CHARACTERIZATION OF ASTROVIRUS VA1, A NOVEL NEUROTROPIC VIRUS

新型神经营养病毒 ASTROVIRUS VA1 的特征

基本信息

批准号：
9527420
负责人：
Andrew Bok Seng Janowski
金额：
$ 16.18万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-05 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9527420
关键词：
Accounting Acute Address Adult Advisory Committees Affect Animals Antibodies Antiviral Agents Antiviral Therapy Astrocytes Astrovirus Award Biological Assay Biology CRISPR/Cas technology Caco-2 Cells Cattle Cell Culture System Cell Culture Techniques Cell Line Cell Lineage Cells Cellular Tropism Central Nervous System Diseases Central Nervous System Infections Cerebrospinal Fluid Chickens Child Clinical Data Development Development Plans Diagnostic Diamond Disease Doctor of Philosophy Encephalitis Enrollment Epidemiology Etiology Experimental Designs Experimental Models Exposure to Expression Profiling Family Family suidae Foundations Future Genes Goals Growth Human Immune Immune response Immunohistochemistry In Situ Hybridization In Vitro Infection Institution Integration Host Factors Interferon Type I Interferon-beta Interferons K-Series Research Career Programs Kinetics Master&apos s Degree Measures Mediating Mentors Mink Modeling Names Neuraxis Neuroglia Neurologic Deficit Neurons Neurotropism Outcome Patients Pattern Pattern recognition receptor Physicians Pleocytosis Prevalence Proteins Publishing Quantitative Reverse Transcriptase PCR RNA RNA Phages Receptor Signaling Reporting Research Research Project Grants Retrospective cohort Role Sampling Scientist Seroprevalences Specimen Supporting Cell Surveys System Technology Training Universities Vertebrates Viral Virus Washington Work base brain tissue burden of illness career career development cell immortalization clinical investigation cohort design exposed human population gastrointestinal infection improved improved outcome in vitro Model in vivo indexing insight loss of function metagenomic sequencing nervous system disorder neurotropic neurotropic virus neutralizing antibody next generation sequencing novel novel virus pathogen prospective receptor repository research clinical testing response skills targeted treatment translational medicine virus host interaction

项目摘要

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.

项目概要/摘要该指导性职业发展奖（K08）提案描述了研究和教育计划对于候选人，医学博士安德鲁·贾诺夫斯基（Andrew Janowski）来说，他的长期目标是发现并描述病毒的病因。中枢神经系统（CNS）感染是脑炎的一种，通常是由传染性病原体引起的。尽管进行了广泛的临床测试，但通常仍无法确定病因。预计是由新型病毒或具有未被认识到的向神经性的病毒引起的。之前使用下一代测序技术发现了一个新的病毒家族，名为 Statoviruses 并将已知的 RNA 噬菌体数量增加了六倍以上。这些技能可识别中枢神经系统感染中的新病原体并表征其生物学特征。星状病毒 VA1 (VA1) 是一种最初由 Janowski 博士的导师 David Wang 博士发现的病毒，然而，该病毒的基础生物学研究已被确定为脑炎的新病因。由于缺乏感染实验模型，Janowski 博士开发出了第一个细胞。 VA1感染的体外培养模型，现在可以利用该模型阐明VA1感染的生物学在目标 1 中，他将开发 VA1 CNS 感染的体外模型，他将定义宿主表达谱。感染后并确定是否存在任何 VA1 菌株特异性细胞向性宿主免疫反应。 VA1 感染的定义也不明确，但初步证据表明干扰素-β 是由 VA1 诱导的对于目标 2，添加外源干扰素 VA1 会减少复制。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在目标 3 中，Janowski 博士将使用中和试验来确定中和反应的血清阳性率。他还将使用 qRT-PCR 来确定 VA1 引起的中枢神经系统疾病负担，因为他拥有急性中枢神经系统疾病患者的临床标本库。描述 VA1 的特征将使 Janowski 博士做好准备，以成为一名独立的医师科学家 K08 奖励期结束后，他的职业发展将由他的共同导师 David Wang 博士负责监督。格雷戈里·斯托奇 (Gregory Storch)，罗宾·克莱因 (Robyn Klein) 博士、赫伯特·“斯基普”·维尔京 (Herbert “Skip” Virgin)、迈克·戴蒙德 (Mike Diamond) 和大卫·亨斯塔德 (David Hunstad) 组成一个杰出的顾问委员会将协助职业发展和实验设计。 Janowski 博士将在华盛顿完成转化医学临床研究硕士学位圣路易斯大学加强了他的正规科学训练，他得到了世界一流机构的支持，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有关 VA1 生物学的独特且具有挑战性的结果。该 K08 提案的结果将不仅建立在中枢神经系统感染中宿主与病毒相互作用的基础上，还将开启独立研究生涯贾诺夫斯基博士。