HPSE in Ocular Herpes Infection

HPSE 在眼部疱疹感染中的应用

• 批准号: 9761531
• 负责人: DEEPAK SHUKLA
• 金额: $ 39.98万
• 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/9761531
• 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; experimental study; heparanase; immunological status; lymph nodes; 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、角膜缺血和单独使用抗病毒药物导致视力丧失。 通常无法纠正问题，因为许多疾病症状可能是由感染发出的信号，但 然后由宿主分子更直接地调节。该提案研究了宿主人乙酰肝素酶（HPSE）。 HPSE 是一种可能调节病毒生长和角膜疾病表现的酶。 硫酸乙酰肝素 (HS) 糖苷内切酶，其水平与上皮细胞和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结合细胞因子和生长因子的释放促进血管生成和炎症 PI 实验室最近观察到 HPSE 表达显着增加， HSV 感染人类角膜细胞、培养的人类角膜和动物时具有更高的酶活性 眼部感染模型并证明了其在小鼠病毒传播和传播中的重要性。 角膜 HPSE 上调直接导致角膜疾病病理，包括溃疡、 此外，我们意外地发现 HPSE 敲除细胞表现出炎症和新生血管形成。 因此，根据我们新产生的初步结果，我们发现病毒生产严重不足。 确立了 HPSE 在 HSV-1 转录中的重要调节作用，并建议将 HPSE 建立为 我们认为激活的 HPSE 会直接增加疱疹疾病的严重程度。 包括组织损伤并促进病毒感染和重新激活。 了解促进 HSV-1 在角膜中生长的 HPSE 驱动机制，以及 证明 HPSE 抑制或染色体缺失的抗病毒/抗炎作用。 提出的第一个目标是建立细胞中 HSV-1 感染消失的分子基础。 它基于我们的假设，HPSE 通过其核定位和复杂的设置。 第二个目标是确定 HPSE 在 HSV-1 中的重要性。 该目标将使用 HPSE 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 HSV-1 感染、导致组织损伤和 疾病病理学，并促进病毒传播到三叉神经节（TG）并从三叉神经节返回 第三个也是最后一个目标将确定 HPSE 抑制剂的治疗效果。 HSV-1 感染的角膜将证明我们的假设，即药物抑制 HPSE。 将带来前所未有的治疗效益，包括快速解决感染和角膜问题 我们研究的成功结束将有助于确立 HPSE 作为宿主的全新角色。 毒力因子并确定有助于促进其在角膜中致病活性的新因子。 将显着增强我们对 HPSE 功能和疱疹疾病机制的理解，并且 加速控制眼部感染的新策略。