Core 1: Osteoimmunology Core

核心1：骨免疫学核心

• 批准号: 10247794
• 负责人: John Linforth Daiss
• 金额: $ 27.89万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-20 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10247794
• 关键词: 3D Print; Abscess; Animals; Antibiotics; Antibodies; Antigens; Behavior; Biological Assay; Biological Markers; Blood; Candidate Disease Gene; Cells; Clinical; Cloning; Collagen; Communicable Diseases; Custom; Defect; Development; Diagnostic; Electron Microscopy; Eligibility Determination; Engineering; Equus caballus; Future; Goals; Hematopoietic; Host Defense; Human; Immune; Immunoglobulin-Secreting Cells; Immunology; In Vitro; Infection; Institution; Intervention; Joint Prosthesis; Libraries; Local Therapy; Membrane; Methods; Microbial Biofilms; Modeling; Mus; Nanoporous; Natural History; Orthopedic Surgery; Osteocytes; Osteomyelitis; Outcome; Pathogenesis; Pathology; Patients; Process; Proteins; Proteome; Protocols documentation; Registries; Reproducibility; Research; Research Personnel; Research Project Grants; Resources; Safety; Sampling; Services; Staphylococcal Infections; Staphylococcus aureus; Staphylococcus aureus infection; Staphylococcus epidermidis; Swab; Technology; Tissue Banks; Tissues; Translating; Translational Research; Trauma; Whole Blood; assay development; bone; calcium phosphate; clinical diagnostics; cost efficient; experience; gene cloning; in vivo; innovative technologies; joint infection; macrophage; man; materials science; microbial; migration; multidisciplinary; mutant; new technology; novel; novel strategies; osteoimmunology; programs; scaffold; screening; septic; technology development; tool; transposon sequencing

Abstract The field of Osteoimmunology was established in 2005 to understand how bone influences immune and hematopoietic cells, as well as how hematopoietic and immune cells influence bone, in clinical conditions like prosthetic joint infection (PJI). However, critical tools and research protocols for discovery, and technologies required to translate discoveries into clinical diagnostics and interventions for osteomyelitis, did not exist at the commencement of our AO Trauma Clinical Priority Program (AO-CPP) 5 years ago. Thus, we assembled a multidisciplinary team of investigators with extensive experience in bone pathology, microbial pathogenesis, immunology, electron microscopy, material science & engineering, infectious disease and orthopaedic surgery, to enable the major advances that serve as the scientific premise of this Center of Research Translation on the Osteoimmunology of Bone Infection (CoRTOBI). Despite these major advances, additional assay and technology development is needed to complete the research projects in this CoRTOBI. Thus, the goals of the Osteoimmunology Research Core are to: 1) provide the CoRTOBI investigators with state-of-the-art research resources that do not exist elsewhere; and 2) develop novel technologies and assays for future osteoimmunology research. Our approaches to achieve these goals are embodied by the following Specific Aims. In Aim 1 we will process the samples and perform all of the electron microscopy on the infected bone and fabricated nanoporous membranes in Project 1; and the “Trojan horse” macrophages and tissue abscesses in Project 2. In Aim 2 we will generate the array of nanoporous membranes varying in pore sizes to assess the haptotaxis, metamorphic and durotaxis behaviors of S. aureus in vitro, and clone the genes involved as novel antibiotic targets using a candidate gene and TnSeq mutant library screening approach. In Aim 3 we will use our 3D-printing technology to assess novel antibiotic spacers in vivo in Project 1. In Aim 4 we will screen all the sera from the animal studies, and the 300 PJI patients in the AO-CPP registry, using our 14-antigen multiplex Luminex assay, and as technology development, we will add additional S. aureus antigens to our multiplex Luminex assay. In Project 2, we will also generate novel multiplex-Luminex assays for S. epidermidis and S. lugdunensis, and use them to screen sera from mice and PJI patients, as we have done for S. aureus. Finally, in Aim 5 we will establish protocols for screening and cloning antibody-secreting cells (ASC) in whole blood from mice and humans infected with S. aureus. We will also develop reproducible methods to expand and clone anti-S. aureus ASC directly from murine and human blood as new technology in the Core. Importantly, it is also our intent to share our resources with investigators outside of this CoRTOBI, and transport our research platforms to other institutions with a research focus on Staphylococcal infections, and osteomyelitis. !

摘要 骨免疫学领域成立于2005年，目的是了解骨骼如何影响免疫和 造血细胞，以及造血细胞和免疫细胞如何影响骨骼，在临床条件下，如 人工关节感染(PJI)。然而，用于发现的关键工具和研究协议以及技术 需要将发现转化为骨髓炎的临床诊断和干预措施，在 5年前，我们的AO创伤临床优先计划(AO-CPP)开始实施。因此，我们组装了一个 多学科研究团队，在骨骼病理学、微生物发病机制、 免疫学、电子显微镜、材料科学与工程、传染病和骨科外科， 为了使作为本研究翻译中心科学前提的主要进展能够 骨感染的骨免疫学(CoRTOBI)尽管取得了这些重大进展，但额外的化验和 需要技术开发来完成这个CoRTOBI中的研究项目。因此，世界银行的目标是 骨免疫学研究的核心是：1)为CoRTOBI研究人员提供最先进的研究 其他地方不存在的资源；以及2)为未来开发新技术和分析方法 骨免疫学研究。我们实现这些目标的方法具体体现在以下几个方面 目标。在目标1中，我们将对样本进行处理，并对感染的骨骼进行所有的电子显微镜检查。 并在项目1中制备了纳米孔膜；以及“特洛伊木马”巨噬细胞和组织 项目2中的脓肿。在目标2中，我们将生成孔径不同的纳米孔膜阵列以 评估金黄色葡萄球菌的体外趋触性、变态和趋多性行为，并克隆其基因 使用候选基因和TnSeq突变体库筛选方法作为新的抗生素靶点。在……里面 目标3在项目1中，我们将使用我们的3D打印技术在体内评估新型抗生素间隔物。 我们将筛选动物研究中的所有血清，以及AO-CPP登记中的300名PJI患者，使用我们的 14抗原多重Luminex检测，随着技术的发展，我们将增加更多的金黄色葡萄球菌 我们的多重Luminex检测的抗原。在项目2中，我们还将生成新的多重Luminex分析 用于表皮葡萄球菌和卢顿葡萄球菌，并使用它们来筛选小鼠和PJI患者的血清，就像我们所做的那样 金黄色葡萄球菌完蛋了。最后，在目标5中，我们将建立用于筛选和克隆抗体分泌的方案 感染金黄色葡萄球菌的小鼠和人的全血中的细胞(ASC)。我们还将开发可重现的 方法扩增、克隆抗S。直接从小鼠和人的血液中提取金黄色葡萄球菌作为新技术 核心。重要的是，我们也打算与CoRTOBI以外的调查人员共享我们的资源， 并将我们的研究平台转移到其他机构，专注于葡萄球菌感染的研究， 和骨髓炎。 好了！