Abstract
Objective
To examine the mechanism for the inhibited differentiation of osteoclasts in rheumatoid arthritis synovial CD14+ osteoclast precursors, the different expressions of the osteoclastogenesis-related genes in rheumatoid arthritis (RA) synovial fluid CD14+ osteoclast precursors were compared with those of normal peripheral blood (PB) CD14+ osteoclast precursors.
Conclusion
The expression of the osteoclastogenesis-re-lated genes in RA synovial CD14+ osteoclast precursors is different from that of the normal PB CD14+ osteoclast precursors. These results suggest that the different expression of osteoclastogenesis-related genes might be involved in the altered osteoclastogenesis in RA synovial osteoclast precursors.
References
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Figure 1.
Healthy volunteer PB and RA synovial CD14+ cells were cultured with M-CSF (20 ng/mL) for 2 days, and then with RANKL (40 ng/mL) and M-CSF (20 ng/mL) for an additional 6, 9 or 12 days. The cells were stained for TRAP expression. TRAP-positive multinucleated (>3 nuclei/cell) cells were counted as osteoclasts. ∗p<0.05 versus RA 6 days.
![jrd-18-11f1.tif](/upload/SynapseXML/1010jrd/thumb/jrd-18-11f1.gif)
Figure 2.
The array data for gene expression was validated by performing quantitative real-time PCR in Healthy volunteer PB and RA synovial CD14+ cells. ∗p<0.05 versus Healthy volunteer PB.
![jrd-18-11f2.tif](/upload/SynapseXML/1010jrd/thumb/jrd-18-11f2.gif)
Table 1.
Differentially expressed mRNAs in the RA synovi CD14+ cells compared to that of the normal PB CD14+ cel (Cytokines and chemokines involved in osteoclastogenesis)
Gene name | Osteoclast differentiation | Fold change | p-value |
---|---|---|---|
BMP2 | Stimulation | ND | ND |
CCL2 (MCP-1)∗ | Stimulation | 8.5 | 0.02 |
CCL3 (MIP-1α) | Stimulation | 1.89 | 0.4 |
CCL4 (MIP-1β) | Stimulation | ND | ND |
CX3CL1 | Stimulation | ND | ND |
CXCL10 | Stimulation | 2.98 | 0.29 |
CXCL11 | Inhibition | ND | ND |
CXCL12 | Stimulation | 2.98 | 0.29 |
GM-CSF | Inhibition/Stimulation | ND | ND |
IFNα | Inhibition | ND | ND |
IFNβ | Inhibition | ND | ND |
IFNg | Inhibition | ND | ND |
IL-10 | Inhibition | −1.13 | 0.68 |
IL-12A | Inhibition | ND | ND |
IL-12B | Inhibition | ND | ND |
IL-17A | Stimulation | ND | ND |
IL-18 | Inhibition | −1.13 | 0.74 |
IL-1β | Inhibition/Stimulation | 1.07 | 1 |
IL-27 | Inhibition | −1.09 | 0.82 |
IL-32∗ | Stimulation | 3.04 | 0.05 |
IL-4 | Inhibition | ND | ND |
IL-6 | Stimulation | ND | ND |
MIF∗ | Inhibition | 2.34 | 0.04 |
TGFβ | Inhibition/Stimulation | ND | ND |
TNFα | Stimulation | 1.43 | 0.63 |
TNFSF11A (RANKL) | Stimulation | ND | ND |
Table 2.
Differentially expressed mRNAs in the RA synovia CD14+ cells compared to that of the normal PB CD14+ cell (Receptors and membrane factors involved in osteoclastogenesis
Gene name | Osteoclast differentiation | Fold change | p-value |
---|---|---|---|
ATP6v0d2 | Positive | ND | ND |
CSF1R∗ | Positive | 1.77 | 0.01 |
FCER1G (FcRγ) | Positive | −1.03 | 0.99 |
ITGAM (CD11b) | Positive | 1.28 | 0.22 |
ITGB3 (CD61) | Positive | ND | ND |
LILRB1 | Negative | 1.33 | 0.11 |
LILRB3 | Negative | 1.14 | 0.67 |
LILRB4∗ | Negative | 2.87 | 0 |
MSR1 (SR-A)∗ | Positive | 4.84 | 0 |
OSCAR | Positive | −1.21 | 0.71 |
Plexin-A1 | Positive | 2.16 | 0.13 |
TM7SF4 (DC STAMP) | Positive | ND | ND |
TNFR11A (RANK) | Positive | ND | ND |
TREM2∗ | Positive | 1.59 | 0.04 |
TYROBP (DAP12) | Positive | −1.29 | 0.54 |
Table 3.
Differentially expressed mRNAs in the RA synovial CD14+ cells compared to that of the normal PB CD14+ cells (Intracellular molecules involved in osteoclastogenesis))
Gene name Osteo | oclast differentia | tion Fold change | p-value |
---|---|---|---|
Bcl-2∗ | Positive | 2.83 | 0.01 |
Btk | Positive | −1.37 | 0.36 |
C/EBPβ | Negative | 1.39 | 0.38 |
CaMKIV | Positive | ND | ND |
c-Fos | Positive | −1.86 | 0.06 |
c-Jun | Positive | 1.29 | 0.54 |
DYRK1A | Negative | ND | ND |
FOSL1 (Fra-1) | Positive | ND | ND |
Gab2 | Positive | −1.29 | 0.24 |
Id1 | Negative | ND | ND |
Id2 | Negative | 1.34 | 0.56 |
Id3 | Negative | 2.59 | 0.06 |
IKBKB (IKK-β) | Positive | ND | ND |
MafB∗ | Negative | 2.63 | 0.01 |
MITF | Positive | 1.38 | 0.29 |
NFATc1 | Positive | −1.01 | 1 |
NFKB1 (NFκ B p50) | Positive | 1.38 | 0.15 |
NFKB2 (NFκ B p52) | Positive | −1.02 | 0.99 |
MAP3K14 (NIK) | Positive | −1.19 | 0.21 |
PIAS3 | Negative | −1.08 | 0.39 |
PLCγ2 | Positive | −1.13 | 0.09 |
PU.1 | Positive | 1.02 | 0.97 |
INPP5D (SHIP)∗ | Negative | −1.67 | 0.01 |
PTPN6 (SHP-1) | Negative | ND | ND |
SOCS1 | Positive | 1.76 | 0.14 |
SOCS3 | Positive | 1.07 | 0.74 |
STAT1 | Negative | 1.49 | 0.49 |
Syk | Positive | 1.08 | 0.8 |
MAP3K7 (TAK1) | Positive | 1.18 | 0.36 |
Tec | Positive | 1.08 | 0.22 |
TRAF6 | Positive | 1.06 | 0.54 |