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种策略，该策略可以在删除的骨科直接的表面上采样材料 超声处理并表明衍生的生物膜采样“ Sonicate Fluids”的产量最高 用于微生物检测的类型，还可以用于评估植入物表面的宿主反应。尽管 对宿主反应的评估已在临床上杠杆化，以区分未感染的骨科 植入物，迄今为止部署的基于宿主的方法很简单，检测α-防御素或分析 中性粒细胞数量，例如，没有定义感染性节肢动物或基础的微生物学 非感染性关节置换术衰竭的病因。我们假设，除了使感染力与 非感染性关节置换术衰竭，使用蛋白质组学捕获宿主反应 方法将识别周围关节感染的微生物病因和非 - 感染性关节置换术失败。考虑到周围关节感染中的病因生物的检测和 定义非感染性关节置换术失败的个人原因，直接管理不需要 通过额外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无偏的蛋白质组学分析，评估了体内人体免疫 与非感染性关节置换术及其子集相比，对传染性的反应。我们进一步提出建议 对周围关节感染中的Sonicate液中微生物蛋白的无偏全球分析。液体 在初步工作中已显示适合提出的蛋白质组学分析。从中使用液体 使用液相色谱串联的新技术将研究总膝关节碎裂的失败。 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， 葡萄球菌表皮，lugdunensis，肠球菌和肠链球菌agalactiae 人假体的关节感染和无菌松动，周围的无感染关节置换术子集子集 断裂，不稳定性和骨溶解/不良组织反应。除了质谱法， 基于下一代测序的接近扩展测定方法，测量3,072的水平 细胞因子和其他宿主蛋白将用于测量低抽象的人类蛋白质遗漏 仅质谱法，就可以更详细地检查宿主反应。最后，微生物 将进行对牙周假体关节感染患者的Sonate液体的蛋白质组学分析，以确定 微生物蛋白质组学分析是否可以区分金黄色葡萄球菌，表皮链球菌，S。lugdunensis，E。Faecalis和 S. agalactiae感染。