Novel Mechanisms in Adenoviral Ocular Pathogenesis

腺病毒眼部发病机制的新机制

基本信息

批准号：
10642924
负责人：
James Chodosh
金额：
$ 38.13万
依托单位：
UNIVERSITY OF NEW MEXICO HEALTH SCIS CTR
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-30 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10642924
关键词：
3-Dimensional ATP phosphohydrolase Accounting Adenosine Adenovirus Infections Adenoviruses Antiviral Therapy Binding Binding Proteins Biological Assay Capsid Capsid Proteins Cell Nucleus Cells Cementation Chemotaxis Chronic Clathrin Clinical Co-Immunoprecipitations Complex Confocal Microscopy Conjunctivitis Cornea Corneal Stroma Cytoplasm Disparate Electron Microscopy Endosomes Epidemic Keratoconjunctivitis Epithelial Cells Epithelium Event Eye Infections Fibroblasts Future Gene Expression Goals HMGB1 gene Human Image Immune response In Vitro Infection Infiltration Inflammation Inflammation Mediators Inflammatory Innate Immune Response Keratitis Kinesin Kinetics Label Leucocytic infiltrate Leukocytes Link Liquid Chromatography Mediating Membrane Microdomains Microtubules Minor Modeling Morbidity - disease rate Nuclear Nuclear Localization Signal Nuclear Pore Complex Proteins Nuclear Translocation Office Visits Pathogenesis Pathway interactions Peptide Hydrolases Persons Phosphorylation Photophobia Play Primary Care Proteins Proteomics Proton Pump Publishing Recurrence Role Signal Pathway Signal Transduction Small Interfering RNA Surface Plasmon Resonance Testing Therapeutic Translating Ubiquitin United States Viral Viral Genes Viral Pathogenesis Viral Proteins Viral Structural Proteins Virion Virulent Virus Virus Diseases Virus Replication Visual impairment Western Blotting Work acute infection caveolin 1 cell stroma cell type confocal imaging corneal epithelium cost design effective therapy experimental study imaging approach in vivo knock-down migration mutant novel novel strategies nucleocytoplasmic transport ocular surface pathogen penton base tandem mass spectrometry trafficking viral DNA

项目摘要

PROJECT SUMMARY Conjunctivitis impacts approximately 6 million people annually in the United States, accounting for 1% of all primary care office visits, and costing 430 million USD annually. Viruses are responsible for ~80% of conjunctivitis cases, and human adenoviruses (HAdVs) are implicated in up to 65% of these. A particularly severe cause of conjunctivitis, epidemic keratoconjunctivitis (EKC) is caused principally by viruses within human adenovirus species D (HAdV-D), for example the highly virulent adenovirus type 37 (HAdV-D37). The clinical manifestations of EKC include severe membranous conjunctivitis and epithelial keratitis, followed by delayed onset of multifocal subepithelial (stromal) corneal infiltrates. The latter cause light sensitivity and reduced vision, and can persist or recur for months to years after the acute infection, long after the virus is gone. We have shown in work published over the last 20+ years that adenovirus binding to human corneal cells activates complex intracellular signaling events that control subsequent cellular entry, intracellular trafficking of virions for delivery of viral DNA into cell nuclei, virion assembly and viral replication, and proinflammatory gene expression by infected cells. In this application, we extend our studies with new revelations on viral trafficking, new functions for a viral structural protein, and new evidence for expression of a novel proinflammatory mediator by infected corneal epithelial cells. The specific aims of this proposal are to test the hypotheses that 1) kinesin-1 and V-ATPase play previously unidentified roles in adenovirus trafficking, 2) adenoviral minor coat protein IIIa promotes adenovirus replication, and 3) corneal epithelial cell HMGB1 promotes inflammation in adenovirus keratitis. We will apply proteomic, siRNA, and imaging approaches to determine final stages of virion uncoating in human corneal epithelial cells and human corneal fibroblasts. A role for viral protein IIIa in nuclear transport of viral proteins for virion assembly will be determined by use of deletion and point mutants of pIIIa through surface plasmon resonance, imaging, PCR, and proteomics experiments. Finally, we will determine a role for human corneal epithelial cell expression of the multi-potent molecule HMGB1 in the formation of subepithelial infiltrates in EKC. These studies are designed to move us toward a more detailed understanding of adenoviral corneal pathogenesis, in order to reach our goal of an information-based therapy to mitigate the morbidity of adenovirus ocular infections.

项目摘要结膜炎每年在美国影响约600万人，占1％在所有初级保健办公室访问中，每年耗资4.3亿美元。病毒负责约80％结膜炎病例和人类腺病毒（HADV）的含量最多为65％。一个特别严重的结膜炎原因，流行角膜结膜炎（EKC）主要是由人类腺病毒物种D（HADV-D）中的病毒，例如高毒性腺病毒类型 37（HADV-D37）。 EKC的临床表现包括严重的膜状结膜炎和上皮角膜炎，然后延迟发作多焦点上皮下（基质）角膜浸润。这后者会引起光灵敏度和视力降低，并且可以在急性感染，病毒消失了很长时间。在过去的20多年中，我们在发表的作品中展示了腺病毒与人角膜细胞的结合激活了复杂的细胞内信号传导事件控制随后的细胞进入，病毒体的细胞内运输将病毒DNA递送到细胞中核，病毒体组装和病毒复制以及感染细胞的促炎基因表达。在在此应用中，我们通过有关病毒贩运的新启示扩展了研究，病毒的新功能结构蛋白，以及被感染的新型促炎介质表达的新证据角膜上皮细胞。该提议的具体目的是检验假设：1）kinesin-1 和V-ATPase在腺病毒贩运中扮演以前未识别的角色，2）腺病毒小外套蛋白IIIA促进腺病毒复制，3）角膜上皮细胞HMGB1促进腺病毒角膜炎的炎症。我们将应用蛋白质组学，siRNA和成像方法确定在人角膜上皮细胞和人角膜中脱膜的最终阶段成纤维细胞。病毒蛋白IIIA在病毒蛋白核转运中的作用将是病毒素组装的作用通过通过表面等离子体共振，成像，成像的删除和点突变体的使用确定 PCR和蛋白质组学实验。最后，我们将确定人角膜上皮细胞的作用在EKC中，多能量分子HMGB1的表达。这些研究旨在使我们迈向对腺病毒角膜的更详细的了解发病机理，以实现我们的基于信息疗法的目标，以减轻腺病毒眼感染。