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：
10202488
负责人：
STEPHEN J WEISS
金额：
$ 33.29万
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/10202488
关键词：
Acids Affect Aging 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 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 macrophage 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过度骨细胞（OC）活性导致多种骨骼疾病，范围从骨质疏松症和类风湿关节炎，可导致肿瘤诱导的骨溶解。尽管发展了几种反标题治疗剂，它们的姑息作用通常受到限制或伴有不必要的副作用。在这方面，鉴于其作为胶原式酶的关键作用，组织蛋白酶K已成为治疗干预的目标 I型胶原蛋白的降解，这是骨外基质的主要蛋白质成分。有趣的是，然而，OC还表达基质金属蛋白酶，其在OC骨头敏感活性中的功能仍然存在在很大程度上不确定。在此，我们利用了无偏的转录组筛选以及井以及离体测定法来识别 MMP9和MMP14作为OC表示的两个主要MMP。但是，生成MMP9 - / - 或髓样特异性CSF1R-CRE/MMP14F/F OC，初步研究表明，蛋白酶在骨吸收。出乎意料的是，我们发现CSF1R-CRE/MMP14F/F/MMP9 - / - 双敲门OCS显示专业体外和体内骨吸收的缺陷。为了定义组合的基础机制 OC功能中MMP9和MMP14的需求，野生型与双基因敲除OC的初步分析基因表达确定了碳水化合物代谢，锌指结合和线粒体。这些发现使我们提出并确认合作，MMP9和MMP14起着至关重要的作用在调节锌指转录阻遏物Zeb1屈光者的代谢途径中的作用来控制OC激活与MMP介导骨胶原式作用的能力同时。因此，我们向i）提出特征 MMP9/MMP14在控制破骨细胞功能和体外和体内骨吸收中的合作作用，利用髓样和OC特异性条件敲除小鼠，ii）确定新型MMP9/MMP14-ZEB1轴在体外和体内调节能量代谢和破骨细胞活性时，以及iii）定义了双重作用 MMP9/MMP14与Zeb1上游的半乳糖素-3膜晶格的共依赖性蛋白水解与骨I型胶原分解在调节破骨细胞骨头敏化功能方面。独特的蛋白水解和代谢该提案中概述的信号通道应为OC介导的骨骼重塑提供新的见解，并提供提高旨在防止病理骨质流失的改进治疗策略的搜索。