Proteomic analysis of S aureus biofilm in vivo

描述（由申请人提供）：金黄色葡萄球菌是一种主要的人类病原体，可引起一系列疾病。葡萄球菌感染的发病率继续上升。目前的治疗方法几乎完全依靠抗生素治疗。生物膜的形成是金黄色葡萄球菌毒力的主要贡献者，因为它保护细菌免受抗生素或宿主免疫系统的侵害。具体来说，生物膜提供了对抗生素的内在抗性和对宿主防御的保护，这导致治疗失败。生物膜由多层细菌细胞组成，细胞外基质使生物膜保持完整。了解生物膜基质的性质可能会导致新的方法来防止生物膜的形成或根除生物膜内的细菌，从而提供一种克服生物膜相关感染的治疗顽固性的方法。先前显示的促进金黄色葡萄球菌生物膜形成的因素包括表面相关蛋白、胞外多糖和细胞外DNA，但迄今为止，这些成分对葡萄球菌生物膜相关感染的贡献尚未确定。更重要的是，我们并不完全了解感染过程中形成的生物膜中生物膜基质的组成，特别是蛋白质成分。该应用程序的目的是通过合适的动物模型从感染部位识别生物膜中的蛋白质。为此，我们最近开发了一种植入物相关骨科感染的大鼠模型，使我们能够从感染部位分离出足够的细菌，从而使用先进的蛋白质组学方法直接鉴定细菌成分。因此，在本应用中，我们建议全面鉴定在体内条件下产生的生物膜基质中的蛋白质（目标1），并验证感染部位存在一组优先的单个成分（目标2）。

DESCRIPTION (provided by applicant): Staphylocuccus aureus is a major human pathogen that causes a range of diseases. The incidence of staphylococcal infections continues to rise. Current methods of treatment rely almost exclusively on antibiotic therapy. Biofilm formation is a major contributor of S. aureus virulence because it protects the bacteria from antibiotics or host immune system. Specifically, biofilms provide intrinsic resistance to antibiotics and protection from host defenses, which leads to treatment failures. Biofilms consist of multiple layers of bacterial cells contained within an extracellular matrix that keeps the biofilm intact. Understanding the nature of biofilm matrix could lead to new methods to prevent biofilm formation or eradicate bacteria within a biofilm, thus providing a way to overcome the therapeutic recalcitrance of biofilm-associated infections. Factors previously shown to contribute to biofilm formation in S. aureus include surface-associated proteins, exopolysaccarides, and, extracellular DNA, but to date the contribution of these components to staphylococcal biofilm-associated infections has not been defined. More importantly, we do not fully understand the composition of biofilm matrix, especially the protein components, in biofilms formed during an infection. The goal of this application is to identify proteins in biofilms from an infection site sing a suitable animal model. To this end, we have recently developed a rat model of implant-associated orthopedics infection allowing us to isolate sufficient bacteria from an infection site o identify bacterial components directly using an advanced proteomic method. Thus, in this application, we propose to comprehensively identify proteins in biofilm matrix produced under in vivo conditions (Aim 1) and to validate the presence of a set of prioritized individual components in the infection site (Aim 2).