Role of Optineurin in Ocular Herpes Infection

描述(申请人提供)：单纯疱疹病毒1型(HSV-1)可对人类角膜造成不可逆转的损害，并能够感染眼睛中几乎所有主要细胞类型。该病毒也被认为是导致继发性青光眼的辅助因素，并被认为在人类大脑的某些退行性疾病中扮演着类似的角色。众所周知，HSV-1的裂解感染会下调宿主蛋白质的合成，从而有利于病毒蛋白质的产生。有关感染期间上调的宿主蛋白的信息非常有限。这项建议的重点是了解一种细胞蛋白--视神经磷酸酶(OPTN)--一种自噬适配蛋白--在单纯疱疹病毒1型眼睛感染中的功能(S)。编码OPTN的基因与青光眼和肌萎缩侧索硬化症有关。我们的初步数据显示，OPTN在人和小鼠的角膜中广泛表达。我们还发现OPTN在人类角膜上皮细胞(HCE)感染HSV-1后上调表达，并且OPTN在HCE细胞中的瞬时过表达导致HSV-1形成的斑块数量增加。这一探索性的建议将检验OPTN是HSV-1有效感染人类角膜细胞的关键这一假设。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感染干扰TANK结合激酶-1(TBK1)的OPTN磷酸化的可能性，这反过来导致抑制自噬诱导和干扰素的产生，从而导致病毒复制。第二个目标将研究OPTN通过与肌球蛋白VI相互作用，在新产生的HSV-1糖蛋白的运输以及携带病毒的囊泡与促进病毒从细胞中释放的质膜融合中的作用。总体而言，我们的研究有望带来新的曙光，并首次将OPTN作为HSV-1感染角膜细胞的重要辅助因素。

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.