Inhibition of Zika Virus by hybrid interferons

寨卡病毒(ZIKV)是一种重新出现的蚊媒黄病毒。在大多数情况下，寨卡病毒感染是轻微的，并自行消失。然而，ZIKV也与胎儿发育异常有关，包括小头畸形和其他严重的神经系统疾病，如成人的格林-巴利综合征。尽管我们被宣布为国际卫生紧急情况，但我们还没有针对ZIKV感染的特定治疗或疫苗。干扰素(IFN)是我们身体对病毒的第一线防御。根据其受体的使用情况，IFN分为三种类型，类型1、2和3型。它们通过诱导干扰素刺激基因(ISGs)编码的抗病毒蛋白的表达来控制病毒感染。宿主对病毒感染的最初反应是诱导1型IFN。有12种人干扰素α(a1型干扰素)亚型，但临床上仅使用IFNA 2a和2b亚型。FDA已批准它们用于治疗某些感染和癌症，包括慢性丙型肝炎、艾滋病相关的卡波西肉瘤、成人毛细胞白血病和乙肝。然而，研究表明，与其他更有效的IFNA亚型相比，IFNA 2亚型的抗病毒活性较弱。例如，IFNA 14已被证明对包括艾滋病毒和甲型流感病毒在内的病毒具有高度活性。与我们的工业合作伙伴ILC Treateutics合作，我们可以获得新型混合干扰素以及基于IFNA 14主干的内部合成和自然产生的IFNA变体。本研究旨在测试IFNA 14及其衍生物是否具有抗ZIKV感染的活性。为了做到这一点，我们的目标是测试不同的IFNA亚型及其合成衍生物对不同谱系ZIKV的抑制作用，利用斑块分析、报告分析和qPCRs。我们还将利用RNA-Seq分析来鉴定在不同IFNA亚型刺激的细胞中差异表达的不同干扰素刺激基因(ISGs)，并通过qPCR和PULSE SILAC蛋白质组学分析来验证所鉴定的ISGs。本研究还将建立ISG过表达细胞系和敲除细胞系，以测试它们对ZIKV的敏感性和抵抗力；并确定ZIKV蛋白NS5在抵消不同IFNA亚型的抗病毒活性中的作用。这将使我们能够了解混合干扰素如何控制ZIKV感染，并揭示不同病毒基因型在治疗反应性方面的差异。

Zika Virus (ZIKV) is a re-emerging mosquito-borne flavivirus. In most cases ZIKV infection is mild and resolve on their own. However, ZIKV is also linked to developmental abnormalities in foetuses including microcephaly and other serious neurological diseases like Guillain-Barre syndrome in adults. Despite being declared an international health emergency, we have no specific treatment or vaccine available for ZIKV infection.Interferons (IFNs) are our bodies first line defence against viruses. Depending on their receptor usage IFNs are divided into three types, type 1, 2 and 3 IFNs. They control viral infection by inducing the expression of antiviral proteins that are encoded by interferon-stimulated genes (ISGs). The initial host response against virus infection is the induction of type 1 IFNs. There are 12 human IFN alpha (a type 1 IFN) subtypes but only IFNa 2a and 2b subtypes are used clinically. They have been approved by the FDA for the treatment of certain infections and cancers including chronic Hepatitis C, AIDS-related Kaposi sarcoma, hairy cell leukaemia, and hepatitis B in adults. However, studies have shown that IFNa 2 subtypes have weaker antiviral activity as compared to other more potent IFNa subtypes. For instance, IFNa 14 has been shown to be highly active against viruses including HIV and influenza A virus. Working with our industrial partner ILC Therapeutics, we have access to novel hybrid interferons alongside in-house synthetic and naturally occurring IFNa variants that are based on the IFNa 14 backbone. This study aims to test if the IFNa 14 and its derivatives have antiviral activity against ZIKV infection. To do this, we aim to test different IFNa subtypes and their synthetic derivatives for their inhibition of different lineage ZIKV utilising plaque assays, reporter assays and qPCRs. We will also aim to identify the different interferon stimulated genes (ISGs) that are differentially expressed in cells stimulated with different IFNa subtypes using RNA-Seq analysis and to validate the identified ISGs with qPCR and pulse SILAC proteomic analyses. This study will also generate ISG overexpression cell lines and knock-out cells lines to test for their sensitivity and resistance to ZIKV; and to determine the role of the ZIKV protein NS5 in counteracting the antiviral activity of the different IFNa subtypes. This will enable us to understand how hybrid interferons might control ZIKV infection and reveal the differences in therapeutic responsiveness according to viral genotype.