Determining the role of host lipids in Zika virus infection

确定宿主脂质在寨卡病毒感染中的作用

• 批准号: 9387983
• 负责人: Fikadu G. Tafesse
• 金额: $ 23.1万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9387983
• 关键词: Adult; Anabolism; Antiviral Agents; Antiviral Therapy; Arthralgia; Autophagocytosis; Biological; Biology; Biosensor; CRISPR/Cas technology; Capsid Proteins; Cell Line; Cells; Complement; Complex; Conjunctivitis; Cytosol; Data; Dendritic Cells; Dengue Virus; Disease; Electron Microscopy; Endoplasmic Reticulum; Enzymes; Exanthema; Fetus; Flavivirus; Foundations; Future; Genes; Genome; Goals; Guillain-Barré Syndrome; Hepatocyte; Host Defense Mechanism; Human; Immune system; Individual; Infant; Infection; Integration Host Factors; Knock-out; Knowledge; Lateral; Life Cycle Stages; Link; Lipid Bilayers; Lipids; Mammals; Membrane; Microcephaly; Microscopy; Myelin Sheath; Nail plate; Nature; Nervous system structure; Neuraxis; Neurons; Neuropathy; Nucleocapsid; Organelles; Pathology; Pathway interactions; Peripheral; Plaque Assay; Play; Process; RNA; Reporter; Role; Route; Site; Skin; Sphingolipids; Stem cells; Technology; Testing; Tropism; Uganda; Vaccine Therapy; Viral Envelope Proteins; Virion; Virus; Virus Assembly; Virus Diseases; Virus Replication; Work; Zika Virus; antimicrobial; brain tissue; combat; design; fetal; forest; genetic approach; genome editing; global health; inhibitor/antagonist; keratinocyte; lipid biosynthesis; lipid metabolism; mutant; nerve stem cell; nervous system disorder; neuroblastoma cell; new therapeutic target; outcome prediction; physical property; small molecule inhibitor; success; tool; transmission process; viral RNA; virus envelope; virus pathogenesis

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.

项目总结/摘要 虽然寨卡病毒（ZIKV）差不多70年前就在乌干达的寨卡森林中被发现，但直到最近 被认为是全球公共威胁。从历史上看，ZIKV感染与无症状性 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，包括婴儿的小头畸形和神经系统疾病 成人GBS到目前为止，还没有疫苗或抗病毒治疗， 准确预测结果以及由于ZIKV感染而导致的并发症。有一个 在了解这种病毒的基本发病机制方面存在知识差距，这对制定战略至关重要 来对抗这种疾病宿主因子特别是宿主脂质在ZIKV发病机制中的作用在很大程度上是由宿主因子引起的。 未知黄病毒如ZIKV在引起疾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， 免疫系统我们对这些过程的理解，特别是在环境中宿主脂质生物学的理解， ZIKV有限。本研究的目的是研究宿主的脂质生物合成在入侵过程中的作用， ZIKV的复制、组装和释放。为此目的，单独敲除Huh 7和SH-SY 5 Y细胞系 将使用CRISPR/Cas9技术产生缺乏参与脂质生物合成的关键基因的细胞。到 作为我们遗传方法的补充，小分子抑制剂将用于阻断这些关键途径， 将研究它们在ZIKV感染中的影响。除了空斑分析，自动化高含量 将在ZIKV感染的突变体和对照细胞中进行显微镜检查，以确定宿主脂质的功能， ZIKV感染的不同阶段。其中一个主要的细胞抗菌过程，自噬，将 在ZIKV感染期间在敲除细胞和对照细胞中评估。使用脂质报告工具，我们还计划 研究ZIKV感染期间脂质的空间和时间（再）分布。了解这些 对于ZIKV的生命周期至关重要的途径或过程是至关重要的，因为它们代表了ZIKV新的潜在靶点。 治疗学