CHARACTERIZATION OF ASTROVIRUS VA1, A NOVEL NEUROTROPIC VIRUS

项目摘要/摘要 这份指导职业发展奖(K08)提案描述了研究和教育计划 候选人安德鲁·雅诺夫斯基医学博士。他的长期目标是发现和表征病毒引起的 中枢神经系统(CNS)感染。脑炎通常是由传染性病原体引起的，但 通常情况下，尽管进行了广泛的临床测试，但无法确定病因。脑炎病例的一个子集是 预计由新病毒或具有未被发现的神经嗜性的病毒引起的。雅诺夫斯基博士已经 之前使用下一代测序技术发现了一种新的病毒家族，名为 并将已知的RNA噬菌体数量增加了六倍以上。他会申请 这些技能有助于在中枢神经系统感染中识别新的病原体并确定其生物学特征。 星状病毒VA1(VA1)，一种最初由Janowski博士的导师David Wang博士发现的病毒，已经 已被确定为脑炎的新病因。然而，对这种病毒的基础生物学的研究 由于缺乏感染的实验模型而被排除在外。雅诺夫斯基博士已经研制出第一个细胞 VA1感染的体外培养模型，现在可以用这个来阐明VA1感染的生物学 系统。在目标1中，他将建立VA1中枢神经系统感染的体外模型。他将定义主持人表达简档 在感染后，确定是否有任何VA1株特有的细胞趋向性。宿主免疫反应 对维生素A_1感染的影响也不明确，但初步证据表明干扰素-β是由维生素A_1诱导的 外源性干扰素的加入减少了VA1的复制。对于目标2，Janowski博士将使用CRISPR/CAS- 9确定模式识别受体和干扰素刺激基因的方法 回应。在目标3中，Janowski博士将使用中和试验来定义中和的血清阳性率 抗VA1抗体。他还将使用qRT-PCR方法来确定VA1引起的中枢神经系统疾病的负担， 因为他有来自急性中枢神经系统疾病患者的临床标本储存库。 VA1的特征将使Janowski博士做好准备，在 K08奖励期结束。他的职业发展将由他的共同导师王大卫博士监督。 格雷戈里·斯托奇，罗宾·克莱恩博士，赫伯特·“跳过”维珍，迈克·戴蒙德和大卫·亨斯塔德组成 一个杰出的咨询委员会，将协助职业发展和实验设计。此外, 亚诺夫斯基博士将在华盛顿完成转化医学临床研究的硕士学位 圣路易斯大学，以加强他的正式科学训练。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VA1生物学的独特和具有挑衅性的结果。这项K08提案的结果将 它不仅建立在中枢神经系统感染的宿主与病毒相互作用的基础上，而且还将启动独立的研究事业 雅诺夫斯基博士的。

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.