Pharmacological inhibition of protein S-palmitoylation suppresses osteoclastogenesis and ameliorates ovariectomy-induced bone loss.
Pharmacological inhibition of protein S-palmitoylation suppresses osteoclastogenesis and ameliorates ovariectomy-induced bone loss.
复制标题
DOI：
10.1016/j.jot.2023.06.002
复制
发表时间：
2023-09
影响因子：
6.6
通讯作者：
Chen, Jianquan
中科院分区：
文献类型：
--
作者：
Ma, Linghui;Zhang, Liwei;Liao, Zirui;Xiu, Chunmei;Luo, Xi;Luo, Na;Zhang, Lei;He, Guangxu;Chen, Jianquan
过多的破骨细胞形成扰乱了骨的动态平衡，从而显著地导致了与各种疾病相关的病理性骨丢失。蛋白质S-棕榈酰化是由棕榈酰基转移酶家族催化的可逆的翻译后脂质修饰，在多种生理和病理过程中发挥重要作用。然而，棕榈酰化在破骨细胞形成中的作用从未被探索过。因此，目前还不清楚这一过程是否可以有针对性地治疗主要由破骨细胞过度形成引起的溶骨性疾病。在本研究中，我们利用酰基-生物素交换(ABE)法研究了S-棕榈酰化蛋白在破骨细胞分化中的作用。用S棕榈酰化蛋白的药理抑制剂2-溴-帕米酸(2-BP)抑制小鼠骨髓来源的巨噬细胞的蛋白棕榈酰化，并通过TRAP染色、鬼臼苷染色、qPCR分析和凹坑形成实验，检测其对核因子κβ配体(RANKL)诱导的破骨细胞分化和活性的影响。我们还用μCT、H&E染色、TRAP染色和ELISA法评价了2-BP对去卵巢小鼠雌激素缺乏所致骨丢失和骨吸收的保护作用。此外，我们还进行了蛋白质印迹分析，以探讨2-BP抑制破骨细胞生成的分子机制。我们发现许多蛋白质在OCs分化过程中被棕榈酰化，药物抑制棕榈酰化抑制RANKL诱导的破骨细胞的形成、破骨细胞特异性基因的表达、F-肌动蛋白环的形成和体外破骨细胞的骨吸收，并在较小程度上阻碍MC3T3-E1细胞的成骨细胞形成。此外，我们在体内证明了2-BP对卵巢切除所致的骨质疏松症和骨吸收的保护作用。在机制上，我们发现2-BP处理部分通过下调c-Fos和NFATc1的表达来抑制破骨细胞的形成，而不明显影响RANKL诱导的破骨细胞生成AKT、MAPK和NF-κB通路的激活。药理抑制棕榈酰化在体外有效地抑制RANKL介导的破骨细胞分化，并在体内保护小鼠免受OVX诱导的骨质疏松。在机制上，棕榈酰化通过促进c-Fos和NFATc1的表达来调节破骨细胞的分化。因此，棕榈酰化在促进破骨细胞分化和活性方面起着关键作用，可能成为治疗骨质疏松症和其他破骨细胞相关疾病的潜在靶点。这篇文章的翻译潜力在于，我们首次揭示了棕榈酰化是调节破骨细胞分化的关键机制，从而为治疗溶骨性疾病提供了一个潜在的治疗靶点。本文的翻译潜力：本文的翻译潜力在于，我们首次揭示了棕榈酰化是调节破骨细胞分化的关键机制，从而为治疗溶骨性疾病提供了一个潜在的治疗靶点。
Excessive osteoclast formation disrupts bone homeostasis, thereby significantly contributing to pathological bone loss associated with a variety of diseases. Protein S-palmitoylation is a reversible post-translational lipid modification catalyzed by ZDHHC family of palmitoyl acyltransferases, which plays an important role in various physiological and pathological processes. However, the role of palmitoylation in osteoclastogenesis has never been explored. Consequently, it is unclear whether this process can be targeted to treat osteolytic bone diseases that are mainly caused by excessive osteoclast formation. In this study, we employed acyl-biotin exchange (ABE) assay to reveal protein S-palmitoylation in differentiating osteoclasts (OCs). We utilized 2-bromopalmitic acid (2-BP), a pharmacological inhibitor of protein S-palmitoylation, to inhibit protein palmitoylation in mouse bone marrow-derived macrophages (BMMs), and tested its effect on receptor activator of nuclear factor κβ ligand (RANKL)-induced osteoclast differentiation and activity by TRAP staining, phalloidin staining, qPCR analyses, and pit formation assays. We also evaluated the protective effect of 2-BP against estrogen deficiency-induced bone loss and bone resorption in ovariectomized (OVX) mice using μCT, H&E staining, TRAP staining, and ELISA assay. Furthermore, we performed western blot analyses to explore the molecular mechanism underlying the inhibitory effect of 2-BP on osteoclastogenesis. We found that many proteins were palmitoylated in differentiating OCs and that pharmacological inhibition of palmitoylation impeded RANKL-induced osteoclastogenesis, osteoclast-specific gene expression, F-actin ring formation and osteoclastic bone resorption in vitro, and to a lesser extent, osteoblast formation from MC3T3-E1 cells. Furthermore, we demonstrated that administration of 2-BP protected mice from ovariectomy-induced osteoporosis and bone resorption in vivo. Mechanistically, we showed that 2-BP treatment inhibited osteoclastogenesis partly by downregulating the expression of c-Fos and NFATc1 without overtly affecting RANKL-induced activation of osteoclastogenic AKT, MAPK, and NF-κB pathways. Pharmacological inhibition of palmitoylation potently suppresses RANKL-mediated osteoclast differentiation in vitro and protects mice against OVX-induced osteoporosis in vivo. Mechanistically, palmitoylation regulates osteoclast differentiation partly by promoting the expression of c-Fos and NFATc1. Thus, palmitoylation plays a key role in promoting osteoclast differentiation and activity, and could serve as a potential therapeutic target for the treatment of osteoporosis and other osteoclast-related diseases. The translation potential of this article is that we first revealed palmitoylation as a key mechanism regulating osteoclast differentiation, and therefore provided a potential therapeutic target for treating osteolytic bone diseases. The translation potential of this article: The translation potential of this article is that we first revealed palmitoylation as a key mechanism regulating osteoclast differentiation, and therefore provided a potential therapeutic target for treating osteolytic bone diseases.
登录
查看更多内容
影响因子：
3.7
作者：
Leong, Wai Fook;Zhou, Tielin;Lim, Gek Liang;Li, Baojie
通讯作者：
Li, Baojie
2-溴棕榈酸酯靶向视黄酸受体α并克服急性早幼粒细胞白血病的全反式视黄酸耐药性
影响因子：
10.1
作者：
Lu Y;Yan JS;Xia L;Qin K;Yin QQ;Xu HT;Gao MQ;Qu XN;Sun YT;Chen GQ
通讯作者：
Chen GQ
影响因子：
3.7
作者：
Song, I-Wen;Li, Wei-Ru;Chen, Yuan-Tsong
通讯作者：
Chen, Yuan-Tsong
DOI：
10.1073/pnas.1912671116
发表时间：
2019-12-17
影响因子：
11.1
作者：
Wegleiter, Thomas;Buthey, Kilian;Jessberger, Sebastian
通讯作者：
Jessberger, Sebastian
影响因子：
4
作者：
Li, Zhiwei;Cheng, Yuan;Jin, Xiangyun;Liu, Shenghe;Zhang, Chao
通讯作者：
Zhang, Chao