Novel Countermeasures to Hemorrhagic Fever Viruses

描述（由申请人提供）：相关性。“A类”出血热病毒（HFV），如裂谷热病毒（RVFV），已经在积极开发中用作BWT剂。尽管作出了广泛的努力，但目前还没有有效的医疗对策来应对这些威胁。因此，快速产生抗HFV治疗剂的能力对于保护美国平民和部队至关重要，从而防止与HFV感染相关的潜在破坏性伤亡。 背景由于目前开发抗病毒化合物的方法非常昂贵和耗时，但很少成功，我们已经开发了一种革命性的技术来快速产生对细胞溶解性病原体的治疗剂。 目标.我们的病原体选择性抗病原体技术利用多种（即，10 e8）细胞驻留组合蛋白质文库（称为衔接蛋白文库）。我们建议筛选这些库，以确定新的广谱治疗，保护细胞免受“A类”HFVs的挑战。这些治疗剂设计用于暴露于BWT制剂之前或之后，以防止平民和军事人员伤亡。 研究设计.为了成功完成这一创新项目，我们组建了一个由病毒学家、微生物学家和结构生物学家组成的多学科团队。复杂的衔接蛋白文库已经整合到哺乳动物细胞中，使得每个细胞表达衔接蛋白文库的独特成员。我们将用RVFV攻击这些异质细胞群。将选择在病毒攻击中存活的细胞，克隆，并通过巢式PCR测序鉴定负责抗病毒活性的保护性衔接蛋白。每种保护性衔接蛋白的纯化的细胞渗透变体将被开发为治疗剂，并且这些衔接蛋白保护细胞和动物免受RVFV的功效将由剂量反应研究确定。在项目第二年，我们新的BSL-4设施将可用于将我们的策略应用于其他HFV，从而测试我们的适应蛋白对这些病毒的效力。以这种方式，将产生提供针对不同HFV的广泛保护的化合物。我们的技术的一个显著优势是，它消除了基于靶标或计算药物发现方法中固有的先验假设，最终使我们的技术能够发现BWT或新出现的病原体的新疗法，而无需详细的病原体表征。

DESCRIPTION (provided by applicant): Relevance. "Category A" hemorrhagic fever viruses (HFVs), such as Rift Valley fever virus (RVFV), have been under active development for use as BWT agents. Despite extensive effort, no effective medical countermeasures are currently available to combat these threats. Thus, the ability to rapidly generate anti-HFV therapeutics is essential to protect U.S. civilians and Forces, thereby preventing potentially devastating casualties associated with HFV infections. Background. Since current approaches to developing antiviral compounds are extremely costly and time-consuming, but rarely successful, we have developed a revolutionary technology to rapidly generate therapeutics to cytolytic pathogens. Objectives. Our pathogen-selected anti-pathogen technology utilizes diverse (i.e., 10e8) cell-resident combinatorial protein libraries (termed adaptein libraries). We propose to screen these libraries to identify novel broad-spectrum therapeutics that protect cells from challenge with "Category A" HFVs. These therapeutics are designed for use before or after exposure to BWT agents to prevent civilian and military casualties. Study design. To successfully accomplish this innovative project, we have assembled a multi-disciplinary team of expert virologists, microbiologists and structural biologists. Complex adaptein libraries have been integrated into mammalian cells, such that each cell expresses a unique member of the adaptein library. We will challenge these heterogeneous cell populations with RVFV. Cells that survive virus challenge will be selected, cloned, and the protective adaptein responsible for antiviral activity identified by nested-PCR sequencing. Purified cell-permeant variants of each protective adaptein will be developed as therapeutic agents, and the efficacy of these adapteins to protect cells and animals from RVFV will be determined from dose-response studies. During project year 2, our new BSL-4 facility will be available to apply our strategy to other HFVs, and thus test the efficacy of our adapteins against these viruses. In this manner, compounds that provide broad protection against different HFVs will be generated. A significant strength of our technology is that it eliminates the a priori assumptions inherent within target-based or computational drug discovery methods, ultimately allowing our technology to discover novel therapeutics to BWT or emergent pathogens without the need for detailed pathogen characterization.