可以用来阻止当前埃博拉病毒疾病的爆发。此外，调查人员还将与公众互动，提供有关埃博拉病毒、其感染过程以及导致埃博拉疫情爆发的因素的信息。技术摘要：目前，还没有可用的筛查工具来直接测试抗融合化合物对埃博拉病毒的疗效。融合步骤是治疗学的一个特别有希望的目标，因为埃博拉病毒尖峰蛋白(GP)的融合结构域进化得比它的结合域慢得多。为了满足国家对识别阻止埃博拉病毒感染的化合物的迫切需求，PI建议(1)创建一个针对埃博拉病毒的抗融合化合物的快速筛选平台；以及(2)识别破坏GP融合功能的抗体和化合物。PI将在她之前工作的基础上采用微流控病毒融合分析平台，使用单粒子跟踪(SPT)方法来研究包膜病毒融合。该平台包括一个多通道微流控装置，在通道壁上覆盖宿主细胞模拟物(称为支持的脂质双层，SLB)，以及含有可与SLB融合的埃博拉病毒GP蛋白的病毒(或病毒样颗粒，VLP)。这项技术以高通量的方式直接探测病毒融合，并提供关于融合机制的定量信息。预计该平台可用于快速筛选和鉴定抗埃博拉病毒的抗融合抗病毒化合物。此外，作为继续教育系列和埃博拉应对计划的一部分，私人投资机构将参与有关病毒的公开讨论，提供有关感染过程的信息，并通过在地区医院(卡尤加医疗中心)向医务人员介绍等活动提供有关疫情的观点。

The Ebola virus infection begins when viruses enter specific cells in the body, through a process involving binding to the cell surface followed by fusion during which the virus releases its genetic material into the cell. If either binding or fusion is interrupted, the virus will no longer be able to cause infection. The main objective of this work is to develop therapeutics that block virus fusion to the cell. To accomplish this, the PI will use a novel in vitro platform that allows her to screen for virus fusion preventing compounds. At the conclusion of this work, the benefit to society will be: 1) a screening tool that can be readily adapted to discover compounds to combat future viral outbreaks, and 2) antibody candidates that can be used to stop the progression of the current Ebola virus disease outbreak. Additionally, the investigators will interact with the public to provide information about the Ebola virus, its infection process, and factors that contribute to outbreaks such as the current one.Technical Abstract: At present, there are no available screening tools that can directly test the efficacy of anti-fusogenic compounds against Ebola. The fusion step is an especially promising target for therapeutics, as the fusion domain of Ebola virus spike protein (GP) evolves much slower than its binding domain. To address the urgent national need to identify compounds that stop Ebola virus infection, the PI proposes to (1) create a rapid screening platform for anti-fusogenic compounds against Ebola virus; and (2) identify antibodies and compounds that disrupt the fusion function of GP. The PI will adapt a microfluidic virus fusion assay platform based on her previous work using single particle tracking (SPT) approaches to investigate enveloped virus fusion. The platform consists of a multi-channel microfluidic device with host cell mimics coated on the channel walls (referred to as a supported lipid bilayer, SLB), and virus (or virus-like particles, VLPs) containing Ebola virus GP protein that can fuse with the SLB. This technology directly probes viral fusion in a high-throughput manner and provides quantitative information on the fusion mechanism. It is anticipated that this platform can be used for the rapid screening and identification of anti-fusogenic antiviral compounds against Ebola virus. In addition, the PIs will participate in public discourse about the virus, providing information about the infection process and providing perspective about the outbreak through such activities as presentations to the medical staff at the regional hospital (Cayuga Medical Center) as part of their Continuing Education Series and Ebola-response plan.