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

全球新冠肺炎大流行对人类健康和经济造成了相当大的影响，给研究界带来了一系列独特的迫切挑战，需要加以应对。其中一个挑战是减缓导致新冠肺炎的SARS-CoV2病毒的传播。该项目的目标是设计简单的技术来制造防止病毒传播的防护材料。为了实现这一目标，病毒颗粒将被设计成使病毒研究更容易、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、空调过滤器和工作表面的材料。通过简单地将植物蛋白质浸泡在蛋白质溶液中，植物蛋白质也很容易附着在棉花和其他天然和合成纤维等表面。新冠肺炎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经过改造，在其表面产生SARS-CoV2的“尖峰”；(2)尖峰蛋白将被插入到合成脂膜液滴中，类似于包裹SARS-CoV2的那些液滴。这两种非致病性模型病毒将允许对冠状病毒和其他新出现的病毒进行广泛研究，而不需要高度保护性的专门设备。这项研究的成功完成将有助于保护公众的努力，并可能导致新的有效的基于自然的保护措施，以防止SARS-CoV2和其他类似结构的病毒的传播。SARS-CoV2(新冠肺炎的病毒)是一种脂质包裹的病毒，具有突出的尖峰蛋白。德克萨斯大学的研究人员最近确定了这种尖峰蛋白的结构。这项拟议的研究的目的是评估特定植物来源的抗菌肽(AMPs)功能化的表面结合这种刺激蛋白的能力。这些AMP包括从辣木籽(MO)水提取物中获得的两种蛋白质：几丁质结合蛋白(MoCBP)和阳离子蛋白(MO2.1)。这项工作的中心假设是：(I)使用MoCBP功能化表面特异性结合SARS-CoV2刺激蛋白受体结合域可以作为一种有效的去除技术，(Ii)功能化表面上的MO2.1将通过破坏病毒的脂膜来灭活SARS-CoV2。最近的模拟和实验结果表明，MoCBP与纯化的SARS-CoV2刺突蛋白具有很强的相互作用。为了促进这项研究，我们将开发一种非致病的SARS-CoV2模型，用于快速实验，而不需要专门的安全设备。为了验证假设并实现本研究的目标，将开展以下工作：1)以SARS-CoV2刺突蛋白和显示SARS-CoV2受体结合结构域的修饰T7噬菌体为替代物，通过过滤实验测试MO包被棉对水和空气中病毒的去除效率；2)以经Spike蛋白修饰的病毒样脂质颗粒为病毒替代物，测试MO2.1与SARS-CoV2脂膜的相互作用；以及3)测试易用医用口罩和涂有MO蛋白的HVAC过滤器对固定化SARS-CoV2病毒的效果。这项研究的成功完成具有巨大的潜力，可以导致开发有效的技术来移除和保护SARS-CoV2病毒。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.