Determining the role of host lipids in Zika virus infection

项目摘要/摘要 虽然寨卡病毒(ZIKV)是近70年前在乌干达的寨卡森林中发现的，但直到最近才发现 被公认为全球公共威胁。从历史上看，ZIKV感染与无症状有关 皮疹、关节痛和结膜炎等疾病。然而，最近几项研究发现，ZIKV 感染会导致严重的胎儿畸形，包括婴儿小头畸形和神经功能障碍 成人中的GBS。到目前为止，还没有疫苗或抗病毒疗法，而且可用于治疗的信息有限。 准确预测ZIKV感染的结果以及由此产生的并发症。有一个 了解这种病毒的基本发病机制方面的知识差距，这对制定策略至关重要 来抗击这种疾病。宿主因素，特别是宿主脂质在ZIKV致病中的作用很大程度上 未知。像ZIKV这样的黄病毒在致病方面的巨大成功取决于它的能力 成功地利用宿主的细胞机制，包括脂质生物合成途径来颠覆 免疫系统。我们对这些过程的理解，特别是对宿主脂质生物学的理解 ZIKV是有限的。这项拟议工作的目的是研究宿主的脂质生物合成在条目中的作用， ZIKV的复制、组装和释放。为此，单个敲除的Huh7和SH-SY5Y细胞系 缺乏参与脂质生物合成的关键基因将使用CRISPR/Cas9技术产生。至 作为我们遗传方法的补充，小分子抑制剂将被用来阻断这些关键途径，并且 他们在ZIKV感染中的影响将被调查。除了斑块分析，自动化的高含量 将在ZIKV感染的突变体和对照细胞中进行显微镜检查，以确定宿主脂类在 ZIKV感染的不同阶段。主要的细胞抗菌过程之一，自噬，将是 在ZIKV感染期间在基因敲除细胞和对照细胞中进行评估。使用脂质报告工具，我们还计划 研究ZIKV感染过程中血脂的时空(再)分布。了解这些 对ZIKV生命周期至关重要的路径或过程至关重要，因为它们代表着新的 治疗学。

PROJECT SUMMARY/ABSTRACT Although Zika virus (ZIKV) was discovered almost 70 years ago in Zika forest in Uganda, it is only recently recognized as a global public threat. Historically, ZIKV infection has been associated with asymptomatic illnesses such as rash, arthralgia, and conjunctivitis. However, recently several studies have found that ZIKV infection causes severe fetal abnormalities that include microcephaly in infants and the neurological disorder GBS in adults. To date, there is no vaccine or antiviral therapy, and limited information is available for accurately predicting the outcome as well as the resulting complications due to ZIKV infection. There is a knowledge gap in understanding the basic pathogenesis of this virus, which is critical for developing strategies to combat this disease. The role of host factors especially that of host lipids in ZIKV pathogenesis is largely unknown. The profound success of flaviviruses such as ZIKV in causing disease depends on its ability to successfully utilize the host's cellular machineries including the lipid biosynthesis pathway to subvert the immune system. Our understanding of these processes especially that of the host lipid biology in the context of ZIKV is limited. The aim of the proposed work is to study the role of host's lipid biosynthesis in the entry, replication, assembly and release of ZIKV. For this purpose, individual knockout Huh7 and SH-SY5Y cell lines that lack key genes involved in the biosynthesis of lipids will be generated using CRISPR/Cas9-technology. To complement our genetic approach, small molecule inhibitors will be used to block these key pathways, and their impact in ZIKV infection will be investigated. In addition to plaque assay, automated high-content microscopy will be performed in ZIKV infected mutant and control cells to define the function of host lipids at different stages of ZIKV infection. One of the major cellular antimicrobial processes, autophagy, will be assessed in both knockout and control cells during ZIKV infection. Using lipid reporter tools we also plan to investigate the spatial and temporal (re-)distribution of lipids during ZIKV infection. Understanding these pathways or processes essential for the life cycle of ZIKV is crucial, as they represent potential targets for new therapeutics.