Core 1: Osteoimmunology Core

核心1：骨免疫学核心

• 批准号: 9370635
• 负责人: John Linforth Daiss
• 金额: $ 29.87万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9370635
• 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; Natural History; Orthopedic Surgery procedures; Osteocytes; Osteomyelitis; Outcome; Pathogenesis; Pathology; Patients; Printing; Process; Proteins; Proteome; Protocols documentation; Registries; Reproducibility; Research; Research Personnel; Research Project Grants; Resources; Safety; Sampling; Services; Staphylococcal Infections; Staphylococcus aureus; 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

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. !

抽象的 骨免疫学领域成立于 2005 年，旨在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 假体关节感染（PJI）。然而，发现和技术的关键工具和研究协议 将发现转化为骨髓炎的临床诊断和干预措施所需的技术，在当时并不存在 我们的 AO 创伤临床优先计划 (AO-CPP) 于 5 年前开始。因此，我们组装了一个 多学科研究团队在骨病理学、微生物发病机制、 免疫学、电子显微镜、材料科学与工程、传染病与骨科、 实现作为该研究翻译中心的科学前提的重大进展 骨感染的骨免疫学（CoRTOBI）。尽管取得了这些重大进展，但额外的检测和 完成该 CoRTOBI 中的研究项目需要技术开发。因此，该项目的目标是 骨免疫学研究核心是：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； 2）为未来开发新技术和检测方法 骨免疫学研究。我们实现这些目标的方法体现为以下具体措施 目标。在目标 1 中，我们将处理样本并对受感染的骨骼进行所有电子显微镜检查 项目1制备纳米多孔膜；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多孔膜阵列，以 体外评估金黄色葡萄球菌的趋触性、变态性和趋向性行为，并克隆基因 使用候选基因和 TnSeq 突变体库筛选方法作为新型抗生素靶点。在 目标 3，我们将在项目 1 中使用我们的 3D 打印技术在体内评估新型抗生素间隔物。目标 4 我们将使用我们的方法对动物研究中的所有血清以及 AO-CPP 登记处的 300 名 PJI 患者进行筛查 14抗原多重Luminex检测，随着技术的发展，我们将添加额外的金黄色葡萄球菌 我们的多重 Luminex 检测的抗原。在项目 2 中，我们还将生成新型多重 Luminex 检测方法 表皮葡萄球菌和路邓葡萄球菌，并用它们筛选小鼠和 PJI 患者的血清，正如我们所研究的 为金黄色葡萄球菌完成。最后，在目标 5 中，我们将建立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类全血中的细胞（ASC）。我们还将开发可复制的 扩展和克隆抗-S的方法。作为新技术，直接从小鼠和人类血液中提取金黄色葡萄球菌 ASC 核心。重要的是，我们还打算与 CoRTOBI 之外的研究人员共享我们的资源， 并将我们的研究平台转移到其他以葡萄球菌感染为研究重点的机构， 和骨髓炎。 ！