Differentially expressed genes in autosomal dominant osteopetrosis type II osteoclasts reveal known and novel pathways for osteoclast biology
Differentially expressed genes in autosomal dominant osteopetrosis type II osteoclasts reveal known and novel pathways for osteoclast biology
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DOI：
10.1038/labinvest.2013.140
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发表时间：
2014-03-01
影响因子：
5
通讯作者：
de Vernejoul, Marie-Christine
中科院分区：
文献类型：
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作者：
Coudert, Amelie E.;Del Fattore, Andrea;de Vernejoul, Marie-Christine
常染色体显性骨硬化症 II 型 (ADO II) 是一种罕见的遗传性骨病，其特征是骨量高和破骨细胞活性不足。据报道，CLCN7 基因突变可导致 ADO II。为了获得对 ADOII 破骨细胞失调途径的新见解，我们鉴定了具有 CLCN7 杂合突变的患者破骨细胞基因表达的变化。为此，我们进行了一项转录组学研究，比较了 ADO II 患者和健康捐赠者破骨细胞中的基因表达。我们的数据显示，根据我们的选择标准，患者和对照组的破骨细胞中有 182 个基因存在差异表达。从微阵列中显示最高变化的 18 个中，我们通过 qPCR 确认了 7 个的差异表达。虽然其中两个先前已被发现在破骨细胞中表达（ITGB5和SERPINE2），但其他五个（CES1（羧基酯酶1）、UCHL1（泛素羧基末端酯酶L1，也称为泛素硫酯酶）、WARS（色氨酰-tRNA合成酶）、GBP4（鸟苷酸结合蛋白4）和PRF1）尚不清楚。在此细胞类型中发挥作用。在蛋白质水平上，我们证实了 ITGB5 表达升高，WARS、PRF1 和 SERPINE2 表达降低。将携带 G215R 突变的 ClC-7 转染至破骨细胞中，导致 ITGB5 表达增加，PRF1 表达降低，具有临界意义。最后，我们观察到 ADO II 患者的骨形成标志物血清水平正常或升高，表明功能失调的破骨细胞和成骨细胞之间存在耦合。鞘氨醇激酶 1 mRNA 在 ADO II 和对照破骨细胞中表达相同水平。总之，这些数据表明，除了 CLCN7 突变引起的酸化功能障碍之外，ITGB5、PRF1、WARS 和 SERPINE2 表达的变化可能是 ADO II 破骨细胞表型的一部分。
Autosomal dominant osteopetrosis type II (ADO II) is a rare, heritable bone disorder characterized by a high bone mass and insufficient osteoclast activity. Mutations in the CLCN7 gene have been reported to cause ADO II. To gain novel insights into the pathways dysregulated in ADOII osteoclasts, we identified changes in gene expression in osteoclasts from patients with a heterozygous mutation of CLCN7. To do this, we carried out a transcriptomic study comparing gene expression in the osteoclasts of patients with ADO II and healthy donors. Our data show that, according to our selection criteria, 182 genes were differentially expressed in osteoclasts from patients and controls. From the 18 displaying the highest change in microarray, we confirmed differential expression for seven by qPCR. Although two of them have previously been found to be expressed in osteoclasts (ITGB5 and SERPINE2), the other five (CES1 (carboxyl esterase 1), UCHL1 (ubiquitin carboxy-terminal esterase L1, also known as ubiquitin thiolesterase), WARS (tryptophanyl-tRNA synthetase), GBP4 (guanylate-binding protein 4), and PRF1) are not yet known to have a role in this cell type. At the protein level, we confirmed elevated expression of ITGB5 and reduced expression of WARS, PRF1, and SERPINE2. Transfection of ClC-7 harboring the G215R mutation into osteoclasts resulted in an increased ITGB5 and reduced PRF1 expression of borderline significance. Finally, we observed that the ADO II patients presented a normal or increased serum level of bone formation markers, demonstrating a coupling between dysfunctional osteoclasts and osteoblasts. Sphingosine kinase 1 mRNA was expressed at the same level in ADO II and control osteoclasts. In conclusion, these data suggest that in addition to an acidification dysfunction caused by the CLCN7 mutation, a change in ITGB5, PRF1, WARS, and SERPINE2 expression could be part of the osteoclastic phenotype of ADO II.