RAPID: Accessible Surfaces for Interrupting Sustained Coronavirus Transmission (ASsIST)

全球COVID-19大流行对人类健康和经济造成了巨大影响，为研究界带来了一系列需要解决的独特紧迫挑战。挑战之一是减缓导致COVID-19的SARS-CoV 2的传播。这个项目的目标是设计简单的技术来制作防止病毒传播的防护材料。为了实现这一目标，病毒粒子将被设计成使病毒研究更容易和更安全。将被测试的材料将被涂上容易获得的植物蛋白，以防止病毒传播。这些材料将包括用于口罩、空调过滤器和工作台面的材料。植物蛋白也很容易附着在表面，如棉花和其他天然和合成纤维，只需将这些材料浸入蛋白质溶液中。COVID-19的模型将通过两种不同的方法创建：（1）通常感染细菌的病毒将被修改，以在其表面产生SARS-CoV 2的“刺突”;（2）刺突蛋白将被插入类似于SARS-CoV 2涂层的合成脂质膜液滴中。这两种非致病性模型病毒将允许对冠状病毒和其他新兴病毒进行广泛研究，而无需高度保护性的专用设备。这项研究的成功完成将为保护公众的努力提供信息，并可能导致新的有效的基于自然的保护措施，以防止SARS-CoV 2和其他类似结构的病毒的传播。SARS-CoV 2（导致COVID-19的病毒）是一种具有突出刺突蛋白的脂质包膜病毒。刺突蛋白的结构最近由德克萨斯大学的研究人员确定。这项研究的目的是评估用特定植物源性抗菌肽（AMP）功能化的表面结合这种刺突蛋白的能力。这些AMP包括从辣木种子（MO）的水性提取物获得的两种蛋白质：几丁质结合蛋白（MoCBP）和阳离子蛋白（MO2.1）。所提出的工作的中心假设是：（i）使用MoCBP功能化表面的SARS-CoV 2刺突蛋白受体结合结构域的特异性结合可以用作有效的去除技术，以及（ii）功能化表面上的MO2.1将通过破坏病毒的脂质包膜来破坏SARS-CoV 2。最近的模拟和实验结果表明，强烈的相互作用MoCBP与纯化的刺突蛋白从SARS冠状病毒2。为了促进这项研究，我们将开发一种非致病性的SARS-CoV 2模型，用于快速实验，而不需要专门的安全设备。本研究的主要目的是：1）以SARS-CoV 2刺突蛋白和修饰的T7噬菌体为替代物，通过过滤实验，检测MO涂层棉对水和空气中病毒的去除效果; 2）用修饰有刺突蛋白的病毒样脂质颗粒作为病毒替代物，检测MO2.1与SARS-CoV 2脂质膜的相互作用，以建立去除/灭活;以及3）测试容易获得的外科口罩和涂覆有MO蛋白的HVAC过滤器在固定SARS-CoV 2中的有效性。这项研究的成功完成有很大的潜力，导致有效的技术，消除和保护对SARS-CoV 2的发展。这一奖项反映了NSF的法定使命，并已被认为是值得的支持，通过评估使用基金会的知识价值和更广泛的影响审查标准。

The global COVID-19 pandemic causing considerable human health and economic impacts presents the research community with a unique set of urgent challenges that need to be addressed. One of the challenges is to slow the transmission of SARS-CoV2, the virus that causes COVID-19. The goal of this project is to design simple techniques to make protective materials that prevent virus transmission. To achieve this goal, virus particles will be designed to make virus research easier and safer. Materials that will be tested will be coated with readily available plant proteins to prevent virus transmission. These materials will include those used for masks, air conditioning filters, and work surfaces. The plant proteins are also easy to attach to surfaces such as cotton and other natural and synthetic fibers by simply immersing these materials in protein solutions. Models of COVID-19 will be created in two different methods: (1) a virus that normally infects bacteria will be modified to produce the “spike” of SARS-CoV2 on its surface; (2)The spike protein will be inserted into synthetic lipid membrane droplets similar to those coating SARS-CoV2. These two non-pathogenic model viruses will allow widespread research into coronavirus and other emerging viruses without the need for highly protective specialized equipment. Successful completion of this research will inform efforts to protect the public and potentially lead to new effective nature-based protective measures against the spread of SARS-CoV2 and other similarly structured viruses.SARS-CoV2 (the virus responsible for COVID-19) is a lipid enveloped virus with protruding spike proteins. The structure of the spike protein was recently determined by researchers at the University of Texas. The goal of this proposed research is to evaluate the ability of surfaces functionalized with specific plant-derived antimicrobial peptides (AMPs) to bind this spike protein. These AMPs include two proteins obtained from aqueous extracts of the Moringa oleifera seed (MO): a chitin binding protein (MoCBP) and a cationic protein (MO2.1). The central hypotheses of the proposed work are: (i) specific binding of the SARS-CoV2 spike protein receptor binding domain using MoCBP-functionalized surfaces can be used as an effective removal technique, and (ii) MO2.1 on functionalized surfaces will inactivate SARS-CoV2 by damaging the lipid envelope of the virus. Recent simulation and experimental results demonstrate strong interactions of MoCBP with the purified spike protein from SARS-CoV2. To facilitate this research, we will develop a non-pathogenic model of SARS-CoV2 for rapid experimentation without the need for specialized safety equipment. The following tasks will be performed to test hypotheses and achieve the goal of this research: 1) test virus removal efficiency of MO-coated cotton from water and air by using filtration experiments with SARS-CoV2 spike protein and modified T7 bacteriophages displaying the receptor binding domain of SARS-CoV2 as surrogates; 2) test the interaction of MO2.1 with the lipid membrane of SARS-CoV2 by using virus-like lipid particles amended with spike protein as virus surrogates to establish removal/inactivation; and 3) test the effectiveness of easily accessible surgical masks and HVAC filters coated with MO proteins in immobilizing SARS-CoV2. Successful completion of this research has great potential to lead to the development of effective technology for removal and protection against SARS-CoV2.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.