Dissecting Dengue Virus Permissiveness using a Stem Cell Differentiation System

 描述（由申请人提供）：我们的研究项目的广泛、长期目标是通过表征病毒感染所必需的细胞辅因子，特别是正链RNA病毒，如登革热病毒（DENV）和丙型肝炎病毒（HCV），来增进对病毒-宿主细胞相互作用的了解。目前的提案侧重于DENV，它感染了全世界数百万人，对人类健康构成了重大的新威胁。了解DENV如何与宿主细胞相互作用对于开发抗病毒药物和预防性疫苗至关重要，这两者目前都缺乏。基于我们先前在体外肝分化和HCV感染方面的工作， HCV易感性的宿主决定因素，我们建议调查有助于单核细胞和肝细胞中DENV容许性的细胞参数。我们已经在初步实验中发现了在多能干细胞的定向分化期间向DENV容许性的离散细胞转变。我们将在这一过渡期内仔细检查基因谱，以确定DENV感染所需的推定宿主因子，这些因子反过来可能被用作抗病毒治疗的新药物靶点。此外，我们将分析下调的抗病毒蛋白对向DENV易感性转变的贡献，并研究有效抑制DENV感染的任何抗病毒蛋白的作用机制。拟议研究的结果不仅将提供重要的见解， DENV如何劫持细胞蛋白质和机制以促进其在人类细胞中的复制;但也可能揭示针对重要疾病的抗病毒干预的新治疗靶点。 人类病原体

 DESCRIPTION (provided by applicant): The broad, long-term goal of our research program is to advance knowledge of virus-host cell interactions by characterizing cellular cofactors essential for viral infections, particularly positive- strand RNA viruses such as dengue virus (DENV) and hepatitis C virus (HCV). The current proposal focuses on DENV, which infects millions of people worldwide and poses major emerging threat to human health. Understanding how DENV interacts with the host cell is of critical importance to the development of antiviral drugs and a prophylactic vaccine, both of which are currently lacking. Building upon our previous work on in vitro hepatic differentiation and HCV infection which led to the identification of host determinants of HCV susceptibility, we propose to investigate the cellular parameters that contribute to DENV permissiveness in both monocytes and hepatic cells. We have uncovered a discrete cellular transition to DENV permissiveness during directed differentiation of pluripotent stem cells in preliminary experiments. We will scrutinize the gene profiles during this transition period to identify putative host factors required for DENV infection which in turn may be exploited as new drug targets for antiviral therapy. In addition, we will analyze the contribution of downregulated antiviral proteins to the transition to DENV susceptibility and investigate the mechanism of action for any antiviral proteins that are effective at suppressing DENV infection. The results of the proposed studies will not only provide significant insights into how DENV hijacks cellular proteins and machineries to facilitate its own replication in human cells; but also may reveal new therapeutic targets for antiviral intervention directed at important human pathogens.