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.

与眼睛的单纯疱疹病毒-1（HSV -1）感染有关的主要临床并发症包括 角膜溃疡，严重的疼痛，感染，角膜血管的丧失和视力丧失。仅抗病毒药物 通常无法纠正问题，因为许多疾病症状可能是通过感染签署的，但是 然后通过宿主分子更直接地调节。该建议研究，人类肝素酶（HPSE），宿主 可能调节角膜中病毒生长和疾病表现的酶。 HPSE是一个 硫酸乙酰肝素（HS）内糖苷酶，其水平与上皮分解直接相关 内皮地下室膜屏障，血管渗透性增加和白细胞渗出以及 HS结合的细胞因子和生长因子的解放，促进了血管生成和炎症 周围地区。 PI的实验室最近观察到HPSE表达和 人角膜细胞，培养的人角膜和动物的HSV感染后，酶活性较高 眼部感染模型，并证明了其在病毒扩散和传播中的重要性。在鼠 角膜HPSE上调直接有助于角膜疾病病理，包括溃疡， 此外，我们出乎意料地发现HPSE敲除细胞显示 病毒生产严重缺乏。因此，根据我们新生成的初步结果 假设HPSE在HSV-1转录中的重要调节作用，并提出建立HPSE为 关键的宿主病毒因子。我们建议激活的HPSE直接增加了疱疹性疾病的严重性 包括组织损伤并促进病毒感染和重新激活。该提议将重点放在 了解HPSE驱动的机制，这些机制有助于角膜中HSV-1的增长 证明了HPSE抑制或染色体缺失的抗病毒/抗炎作用。三 提出了目标。第一个目标将在细胞中HSV-1感染丧失背后建立分子基础 缺乏HPSE。它是基于我们的假设，即HPSE通过其核定位和复杂的集合。 相互作用促进HSV-1转录。第二个目标将确定HPSE在HSV-1中的重要性 鼠角膜的感染。此目标将使用HPSE敲除鼠标模型来证明我们的假设 HPSE是促进角膜中HSV-1感染的宿主病毒因子，驱动组织损伤和 疾病的病理，并促进病毒传播到三叉神经节（TG）期间的病毒传播 重新激活。第三个也是最后一个目标将决定HPSE抑制剂对抗的治疗益处 角膜的HSV-1感染。这个目标将证明我们的假设是HPSE的药物抑制 将带来前所未有的治疗益处，包括快速解决感染和角膜 炎。我们的研究的成功结论将有助于为HPSE担任全新角色 毒力因子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理解，以及 加速控制眼部感染的新策略。