Upregulation of ZAP expression as a novel approach for anti-alphavirus therapy

描述(申请人提供)：甲型病毒(Togaviridae科)可导致相当大的人类和动物发病率和死亡率。目前还没有针对这些病毒引起的疾病的特效药。锌指抗病毒蛋白(ZAP)是一种宿主蛋白，当过度表达时，可以有效地抑制甲型病毒，包括原型Sindbis病毒，以及逆转录病毒科和丝状病毒科的病毒。最近发现，小RNA类似于微小或小干扰RNA，但针对基因启动子序列(反基因，或agRNAs)，介导转录的上调和下调。该项目的目标是上调内源性ZAP的表达，并在体内模型中测试这是否改善了甲病毒感染引起的症状。这些目标将通过两个具体目标来实现。在目标1中，针对ZAP启动子序列的agRNAs将在细胞培养中测试ZAP转录上调活性。活性序列将表征ZAP mRNA和蛋白质上调的有效性和特异性，以及它们对辛德比斯病毒复制的影响。在目标2中，活性agRNAs将在具有良好特性的小鼠模型中测试其上调ZAP表达和减少神经毒力Sindbis病毒感染引起的疾病的能力。如果成功，这项工作将产生很大的影响，为最近发现的转录控制途径增加知识，并开辟治疗甲型病毒感染的全新方法。这项工作将更广泛地适用于治疗逆转录病毒和丝状病毒家族成员的感染。此外，概念方法可以应用于治疗任何传染病，以及任何通过上调特定基因或一组基因的调控而受益的疾病或疾病。 公共卫生相关性：人类感染Togaviridae家族的甲型病毒属病毒可引起发烧、皮疹、关节炎、脑炎和死亡，目前尚无抗击这些疾病的具体治疗方法。该提案中描述的研究将测试一种创新治疗方法的可行性。强大的抗甲病毒宿主因子ZAP的表达将不受调控，其对甲病毒感染的影响将在小鼠模型中进行测试。

DESCRIPTION (provided by applicant): Alphaviruses (Togaviridae family) cause considerable human and animal morbidity and mortality. There are currently no specific treatments for diseases caused by these viruses. The zinc-finger antiviral protein (ZAP) is a host protein that when over expressed mediates potent inhibition of alphaviruses, including the prototype Sindbis virus, as well as viruses in the Retroviridae and Filoviridae families. Recently, small RNAs similar to micro or small interfering RNAs, but directed to gene promoter sequences (anti-gene, or agRNAs), have been found to mediate transcriptional up- and down regulation. The objectives of this project are to up regulate expression of endogenous ZAP and test whether this ameliorates symptoms due to alphavirus infection in an in vivo model. The objectives will be accomplished in two specific aims. In Aim 1, agRNAs directed to ZAP promoter sequences will be tested in cell culture for ZAP transcriptional up regulation activity. Active sequences will be characterized for efficacy and specificity of the ZAP mRNA and protein up regulation and their effect on Sindbis virus replication. In Aim 2, the active agRNAs will tested in a well characterize murine model for their ability to up regulate ZAP expression and reduce disease caused by neurovirulent Sindbis virus infection. If successful, the work will be of high impact, adding knowledge to this recently discovered transcriptional control pathway, and opening up a whole new approach to the treatment of alphavirus infection. The work will be more broadly applicable to the treatment of infection by members of the Retrovirus and Filovirus families. Moreover, the conceptual approach could be applied to treat any infectious disease, as well as any disease or condition that would benefit by up regulation of a specific gene or set of genes. PUBLIC HEALTH RELEVANCE: Infection of humans by viruses in the Alphavirus genus of the Togaviridae family can cause fever, rash, arthritis, encephalitis and death and no specific treatments are available to combat these diseases. The studies described in this proposal will test the feasibility of an innovative therapeutic approach. Expression of the potent anti-alphavirus host factor called ZAP will be unregulated and its effect on alphavirus infection will b tested in a murine model.