Braking Macrophage Cell-Cell Fusion in the Foreign Body Response

在异物反应中阻止巨噬细胞细胞融合

• 批准号: 10373536
• 负责人: Mallika Ghosh
• 金额: $ 20.5万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-01 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10373536
• 关键词: Abnormal Cell; Actins; Address; Adhesions; Antibodies; Area; Arthritis; Artificial Implants; Bacterial Infections; Biocompatible Materials; Biological Process; Blocking Antibodies; Bone Marrow; Bone Surface; Cell Line; Cell Nucleus; Cell fusion; Cell membrane; Cell physiology; Cell surface; Cell-Cell Adhesion; Cells; Characteristics; Chimeric Proteins; Clustered Regularly Interspaced Short Palindromic Repeats; Cues; Cytoplasm; Cytoskeleton; Data; Defect; Disease; Dynamin; Electron Microscopy; Endocytosis; Endothelial Cells; Epitopes; Event; Failure; Foreign Bodies; Foreign-Body Giant Cells; Foreign-Body Reaction; Giant Cells; Hematopoietic; Human; Immune; Implant; In Vitro; Individual; Infection Control; Inflammatory; Integral Membrane Protein; Interleukin-13; Interleukin-4; Investigation; Knock-out; Knockout Mice; Lead; Mediating; Mediator of activation protein; Medical Device; Membrane; Membrane Fusion; Metalloproteases; Microscopy; Molecular; Monoclonal Antibodies; Mus; Myelogenous; Myeloid Cells; Normal Statistical Distribution; Osteoclasts; Osteogenesis; Osteolysis; Osteolytic; Pain; Parasitic infection; Pathologic; Pathology; Pathway interactions; Phenotype; Physiological Processes; Process; Prosthesis Loosenings; Proteins; Recycling; Replacement Arthroplasty; Scheme; Signal Pathway; Structure; System; Therapeutic; Therapeutic Intervention; Time; Tissues; Tumor Cell Invasion; Work; alanine aminopeptidase; bone; bone mass; cell type; cytokine; implant material; implantation; in vivo; macrophage; medical implant; membrane model; migration; mouse model; new therapeutic target; novel; particle; progenitor; protein expression; receptor mediated endocytosis; recruit; response; stem cells; syncytin; trafficking; wound healing

By virtue of their versatility and plasticity, macrophages have the ability to undergo phenotypic and functional changes in response to microenvironmental cues. One such change is the formation of multinucleated giant cells via intercellular fusion triggered by certain bacterial or parasitic infections or implantation of medical devices to induce a foreign body response. We and others have shown that the transmembrane protein CD13 is a multifunctional molecule that regulates diverse processes such as tumor cell invasion, immune cell trafficking, receptor mediated endocytosis and recycling and organization of the actin cytoskeleton. In our preliminary data, we demonstrate that despite a relatively normal distribution of hematopoietic components in bone marrow and periphery, CD13KO mice have reduced bone mass with increased osteoclast (OC) numbers per bone surface area but normal bone formation parameters. In vitro induction of CD13-deficient myeloid progenitors generated from bone marrow resulted in hyperfusion to generate multinucleated giant cells (MGCs) or OCs that were considerably larger in size, contained many more nuclei than those from wild type progenitors, suggesting that CD13 is a component of common fusion pathways shared by MGC and OC. We observed that while expression of the key fusion proteins, dynamin and DC-STAMP, are typically downregulated in mature cells post-fusion, expression of these proteins is sustained at high levels in multinucleated cells lacking CD13. Thus, CD13 acts as a brake to restrain a common cell-cell fusion pathway and may be a novel therapeutic target in pathological conditions mediated by abnormal cell fusion. In this proposal, we will identify the CD13-dependent mechanisms, molecules and signaling pathways involved in cell- cell fusion in macrophages undergoing giant cell fusion (Aim 1) and the potential for fusion-blocking anti-CD13 monoclonal antibodies as therapeutics in vitro and in vivo (Aim 2). We will use our extensive panels of wild type and CD13 knock out primary cells of mouse and human origin, CD13 blocking or activating antibodies, novel CRISPR-deleted CD13KO myeloid cell lines along with confocal and spinning disk microscopy and CD13 knockout mouse models to address these questions.

由于它们的多功能性和可塑性，巨噬细胞具有表型的能力 以及响应微环境线索的功能变化。一个这样的改变是 通过某些细菌或 寄生虫感染或植入医疗设备以诱导异物反应。我们 其他人则表明，跨膜蛋白CD13是多功能分子 调节各种过程，例如肿瘤细胞侵袭，免疫细胞运输，受体 肌动蛋白细胞骨架的介导的内吞和回收和组织。在我们的 初步数据，我们证明，尽管造血的分布相对正常 骨髓和外围的成分，CD13KO小鼠的骨骼质量减少了 每个骨表面积增加破骨细胞（OC）数量，但骨形成正常 参数。从骨骼产生的CD13缺陷髓样祖细胞的体外诱导 骨髓导致过度灌注产生多核巨细胞（MGC）或OCS 尺寸大得多，包含比野生型的核更大的核 祖细胞，表明CD13是共享的常见融合途径的组成部分 MGC和OC。我们观察到，虽然键融合蛋白的表达，但动力蛋白和 DC stamp通常在融合后成熟细胞中下调，这些表达 蛋白质在缺乏CD13的多核细胞中持续高水平。因此，CD13充当 制动以限制常见的细胞融合途径，可能是一个新型的治疗靶标 在病理条件下，由异常细胞融合介导。在此提案中，我们将确定 CD13依赖性机制，分子和信号通路参与细胞 - 巨噬细胞中经历巨细胞融合的巨噬细胞融合（AIM 1）和潜力 融合阻滞抗CD13单克隆抗体作为体外和体内的疗法（AIM 2）。我们将使用广泛的野生型面板和CD13敲除 鼠标和人类起源，CD13阻断或激活抗体，新颖的CRISPRETED CD13KO髓样细胞系以及共聚焦和旋转磁盘显微镜以及 CD13敲除鼠标模型以解决这些问题。