Flavivirus cellular tropisms driving dissemination and transmission in mosquito vectors

项目摘要/摘要 该研究会的研究目标是确定三种蚊媒的细胞取向。 伊蚊体内对公众健康有重要影响的黄病毒--寨卡病毒、登革热病毒和黄热病病毒 埃及伊蚊载体，并确定特定细胞群体在病毒传播和传播中的作用。这个 目的是1)确定病毒从中肠传播到唾液的关键细胞群 2)确定特定的组织取向对蚊子取食行为和病毒传播的影响。 目标1将通过生成中肠、血细胞和 神经细胞群体和选择在每个群体中高度差异表达的miRNAs。我们 将确认表达这些miRNA的细胞受到感染，并产生细胞特异性的miRNA限制性病毒 包含细胞特异性miRNAs的目标序列，以探索每种细胞类型在病毒传播中的作用。 目标2将通过使用miRNA限制性病毒来确定神经细胞和唾液的影响来实现。 腺感染对蚊虫取食行为及后续病毒传播的观察 以老鼠为食并检测老鼠是否携带病毒的蚊子。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的病毒复制的传播控制策略 媒介内的种群。研究和博士后培训活动将在这两个科罗拉多州进行 科罗拉多州柯林斯堡的州立大学和疾病控制与预防中心。赞助商和联席- 赞助实验室具有互补的研究兴趣和设施，将促进这项研究的成功 项目。除了研究，培训计划还包括完成下一代课程的学习 对实验和分析进行排序，撰写拨款，并负责进行研究；参与 监督培训；参加各种讲习班、研讨会和会议，以提高写作水平， 演示和职业发展技能。

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.