The Sumoylation System as a Novel Target for Anti-Influenza Therapies

描述(申请人提供)：流感仍然是人类最可怕的祸害之一。据计算，每年约有36,000名美国人死于与流感感染相关的并发症。事实证明，疫苗接种和目前的抗病毒药物在预防流感感染引起的致命并发症方面是有效的。然而，新的大流行性流感毒株可能对当前的抗病毒药物产生抗药性的迫在眉睫的演变，以及与当前疫苗生产方法相关的内在技术限制，突显了开发新的抗流感疗法的迫切需要。阻断或加强对病毒复制至关重要或有害的细胞过程的抗病毒疗法，极大地减少了产生抗药性病毒的可能性。此外，通过影响病毒使用的细胞途径，这种疗法更有可能独立于入侵病毒的类型、毒株和抗原性而发挥作用。因此，这些疗法构成了理想的抗流感策略。 在这里，我们假设增强相扑系统的活性，这是一个细胞翻译后修饰系统，将抑制流感病毒的复制。这项研究的目的是评估苏莫化与流感病毒感染的相关性，最终目标是确定苏莫化系统是否为新的抗流感疗法提供新的靶点。为此，我们将追求两个特定的目标：(1)表征感染过程中被总甲基化的流感病毒蛋白的全序列，以及(2)确定调节宿主细胞总甲基化系统的活性对流感病毒增殖的影响。 为了实现这些目标，我们将利用蛋白质组学方法、瞬时和稳定的转染法和腺病毒表达系统相结合。所提出的检测方法将为流感感染过程中调节和甲基化的作用提供确凿的数据，并为病毒的生命周期中的效应提供有意义的提示。此外，在拟议的研究期间开发的工具将允许在其他病毒上测试调节相扑系统的效果。这项研究具有很高的创新性，因为相扑系统与流感和其他病毒感染的相关性仍未被探索。这也是非常紧迫的，因为它可能导致开发可能适用于治疗大流行性流感和其他病毒疾病的新型抗流感疗法。此外，实现提议的目标还将为PI提供制定具有竞争力的NIH R01提案所需的基础和初步数据。

DESCRIPTION (provided by applicant): Influenza still represents one of the most dreadful human scourges. Complications associated to influenza infections are calculated to kill approximately 36,000 Americans every year. Vaccination and current anti-viral drugs have proven useful in preventing fatal complications upon influenza infection. However, the imminent evolution of new pandemic influenza strains likely to be resistant to current anti-viral drugs and the intrinsic technical limitations associated to current vaccine production methods underscore the urgent need to develop novel anti-influenza therapies. Anti-viral therapies that block or enhance cellular processes essential or detrimental for viral replication are significantly less prone to development of resistant viruses. Additionally, by affecting cellular pathways used by the virus, such therapies are more likely to work independently of the type, strain, and antigenic properties of the invading virus. Therefore, such therapies constitute ideal anti-influenza strategies. Here, we hypothesize that augmenting the activity of the sumoylation system, a cellular post-translational modification system, will inhibit influenza virus multiplication. The objective of this study is to evaluate the relevance of sumoylation for influenza virus infection, with the ultimate goal of determining whether the sumoylation system provides new targets for novel anti-influenza therapies. To this end, we will pursue two specific aims: (1) characterize the full array of influenza viral proteins that are sumoylated during infection, and (2) determine the effect of modulating the activity of the host cell sumoylation system on influenza virus multiplication. To achieve these goals, we will utilize a combination of proteomic methods, transient and stable transfection approaches, and adenovirus expression systems. The proposed assays will provide conclusive data on the effect of modulating sumoylation during influenza infections, and give meaningful hints to the effect of sumoylation on the life cycle of the virus. Additionally, the tools developed during the proposed studies will allow the effects of modulating the sumoylation system to be tested on other viruses. This research is highly innovative because the relevance of the sumoylation system for influenza and other viral infections remains unexplored. It is also of great urgency as it could lead to the development of novel anti-influenza therapies potentially applicable to the treatment of pandemic flu and other viral diseases. Furthermore, attaining the proposed goals will also provide the PI with the foundation and preliminary data required for crafting competitive NIH R01 proposals.