HPSE in Ocular Herpes Infection

HPSE 在眼部疱疹感染中的应用

• 批准号: 10475706
• 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/10475706
• 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(HSV-1)感染相关的主要临床并发症包括 角膜溃疡、剧烈疼痛、炎症、角膜血管丧失和视力丧失。单独使用抗病毒药物 通常无法纠正问题，因为许多疾病症状可能是感染的信号，但 然后更直接地受到宿主分子的调节。这项建议研究的是，人乙酰肝素酶(HPSE)的一种宿主 这种酶可能潜在地调节角膜中病毒的生长和疾病的表现。HPSE是一种 硫酸乙酰肝素(HS)内切糖苷酶的水平与上皮细胞和 内皮细胞基底膜屏障，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表达显著增加， 人角膜细胞、培养的人角膜和动物对HSV感染的较高酶活性 建立了眼部感染模型，并论证了其在病毒传播和传播中的意义。在小鼠体内 角膜HPSE上调直接导致包括溃疡在内的角膜疾病病理， 炎症和新生血管。此外，我们意外地发现，HPSE基因敲除细胞显示出 病毒生产严重不足。因此，根据我们新产生的初步结果，我们 假设HPSE在HSV-1转录中的重要调节作用，并建议将HPSE建立为 一个关键的寄主毒力因子。我们认为激活的HPSE直接增加了疱疹疾病的严重程度 包括组织损伤和促进病毒感染和重新激活。这项提案将重点放在 了解HPSE驱动的促进HSV-1在角膜和 展示抑制HPSE或染色体缺失的抗病毒/抗炎作用。三 提出了目标。第一个目标是建立细胞中HSV-1感染丧失的分子基础 缺乏HPSE。它是基于我们的假设，即HPSE通过其核局部化和复杂的 相互作用促进HSV-1转录。第二个目标将确定HPSE在HSV-1中的意义 小鼠角膜感染。这一目标将使用HPSE基因敲除小鼠模型来验证我们的假设 HPSE是一种宿主毒力因子，可促进角膜中HSV-1感染，推动组织损伤和 疾病病理，并促进病毒在三叉神经节(TG)的传播和返回 重新激活。第三个也是最后一个目标将确定HPSE抑制剂的治疗效果。 角膜的单纯疱疹病毒1型感染。这一目的将证明我们的假设，即HPSE的药理抑制 将带来前所未有的治疗益处，包括感染和角膜的快速消退 发炎。我们研究的成功完成将有助于建立HPSE作为东道主的全新角色 并确定有助于促进其在角膜中的致病活性的新因素。我们的结果 将大大增强我们对HPSE功能和疱疹疾病机制的了解，以及 加快制定控制眼部感染的新战略。