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.

在过去的一年里，寨卡病毒由于迅速传播感染而引起了全世界的关注，其 与格林-巴利综合征的关系，其通过性传播和在子宫内传播给胎儿的能力， 以及患有小头畸形和其他严重脑缺陷的婴儿数量增加。的基因组 目前的流行性病毒相对于最初分离自 1947年在乌干达的恒河猴（MR 766），它似乎只引起了零星的和轻度的 疾病这些取代中的任何一种是否对病毒复制适应性、发病机制或 人类之间的传播是未知的。因此，利用非洲分离株进行研究是很重要的 以及最近流行的菌株。本提案的目的是阐明 控制流行性病毒严重神经发病机制，并验证我们的假设 自噬的诱导会抑制病毒感染。该提案汇集了以下知识： 两名在相关领域地位稳固、经验丰富的调查员。通过结合专业知识， Spector病毒学实验室，特别是分子/细胞生物学和人类 巨细胞病毒（目前神经出生缺陷的主要病毒原因）和干细胞分化，以及 在Yeo人类干细胞生物学实验室（包括人类诱导多能干细胞的分化）， 细胞[iPSC]到大脑皮层神经元），基因表达和RNA的高通量分析 处理（包括单细胞RNA测序）和尖端的计算技术，我们有一个 这是一个独特的机会，可以更好地了解这一重大的医疗问题。康贝特人将以 当细胞在不同分化阶段感染时，病毒感染和细胞死亡的动力学 iPSC转化为皮层神经元。我们还将利用我们在全基因组技术方面的优势， 宿主基因（包括编码和非编码RNA和microRNA）表达的变化 感染重要的是，我们将比较感染早期非洲寨卡病毒之间的差异， 菌株MR 766和最近流行分离株。也是一种新颖的潜在的治疗方法， 重要的是我们提出的诱导自噬以抑制ZIKV感染并促进细胞增殖的策略。 生存和分化。具体来说，我们将测试用无毒的 二糖海藻糖通过一种新的mTOR非依赖性途径诱导自噬， 感染，正如我们在人类巨细胞病毒中发现的那样。我们还将测试其他2种自噬诱导剂， mTOR非依赖性小分子SMER 28和tat-beclin 1。实现这一目标 该提案将有助于制定预防和治疗寨卡病毒的新策略 感染