Liposomal antagomiR-155-5p restores anti-inflammatory macrophages and improves arthritis in preclinical models of rheumatoid arthritis

Objective: We previously reported an increased expression of microRNA‐155 (miR‐155) in the blood monocytes of patients with rheumatoid arthritis (RA) that could be responsible for impaired monocyte polarization to anti‐inflammatory M2‐like macrophages. In this study, we employed two preclinical models of RA, collagen‐induced arthritis and K/BxN serum transfer arthritis, to examine the therapeutic potential of antagomiR‐155‐5p entrapped within PEGylated (polyethylene glycol [PEG]) liposomes in resolution of arthritis and repolarization of monocytes towards the anti‐inflammatory M2 phenotype. Methods: AntagomiR‐155‐5p or antagomiR‐control were encapsulated in PEG liposomes of 100 nm in size and −10 mV in zeta potential with high antagomiR loading efficiency (above 80%). Mice were injected intravenously with 1.5 nmol/100 μL PEG liposomes containing antagomiR‐155‐5p or control after the induction of arthritis. Results: We demonstrated the biodistribution of fluorescently tagged PEG liposomes to inflamed joints one hour after the injection of fluorescently tagged PEG liposomes, as well as the liver's subsequent accumulation after 48 hours, indicative of hepatic clearance, in mice with arthritis. The injection of PEG liposomes containing antagomiR‐155‐5p decreased arthritis score and paw swelling compared with PEG liposomes containing antagomiR‐control or the systemic delivery of free antagomiR‐155‐5p. Moreover, treatment with PEG liposomes containing antagomiR‐155‐5p led to the restoration of bone marrow monocyte defects in anti‐inflammatory macrophage differentiation without any significant functional change in other immune cells, including splenic B and T cells. Conclusion: The injection of antagomiR‐155‐5p encapsulated in PEG liposomes allows the delivery of small RNA to monocytes and macrophages and reduces joint inflammation in murine models of RA, providing a promising strategy in human disease.imag

Serum IL-33, a new marker predicting response to rituximab in rheumatoid arthritis

Background. Recent works have suggested a possible link between IL-33 and B-cell biology. We aimed to study in different cohorts and with an accurate ELISA assay the possible association between serum IL-33 detection and response to rituximab (RTX) in rheumatoid arthritis (RA) patients. Method. Serum IL-33, rheumatoid factor (RF), anti-citrullinated cyclic peptide antibodies (anti-CCP), high serum IgG level were assessed in 111 RA patients receiving a first course of 2 grams RTX (cohort 1) in an observational study and in 74 RA patients treated with the same schedule in routine care (cohort 2). Uni and multivariate analyzes identified factors associated with a European League Against Rheumatism response at 24 weeks. Results. At week 24, 84/111 (76%) and 54/74 (73%) patients reached EULAR response in the cohorts 1 and 2, respectively. Serum IL-33 was detectable in only 33,5% of the patients. In the combined cohorts, presence of RF or anti-CCP (OR 3.27, 95%CI [1.13-9.46]; p=0.03), high serum IgG (OR 2.32, 95%CI [1.01-5.33]; p=0.048) and detectable serum IL-33 (OR 2.40, 95%CI [1.01-5.72]; p=0.047) were all associated with RTX response in multivariate analysis. Combination of these 3 factors increased the likelihood to response to RTX. When serum IL-33 detection was added to seropositivity and serum IgG level, 100% of the patients with the 3 risk factors (corresponding to 9% of the population) responded to RTX (OR versus patients with none of the 3 risk factors = 29.61; 95% CI [1.30-674.79] p=0.034) Conclusion. Detectable serum IL-33 may predict clinical response to RTX, independently of and synergistically with autoantibodies and serum IgG level

Hyperosmolar environment and salivary gland epithelial cells increase extra-cellular matrix remodeling and lymphocytic infiltration in Sjögren’s syndrome.

info:eu-repo/semantics/inPres

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

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

Salivary gland epithelial cells from patients with Sjögren's syndrome induce B-lymphocyte survival and activation

International audienceObjectives: Primary Sjögren's syndrome (pSS) is characterized by chronic hyperactivation of Blymphocytes. Salivary gland epithelial cells (SGECs) could play a role in promoting Blymphocyte activation within the target tissue. We aimed to study the interactions between SGECs from pSS patients or controls and B-lymphocytes. Methods: Patients had pSS according to 2016 EULAR/ACR criteria. Gene expression analysis of SGECs and B-lymphocytes from pSS and controls isolated from salivary gland biopsies and blood was performed by RNA-seq. SGECs from pSS and controls were co-cultured with Blymphocytes sorted from healthy donor blood and stimulated. Transwell and inhibition experiments were performed. Results: Gene expression analysis of SGECs identified an upregulation of interferon signaling pathway and genes involved in immune responses (HLA-DRA, IL7, BAFFR) in pSS. Activation genes CD40 and CD48 were upregulated in salivary gland sorted B-lymphocytes from pSS patients. SGECs induced an increase in B-lymphocyte survival, which was higher for SGECs from pSS patients than controls. Moreover, when stimulated with Poly(I:C), SGECs from pSS patients induced higher activation of B-lymphocytes than those from controls. This effect depended on soluble factors. Inhibition with anti-BAFF, anti-APRIL, anti-IL6-R antibodies JAK1/3 inhibitor, or hydroxychloroquine had no effect, conversely to leflunomide, BTK or PI3K inhibitors. Conclusions: SGECs from patients with pSS had better ability than those from controls to induce survival and activation of B-lymphocytes. Targeting a single cytokine did not inhibit this effect, whereas, leflunomide, BTK or PI3K inhibitors partially decreased B-lymphocytes viability in this model. This gives indications for future therapeutic options in pSS

Long-term exposure to monoclonal anti-TNF is associated with an increased risk of lymphoma in BAFF-transgenic mice

International audienceSummary The impact of treatment on the risk of lymphoma in patients with rheumatoid arthritis (RA) is unclear. Here, we aimed to assess if the risk of lymphoma differs according to the type of tumor necrosis factor inhibitor (TNFi), comparing monoclonal anti-TNF antibodies to the soluble TNF receptor. We used B cell activating factor belonging to the TNF family (BAFF)-transgenic (Tg) mice as a model of autoimmunity-associated lymphoma. Six-month-old BAFF-Tg mice were treated with TNFi for 12 months. Histological examination of the spleen, assessment of the cellular composition of the spleen by flow cytometry and assessment of B cell clonality were performed at euthanasia. Crude mortality and incidence of lymphoma were significantly higher in mice treated with monoclonal anti-TNF antibodies compared to both controls and mice treated with the soluble TNF receptor, even at a high dose. Flow cytometry analysis revealed decreased splenic macrophage infiltration in mice treated with monoclonal anti-TNF antibodies. Overall, this study demonstrates, for the first time, that a very prolonged treatment with monoclonal anti-TNF antibodies increase the risk of lymphoma in B cell-driven autoimmunity. These data suggest a closer monitoring for lymphoma development in patients suffering from B cell-driven autoimmune disease with long-term exposure to monoclonal anti-TNF antibodies

Interleukin‐7/Interferon axis drives T‐cell and salivary gland epithelial cell interactions in Sjögren’s syndrome

International audienceObjectivePrimary Sjögren’s syndrome (pSS) is characterized by a lymphocytic infiltration of salivary glands and the presence of an interferon (IFN) signature. Salivary gland epithelial cells (SGECs) play an active role in pSS pathophysiology. The aim of this work was to study the interactions between SGECs and T cells in pSS and the role of the IL‐7/IFN axis.MethodsPrimary cultured SGECs from pSS and controls were stimulated with poly(I:C), IFNα or IFNγ. T cells were sorted from blood and stimulated with IL‐7. CD25 expression was assessed by flow cytometry. Salivary gland explants were cultured for 4 days with anti‐IL‐7R antagonist antibody (OSE‐127) and transcriptomic analysis was performed by using nanostring.ResultsIL‐7 serum level was increased in pSS patients versus controls and was associated with B‐cell biomarkers. IL7R expression was decreased in T cells from pSS patients versus controls. SGECs stimulated with poly(I:C), IFNα, or IFNγ secreted IL‐7. IL‐7 stimulation increased the activation of T cells, as well as IFNγ secretion. Transcriptomic analysis of salivary gland explants showed a correlation between IL7 and IFN expression. Lastly, explants cultured with anti‐IL‐7R antibody showed decreased IFN‐stimulated gene expression.ConclusionThese results suggest an IL‐7‐IFNγ amplification loop involving SGECs and T cells in pSS. IL‐7 was secreted by SGECs stimulated with type 1 or type 2 IFN and, in turn, activate T cells that secrete type 2 IFN. An anti‐IL‐7R antibody decreased the IFN signature in T cells during pSS and could be of therapeutic interest

Interleukin‐7/Interferon axis drives T‐cell and salivary gland epithelial cell interactions in Sjögren’s syndrome

International audienceObjectivePrimary Sjögren’s syndrome (pSS) is characterized by a lymphocytic infiltration of salivary glands and the presence of an interferon (IFN) signature. Salivary gland epithelial cells (SGECs) play an active role in pSS pathophysiology. The aim of this work was to study the interactions between SGECs and T cells in pSS and the role of the IL‐7/IFN axis.MethodsPrimary cultured SGECs from pSS and controls were stimulated with poly(I:C), IFNα or IFNγ. T cells were sorted from blood and stimulated with IL‐7. CD25 expression was assessed by flow cytometry. Salivary gland explants were cultured for 4 days with anti‐IL‐7R antagonist antibody (OSE‐127) and transcriptomic analysis was performed by using nanostring.ResultsIL‐7 serum level was increased in pSS patients versus controls and was associated with B‐cell biomarkers. IL7R expression was decreased in T cells from pSS patients versus controls. SGECs stimulated with poly(I:C), IFNα, or IFNγ secreted IL‐7. IL‐7 stimulation increased the activation of T cells, as well as IFNγ secretion. Transcriptomic analysis of salivary gland explants showed a correlation between IL7 and IFN expression. Lastly, explants cultured with anti‐IL‐7R antibody showed decreased IFN‐stimulated gene expression.ConclusionThese results suggest an IL‐7‐IFNγ amplification loop involving SGECs and T cells in pSS. IL‐7 was secreted by SGECs stimulated with type 1 or type 2 IFN and, in turn, activate T cells that secrete type 2 IFN. An anti‐IL‐7R antibody decreased the IFN signature in T cells during pSS and could be of therapeutic interest