Restoration of Default Blood Monocyte-Derived Macrophage Polarization With Adalimumab But Not Etanercept in Rheumatoid Arthritis

Abstract

Introduction: We previously reported a specific defect of rheumatoid arthritis (RA) monocyte polarization to anti-inflammatory M2-like macrophages related to increased miR-155 expression in all RA patients except those receiving adalimumab (ADA). In this longitudinal study, we examined whether different tumor necrosis factor inhibitors were able to restore monocyte polarization to M2-like macrophages and their effect on the transcriptomic signature.

Methods: M2-like polarization induced by human serum AB was studied in 7 healthy donors and 20 RA patients included in the ABIRA cohort before and 3 months after starting ADA or etanercept (ETA). The differential gene expression of M2- and M1-related transcripts was studied in macrophage-derived monocytes after differentiation.

Results: At baseline, RA monocytes showed a defect of polarization to M2-like macrophages as compared with healthy donor monocytes, which was negatively correlated with disease activity. M2-like polarization from circulating monocytes was restored only with ADA and not ETA treatment. The transcriptomic signature demonstrated downregulation of M2-related transcripts and upregulation of M1-related transcripts in active RA. In patients receiving ADA, the transcriptomic signature of M2-related transcripts was restored.

Conclusion: This longitudinal study demonstrates that ADA but not ETA is able to restore the M2-like polarization of monocytes that is defective in RA.

Trial registration: ClinicalTrials.gov NCT02116504.

Keywords: Adalimumab; Etanercept; M2-like macrophages; monocyte-derived macrophages; rheumatoid arthritis.

Conflict of interest statement

XM has served on scientific advisory boards for BMS, Galapagos, GSK, Janssen, Novartis, Pfizer, and UCB. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2022 Paoletti, Ly, Bitoun, Nocturne, Rivière, Manson, Matucci, Pallardy, De Vries and Mariette.

Figures

Figure 1

M2-like macrophage polarization defect in rheumatoid arthritis (RA) monocytes under serum AB differentiation. The differentiation of frozen peripheral blood mononuclear cells (PBMCs) to M2-like macrophages was assessed in healthy donors (HDs) (n = 7) and RA patients (n = 20) by flow cytometry with anti-CD11b and anti-CD71 antibodies (A). Specific markers of M2-like macrophage polarization were assessed: CD206 (B) and CD163 (C). Spearman’s correlation analysis of macrophage CD206 expression in RA patients (geometric mean [gMFI]) and disease activity (Disease Activity Score in 28 joints [DAS28]) (D). Data are shown as symbols and mean ± SEM and were compared by Mann–Whitney t-test. ***p < 0.001 and ****p < 0.0001.

Figure 2

M2-like macrophage polarization defect in rheumatoid arthritis (RA) monocytes is restored with adalimumab (ADA) but not etanercept (ETA) treatment. The differentiation of frozen peripheral blood mononuclear cells (PBMCs) to M2-like macrophages was assessed in healthy donors (HDs) (n = 7) and RA patients at baseline and divided upstream between patients who would receive ADA (A–C) or ETA (D–F) (n = 20), RA at 3 months after ADA (n = 10, red dots), and RA at 3 months after ETA (n = 10, blue dots) by flow cytometry analysis using anti-CD11b and anti-CD71 antibodies (A, D). Specific markers of M2-like macrophage polarization were assessed: CD206 (B, E) and CD163 (C, F). Results are shown as symbols, lines, and mean ± SEM and were compared by Kruskal–Wallis test with Dunn’s multiple comparisons. *p < 0.05, **p < 0.01, and ***p < 0.001. ns, not significant.

Figure 3

Monocyte subpopulations and activation. Ex vivo CD14 and/or CD16 monocytes (A) and HLA-DR (B) expression in rheumatoid arthritis (RA) at baseline (n = 17), RA at 3 months after adalimumab (ADA) (n = 9), and RA at 3 months after ETA (n = 8) determined by flow cytometry analyses of frozen peripheral blood mononuclear cells (PBMCs with anti-HLA-DR, anti-CD14, anti-CD16, anti-CD2, anti-CD19, and anti-CD56 antibodies. Results are shown as symbols, lines, and mean ± SEM and were compared by Kruskal–Wallis test with Dunn’s multiple comparisons and the two-tailed nonparametric test.

Figure 4

The defect in M2-like polarization leads to an M1-like macrophage phenotype. Heatmap of a predicted upstream regulator effect on M2-like macrophages after monocyte polarization with serum AB (SAB) differentiation in rheumatoid arthritis-methotrexate (RA-MTX) patients versus healthy donors (HDs) or RA-adalimumab (RA-ADA) versus RA-MTX patients. The color of each square represents the activation z-score: activated = yellow (+5), inhibited = dark blue (−5) (A). Heatmap of the ratio of the normalized count of M2-related genes in RA-MTX patients versus HDs (HD = 1 yellow) (B) or RA-ADA patients versus RA-MTX patients (RA = 1 dark blue) (C). Ratio of normalized counts of M1-related genes in RA patients versus HDs (HD = 1 dark blue) (D) or RA-ADA patients versus RA patients (RA = 1 yellow) (E). MRC-1 gene = CD206 receptor.