Connectivity Map-based discovery of parbendazole reveals targetable human osteogenic pathway
Connectivity Map-based discovery of parbendazole reveals targetable human osteogenic pathway
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DOI：
10.1073/pnas.1501597112
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发表时间：
2015-10-13
影响因子：
11.1
通讯作者：
van Leeuwen, Johannes P. T. M.
中科院分区：
文献类型：
--
作者：
Brum, Andrea M.;van de Peppel, Jeroen;van Leeuwen, Johannes P. T. M.
骨质疏松症是一种常见的骨骼疾病，其特征在于低骨量导致骨脆性增加和骨折易感性增加。在这项研究中，我们已经确定了刺激人间充质基质细胞（hMSCs）分化成骨细胞的途径。在向成骨细胞分化的hMSC中进行基因表达谱分析（在6小时）。通过计算机模拟分析了显著调控的基因，并使用连接图（CMap）来鉴定候选骨刺激化合物。帕苯达唑的特征与观察到的成骨hMSC的表达变化相匹配。帕苯达唑刺激成骨细胞分化，如在对帕苯达唑的凋亡作用有抵抗力的hMSC群体的亚群中碱性磷酸酶活性增加、矿化和骨标记基因（碱性磷酸酶/ALPL、骨桥蛋白/SPP 1和骨唾液蛋白II/IBSP）上调所示。这些成骨作用不依赖于糖皮质激素，因为帕苯达唑不上调糖皮质激素受体（GR）靶基因，也不受GR拮抗剂米非司酮的抑制。帕苯达唑引起深刻的细胞骨架变化，包括微管降解和局灶性粘连增加。紫杉醇预处理稳定微管抑制成骨细胞分化。帕苯达唑上调骨形态发生蛋白2（BMP-2）基因表达和活性。与BMP-2拮抗剂DMH 1的共处理限制，但不阻断，帕苯达唑诱导的矿化。使用CMap，我们已经确定了一个以前未确定的谱系特异性，骨合成代谢化合物，帕苯达唑，诱导成骨分化，通过细胞骨架的变化和增加BMP-2活性的组合。
Osteoporosis is a common skeletal disorder characterized by low bone mass leading to increased bone fragility and fracture susceptibility. In this study, we have identified pathways that stimulate differentiation of bone forming osteoblasts from human mesenchymal stromal cells (hMSCs). Gene expression profiling was performed in hMSCs differentiated toward osteoblasts (at 6 h). Significantly regulated genes were analyzed in silico, and the Connectivity Map (CMap) was used to identify candidate bone stimulatory compounds. The signature of parbendazole matches the expression changes observed for osteogenic hMSCs. Parbendazole stimulates osteoblast differentiation as indicated by increased alkaline phosphatase activity, mineralization, and up-regulation of bone marker genes (alkaline phosphatase/ALPL, osteopontin/SPP1, and bone sialoprotein II/IBSP) in a subset of the hMSC population resistant to the apoptotic effects of parbendazole. These osteogenic effects are independent of glucocorticoids because parbendazole does not up-regulate glucocorticoid receptor (GR) target genes and is not inhibited by the GR antagonist mifepristone. Parbendazole causes profound cytoskeletal changes including degradation of microtubules and increased focal adhesions. Stabilization of microtubules by pretreatment with Taxol inhibits osteoblast differentiation. Parbendazole up-regulates bone morphogenetic protein 2 (BMP-2) gene expression and activity. Cotreatment with the BMP-2 antagonist DMH1 limits, but does not block, parbendazole-induced mineralization. Using the CMap we have identified a previously unidentified lineage-specific, bone anabolic compound, parbendazole, which induces osteogenic differentiation through a combination of cytoskeletal changes and increased BMP-2 activity.