Genetic determinants of HSV-1 virulence in neurons

HSV-1 神经元毒力的遗传决定因素

基本信息

批准号：
8681309
负责人：
MORIAH SZPARA
金额：
$ 10万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-17 至 2015-05-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): My career goal is to establish an academic research laboratory devoted to understanding how pathogens affect the nervous system, and what features of neurons could provide useful therapeutic targets to combat these infections. I am well-prepared to contribute significantly to our understanding of neurotropic pathogens, based on the strong foundations of my graduate training in neurobiology and postdoctoral training in virology. This award will facilitate my career development by allowing me to gain training in bioinformatics and immunology, and also by providing funding and time to generate data and publications to facilitate a future R01 application. The overall goal of this research proposal is to understand how pathogens interact with the host nervous system, specifically to grasp what genetic features can make a neurotropic pathogen more or less virulent. Herpes simplex virus 1 (HSV-1) is a widespread human pathogen that establishes latency in human peripheral neurons and reactivates repeatedly throughout a patient's lifetime. HSV-1 is a leading cause of infectious blindness in the United States, and the main cause of sporadic, fatal encephalitis. It is unknown why certain HSV-1 infections progress to the level of encephalitis, but it is almost certainly influenced by both viral and host factors. Mouse models of this disease have demonstrated that viral strains differ in their virulence and ability to cause encephalitis. However the ability to map these phenotypic differences to specific viral genes has been limited by the lack of comparative genomic information about this virus - only one wild type HSV-1 genome has been known for the last two decades. I recently demonstrated that high-throughput sequencing (HTS) and bioinformatic assembly can be used to obtain further HSV-1 genomes. I propose to use HTS to test this hypothesis: that genetic loci which are enriched in neurovirulent strains, and lacking in strains that do not cause encephalitis in mice, are highly likely to be causally associated with the neurovirulence phenotype. To achieve this, my specific aims are: (1) to use HTS, genome assembly, & bioinformatic comparison of diverse clinical isolates of HSV-1, to find loci that are enriched in highly neurovirulent strains versus non-virulent ones; and (2) to generate viral recombinants, by either adding or removing putative neurovirulence loci, and test for associated changes in phenotype using an in vivo model of viral encephalitis. These aims will lead to future research directions, using standard molecular and cell biological approaches to explore how these neurovirulence loci affect neuronal function and the viral infectious cycle. This proposal will shed light on mechanisms of neurovirulence in this pervasive human virus, and highlight aspects of virulence to consider in other neurotropic pathogens. NARRATIVE: Herpes simplex virus 1 (HSV-1) is a widespread human virus. This research will investigate how HSV-1 virus isolated from one patient may differ from that of another. This will help us understand why some patients suffer more severe symptoms from HSV-1 infection than others.

描述（由申请人提供）：我的职业目标是建立一个致力于了解病原体如何影响神经系统的学术研究实验室，以及哪些神经元的特征可以提供有用的治疗靶标，以抗击这些感染。基于我在病毒学领域的神经生物学和博士后培训的强大基础上，我为我们对神经病原体的理解做出了重大贡献。该奖项将通过允许我获得生物信息学和免疫学培训，并提供资金和时间来生成数据和出版物以促进未来的R01应用程序，从而促进我的职业发展。该研究建议的总体目标是了解病原体如何与宿主神经系统相互作用，特别是要掌握哪些遗传特征会或多或少地使神经性病原体具有更高的毒性。单纯疱疹病毒1（HSV-1）是一种广泛的人类病原体，它在人类周围神经元中的潜伏期建立了潜伏期，并在患者的一生中反复激活。 HSV-1是美国传染失明的主要原因，也是零星，致命性脑炎的主要原因。尚不清楚为什么某些HSV-1感染发展到脑炎水平，但几乎可以肯定地受到病毒和宿主因素的影响。该疾病的小鼠模型表明，病毒株的毒力和引起脑炎的能力有所不同。但是，由于缺乏有关该病毒的比较基因组信息，将这些表型差异映射到特定病毒基因的能力受到限制 - 在过去的二十年中，只有一种野生型HSV -1基因组闻名。我最近证明，高通量测序（HTS）和生物信息学可以用于获得进一步的HSV-1基因组。我建议使用HTS检验这一假设：富含神经毒动菌株的遗传基因座，并且缺乏不会导致小鼠脑炎的菌株，很可能与神经动力毒素的表型有关。为此，我的具体目的是：（1）使用HTS，基因组组装和HSV-1各种临床分离株的生物信息学比较，以找到富含高度神经毒性菌株的基因座；（2）通过添加或去除假定的神经电动毒素基因座来产生病毒重组剂，并使用病毒性脑炎的体内模型测试表型的相关变化。这些目的将使用标准分子和细胞生物学方法来导致未来的研究方向，以探讨这些神经毒化基因座如何影响神经元功能和病毒感染周期。该提议将阐明这种普遍的人类病毒中神经动力的机制，并强调在其他神经性病原体中要考虑的毒力方面。叙事：单纯疱疹病毒1（HSV-1）是一种广泛的人类病毒。这项研究将研究从一个患者中分离出的HSV-1病毒与另一例患者的病毒有何不同。这将有助于我们理解为什么有些患者从HSV-1感染中遭受比其他患者更严重的症状。