Comparison of secretome from osteoblasts derived from sclerotic versus non-sclerotic subchondral bone in OA: A pilot study.
Comparison of secretome from osteoblasts derived from sclerotic versus non-sclerotic subchondral bone in OA: A pilot study.
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DOI：
10.1371/journal.pone.0194591
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Henrotin Y
中科院分区：
文献类型：
--
作者：
Sanchez C;Mazzucchelli G;Lambert C;Comblain F;DePauw E;Henrotin Y
骨关节炎（OA）的特征是软骨退化，但也有其他关节组织的变化，如软骨下骨硬化。本研究采用蛋白质组学方法比较了OA软骨下骨成骨细胞分泌组的差异。在nanoUPLC G2 HDMS系统上使用差异定量和相对无标记分析来分析分泌组。通过RT-PCR在来自其他5名患者的细胞培养物中定量表达更多差异分泌蛋白的mRNA。最后，骨调节蛋白和fibulin-3序列定量蛋白质印迹和免疫测定血清和培养上清液。在NSC成骨细胞分泌组中鉴定出175种蛋白质。数据可通过ProteomeXchange获得，标识符为PXD 008494。与NSC成骨细胞分泌组相比，12种蛋白质的分泌明显减少（骨调节蛋白、IGFBP5、VCAM-1、IGF2、78 kDa葡萄糖调节蛋白、多功能蛋白聚糖、钙调蛋白、IGFBP2、血小板反应蛋白-4、骨膜蛋白、网织红蛋白1和骨粘连蛋白），并且13种蛋白质由SC成骨细胞显著更多地分泌（CHI3L1、纤蛋白-3、SERPINE2、IGFBP6、SH3BGRL 3、SERPINE1、网织红蛋白3、α-2-HS-糖蛋白、TIMP-2、IGFBP3、TIMP-1、SERPINF 1、CSF-1）。观察到骨调节蛋白、IGF2、SERPINE1、fibulin-3和CHI3L1 mRNA水平的类似变化。ELISA测定证实，与NSC相比，SC成骨细胞上清液中的骨调节蛋白蛋白减少了一半，与健康受试者相比，OA患者血清中的骨调节蛋白减少了一半。与NSC相比，SC成骨细胞上清液中的Fibulin-3表位Fib3 - 1、Fib3 - 2和Fib3 - 3也增加。我们重点介绍了OA软骨下骨硬化成骨细胞差异分泌的一些蛋白质。这些变化有助于解释在OA软骨下骨中观察到的一些特征，如骨重建增加或骨基质矿化异常。在所鉴定的蛋白质中，发现骨调节蛋白在OA患者血清中减少，而fibulin-3在OA患者血清中增加。这些结果表明，骨调节蛋白和fibulin-3片段可能是生物标志物，以监测早期变化的软骨下骨代谢的OA。
Osteoarthritis (OA) is characterized by cartilage degradation but also by other joint tissues modifications like subchondral bone sclerosis. In this study, we used a proteomic approach to compare secretome of osteoblast isolated from sclerotic (SC) or non sclerotic (NSC) area of OA subchondral bone. Secretome was analyzed using differential quantitative and relative label free analysis on nanoUPLC G2 HDMS system. mRNA of the more differentially secreted proteins were quantified by RT-PCR in cell culture from 5 other patients. Finally, osteomodulin and fibulin-3 sequences were quantified by western blot and immunoassays in serum and culture supernatants. 175 proteins were identified in NSC osteoblast secretome. Data are available via ProteomeXchange with identifier PXD008494. Compared to NSC osteoblast secretome, 12 proteins were significantly less secreted (Osteomodulin, IGFBP5, VCAM-1, IGF2, 78 kDa glucose-regulated protein, versican, calumenin, IGFBP2, thrombospondin-4, periostin, reticulocalbin 1 and osteonectin), and 13 proteins were significantly more secreted by SC osteoblasts (CHI3L1, fibulin-3, SERPINE2, IGFBP6, SH3BGRL3, SERPINE1, reticulocalbin3, alpha-2-HS-glycoprotein, TIMP-2, IGFBP3, TIMP-1, SERPINF1, CSF-1). Similar changes in osteomodulin, IGF2, SERPINE1, fibulin-3 and CHI3L1 mRNA levels were observed. ELISAs assays confirm the decrease by half of osteomodulin protein in SC osteoblasts supernatant compared to NSC and in OA patients serum compared to healthy subjects. Fibulin-3 epitopes Fib3-1, Fib3-2 and Fib3-3 were also increased in SC osteoblasts supernatant compared to NSC. We highlighted some proteins differentially secreted by the osteoblasts coming from OA subchondral bone sclerosis. These changes contribute to explain some features observed in OA subchondral bone, like the increase of bone remodeling or abnormalities in bone matrix mineralization. Among identified proteins, osteomodulin was found decreased and fibulin-3 increased in serum of OA patients. These findings suggest that osteomodulin and fibulin-3 fragments could be biomarkers to monitor early changes in subchondral bone metabolism in OA.
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影响因子：
7
作者：
Silva, JC;Gorenstein, MV;Geromanos, SJ
通讯作者：
Geromanos, SJ
影响因子：
3.5
作者：
McLaughlin, Precious J.;Bakall, Benjamin;Marmorstein, Lihua Y.
通讯作者：
Marmorstein, Lihua Y.
影响因子：
27.4
作者：
COLLINS, DH;MCELLIGOTT, TF
通讯作者：
MCELLIGOTT, TF
影响因子：
7.6
作者：
Lucchini, M;Couble, ML;Farges, JC
通讯作者：
Farges, JC
影响因子：
81.5
作者：
Martel-Pelletier, Johanne;Barr, Andrew J.;Pelletier, Jean-Pierre
通讯作者：
Pelletier, Jean-Pierre