CHARACTERIZATION OF ASTROVIRUS VA1, A NOVEL NEUROTROPIC VIRUS

项目总结/摘要 这份指导性职业发展奖（K 08）提案描述了研究和教育计划 安德鲁·雅诺夫斯基医学博士他的长期目标是发现和描述 中枢神经系统（CNS）感染。一个实体，脑炎，往往是由传染性病原体引起的，但 尽管进行了广泛的临床试验，但常常不能确定病因。脑炎病例的一个子集是 预期由新病毒或具有未被认识到的嗜神经性的病毒引起。杰诺夫斯基医生 之前使用下一代测序技术发现了一个名为 目前已知的RNA噬菌体的数量已经增加了六倍以上。他会申请 这些技能，以确定新的病原体在中枢神经系统感染和表征其生物学。 星状病毒VA 1（VA 1），最初由Janowski博士的导师大卫王博士发现的一种病毒， 被确认为脑炎的一种新病因然而，对这种病毒的基本生物学研究 由于缺乏感染的实验模型而被排除在外。雅诺夫斯基博士研制出了第一个细胞 VA 1感染的体外培养模型，现在可以使用该模型阐明VA 1感染的生物学 系统在目标1中，他将开发VA 1 CNS感染的体外模型。他将定义宿主表达谱 并确定是否存在任何VA 1菌株特异性细胞向性。宿主免疫应答 对VA 1感染的定义也不明确，但初步证据表明，干扰素-β是由VA 1诱导的， 外源性干扰素的加入减少了VA 1的复制。对于目标2，Janowski博士将使用CRISPR/Cas- 9定义模式识别受体和介导干扰素的干扰素刺激基因的方法 反应在目标3中，Janowski博士将使用中和试验来确定中和抗体的血清阳性率。 VA 1抗体。他还将使用qRT-PCR测定来确定VA 1引起的CNS疾病的负担， 因为他有急性中枢神经系统疾病患者的临床标本库。 表征VA 1将为Janowski博士过渡为独立的物理学家-科学家做好准备， K 08奖励期结束。他的职业发展将由他的共同导师大卫王博士监督 和格雷戈里斯托奇，博士罗宾克莱因，赫伯特“跳过”维珍，迈克钻石，和大卫亨斯塔德形成 一个杰出的咨询委员会，将协助职业发展和实验设计。此外，本发明还提供了一种方法， 博士Janowski将在华盛顿完成转化医学临床研究硕士学位 圣刘易斯的一所大学，以加强他的正规科学训练。他得到了世界级机构的支持 优秀的导师和顾问团队，完善的教育计划和研究项目， 将提供关于VA 1生物学的独特和挑衅性的结果。K 08提案的结果将 它不仅建立在中枢神经系统感染中宿主-病毒相互作用的基础上， Janowski博士的

Project Summary/Abstract This mentored career development award (K08) proposal describes the research and educational plans for the candidate, Andrew Janowski MD. His long-term goal is to discover and characterize viral causes of central nervous system (CNS) infection. One entity, encephalitis, is often caused by infectious pathogens, but frequently an etiology cannot be identified despite extensive clinical testing. A subset of encephalitis cases are anticipated to be caused by novel viruses or viruses with an unappreciated neurotropism. Dr. Janowski has previously used next-generation sequencing technology to discover a novel family of viruses named statoviruses and has also increased the number of known RNA bacteriophages by over six-fold. He will apply these skills to identify novel pathogens in CNS infections and to characterize their biology. Astrovirus VA1 (VA1), a virus originally discovered by Dr. Janowski's mentor, David Wang PhD, has been identified as an emerging cause of encephalitis. However, studies of the fundamental biology of this virus has been precluded by the lack of an experimental model of infection. Dr. Janowski has developed the first cell culture model of VA1 infection in vitro, and now the biology of VA1 infection can be elucidated using this system. In Aim 1, he will develop an in vitro model of VA1 CNS infection. He will define host expression profiles upon infection and determine if there are any VA1 strain specific cellular tropisms. The host immune response to VA1 infection is also poorly defined, but preliminary evidence suggests that interferon-β is induced by VA1 and addition of exogenous interferon reduces VA1 replication. For Aim 2, Dr. Janowski will use a CRISPR/Cas- 9 approach to define pattern recognition receptors and interferon stimulated genes that mediate the interferon response. In Aim 3, Dr. Janowski will use a neutralization assay to define the seroprevalence of neutralizing antibodies to VA1. He will also use a qRT-PCR assay to define the burden of CNS diseases caused by VA1, as he has a repository of clinical specimens from patients with acute CNS diseases. Characterizing VA1 will prepare Dr. Janowski to transition as an independent physician-scientist by the end of the K08 award period. His career development will be overseen by his co-mentors, Drs. David Wang and Gregory Storch, with Drs. Robyn Klein, Herbert “Skip” Virgin, Mike Diamond, and David Hunstad forming an illustrious advisory committee that will assist in career development and experimental design. In addition, Dr. Janowski will complete a master's degree in clinical investigation in translational medicine at Washington University in St Louis to enhance his formal scientific training. He has the support of a world-class institution, an outstanding group of mentors and advisors, a well-developed educational plan, and a research project that will provide unique and provocative results regarding the biology of VA1. The results of this K08 proposal will not only build upon host-virus interactions in CNS infections but it will launch the independent research career of Dr. Janowski.