Zika Virus Pathogenesis and Selective Autophagy Induction to Inhibit Virus Production

寨卡病毒发病机制和选择性自噬诱导抑制病毒产生

• 批准号: 9277152
• 负责人: DEBORAH Hye SPECTOR
• 金额: $ 23.25万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-04 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9277152
• 关键词: Address; Africa; African; Amino Acid Substitution; Antibodies; Asians; Attention; Autophagocytosis; Autophagosome; Brain; Brazil; Cell Count; Cell Death; Cell Differentiation process; Cell Survival; Cells; Cellular biology; Cerebral cortex; Cerebrum; Code; Color; Congenital Abnormality; Cytomegalovirus; Defect; Development; Disaccharides; Disease; Dissociation; Endothelial Cells; Epidemic; FGF2 gene; FRAP1 gene; Fetus; Fibroblasts; Flow Cytometry; Foundations; Future; Gene Expression Profiling; Genes; Genome; Goals; Growth; Guillain-Barré Syndrome; Health; Human; Indirect Immunofluorescence; Infant; Infection; Kinetics; Knowledge; Macaca mulatta; Medical; MicroRNAs; Microcephaly; Molecular; Neurons; Neurotropism; Pathogenesis; Pathway interactions; Population; Production; Protein Biosynthesis; Puerto Rican; RNA; RNA Processing; RNA chemical synthesis; Research; Research Personnel; South America; Southeastern Asia; Stem cells; Technology; Testing; Therapeutic; Time; Trehalose; Uganda; Undifferentiated; Untranslated RNA; Viral; Virus; Virus Diseases; Virus Replication; Withdrawal; Zika Virus; base; cell fixing; cell type; cytotoxicity; design; experience; experimental study; fitness; genome-wide; high throughput analysis; human stem cells; in utero; induced pluripotent stem cell; insight; interest; nestin protein; neurogenesis; novel; novel strategies; prevent; progenitor; relating to nervous system; small molecule; stem cell differentiation; transcriptome sequencing; transmission process; viral RNA; viral transmission; virology; virus pathogenesis

In the past year, Zika virus has gained worldwide attention due to the rapidly spreading infection, its association with Guillain-Barré syndrome, its ability to be transmitted sexually and in utero to the fetus, and the increased number of infants with microcephaly and other severe brain defects. The genome of the current epidemic virus has 75 amino acid substitutions relative to the virus originally isolated from a rhesus monkey in Uganda in 1947 (MR766), which appears to have caused only sporadic and mild disease. Whether any of these substitutions has effects on viral replication fitness, pathogenesis, or spread between humans is unknown. Thus it is important that studies be done using the African isolate as well as more recent epidemic strains. The goal of this proposal to elucidate the underlying mechanisms governing the severe neural pathogenesis of the epidemic virus and to test our hypothesis that induction of autophagy will inhibit the virus infection. This proposal brings together the knowledge of two well-established and experienced investigators in related fields. By combining the expertise in the Spector lab in virology, specifically the molecular/cell biology and pathogenesis of human cytomegalovirus (currently the major viral cause of neural birth defects), and stem cell differentiation, and in the Yeo lab on human stem cell biology (including differentiation of human induced pluripotent stem cells [iPSCs] to cerebral cortex neurons), high-throughput analysis of gene expression and RNA processing (including single cell RNA seq), and cutting edge computational technologies, we have a unique opportunity to gain greater understanding of this major medical problem. We will determine the kinetics of viral infection and cell death when cells are infected at different stages of differentiation of iPSCs to cortical neurons. We will also leverage our strengths in genome-wide technologies to identify changes in expression of host genes (including coding and noncoding RNAs and microRNAs) during infection. Importantly, we will compare the differences between infection with the early African Zika virus strain MR766 and a more recent epidemic isolate. Also novel and potentially of great therapeutic significance is our proposed strategy of inducing autophagy to inhibit ZIKV infection and promote cell survival and differentiation. Specifically, we will test whether treatment of cells with the nontoxic disaccharide trehalose, which induces autophagy via a novel mTOR-independent pathway, will inhibit the infection, as we have found for human cytomegalovirus. We will also test 2 other autophagy inducers, a mTOR-independent small molecule SMER28 and tat-beclin 1. Accomplishment of the goals of this proposal will facilitate the development of new strategies designed to prevent and treat Zika virus infection.

在过去的一年中，寨卡病毒由于迅速传播的感染而引起了全球关注 与Guillain-Barré综合征的关联，其在性行为和子宫内传播到胎儿的能力， 以及小头畸形和其他严重大脑缺陷的婴儿数量增加。基因组 当前的流行病病毒具有75种氨基酸取代，相对于最初从A的病毒分离的病毒 1947年在乌干达的豪斯猴（MR766），这似乎仅造成零星和中间 疾病。这些替代中的任何一个是否对病毒复制适应性，发病机理或 在人之间传播是未知的。重要的是要使用非洲分离株进行研究 以及最近的流行病。该提议的目的是阐明基础 控制流行病病毒严重神经发病机理的机制并检验我们的假设 自噬的诱导将抑制病毒感染。该提议汇集了 相关领域的两个成熟且经验丰富的调查员。通过结合专业知识 Spector Lab在病毒学中，特别是人类的分子/细胞生物学和发病机理 巨细胞病毒（目前是神经birth缺陷的主要病毒），干细胞分化和 在人类干细胞生物学的YEO实验室中（包括人类诱导多能茎的分化 细胞[IPSC]至脑皮质神经元），基因表达和RNA的高通量分析 处理（包括单细胞RNA SEQ）和尖端的计算技术，我们有一个 独特的机会，可以更了解这一主要医学问题。我们将确定 当细胞在不同的分化阶段感染细胞时，病毒感染和细胞死亡的动力学 iPSC到皮质神经元。我们还将利用全基因组技术的优势来识别 宿主基因表达（包括编码和非编码RNA和microRNA）的表达变化 感染。重要的是，我们将比较感染之间的差异与早期的非洲寨卡病毒 菌株MR766和最近的流行病分离物。也是新颖的，可能是出色的治疗 意义是我们提出的诱导自噬抑制ZIKV感染并促进细胞的策略 生存和分化。具体而言，我们将测试是否使用无毒细胞处理细胞 二糖海藻糖通过新型MTOR独立途径诱导自噬，将抑制 正如我们发现的人类巨细胞病毒所发现的那样。我们还将测试其他2个自噬影响者，一个 独立于m的小分子smer28和tat-beclin 1。实现目标的实现 提案将促进旨在预防和治疗寨卡病毒的新策略 感染。