HPSE in Ocular Herpes Infection

HPSE 在眼部疱疹感染中的应用

• 批准号: 10242774
• 负责人: DEEPAK SHUKLA
• 金额: $ 38.78万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242774
• 关键词: Adverse effects; Animal Model; Antiinflammatory Effect; Antiviral Agents; Area; Basement membrane; Binding; Biological Assay; Blindness; CRISPR/Cas technology; Cell Nucleus; Cells; Chromosome Deletion; Clinical; Complex; Cornea; Corneal Diseases; Corneal Ulcer; Data; Development; Disease; Endothelium; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Epithelial; Epithelial Cells; Extravasation; Eye Infections; Flow Cytometry; Genetic Transcription; Growth; Growth Factor; Heparanase inhibitors; Heparitin Sulfate; Herpesviridae Infections; Herpesvirus 1; Herpetic Keratitis; Human; Immune; Immunofluorescence Immunologic; Immunohistochemistry; Infection; Inflammation; Investigation; Keratitis; Knock-out; Knockout Mice; Laboratories; Leukocytes; Lymphoid; Molecular; Mus; Myelogenous; NF-kappa B; Nuclear; Pain; Pathogenicity; Pathology; Pharmacology; Plaque Assay; Population; Production; Protein Biosynthesis; Proteins; Proteomics; Publishing; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; Severities; Signal Transduction; Simplexvirus; Structure of trigeminal ganglion; Symptoms; Systems Biology; Therapeutic; Tissues; Ulcer; Up-Regulation; Vascular Permeabilities; Viral; Viral load measurement; Virulence Factors; Virus; Virus Diseases; Vision; Western Blotting; angiogenesis; base; corneal epithelium; cytokine; draining lymph node; experimental study; heparanase; immunological status; mouse model; neovascularization; novel drug class; promoter; response; sugar; sulfurtransferase; transcription factor; transmission process

The major clinical complications associated with herpes simplex virus -1 (HSV-1) infection of the eye include corneal ulcers, severe pain, inflammation, loss of corneal avascularity, and vision loss. Antiviral agents alone often fail to correct the problems because many of the disease symptoms may be signaled by infection but then regulated more directly by host molecules. This proposal studies, human heparanase (HPSE), a host enzyme that may potentially regulate both, virus growth and disease manifestations in the cornea. HPSE is a heparan sulfate (HS) endoglycosidase whose levels correlate directly with the breakdown of epithelial and endothelial basement membrane barriers, increased vascular permeability and leukocyte extravasation, and liberation of HS-bound cytokines and growth factors promoting angiogenesis and inflammation in the surrounding areas. The PI’s laboratory has recently observed a significant increase in HPSE expression and higher enzymatic activity upon HSV infection of human corneal cells, cultured human corneas, and animal models of ocular infection and demonstrated its significance in viral spread and transmission. In murine corneas HPSE upregulation directly contributes to corneal disease pathologies including ulceration, inflammation and neovascularization. In addition, we unexpectedly found that HPSE knockout cells show a severe deficiency in virus production. Therefore, based on our newly generated preliminary results we hypothesize an important regulatory role for HPSE in HSV-1 transcription and propose to establish HPSE as a key host virulence factor. We propose that activated HPSE directly adds to the severity of herpetic diseases including tissue damage and promotes viral infection and reactivation. This proposal will focus on understanding the HPSE driven mechanisms that contribute to HSV-1 growth in the cornea and demonstrating the antiviral/anti-inflammatory effects of HPSE inhibition or chromosomal deletion. Three Aims are proposed. First Aim will establish the molecular basis behind the loss of HSV-1 infection in cells lacking HPSE. It is based on our hypothesis that HPSE through its nuclear localization and complex set of interactions promote HSV-1 transcription. Second Aim will determine the significance of HPSE in HSV-1 infection of the murine cornea. This Aim will use a HPSE knockout mouse model to prove our hypothesis that HPSE is a host virulence factor that promotes HSV-1 infection in the cornea, drives tissue damage and disease pathologies, and facilitates viral spread to and return from the trigeminal ganglia (TG) during reactivation. The third and final Aim will determine the therapeutic benefits of an HPSE inhibitor against HSV-1 infection of the cornea. This Aim will prove our hypothesis that pharmacological inhibition of HPSE will result in unprecedented therapeutic benefits including quick resolution of infection and corneal inflammation. Successful conclusion of our study will help establish a brand new role for HPSE as a host virulence factor and identify new factors that help promote its pathogenic activities in the cornea. Our results will significantly enhance our understanding of HPSE functions and herpetic disease mechanisms, and accelerate new strategies to control ocular infection.

与眼部单纯疱疹病毒-1（HSV-1）感染相关的主要临床并发症包括 角膜溃疡、严重疼痛、炎症、角膜无血管性丧失和视力丧失。单独使用抗病毒药物 通常不能纠正这些问题，因为许多疾病症状可能是由感染引起的， 然后由宿主分子更直接地调节。本研究以人乙酰肝素酶（HPSE）为研究对象， 这种酶可能潜在地调节角膜中的病毒生长和疾病表现。HPSE是一个 硫酸乙酰肝素（HS）糖苷内切酶，其水平与上皮细胞的分解直接相关， 内皮基底膜屏障，血管通透性增加和白细胞外渗，以及 HS结合的细胞因子和生长因子的释放促进血管生成和炎症， 周边地区PI的实验室最近观察到HPSE表达的显著增加， 在HSV感染人角膜细胞、培养的人角膜和动物角膜时， 眼部感染模型，并证明了其在病毒传播和传播中的重要性。小鼠 角膜HPSE上调直接导致角膜疾病病理包括溃疡， 炎症和新血管形成。此外，我们意外地发现HPSE敲除细胞表现出一种 病毒产量严重不足。因此，根据我们新产生的初步结果， 假设HPSE在HSV-1转录中具有重要的调节作用，并建议将HPSE确立为 一个关键的宿主毒力因子。我们认为激活的HPSE直接增加了疱疹性疾病的严重程度 包括组织损伤并促进病毒感染和再活化。该提案将重点关注 了解有助于HSV-1在角膜中生长的HPSE驱动机制， 证明HPSE抑制或染色体缺失的抗病毒/抗炎作用。三 提出了目标。第一个目标将建立细胞中HSV-1感染丧失背后的分子基础 缺少HPSE。这是基于我们的假设，HPSE通过其核定位和一组复杂的 相互作用促进HSV-1转录。第二个目的是确定HPSE在HSV-1中的意义 小鼠角膜的感染。本研究将使用HPSE基因敲除小鼠模型来证明我们的假设 HPSE是一种宿主毒力因子，可促进角膜中的HSV-1感染，导致组织损伤， 疾病病理，并促进病毒传播和返回三叉神经节（TG）期间 重新激活第三个也是最后一个目标将确定HPSE抑制剂对以下疾病的治疗益处： HSV-1角膜感染。这一目的将证明我们的假设，药物抑制HPSE， 将带来前所未有的治疗益处，包括快速解决感染和角膜 炎症本研究的成功完成将有助于建立一个全新的角色HPSE作为主机 毒力因子，并确定新的因素，有助于促进其致病活动的角膜。我们的结果 将显著增强我们对HPSE功能和疱疹疾病机制的理解， 加快制定控制眼部感染的新战略。