The role of exosomes in Pseudomonas Aeruginosa Corneal Infection

外泌体在铜绿假单胞菌角膜感染中的作用

• 批准号: 10166851
• 负责人: DANIELLE M. ROBERTSON
• 金额: $ 39.29万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10166851
• 关键词: Adrenal Cortex Hormones; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antibiotic Resistance; Antimicrobial Effect; Autologous; Bioenergetics; Biology; Blindness; Blood; Body Fluids; Cells; Cellular biology; Chemotaxis; Communicable Diseases; Contact Lenses; Core Facility; Cornea; Corneal Diseases; Country; Coupled; Data; Disease; Epithelial Cells; Exhibits; Eye; Fluoroquinolones; Genetic Transcription; Goals; Host Defense; Host Defense Mechanism; Human; Human body; Immune; In Vitro; Infection; Infectious Agent; Inflammation; Inflammation Mediators; Invaded; Keratitis; Laboratories; Mass Spectrum Analysis; Mediating; Modeling; Neutrophil Activation; Ophthalmology; Oryctolagus cuniculus; Parotid Gland; Pathologic; Peptide Hydrolases; Phagocytosis; Pilot Projects; Play; Property; Proteins; Proteomics; Pseudomonas; Pseudomonas Infections; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Publishing; Resolution; Resources; Role; Saliva; Salivary; Salivary Glands; Serum; Small Interfering RNA; Sterility; System; Testing; Therapeutic; Therapeutic Agents; Therapeutic Uses; Tissues; Topical Antibiotic; Trauma; antimicrobial; base; combat; comparative efficacy; corneal epithelial wound healing; corneal epithelium; design; exosome; extracellular vesicles; immunoregulation; in vivo; in vivo Model; inhibitor/antagonist; innovation; intercellular communication; microbial; multidisciplinary; neutrophil; novel; novel strategies; novel therapeutics; pathogen; prevent; response; transcriptome sequencing; treatment strategy; uptake; vesicular release

PROJECT SUMMARY Pseudomonas aeruginosa (PA) is the leading causative agent in microbial keratitis. During contact lens wear, host defense mechanisms are compromised. This allows PA to breach the tight corneal barrier and infect the otherwise healthy eye. Recently, exosomes have been implicated as major players in inflammation and infection. In addition, exosomes are becoming increasingly recognized as potential therapeutic agents. Our preliminary data shows that there is massive exosome release from corneal epithelial cells during infection by PA. These exosomes contain a unique mixture of proteases, transcriptional regulators and proteins involved in immune regulation. We further provide data that indicates that these exosomes both promote neutrophil activation and convey protection to corneal epithelial cells against further invasion. Moreover, we have found that exosomes isolated from autologous body fluids have antimicrobial and immunomodulatory properties. Based on our findings, we propose the primary hypothesis that exosomes released from PA-infected corneal epithelial cells promote PA clearance by innate immune cells and prime non-infected corneal epithelial cells to defend against PA infection. We further propose the secondary hypothesis that exosomes derived from exosome rich body fluids contain potent antimicrobial and anti-inflammatory mediators that can be harnessed to promote PA clearance and disease resolution in the cornea. We will test these hypotheses as follows: Aim 1. Establish how exosomes isolated from PA-infected corneal epithelial cells and human body fluids impact innate immune cells in vitro. Aim 2. Determine how exosomes isolated from PA-infected corneal epithelial cells and human body fluids impact the corneal epithelial response to PA in vitro. Aim 3. Determine whether exosomes isolated from autologous human body fluids exhibit protective antimicrobial and immunomodulatory properties in the rabbit contact lens model in vivo. To accomplish these studies, we have compiled a highly collaborative multidisciplinary team and have access to state of the art resources in the Department of Ophthalmology and UT Southwestern core facilities. These studies are significant and innovative because they are the first of their kind for PA infection in any tissue or cell system. The potential therapeutic use of exosomes from human body fluids represents a major paradigm shift for treating corneal infections and has broad therapeutic implications.

项目摘要 铜绿假单胞菌（PA）是微生物性角膜炎的主要病原体。在接触透镜佩戴期间， 宿主的防御机制就被破坏了这使得PA能够突破紧密的角膜屏障并感染 健康的眼睛。最近，外泌体被认为是炎症的主要参与者， 感染此外，外泌体越来越被认为是潜在的治疗剂。 我们的初步数据显示，在感染过程中，角膜上皮细胞释放大量的外泌体， 的PA。这些外泌体含有蛋白酶、转录调节因子和相关蛋白质的独特混合物 在免疫调节中。我们进一步提供的数据表明，这些外来体都促进中性粒细胞 激活并保护角膜上皮细胞免受进一步侵袭。此外，我们还发现， 从自体体液中分离的外来体具有抗微生物和免疫调节特性。 基于我们的发现，我们提出了初步假设，即从PA感染的角膜释放的外泌体， 上皮细胞促进先天免疫细胞对PA的清除， 预防PA感染。我们进一步提出了第二个假设，即外来体来源于 富含外泌体的体液含有有效的抗菌和抗炎介质， 以促进角膜中的PA清除和疾病消退。 我们将测试这些假设如下：目标1。确定从PA感染的 角膜上皮细胞和人体体液在体外影响先天免疫细胞。目标2.确定 从PA感染的角膜上皮细胞和人体体液中分离的外泌体如何影响 角膜上皮对PA的体外反应。目标3.确定是否从 自体人体液显示出保护性抗微生物和免疫调节特性， 兔角膜接触透镜体内模型。 为了完成这些研究，我们组建了一个高度协作的多学科团队， 在眼科和UT西南部的核心设施的最先进的资源。这些 研究是重要的和创新的，因为它们是第一个在任何 组织或细胞系统。来自人体液的外泌体的潜在治疗用途代表了一种新的治疗方法。 这是治疗角膜感染的主要范式转变，具有广泛的治疗意义。