Proteomic Analysis of Implant Surfaces in Athroplasty Failure

关节置换术失败中植入物表面的蛋白质组学分析

• 批准号: 10623873
• 负责人: Robin Patel
• 金额: $ 23.85万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-18 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10623873
• 关键词: Biological Assay; Biological Markers; Clinical; Detection; Development; Diagnostic; Diagnostic Procedure; Diagnostic tests; Enterococcus faecalis; Etiology; Failure; Fracture; Future; Gene Expression; Genus staphylococcus; Goals; Guidelines; Human; Immune response; Implant; Individual; Infection; Knee; Liquid Chromatography; Liquid substance; Mass Spectrum Analysis; Measures; Microbial Biofilms; Microbiology; Organism; Orthopedics; Osteolysis; Pathogenesis; Patients; Performance; Periprosthetic joint infection; Prevention; Prevention approach; Protein Analysis; Proteins; Proteome; Proteomics; Reaction; Replacement Arthroplasty; Ringer' s solution; Sampling; Site; Sonication; Specimen; Staphylococcus aureus; Staphylococcus epidermidis; Sterility; Streptococcus Group B; Surface; Technology; Testing; Tissues; Work; alpha-Defensins; cytokine; design; diagnostic strategy; improved; in vivo; insight; joint infection; knee replacement arthroplasty; lateral flow assay; microbial; microbial host; microorganism; neutrophil; new technology; next generation sequencing; novel diagnostics; protein expression; response; response biomarker; tandem mass spectrometry

PROJECT SUMMARY Our team has developed a strategy that samples materials on surfaces of removed orthopedic implants using sonication and shown that the derived biofilm-sampling “sonicate fluids” have the highest yield of any specimen type for microbial detection and can furthermore be used to assess host response at implant surfaces. While assessment of host response has been clinically leveraged to differentiate infected from non-infected orthopedic implants, host-based approaches deployed to date have been simplistic, detecting alpha-defensin or profiling neutrophil numbers, for example, with none defining the microbiology of infected arthroplasties or underlying etiologies of non-infectious arthroplasty failure. We hypothesize that, beyond differentiating infectious from non-infectious arthroplasty failure, capturing host response in sonicate fluid using a proteomic approach will discern microbial etiologies of periprosthetic joint infection and individual causes of non- infectious arthroplasty failure. Given that detection of causative organisms in periprosthetic joint infection and definition individual causes of non-infectious arthroplasty failure, needed to direct management, are not universally realized with extant diagnostic approaches, this will be clinically useful. We propose the first-ever global, unbiased proteomic analysis of sonicate fluid, assessing in vivo human immune response to infectious compared to non-infectious arthroplasty failure and subsets thereof. We further propose an unbiased global analysis of microbial proteins in sonicate fluid in periprosthetic joint infection. Sonicate fluid has been shown in preliminary work to be suitable for the proposed proteomic analyses. Sonicate fluid from failed total knee arthroplasties will be studied using the new technology of liquid chromatography tandem mass spectrometry to determine whether host proteomic analysis can differentiate Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus lugdunensis, Enterococcus faecalis and Streptococcus agalactiae periprosthetic joint infection, and non-infectious arthroplasty failure subsets of aseptic loosening, periprosthetic fracture, instability, and osteolysis/adverse tissue reaction. In addition to mass spectrometry, a complementary next generation sequencing-based proximity extension assay approach that measures levels of 3,072 cytokines and other host proteins will be used to measure low abundance human proteins missed using mass spectrometry alone, allowing examination of host response in even greater detail. Finally, microbial proteomic analysis of sonicate fluid from patients with periprosthetic joint infection will be performed, to determine whether microbial proteomic analysis can differentiate S. aureus, S. epidermidis, S. lugdunensis, E. faecalis and S. agalactiae infections.

项目摘要 我们的团队已经制定了一项策略，使用以下方法对取出的骨科植入物表面的材料进行采样： 超声处理，并表明衍生的生物膜取样“超声处理液”具有任何标本的最高产量 用于微生物检测，并可进一步用于评估植入物表面的宿主反应。而 在临床上，对宿主反应的评估已被用于区分感染和非感染的骨科 植入物，迄今为止部署的基于宿主的方法过于简单，检测α-防御素或分析 例如，中性粒细胞数量，没有定义感染性关节成形术或潜在的微生物学 非感染性关节置换术失败的病因。我们假设，除了区分传染性和 非感染性关节置换术失败，使用蛋白质组学在超声处理液中捕获宿主反应 方法将辨别假体周围关节感染的微生物病因和非假体周围关节感染的个体原因。 感染性关节成形术失败。鉴于在假体周围关节感染和 定义非感染性关节置换术失败的个体原因，需要直接管理， 通过现有的诊断方法普遍实现，这将在临床上有用。 我们提出了有史以来第一个全球性的，公正的蛋白质组学分析的超声处理液，评估在体内人体免疫 与非感染性关节成形术失败及其子集相比，对感染性关节成形术失败的反应。我们进一步建议 假体周围关节感染超声处理液中微生物蛋白质的无偏倚全球分析。超声处理液 已在初步工作中被证明适合于所提出的蛋白质组学分析。超声处理液体， 失败的全膝关节置换术将使用液相色谱串联的新技术进行研究 质谱法以确定宿主蛋白质组学分析是否可以区分金黄色葡萄球菌， 表皮葡萄球菌、路邓葡萄球菌、粪肠球菌和无乳链球菌 假体周围关节感染和无菌性松动的非感染性关节置换术失败子集，假体周围 骨折、不稳定和骨质溶解/不良组织反应。除了质谱分析， 下一代基于测序的邻近延伸测定方法， 细胞因子和其他宿主蛋白将用于测量使用 质谱单独，允许更详细地检查宿主反应。最后，微生物 将对假体周围关节感染患者的超声处理液进行蛋白质组学分析，以确定 微生物蛋白质组学分析能否区分S.金黄色葡萄球菌、表皮葡萄球菌S. lugdunensis、艾美耳球虫E.粪肠球菌和 S.无乳感染。