Disruption of Conserved RNA Stem-Loops in Filovirus RNA

描述（由申请方提供）：丝状病毒科由埃博拉病毒和马尔堡病毒组成。感染这些出血热病毒会导致胃肠道大量出血，以及穿刺伤口和粘膜出血。高死亡率和未解决的感染模式，加上缺乏有效的疫苗或抗病毒药物，导致这些病毒被疾病控制中心列为A类。这些病毒作为生物武器的潜力需要继续设计，合成和评估新的治疗策略。丝状病毒是负链RNA病毒。病毒复制周期的主要特征已经阐明。在病毒进入细胞并脱壳后，负链基因组RNA作为mRNA合成的模板。埃博拉病毒和马尔堡病毒各编码七种从单个mRNA翻译而来的病毒蛋白质。七种mRNA中的每一种在mRNA的5'末端含有RNA茎环。预期该RNA茎环的破坏将影响病毒蛋白质的表达和复制。因此，RNA茎-环代表了用于开发设计用于抑制病毒基因表达和复制的RNA结合配体的优良靶标。拟议实验的长期目标是开发用于干预丝状病毒感染的新治疗策略。为此，我们建议首先开发与RNA茎环结合以抑制基因表达的配体。提出了以下具体目的：具体目的1：鉴定与埃博拉病毒和马尔堡病毒RNA的RNA茎环结合的RNA和蛋白配体。具体目的2：确定肽和蛋白质配体阻断埃博拉和马尔堡RNA茎环mRNA翻译的能力。这些研究的结果将确立RNA茎环的重要性，并为开发旨在破坏埃博拉病毒和马尔堡病毒复制的新疗法提供基础。

DESCRIPTION (provided by applicant): The family Filoviridae consists of the Ebola and Marburg viruses. Infections with these hemorrhagic fever viruses results in massive bleeding into the gastrointestinal tract as well as hemorrhages from puncture wounds and mucus membranes. The high mortality rate and unresolved mode of infection, coupled with the lack of effective vaccine or antivirals, have resulted in these viruses being classified in a Category A by the Center for Disease Control. The potential for the use of these viruses as bio-weapons necessitates the continued design, synthesis, and evaluation of new therapeutic strategies. Filoviruses are negative stranded RNA viruses. The major features of the viral replication cycle have been elucidated. After the virus enters the cell and uncoats, the negative strand genome RNA serves as a template for the synthesis of mRNAs. Ebola and Marburg viruses each encode seven viral proteins translated from individual mRNAs. Each of the seven mRNAs contains an RNA stem-loop at the 5' end of the mRNA. Disruption of this RNA stem-loop would be expected to effect both on the expression of viral proteins and replication. Thus, the RNA stem-loops represent excellent targets for the development of RNA binding ligands designed to inhibit virus gene expression and replication. The long-term goal of the proposed experiments is to develop new therapeutic strategies for intervention of Filovirus infections. To do this, we propose to first develop ligands that bind to the RNA stem-loops to inhibit gene expression. The following Specific Aims are proposed: Specific Aim 1: To identify RNA and protein ligands that bind to RNA stem-loops of Ebola and Marburg virus RNAs. Specific Aim 2: To determine the capacity of peptide and protein ligands to block translation of mRNAs with the Ebola and Marburg RNA stem loops. The results of these studies will establish the importance of the RNA stem-loops and provide the foundation for development of new therapeutics designated to disrupt both Ebola and Marburg virus replication.