Elucidating How Tri-phosphatase DUSP11 Controls HCV Infection and Hepatocyte Inflammation

项目摘要 丙型肝炎病毒（HCV）的清除及其疾病表现与 炎症反应。然而，关于引发炎症的因素存在很大的知识差距， 控制HCV复制。迫切需要填补这些空白，因为确定 控制HCV感染和相关炎症对于理解HCV相关疾病是必要的。 这个合作团队的长期目标是了解HCV先天控制的机制， 发病机制与这一目标相一致，本提案的总体目标是确定RNA 三磷酸酶DUSP 11与HCV感染和促炎RNA的控制相关 成绩单核心假设是DUSP 11促进促炎宿主和病毒的周转。 三磷酸化的转录物，从而减少HCV复制和RNA相关的炎症。的 这项研究的基本原理是，一旦知道DUSP 11如何控制病毒， 复制和宿主促炎转录，这将促进对先天防御的理解， 这对于指导未来合理的疫苗设计和开发治疗策略至关重要， 解决HCV引起的肝脏发病机制。检验中心假设并完成目标 本提案中概述的将通过以下两个具体目标来实现：1）确定抗HCV DUSP 11在肝细胞中的作用机制，以及2）确定DUSP 11在控制炎症中的功能 与促炎病毒和宿主RNA相关。在第一个目标下，击倒方法结合了 通过细胞、分子和生物化学实验，将揭示DUSP 11及其伴侣如何抑制HCV病毒 感染以及病毒如何通过结合细胞microRNA，miR-122来对抗这种限制。下 第二个目标，专门的高通量三磷酸特异性RNA测序技术， 敲除小鼠将确定DUSP 11如何控制与病毒和宿主前体相关的炎症。 在培养的细胞和体内的炎症RNA。这里的贡献预计将是一个详细的 了解DUSP 11如何限制HCV复制和调节促炎性因子的机制 病毒和宿主RNA在感染过程中的活性。这些贡献将是重要的，因为它们是 预期对于理解活性和可能的病毒抗性具有广泛的翻译重要性 目前的II期抗miR-122 HCV药物的机制以及告知先天和 适应性免疫反应-合理疫苗设计的关键。本申请中提出的研究是 创新，因为它代表了一个新的和实质性的脱离现状， DUSP 11，一种促进HCV限制和控制宿主引发的炎症的单一蛋白 和病毒RNA。这项拟议工作的结果将产生积极影响，因为它们将扩大 了解HCV疾病，并可能指导预防和治疗策略的设计。

PROJECT SUMMARY Both the clearance of hepatitis C virus (HCV) and its disease manifestations are associated with the inflammatory response. Yet, large gaps in knowledge exist about the factors that initiate inflammation and control HCV replication. There is an urgent need to fill these gaps because determining the mechanisms that control HCV infection and associated inflammation is imperative for understanding HCV-associated disease. The long-term goal of this collaborative team is to understand the mechanisms of HCV innate control and pathogenesis. Consistent with this goal, the overall objective in this proposal is to determine how the RNA triphosphatase DUSP11 is associated with control of both HCV infection and pro-inflammatory RNA transcripts. The central hypothesis is that DUSP11 promotes turnover of pro-inflammatory host and viral triphosphorylated transcripts, thereby reducing HCV replication and RNA-associated inflammation. The rationale for this proposed research is that, once it is known how DUSP11 functions in control of virus replication and host pro-inflammatory transcripts, this will advance understanding of the innate defenses against HCV, imperative for guiding future rational vaccine designs and for developing therapeutic strategies to address HCV-induced liver pathogenesis. Testing the central hypothesis and completing the objectives outlined in this proposal will be accomplished via the following two specific aims: 1) Determine the anti-HCV mechanism of DUSP11 in liver cells, and 2) Determine the function of DUSP11 in controlling inflammation associated with pro-inflammatory viral and host RNAs. Under the first aim, knockout approaches combined with cell, molecular and biochemical experiments will reveal how DUSP11 and its partners inhibit HCV virus infection and how the virus counters this restriction by binding the cellular microRNA, miR-122. Under the second aim, specialized high-throughput tri-phosphate-specific RNA sequencing technology combined with knockout mice will determine how DUSP11 controls inflammation associated with viral and host pro- inflammatory RNAs in cultured cells and in vivo. The contribution here is expected to be a detailed understanding of the mechanisms of how DUSP11 restricts HCV replication and modulates pro-inflammatory activities of viral and host RNAs during infection. These contributions will be significant because they are expected to have broad translational importance for understanding the activity and possible viral resistance mechanisms of current phase II anti-miR-122 HCV drugs as well as informing the interface of innate and adaptive immune response – key for rational vaccine design. The research proposed in this application is innovative because it represents a new and substantive departure from the status quo by focusing on DUSP11, a single protein that promotes both restriction of HCV and control of inflammation triggered by host and viral RNAs. The results from this proposed work will have a positive impact because they will expand understanding of HCV disease and likely guide the design of preventative and therapeutic strategies.