Virulence of Staphylococcus aureus Corneal Infections

金黄色葡萄球菌角膜感染的毒力

• 批准号: 10656985
• 负责人: Rachel A F Wozniak
• 金额: $ 41.71万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656985
• 关键词: Address; Adherence; Antibiotic Resistance; Antibiotics; Area; Attention; Automobile Driving; Bacteria; Bacterial Adhesins; Bacterial Proteins; Binding; Binding Proteins; Bioinformatics; Biological; Biological Assay; Blindness; Blinking; Cells; Cellular Assay; Clinical; Collagen; Cornea; Data; Disease; Endocarditis; Enterotoxins; Environment; Epithelial Cells; Evaluation; Eye Infections; Film; Genes; Genetic; Genetic Determinism; Genome; Genomic approach; Genomics; Goals; Healthcare; Hemolysin; Immune; Immune Evasion; Immune response; Immune system; Immunohistochemistry; In Vitro; Individual; Infection; Infectious Skin Diseases; Invaded; Keratitis; Knowledge; Laboratories; Link; Mediating; Nutrient; Osteomyelitis; Pathogenesis; Pathogenicity Island; Physiological; Pilot Projects; Pneumonia; Proteins; Regulation; Research; Role; Sepsis; Severity of illness; Site; Staphylococcus aureus; Staphylococcus aureus infection; Surface; Therapeutic; Therapeutic Intervention; Time; Tissues; Toxic effect; Toxin; Validation; Virulence; Virulence Factors; Vision; Work; body system; comparative genomics; corneal epithelium; cytokine; cytotoxicity; drug discovery; fitness; genome sequencing; human pathogen; in vivo; in vivo Model; innovation; interest; mastitis; mouse model; mutant; novel; novel strategies; novel therapeutics; ocular surface; prospective; rational design; tool; transcriptome; transcriptome sequencing; transcriptomics; transposon sequencing; virtual; whole genome

ABSTRACT Bacterial keratitis (corneal infection) is a vision threatening disease and contributes significantly to world blindness. The important human pathogen, Staphylococcus aureus, is a predominant cause of keratitis, causing aggressive infections that often lead to irreparable ocular tissue damage. While much attention has been given to understanding the pathogenesis of S. aureus in non-ocular sites of infection such as sepsis, endocarditis or osteomyelitis, surprisingly, relatively little is yet known about the bacterial virulence factors that govern S. aureus keratitis. In the absence of this knowledge, it is impossible to understand the fundamental pathogenesis of S. aureus keratitis or rationally design therapeutics to treat this blinding disease. Accordingly, the central goal of this proposal is to identify and characterize S. aureus genetic determinants that modulate keratitis. Given the unique environment of the ocular surface and cornea with respect to tear film composition, sheer forces generated from blinking, and relative immune privilege, we hypothesize that there will be a unique set of virulence factors that drive keratitis. Our laboratory has developed innovative tools that will allow, for the first time, the ability to prospectively identify and validate the comprehensive set of S. aureus virulence factors that play a role in BK pathogenesis. Our discovery pipeline leverages state of the art genomics and bioinformatics including whole genome sequencing, Tn-seq and RNA-seq to generate high priority leads for mechanism of action studies, as well as the evaluation of biologic significance in our murine model of keratitis. We have successfully used this approach to identify and validate enterotoxins, secreted bacterial proteins well known for their cytotoxicity and ability to modulate the host immune response, as important in mediating keratitis. Thus, this proposal will build on this exciting preliminary data to develop an expanded list of the key drivers of keratitis. Ultimately this work will directly advance our understanding of S. aureus keratitis pathogenesis and as such, may be foundational in supporting strategies for the therapeutic intervention of this debilitating disease.

抽象的 细菌性角膜炎（角膜感染）是一种威胁视力的疾病，对世界造成重大影响 失明。重要的人类病原体金黄色葡萄球菌是角膜炎的主要原因，可导致 侵袭性感染通常会导致无法修复的眼组织损伤。虽然受到了很多关注 了解脓毒症、心内膜炎或非眼部感染部位金黄色葡萄球菌的发病机制 令人惊讶的是，对于骨髓炎，人们对控制金黄色葡萄球菌的细菌毒力因子知之甚少 角膜炎。如果缺乏这方面的知识，就不可能了解金黄色葡萄球菌的基本发病机制。 金黄色葡萄球菌角膜炎或合理设计疗法来治疗这种致盲疾病。据此，中心目标 该提案的目的是识别和表征调节角膜炎的金黄色葡萄球菌遗传决定因素。 鉴于眼表和角膜在泪膜成分方面的独特环境，纯粹 眨眼产生的力量和相对免疫特权，我们假设将会有一组独特的 导致角膜炎的毒力因子。我们的实验室开发了创新工具，首次 时间，前瞻性地识别和验证金黄色葡萄球菌毒力因子的综合集的能力 在 BK 发病机制中发挥作用。我们的发现管道利用最先进的基因组学和生物信息学 包括全基因组测序、Tn-seq 和 RNA-seq，为机制生成高优先级线索 行动研究，以及在我们的小鼠角膜炎模型中评估生物学意义。我们有 成功地使用这种方法来识别和验证肠毒素，众所周知的分泌细菌蛋白 它们的细胞毒性和调节宿主免疫反应的能力，对于介导角膜炎很重要。因此， 该提案将以这一令人兴奋的初步数据为基础，制定一份扩大的角膜炎关键驱动因素清单。 最终，这项工作将直接增进我们对金黄色葡萄球菌角膜炎发病机制的理解，因此， 可能是支持这种使人衰弱的疾病的治疗干预策略的基础。