The Role of MMP13 in Multiple Myeloma Bone Disease

MMP13 在多发性骨髓瘤骨病中的作用

• 批准号: 8968822
• 负责人: Suzanne Lentzsch
• 金额: $ 33.11万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-04 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8968822
• 关键词: Achievement; Active Sites; Biological Assay; Bone Diseases; Bone Marrow; Bone Resorption; Bone remodeling; Cell Nucleus; Cell Size; Cell fusion; Cells; Coculture Techniques; Defect; Dendritic Cells; Detection; Development; Disease; Electrophoretic Mobility Shift Assay; Endopeptidases; Enzyme-Linked Immunosorbent Assay; Exhibits; Extracellular Matrix; Extracellular Matrix Degradation; FOS gene; Family; Figs - dietary; Genetic Transcription; Goals; Health; Hematologic Neoplasms; In Vitro; Integral Membrane Protein; Interleukin-6; JUN gene; Lesion; Lytic; Lytic Lesion; Lytic Metastatic Lesion; Malignant Neoplasms; Matrix Metalloproteinases; Mediating; Modeling; Mononuclear; Multiple Myeloma; Mus; Neoplasm Metastasis; Nuclear; Osteoclasts; Osteogenesis; Osteolytic; Pathway interactions; Patients; Plasma Cells; Play; Process; Protein Overexpression; Proteins; Role; STAT3 gene; Serum; Stromal Cells; Testing; Time; Tissue Array Analysis; Transcription Factor AP-1; Tumor Cell Invasion; Tumor Tissue; Up-Regulation; Work; autocrine; bone; bone mass; collagenase 3; in vivo; insight; knock-down; neutralizing antibody; novel; novel strategies; novel therapeutics; paracrine; recombinase; repaired; response; substantia spongiosa; tumor; tumor growth

DESCRIPTION (provided by applicant): Multiple myeloma (MM) is characterized by increased osteoclast (OCL) activity that results in bone destruction and purely lytic lesions in ~80% patients. The excessive bone resorption is mediated by OCLs, which are activated by factors secreted by MM cells and their microenvironment. This is supported by the finding that MM cells are always located in close association with sites of active bone resorption. Matrix metalloproteinase 13 (MMP13) belongs to a family of endopeptidases capable of degrading and remodeling all extracellular matrix (ECM) components. We recently found that secretion of MMP13 by MM cells is up to 400-fold higher than other MMPs. Primary tissue array analysis showed that MMP13 is highly expressed in MM cells, but not in normal plasma cells. ELISA of MM patient sera revealed a 100% correlation between detection of MMP13 protein and bone disease while MMP13 was undetectable in healthy donors. In vitro MMP13 increased bone resorption, where its enhancing effects were associated with dramatically increased OCL size and nuclear number/OCL, suggesting that MMP13 induces fusion of OCL precursors. Further, OCL generated from Mmp13-/- mice showed a significantly decreased number of nuclei and average OCL cell size and bone resorption capacity compared to WT mice. Addition of MMP13 reversed the fusion defect of Mmp13-/- MNCs. Further, MMP13 was strongly upregulated in MM cells in response to IL-6 and EMSA revealed that IL-6-mediated AP-1 activation promoted MMP13 transcription. Dendritic cell-specific transmembrane protein (DC-STAMP), essential for cell-cell fusion of preosteoclasts, was upregulated by exogenous MMP13. Taken together, we hypothesize that myeloma cell-derived MMP13 plays a pivotal role in MM-induced bone destruction. We contend that the upregulation of IL-6 by the MM microenvironment triggers the secretion of MMP13 in MM cells. In turn, MMP13 induces DC-STAMP, resulting in increased OCL activity, bone resorption and ECM degradation. As such, targeting MMP13 may represent an effective and new approach to treat myeloma bone disease (MMBD). To test these hypotheses, we will 1) investigate the autocrine and paracrine mechanisms of MMP13 induction in MM cells, 2) investigate the mechanisms of enhancing of OCL formation and activity by MMP13 and 3) confirm in vivo the role of MMP13 in the development of myeloma-induced osteolytic bone lesions. A successful completion of this work will be crucial for the achievement of our overall goal to identify novel therapies for myeloma bone disease.

描述（由申请人提供）：多发性骨髓瘤 (MM) 的特点是破骨细胞 (OCL) 活性增加，导致约 80% 的患者出现骨质破坏和纯粹的溶解性病变。过度骨吸收是由 OCL 介导的，OCL 是由 MM 细胞及其微环境分泌的因子激活的。这一发现得到了支持，即 MM 细胞总是与活跃的骨吸收部位密切相关。 基质金属蛋白酶 13 (MMP13) 属于肽链内切酶家族，能够降解和重塑所有细胞外基质 (ECM) 成分。我们最近发现MM细胞分泌的MMP13比其他MMP高出400倍。原代组织芯片分析表明，MMP13 在 MM 细胞中高表达，但在正常浆细胞中不表达。 MM 患者血清 ELISA 显示 MMP13 蛋白检测与骨病之间存在 100% 相关性，而在健康供体中检测不到 MMP13。体外 MMP13 增加骨吸收，其增强作用与显着增加的 OCL 大小和核数量/OCL 相关，表明 MMP13 诱导 OCL 前体的融合。此外，与 WT 小鼠相比，Mmp13-/- 小鼠产生的 OCL 显示出细胞核数量、平均 OCL 细胞大小和骨吸收能力显着减少。添加 MMP13 逆转了 Mmp13-/- MNC 的融合缺陷。此外，MMP13 在 MM 细胞中响应 IL-6 强烈上调，EMSA 揭示 IL-6 介导的 AP-1 激活促进 MMP13 转录。树突状细胞特异性跨膜蛋白 (DC-STAMP) 对于破骨细胞前细胞与细胞的融合至关重要，可被外源 MMP13 上调。 综上所述，我们假设骨髓瘤细胞衍生的 MMP13 在 MM 诱导的骨破坏中发挥关键作用。我们认为，MM 微环境对 IL-6 的上调触发了 MM 细胞中 MMP13 的分泌。反过来，MMP13 诱导 DC-STAMP，导致 OCL 活性增加、骨吸收和 ECM 降解。因此，靶向 MMP13 可能代表一种治疗骨髓瘤骨病 (MMBD) 的有效新方法。为了检验这些假设，我们将 1) 研究 MM 细胞中 MMP13 诱导的自分泌和旁分泌机制，2) 研究 MMP13 增强 OCL 形成和活性的机制，3) 体内确认 MMP13 在骨髓瘤引起的溶骨性骨病变。这项工作的成功完成对于实现我们确定骨髓瘤骨病新疗法的总体目标至关重要。