Role of Zika virus (ZIKV) infection in glaucoma pathobiology

项目总结 该项目的总体目标是研究寨卡病毒(ZIKV)在青光眼病理生物学中的作用。ZIKV是 一种新出现的病毒病原体，可导致小头畸形并导致新生儿严重的眼部并发症 感染寨卡病毒的母亲所生。虽然ZIKV的眼部表现主要报道会影响 眼后段导致脉络膜视网膜萎缩，视网膜和脉络膜萎缩，以及视神经 神经异常，几个临床病例报告显示前段受累导致 青光眼。我们实验室以及其他实验室的研究表明，ZIKV可以导致 青光眼的病理包括眼压升高，视网膜神经节细胞丢失， 和视神经损伤。感染ZIKV的后代表现出眼压和RGC损失增加， 这些小鼠中抗黄病毒抗体的存在与青光眼显著增加有关。 抗体依赖性增强所致的病理学改变。直到最近的ZIKV疫情，青光眼一直是 主要被认为是一种遗传和年龄相关的疾病，在接触过病毒的婴儿中尚未报告 在怀孕期间受到感染。现在已有多项研究报道，ZIKV可导致先天性青光眼 怀孕期间被感染的母亲所生的婴儿。考虑到有一种地方病 寨卡病毒在84个国家的传播，有必要调查寨卡病毒与青光眼之间的联系，以 开发新的预测和治疗工具来对抗这一全球健康威胁。我们的实验室已经研制出 几种体外和体内模型来研究眼ZIKV感染的病理生物学。在我们最近的研究中，我们 据报道，ZIKV可在人原代小梁细胞(HTMC)中感染和复制。更多 最近，我们进行了RNAseq分析，发现ZIKV感染HTMC导致转录下调 包括调节内质网应激反应的几个通路的改变和失调， 自噬、缺氧和细胞外基质组织。此外，感染ZIKV的小鼠表现出高眼压、ER 压力，以及眼球前段的自噬。ZIKV感染也导致RGC死亡和损失 RGC和视神经损伤导致顺行轴突运输中断。基于这些小说 结果，我们假设ZIKV诱导内质网应激和自噬导致TM死亡和功能障碍， 眼压升高，以及青光眼的发展。为了检验这一假说，本文提出了两个具体目标。目标 1将确定ZIKV诱导的内质网应激在TM功能障碍和青光眼病理生物学中的作用 C57BL/6(WT)和IFNAR1-/-小鼠/仔鼠以及内质网应激的降低是否减轻ZIKV诱导的 青光眼病理学。目标2将研究使用HTMC的自噬的作用，以及小鼠模型和 FDA批准的药物羟基氯喹(HCQ)对ZIKV感染的疗效评价 青光眼。预期的结果将确定ZIKV感染在青光眼和青光眼的发病机制中的作用。 阐明分子机制和途径介导的治疗靶点，为未来的治疗。

PROJECT SUMMARY The overall goal of this project is to investigate the role of Zika virus (ZIKV) in glaucoma pathobiology. ZIKV is an emerging viral pathogen that causes microcephaly and leads to severe ocular complications in newborns born to ZIKV infected mothers. Although the ocular manifestations of ZIKV are primarily reported to affect the posterior segment of the eye resulting in chorioretinal atrophy, withering of the retina and choroid, and optic nerve abnormalities, several clinical case reports showed the involvement of the anterior segment resulting in glaucoma. Studies from our laboratory, as well as those of others, have shown that ZIKV can cause glaucomatous pathology including an increase in intraocular pressure (IOP), retinal ganglion cell (RGC) loss, and optic nerve damage. The offspring of ZIKV infected dams have shown increased IOP and RGC loss and the presence of anti-flavivirus-antibody in these mice correlates with significantly enhanced glaucoma pathology due to antibody-dependent enhancement. Until the recent ZIKV epidemics, glaucoma has been primarily considered as a genetic and age-related disease and has not been reported among infants exposed to infection during gestation. Several studies have now reported that ZIKV can cause congenital glaucoma in infants born from mothers who were infected during pregnancy. Considering the fact that there is an endemic transmission of ZIKV in >84 countries, it is imperative to investigate the link between ZIKV and glaucoma to develop new prognostic and therapeutic tools to combat this global health threat. Our laboratory has developed several in vitro and in vivo models to study the pathobiology of ocular ZIKV infections. In our recent study, we reported that ZIKV can infect and replicate in human primary Trabecular Meshwork cells (HTMC). More recently, we performed RNAseq analysis and discovered that ZIKV infection of HTMC leads to transcriptomic alteration and dysregulation of several pathways including those that modulate ER stress response, autophagy, hypoxia, and ECM organization. Furthermore, ZIKV-infected mice exhibited increased IOP, ER stress, and autophagy in the anterior segment of the eye. ZIKV infection also caused RGC death and loss of RGC and optic nerve damage leading to disruption of anterograde axonal transport. Based on these novel findings, we hypothesize that ZIKV induces ER stress and autophagy resulting in TM death and dysfunction, increased IOP, and the development of glaucoma. Two specific aims are proposed to test this hypothesis. Aim 1 will determine the role of ZIKV induced ER stress in TM dysfunction and the pathobiology of glaucoma using C57BL/6 (WT) and IFNAR1-/- mice/pups and whether the reduction of ER stress alleviates ZIKV induced glaucomatous pathology. Aim 2 will investigate the role of autophagy using HTMC, and mouse models and evaluate the therapeutic efficacy of an FDA approved drug, hydroxychloroquine (HCQ) in ZIKV induced glaucoma. The anticipated results will establish the role of ZIKV infection in the pathogenesis of glaucoma and elucidate the molecular mechanisms and pathway-mediated therapeutic targets for future treatments.