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

项目总结 丙型肝炎病毒(HCV)的清除及其疾病表现都与 炎症反应。然而，关于引发炎症和炎症的因素，知识存在很大差距。 控制丙型肝炎病毒复制。迫切需要填补这些空白，因为确定 控制丙型肝炎病毒感染和相关炎症是了解丙型肝炎病毒相关疾病的关键。 这个合作小组的长期目标是了解丙型肝炎病毒先天控制和 发病机制。与这一目标一致，该提案的总体目标是确定RNA如何 三磷酸酶DUSP11与控制丙型肝炎病毒感染和促炎RNA有关 成绩单。中心假说是DUSP11促进促炎宿主和病毒的周转 三磷酸化转录本，从而减少丙型肝炎病毒复制和RNA相关炎症。这个 这项拟议研究的基本原理是，一旦了解了DUSP11如何在病毒控制中发挥作用 复制和宿主促炎转录本，这将促进对固有防御的理解 抗丙型肝炎病毒，对于指导未来合理的疫苗设计和开发治疗策略势在必行 解决丙型肝炎病毒诱导的肝脏发病机制。检验中心假设并完成目标 这项建议将通过以下两个具体目标来实现：1)检测抗-丙型肝炎病毒 DUSP11在肝细胞中的作用机制；2)确定DUSP11在控制炎症中的作用 与促炎病毒和宿主RNA相关。在第一个目标下，淘汰赛方法结合在一起 细胞、分子和生化实验将揭示DUSP11及其合作伙伴如何抑制丙型肝炎病毒 感染，以及病毒如何通过结合细胞microRNA，miR-122来对抗这一限制。在.之下 第二个目标，专业的高通量三磷酸特异RNA测序技术与 基因敲除小鼠将决定DUSP11如何控制与病毒和宿主亲本相关的炎症 培养细胞和体内的炎性RNA。这里的贡献预计将是一个详细的 了解DUSP11抑制丙型肝炎病毒复制和调节促炎作用的机制 病毒和宿主RNA在感染过程中的活性。这些贡献将是重大的，因为它们是 预期对了解其活性和可能的病毒耐药性具有广泛的翻译重要性 当前抗miR-122丙型肝炎病毒II期药物的作用机制以及天然和天然的界面信息 适应性免疫反应--合理疫苗设计的关键。这项申请中提出的研究是 创新是因为它代表了对现状的新的实质性偏离，通过专注于 DUSP11，一种同时促进限制丙型肝炎病毒和控制宿主引发的炎症的单一蛋白 和病毒核糖核酸。这项拟议工作的结果将产生积极影响，因为它们将扩大 了解丙型肝炎病毒疾病，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.