Flavivirus cellular tropisms driving dissemination and transmission in mosquito vectors

项目总结/摘要 这项研究的目的是确定三种蚊媒的细胞向性。 寨卡病毒、登革热病毒和黄热病病毒等具有重要公共卫生意义的黄病毒 埃及病毒载体，并确定特定细胞群在病毒传播和传播中的作用。的 目的是1）鉴定对病毒从中肠传播到唾液至关重要的特定细胞群 腺体，和2）确定特定组织嗜性对蚊子进食行为和病毒传播的影响。 目的1将通过生成中肠、血细胞和肠上皮细胞的microRNA（miRNA）表达谱来实现。 神经元细胞群体和选择在每个群体中高度差异表达的miRNA。我们 将证实表达这些miRNA的细胞的感染，并产生细胞特异性的miRNA限制性病毒 含有细胞特异性miRNA的靶序列，以探测每种细胞类型在病毒传播中的作用。 目的2将通过使用miRNA限制性病毒来确定神经细胞和唾液的作用来实现 腺感染对蚊虫取食行为和随后的病毒传播的影响， 蚊子以老鼠为食，并测试老鼠是否有病毒。这项工作产生的数据和工具 可以帮助开发针对特定细胞中病毒复制的传播控制策略， 载体内的种群。研究和博士后培训活动将在两个科罗拉多 州立大学和位于科罗拉多州柯林斯堡的疾病控制和预防中心。 赞助实验室有免费的研究兴趣和设施，将促进这一成功 项目除了研究，培训计划还包括完成下一代课程 测序实验和分析，赠款写作，并负责进行研究;参与 监督培训;以及参加各种讲习班、研讨会和会议，以提高写作水平， 演讲和职业建设技能。

Project Summary/Abstract The research objectives of this fellowship are to determine the cellular tropisms of three mosquito-borne flaviviruses of public health importance – Zika virus, dengue virus and yellow fever virus – within their Aedes aegypti vectors and to define the roles of specific cell populations in viral dissemination and transmission. The aims are to 1) identify the specific cell populations critical for viral dissemination from the midgut to the salivary glands, and 2) define the impact of specific tissue tropisms on mosquito feeding behavior and viral transmission. Aim 1 will be accomplished by generating microRNA (miRNA) expression profiles for midgut, hemocyte and neuronal cell populations and selecting miRNAs that are highly differentially expressed in each population. We will confirm infection of cells expressing these miRNAs and generate cell-specific, miRNA-restricted viruses containing target sequences for cell-specific miRNAs to probe the roles of each cell type in viral dissemination. Aim 2 will be accomplished by using miRNA-restricted viruses to determine the effect of neuronal cell and salivary gland infection on mosquito feeding behaviors and subsequent transmission of the virus by observing infected mosquitoes feeding on mice and testing mice for the presence of virus. The data and tools produced by this work could aid the development of transmission control strategies that target viral replication in specific cell populations within the vector. Research and postdoctoral training activities will be undertaken at both Colorado State University and the Centers for Disease Control and Prevention in Fort Collins, CO. The sponsor and co- sponsor laboratories have complimentary research interests and facilities that will promote the success of this project. In addition to research, the training plan includes completion of coursework in next generation sequencing experiment and analysis, grant writing, and responsible conduct of research; participation in supervisory training; and attendance at various workshops, seminars and conferences to improve writing, presentation and career building skills.