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

用于快速开发和筛选埃博拉病毒抗融合抗病毒药物的病毒外壳蛋白阵列

• 批准号: 1504846
• 负责人: Susan Daniel
• 金额: $ 20万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1504846&HistoricalAwards=false
• 关键词: Viral; coat; protein; arrays; rapid

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.

埃博拉病毒感染开始于病毒进入体内的特定细胞，通过一个过程，包括与细胞表面结合，然后融合，在此过程中病毒将其遗传物质释放到细胞中。如果结合或融合被中断，病毒将不再能够引起感染。这项工作的主要目的是开发阻断病毒与细胞融合的治疗方法。为了实现这一目标，PI将使用一种新的体外平台，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)一种筛选工具，可以很容易地用于发现对抗未来病毒爆发的化合物，以及2)可用于阻止当前埃博拉病毒疾病爆发进展的候选抗体。此外，调查人员将与公众互动，提供有关埃博拉病毒、其感染过程以及导致当前疫情的因素的信息。技术摘要：目前，还没有一种可以直接检测抗促发性化合物对埃博拉病毒疗效的筛选工具。融合步骤是一个特别有希望的治疗靶点，因为埃博拉病毒刺突蛋白（GP）的融合结构域比其结合结构域进化得慢得多。为了解决国家对鉴定阻止埃博拉病毒感染的化合物的迫切需要，PI建议(1)创建一个针对埃博拉病毒的抗融合性化合物的快速筛选平台；(2)鉴定破坏GP融合功能的抗体和化合物。PI将根据她以前使用单粒子跟踪（SPT）方法研究包膜病毒融合的工作，采用微流控病毒融合分析平台。该平台由一个多通道微流控装置组成，该装置在通道壁上包裹有宿主细胞模拟物（称为支持脂质双分子层，SLB），以及含有埃博拉病毒GP蛋白的病毒（或病毒样颗粒，vlp），该病毒可与SLB融合。该技术以高通量的方式直接探测病毒融合，并提供融合机制的定量信息。预计该平台可用于埃博拉病毒抗融合性抗病毒化合物的快速筛选和鉴定。此外，pi将参与有关该病毒的公开讨论，提供有关感染过程的信息，并通过向地区医院（卡尤加医疗中心）的医务人员作报告等活动，提供有关疫情爆发的观点，作为其继续教育系列和埃博拉应对计划的一部分。