Regulation of Osteoclast Differentiation at Multiple Stages by Protein Kinase D Family Kinases
Regulation of Osteoclast Differentiation at Multiple Stages by Protein Kinase D Family Kinases
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DOI：
10.3390/ijms21031056
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发表时间：
2020-02-01
影响因子：
5.6
通讯作者：
Jensen, Eric D.
中科院分区：
文献类型：
--
作者：
Leightner, Amanda C.;Meyers, Carina Mello Guimaraes;Jensen, Eric D.
平衡的破骨细胞和成骨细胞活性对于骨骼健康是必要的，而不平衡的破骨细胞活性在许多骨骼条件下导致骨丢失。更好地了解调节破骨细胞分化和活性的途径对于开发新的治疗方法以更好地管理骨吸收是必要的。蛋白激酶D（PKD）家族丝氨酸/苏氨酸激酶在破骨细胞中的作用尚未得到很好的表征。在这项研究中，我们使用免疫荧光分析揭示，PKD 2和PKD 3，在破骨细胞中表达的亚型，被发现在细胞核和细胞质，有丝分裂纺锤体和中间体，并与肌动蛋白带。我们发现PKD抑制剂CRT 0066101和CID 755673抑制破骨细胞形成的几个不同方面。用较低剂量的PKD抑制剂处理骨髓巨噬细胞对M-CSF + RANKL依赖性诱导成定向破骨细胞前体的影响很小，但抑制其运动性和随后分化成多核成熟破骨细胞，而较高剂量的PKD抑制剂诱导前破骨细胞凋亡。用抑制剂处理融合后的多核破骨细胞破坏了破骨细胞的肌动蛋白带，损害了它们的吸收活性。总之，这些数据暗示PKD激酶作为破骨细胞的正调节因子，其在其形成和功能的多个不同过程中是必不可少的。
Balanced osteoclast and osteoblast activity is necessary for skeletal health, whereas unbalanced osteoclast activity causes bone loss in many skeletal conditions. A better understanding of pathways that regulate osteoclast differentiation and activity is necessary for the development of new therapies to better manage bone resorption. The roles of Protein Kinase D (PKD) family of serine/threonine kinases in osteoclasts have not been well characterized. In this study we use immunofluorescence analysis to reveal that PKD2 and PKD3, the isoforms expressed in osteoclasts, are found in the nucleus and cytoplasm, the mitotic spindle and midbody, and in association with the actin belt. We show that PKD inhibitors CRT0066101 and CID755673 inhibit several distinct aspects of osteoclast formation. Treating bone marrow macrophages with lower doses of the PKD inhibitors had little effect on M-CSF + RANKL-dependent induction into committed osteoclast precursors, but inhibited their motility and subsequent differentiation into multinucleated mature osteoclasts, whereas higher doses of the PKD inhibitors induced apoptosis of the preosteoclasts. Treating post-fusion multinucleated osteoclasts with the inhibitors disrupted the osteoclast actin belts and impaired their resorptive activity. In conclusion, these data implicate PKD kinases as positive regulators of osteoclasts, which are essential for multiple distinct processes throughout their formation and function.