A dual MMP9/MMP14 Axis Regulates Osteoclast Bone Resorptive Function

双 MMP9/MMP14 轴调节破骨细胞骨吸收功能

基本信息

批准号：
10663883
负责人：
STEPHEN J WEISS
金额：
$ 34.32万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10663883
关键词：
Acids Affect Aging Animal Experiments Animal Model Arbitration Binding Biological Assay Bone Diseases Bone Resorption Bone Surface Bone remodeling Caspase Cells Collagen Type I Complement Defect Deposition Development Disease Energy Metabolism Enzymes Event Extracellular Matrix Extracellular Matrix Degradation Family Galectin 3 Gene Expression Gene Expression Profiling Genetic Transcription Human In Vitro Knock-out Knockout Mice Light MMP14 gene MMP9 gene Macrophage Matrix Metalloproteinases Mediating Membrane Metabolic Metabolic Pathway Metabolism Metalloproteases Metastatic Neoplasm to the Bone Myelogenous Osteoclasts Osteogenesis Osteolysis Osteolytic Osteoporosis Osteoporotic Outcome Pathologic Peptide Hydrolases Phenotype Physiological Play Proteins Proteolysis Reporting Research Rheumatoid Arthritis Role Route Series Signal Transduction Surface System Therapeutic Therapeutic Intervention Transcription Repressor Transgenic Mice Zinc Fingers bone bone loss carbohydrate metabolism cathepsin K conditional knockout demineralization design improved in vitro activity in vivo insight loss of function mutation member mitochondrial metabolism monocyte mouse model new therapeutic target novel novel therapeutic intervention osteoclastogenesis palliative prevent programs screening seal side effect targeted treatment transcriptome tumor

项目摘要

A dual MMP9/MMP14 Axis Regulates Osteoclast Bone Resorptive Function Abstract Excessive osteoclast (OC) activity is responsible for a wide range of bone diseases, ranging from osteoporosis and rheumatoid arthritis to tumor-induced osteolysis. Despite the development of several anti-resorptive therapeutics, their palliative effects are often limited or accompanied by unwanted side effects. In this regard, cathepsin K has been targeted for therapeutic intervention given its key role as a collagenolytic enzyme capable of degrading type I collagen, the dominant protein component of the bone extracellular matrix. Interestingly, however, OCs also express matrix metalloproteinases whose function in OC bone-resorptive activity remains largely undefined. Herein, we utilized unbiased transcriptome screening as wells as ex vivo assays to identify MMP9 and MMP14 as the two dominant MMPs expressed by OCs. However, after generating Mmp9-/- or myeloid-specific Csf1r-Cre/Mmp14f/f OCs, preliminary studies suggest that neither proteinase plays a key role in bone resorption. Unexpectedly, we find that Csf1r-Cre/Mmp14f/f/Mmp9-/- double knockout OCs display major defects in bone resorption in vitro and in vivo. In an effort to define the mechanisms underlying a combined requirement for MMP9 and MMP14 in OC function, preliminary analysis of wild-type versus double knockout OC gene expression identified unexpected alterations in carbohydrate metabolism, zinc finger binding and mitochondrion. These findings led us to posit and confirm that working together, MMP9 and MMP14 play a critical role in regulating a zinc finger transcription repressor Zeb1-arbitrated metabolic pathway to govern OC activation in tandem with the ability of the MMPs to mediate bone collagenolytic effects. Thus, we propose to i) characterize the cooperative role of MMP9/MMP14 in controlling osteoclast function and bone resorption in vitro and in vivo, utilizing both myeloid- and OC-specific conditional knockout mice, ii) identify a novel MMP9/MMP14-Zeb1 axis in regulating energy metabolism and osteoclast activity in vitro and in vivo, and iii) Define the dual roles of the MMP9/MMP14 co-dependent proteolysis of galectin-3 membrane lattice as upstream of Zeb1, in parallel with bone type I collagenolysis in regulating osteoclast bone-resorptive function. The unique proteolytic and metabolic signaling route outlined in this proposal should provide new insights into OC-mediated bone remodeling, and advance the search for improved therapeutic strategies designed to prevent pathologic bone loss.

双 MMP9/MMP14 轴调节破骨细胞骨吸收功能抽象的破骨细胞 (OC) 过度活动会导致多种骨疾病，包括骨质疏松症和类风湿性关节炎到肿瘤引起的骨溶解。尽管开发了多种抗再吸收药物治疗，其姑息治疗效果往往有限或伴有不良副作用。对此，鉴于组织蛋白酶 K 作为一种胶原蛋白分解酶的关键作用，它已成为治疗干预的目标降解 I 型胶原蛋白，骨细胞外基质的主要蛋白质成分。有趣的是，然而，OC 也表达基质金属蛋白酶，其在 OC 骨吸收活性中的功能仍然存在很大程度上未定义。在此，我们利用无偏转录组筛选以及离体测定来鉴定 MMP9 和 MMP14 是 OC 表达的两个主要 MMP。然而，在生成 Mmp9-/- 或骨髓特异性 Csf1r-Cre/Mmp14f/f OC，初步研究表明，这两种蛋白酶都在骨吸收。出乎意料的是，我们发现Csf1r-Cre/Mmp14f/f/Mmp9-/-双敲除OCs显示主要体外和体内骨吸收缺陷。为了定义组合背后的机制 OC功能对MMP9和MMP14的要求，野生型与双敲除OC的初步分析基因表达发现碳水化合物代谢、锌指结合和线粒体。这些发现使我们假设并确认，MMP9 和 MMP14 共同发挥着关键作用。在调节锌指转录阻遏物 Zeb1 仲裁代谢途径以控制 OC 激活中的作用与 MMP 介导骨胶原溶解作用的能力相结合。因此，我们建议 i) 表征 MMP9/MMP14 在体外和体内控制破骨细胞功能和骨吸收中的协同作用，利用骨髓特异性和 OC 特异性条件敲除小鼠，ii) 鉴定新的 MMP9/MMP14-Zeb1 轴调节体外和体内的能量代谢和破骨细胞活性，以及 iii) 定义作为 Zeb1 上游的半乳糖凝集素 3 膜晶格的 MMP9/MMP14 共依赖性蛋白水解，与骨 I 型胶原溶解调节破骨细胞骨吸收功能。独特的蛋白水解和代谢该提案中概述的信号通路应该为 OC 介导的骨重塑提供新的见解，并且推进寻找旨在预防病理性骨质流失的改进治疗策略。