Role of Optineurin in Ocular Herpes Infection

描述（由申请人提供）：1型单纯疱疹病毒（HSV-1）可对人角膜造成不可逆的损伤，并能够感染眼睛中几乎所有主要细胞类型。该病毒也被认为是继发性青光眼的辅助因子，并被认为在人类大脑的某些退行性疾病中发挥类似的作用。已知HSV-1的裂解性感染下调宿主蛋白质合成以有利于病毒蛋白质产生。关于在感染期间上调的宿主蛋白质的信息非常有限。该提案的重点是了解细胞蛋白optineurin（OPTN）（一种自噬衔接蛋白）在眼睛HSV-1感染中的功能。编码OPTN的基因与青光眼和肌萎缩侧索硬化症有关。我们的初步数据表明，OPTN在人类和小鼠角膜中广泛表达。我们还发现OPTN响应于人角膜上皮（HCE）细胞的HSV-1感染而上调，并且OPTN在HCE细胞中的瞬时过表达导致由HSV-1形成的斑块数量增加。这个探索性的提议将检验OPTN是HSV-1对人角膜细胞进行有效感染的关键的假设。我们建议探索OPTN在HSV-1感染中的两个独立作用。第一个目标将集中在HSV-1感染通过TANK结合激酶-1（TBK 1）干扰OPTN磷酸化的可能性，这反过来导致自噬诱导和干扰素产生的抑制，以及病毒复制抑制。第二个目标将检查OPTN的作用，通过与肌球蛋白VI的相互作用，在运输新制造的HSV-1糖蛋白和融合的病毒载囊泡与质膜，促进病毒从细胞释放。总的来说，我们的研究有望带来新的发现，并首次表明OPTN是HSV-1感染角膜细胞的重要辅助因子。

DESCRIPTION (provided by applicant): Herpes simplex virus type-1 (HSV-1) can cause irreversible damage to the human cornea and is capable of infecting virtually all major cell types in the eye. The virus is also considered a cofactor in causation of secondary glaucoma and is thought to play similar roles in certain degenerative diseases of the human brain. A lytic infection by HSV-1 is known to down regulate host protein synthesis to favor viral protein production. Very limited information is available on the host proteins that are up-regulated during the infection. This proposal is focused on understanding the function(s) of a cellular protein, optineurin (OPTN), an autophagy adapter protein, in HSV-1 infection of the eye. The gene encoding OPTN has been implicated in glaucoma and amyotrophic lateral sclerosis. Our preliminary data shows that OPTN is widely expressed in human and murine corneas. We have also found that OPTN is up-regulated in response to HSV-1 infection of human corneal epithelial (HCE) cells and transient over-expression of OPTN in HCE cells results in an increased number of plaques formed by HSV-1. This exploratory proposal will test the hypothesis that OPTN is a key to a productive infection of the human corneal cells by HSV-1. We propose to explore two independent roles for OPTN in HSV-1 infection. The first Aim will focus on the possibility that HSV-1 infection interferes with OPTN phosphorylation by TANK binding kinase-1 (TBK1), which in turn results in the inhibition of autophagy induction and interferon production, and viral replication ensues. The second Aim will examine a role for OPTN, via interaction with myosin VI, in the transport of newly made HSV-1 glycoproteins and fusion of virus-laden vesicles with the plasma membrane that facilitate the virus release from cells. Overall, our studies are expected to shed new lights, and for the first time, implicate OPTN as an important co-factor in HSV-1 infection of the cells in the cornea.