Pathogenesis of Fungal Keratitis

真菌性角膜炎的发病机制

基本信息

批准号：
10212713
负责人：
Eric Pearlman
金额：
$ 6.5万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-02-08 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10212713
关键词：
Aspergillus Binding Blindness Bone Marrow Cell Nucleus Cells Cleaved cell Complication Contact Lenses Cornea Corneal Stroma Cytoplasmic Granules Cytoplasmic Protein Data Developing Countries Disease Elastases Environment Enzyme Reactivation Enzyme-Linked Immunosorbent Assay Enzymes Flow Cytometry Funding Fusarium Gene Expression Genes Genomic DNA Grant Growth Harvest Histones Human Hyphae Immune Targeting Immune response Immunology Incidence Infection Inflammatory Interleukin-1 alpha Interleukin-1 beta Interleukin-17 Invaded Journals Keratitis Knock-in Mouse Knockout Mice Leukocyte L1 Antigen Complex Macrophage-1 Antigen Mediating Mediator of activation protein Membrane Molds Molecular Mus Nature Neutrophil Infiltration Nitrogen Nuclear Organism Oxygen Pain Pathogenesis Pathogenicity Pathway interactions Peptide Hydrolases Play Production Proteins Publishing RNA analysis Reporting Reproduction spores Risk Factors Role Seasons Signal Transduction Testing Therapeutic Intervention Tissues Trauma Virulence Factors Vision Visual impairment anterior chamber antimicrobial peptide base beta-Glucans chemokine clinical investigation cytokine cytotoxic dectin 1 experimental study extracellular fungus immunological intervention inhibitor/antagonist mouse model nano-string neutrophil novel ocular surface osteopontin pathogen prevent response transcriptome sequencing

项目摘要

Abstract Pathogenic Fusarium and Aspergillus molds are an important cause of vision loss in the USA and worldwide. Preliminary data from an unbiased gene array analyses (Nanostring and RNA sequencing) showed an anticipated increase in expression of multiple genes in neutrophils isolated from Fusarium and Aspergillus infected mouse corneas compared with bone marrow neutrophils from the same mice. However, we found an unexpected significant increase in expression of the pro-inflammatory cytokine IL-1 alpha, and of Spp1/osteopontin (OPN), which has known pro-inflammatory activity, and which exists as a secreted (sOPN) and intracellular (iOPN). Elevated IL-1 alpha and OPN was confirmed by Q-PCR, ELISA and intracellular flow cytometry. Aim 1 will characterize the membrane, secreted and intracellular activities the pro- and cleaved forms of IL-1 alpha in murine and human neutrophils, and in Fusarium and Aspergillus keratitis. Aim 2 will identify regulators of OPN expression, and use OPN-/- and iOPN knock-in mice to examine the role of the intracellular versus secreted forms of this protein in human neutrophils and in fungal keratitis. Aim 3 will characterize the role of neutrophil extracellular traps (NETs), which can limit hyphal growth, but also have the potential to contribute to tissue damage. Proposed experiments will identify the molecular pathways leading to NET formation in infected corneas, and using mice that lack the enzyme required for histone citrullinatation and decondensation (PAD4), we will examine the role of NETs in Fusarium and Aspergillus keratitis. Collectively, the results of these proposed studies will increase our understanding of the pathogenesis of this blinding disease, and will potentially identify novel targets for immune intervention to prevent neutrophil mediated tissue damage and loss of corneal clarity.

抽象的致病性镰刀菌和曲霉霉菌是美国视力丧失的重要原因，全世界。来自公正的基因阵列分析（纳米串和RNA测序）的初步数据显示从镰刀菌和曲霉分离的中性粒细胞中多个基因表达的预期增加与来自同一小鼠的骨髓性嗜中性粒细胞相比，感染的小鼠角膜。但是，我们发现了促炎性细胞因子IL-1α和 SPP1/骨桥（OPN），已知促炎活性，并且作为分泌的（SOPN）存在和细胞内（IOPN）。 Q-PCR，ELISA和细胞内流动证实了升高的IL-1α和OPN 细胞仪。 AIM 1将表征膜，分泌和细胞内活动鼠和人类中性粒细胞中的IL-1α的形式，镰刀菌和曲曲霉的形式。 AIM 2意志识别OPN表达的调节剂，并使用OPN - / - 和IOPN敲击小鼠检查该蛋白质的细胞内与分泌形式在人类嗜中性粒细胞和真菌角膜炎中。目标3意志表征中性粒细胞外陷阱（网）的作用，这可能限制菌丝的生长，但也具有潜力有助于组织损伤。提出的实验将确定导致的分子途径感染角膜中的净形成，并使用缺乏组蛋白柠檬磷脂所需的酶的小鼠反应（PAD4），我们将检查网中网中的作用在镰刀菌和曲曲霉角膜炎中。共同这些提出的研究的结果将增加我们对这种盲的发病机理的理解疾病，并有可能识别出免疫干预的新靶标，以防止中性粒细胞介导的组织损坏和角膜清晰度的损失。