CAP37 and ocular inflammation

CAP37 和眼部炎症

• 批准号: 7322473
• 负责人: Heloise. ANNE Pereira
• 金额: $ 36.63万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7322473
• 关键词: Acute; Anti-Bacterial Agents; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Bacteria; Bacterial Eye Infections; Bacterial Infections; Biological; Blindness; Cell Adhesion Molecules; Cell Line; Cell physiology; Cells; Chemotaxis; Chronic; Cicatrix; Ciliary epithelium; Cornea; Corneal Stroma; Cytoplasmic Granules; Data; Data Analyses; Development; Disclosure; Emigrations; Enzyme-Linked Immunosorbent Assay; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Event; Extracellular Matrix; Eye; Eye Infections; Flow Cytometry; Healed; Host Defense; Human; Immune; Immunohistochemistry; Infection; Inflammation; Inflammatory; Inflammatory Response; Invaded; Keratitis; Laboratories; Lead; Leukocytes; Location; Measures; Mediating; Mediator of activation protein; Microglia; Modeling; Molecular; Mus; Oryctolagus cuniculus; Peptides; Perforation; Play; Property; Protein Kinase C; Proteins; Pseudomonas aeruginosa; Public Health; RNA Interference; Reaction; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Signaling Molecule; Site; Smooth Muscle Myocytes; Staining method; Stains; Staphylococcus aureus; Stimulus; System; Techniques; Testing; Tissues; Ulcer; Up-Regulation; Vascular Endothelial Cell; Vascular Endothelium; Vision; Wound Infection; antimicrobial; bactericide; base; bulbar conjunctiva; corneal epithelial cell adhesion; corneal epithelium; cytokine; design; healing; in vivo; inhibitor/antagonist; limbal; macrophage; migration; monocyte; mouse model; neutralizing antibody; neutrophil; novel; novel therapeutics; pathogen; response; ward

DESCRIPTION (provided by applicant): This application is designed to investigate the role of the inflammatory mediator, CAP37, in ocular inflammation. CAP37 was recently identified in the eye in response to bacterial infection. Surprisingly, its localization was not confined to the neutrophil (PMN), where it is known to be constitutively expressed, but it was also found induced in corneal epithelial cells, and vascular endothelial cells of the limbal and ciliary vessels. There are many inflammatory-associated biological activities regulated by CAP37 that confer on this molecule a role in innate host defense against invading pathogens. CAP37 is a potent antibiotic, promotes monocyte chemotaxis, and has immunomodulatory effects on host cells including corneal epithelial cells. The induction of a host defense molecule in the outermost layers of the eye and in the vascular endothelium in response to infection, suggests that it could play a key role in regulating inflammation and healing in the cornea and in the long term form the basis for developing new therapeutics for certain eye infections. It is our hypothesis that CAP37, either released by PMNs or induced in corneal epithelial cells at the site of infection, plays a pivotal role in the inflammatory response of the cornea to infection. We hypothesize that it accomplishes this through a novel, dual mechanism of action based on its antibiotic/anti-inflammatory and immunomodulatory properties. Aim 1: To test the hypothesis that the regulation of proliferation, chemotaxis, and adhesion of corneal epithelial cells in ocular inflammation is dependent on CAP37 and is mediated through the intracellular signaling molecule protein kinase C, we will use techniques that include immunohistochemistry, pharmacological inhibitors, and RNA interference. Aim 2: To test the hypothesis that CAP37 is a biologically relevant mediator in host defense in the eye, we will measure the induction and identity of inflammatory cells, mediators, cytokines, and adhesion molecules in response to the administration of CAP37, its bioactive peptides, and neutralizing antibodies into the corneal stroma. Flow cytometry, ELISA, immunohistochemistry, and RT-PCR will be used to analyze data. Aim 3: To test the hypothesis that the anti-inflammatory and immunomodulatory properties of CAP37 are important for host defense in ocular infection we will use CAP37 and its bioactive peptides in a murine model of Pseudomonas aeruginosa keratitis to determine in vivo efficacy. Relevance to public health: Once corneal integrity is breached, prompt treatment with antibiotics is required to halt the development of infection and harmful sequelae such as ulceration, perforation, scarring, and loss of vision. Unfortunately, the antibiotics currently available for ophthalmic use have serious limitations. We believe our findings will help develop new therapies for treating wounds and infections of the eye based on the biological molecule CAP37 and its bioactive peptides.

描述（由申请人提供）：本申请旨在研究炎症介质 CAP37 在眼部炎症中的作用。最近在眼睛中发现了针对细菌感染的 CAP37。令人惊讶的是，它的定位并不局限于已知其组成型表达的中性粒细胞（PMN），而且还发现它在角膜上皮细胞以及角膜缘和睫状血管的血管内皮细胞中被诱导。 CAP37 调节许多与炎症相关的生物活性，使该分子在宿主针对入侵病原体的先天防御中发挥作用。 CAP37 是一种强效抗生素，可促进单核细胞趋化性，并对包括角膜上皮细胞在内的宿主细胞具有免疫调节作用。在眼睛最外层和血管内皮中对感染做出反应时诱导宿主防御分子表明，它可能在调节角膜炎症和愈合中发挥关键作用，并从长远来看为开发某些眼部感染的新疗法奠定了基础。我们假设 CAP37，无论是由 PMN 释放还是在感染部位的角膜上皮细胞中诱导，在角膜对感染的炎症反应中发挥着关键作用。我们假设它通过基于其抗生素/抗炎和免疫调节特性的新型双重作用机制来实现这一目标。目标 1：为了检验眼部炎症中角膜上皮细胞增殖、趋化性和粘附的调节依赖于 CAP37 并通过细胞内信号分子蛋白激酶 C 介导的假设，我们将使用包括免疫组织化学、药理学抑制剂和 RNA 干扰在内的技术。目标 2：为了检验 CAP37 是眼睛宿主防御中生物相关介质的假设，我们将测量炎症细胞、介质、细胞因子和粘附分子响应于将 CAP37、其生物活性肽和中和抗体注入​​角膜基质而产生的诱导和识别。将使用流式细胞术、ELISA、免疫组织化学和 RT-PCR 来分析数据。目标 3：为了检验 CAP37 的抗炎和免疫调节特性对于眼部感染的宿主防御很重要这一假设，我们将在铜绿假单胞菌角膜炎的小鼠模型中使用 CAP37 及其生物活性肽来确定体内功效。与公共卫生的相关性：一旦角膜完整性遭到破坏，就需要立即使用抗生素进行治疗，以阻止感染和有害后遗症的发展，例如溃疡、穿孔、疤痕和视力丧失。不幸的是，目前可用于眼科的抗生素有严重的局限性。我们相信我们的发现将有助于开发基于生物分子 CAP37 及其生物活性肽的治疗伤口和眼部感染的新疗法。