G Model BONSOI-3994; No. of Pages 2
ARTICLE IN PRESS Joint Bone Spine xxx (2014) xxx–xxx
Available online at www.sciencedirect.com
Letter to the Editor Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis
a r t i c l e
i n f o
(Fig. 1c), and nuclear activity was higher in patients treated with corticosteroids (P = 0.03). After resveratrol treatment [6], no significant changes in TNF, IL-6 or IL-8 levels were found (data not shown).
Keywords: Epigenetics Rheumatoid arthritis Sirtuin 1 HDAC IL-6
Sirtuin 1 (Sirt1) is mostly a nuclear enzyme from the class III histone deacetylases (HDACs) modulating gene expression, involved in the regulation of various biological processes (cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling [1,2]. In RA synovial cells, TNF induced over-expression of Sirt1 contributes to chronic inflammation by promoting inflammatory cytokine production and inhibiting apoptosis [3,4]. The main objective of the study was to evaluate Sirt1 activity in peripheral blood mononuclear cells (PBMCs) by venous blood aspiration in rheumatoid arthritis (RA) patients, compared to those of control patients, and to analyze the relationship between Sirt1 activity, disease activity and production of mediators of inflammation and cytokines (TNF alpha, IL-6, IL-8) by the cells, before and after ex vivo treatment with a Sirtuin activator, resveratrol. A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with RA (according to ACR criteria) and controls. Disease activity was assessed by DAS28 (ESR) and CRP. PBMCs were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorimetric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 min point. Culture supernatant levels of TNF alpha, IL-6, IL8 were quantified before and after resveratrol (1 mol and 5 mol) ex vivo treatment, with commercial kits, as previously described [5]. Statistical analysis used Wilcoxon and t tests; significance: P less than 0.05. Twenty-two patients with RA (age 55 ± 12 years, mean DAS 28: 4.27; disease duration 15 ± 10 years; 75% ACPA + mean CRP: 12.7 mg/l) and 18 controls (age 54 ± 13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activities were correlated in patients and controls (Fig. 1a). Sirt1 activity (nuclear and cytoplasmic) was not correlated with DAS28, CRP or ESR, but cytoplasmic Sirt1 activity was correlated to baseline IL-6 (P = 0.02) (Fig. 1b) and baseline TNF (P = 0.04) in RA and controls, but not with IL-8. Sirt1 activity (cytoplasmic and nuclear) was lower in ACPA-positive RA compared to ACPA-negative (P = 0.01)
Fig. 1. a: correlation in rheumatoid arthritis (RA) patients between nuclear and cytoplasmic Sirt1 activity; b: correlation between cytoplasmic Sirt1 activity and IL6 levels in RA patients; c: comparison of cytoplasmic Sirt1 activity according to ACPA positivity (1: ACPA+, 0: ACPA–).
1297-319X/$ – see front matter © 2014 Société franc¸aise de rhumatologie. Published by Elsevier Masson SAS. All rights reserved. doi:10.1016/j.jbspin.2014.02.006
Please cite this article in press as: Wendling D, et al. Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis. Joint Bone Spine (2014), doi:10.1016/j.jbspin.2014.02.006
G Model BONSOI-3994; No. of Pages 2
ARTICLE IN PRESS Letter to the Editor / Joint Bone Spine xxx (2014) xxx–xxx
2
Sirt1 activity (cytoplasmic and nuclear) from PBMCs was not different between RA patients and controls in this study, whereas HDAC activity was found increased in PBMCs from RA patients compared to controls in another study [7]. Nevertheless, the correlation in RA between Sirt1 activity and TNF and IL-6 levels, as well as reduced Sirt1 activity in ACPA-positive patients may suggest an implication of Sirt1-mediated epigenetic regulation in RA. Epigenetics, and particularly Sirtuins represent a matter of investigation in osteoarticular diseases [8–10]. Disclosure of interest The authors declare that they have no conflicts of interest concerning this article. References [1] Herbein G, Wendling D. Histone deacetylases in viral infections. Clin Epigenetics 2010;1:13–24. [2] Viatte S, Plant D, Raychaudhuri S. Genetics and epigenetics of rheumatoid arthritis. Nat Rev Rheumatol 2013;9:141–53. [3] Niederer F, Ospelt C, Brentano F, et al. SIRT1 over-expression in the rheumatoid arthritis synovium contributes to proinflammatory cytokine production and apoptosis resistance. Ann Rheum Dis 2011;70:1866–73. [4] Grabiec AM, Krausz S, de Jager W, et al. Histone deacetylase inhibitors suppress inflammatory activation of rheumatoid arthritis patient synovial macrophages and tissue. J Immunol 2010;184:2718–28. [5] Wendling D, Abbas W, Godfrin-Valnet M, et al. Resveratrol, a sirtuin 1 activator, increases IL-6 production by peripheral blood mononuclear cells of patients with knee osteoarthritis. Clin Epigenetics 2013;5:10, doi:10.1186/1868-7083-5-10. [6] Nakayama H, Yaguchi T, Yoshiya S, et al. Resveratrol induces apoptosis MH7A human rheumatoid arthritis synovial cells in a sirtuin 1-dependent manner. Rheumatol Int 2012;32:151–7. [7] Gillespie J, Savic S, Wong C, et al. Histone deacetylases are dysregulated in rheumatoid arthritis and a novel histone deacetylase 3-selective inhibitor
reduces interleukin-6 production by peripheral blood mononuclear cells from rheumatoid arthritis patients. Arthritis Rheum 2012;64:418–22. [8] Weitzman J. Epigenetics in health and disease. Joint Bone Spine 2012; 79:565. [9] Gabay O, Sanchez C. Epigenetics, sirtuins and osteoarthritis. Joint Bone Spine 2012;79:570–3. [10] Wendling D, Herbein G. Sirt1 and osteoarthritis. Comments on the paper by Gabay et al.: “Sirt1-deficient mice exhibit an altered cartilage phenotype”. Joint Bone Spine 2013. Joint Bone Spine 2014;81:104–5.
Daniel Wendling a,b,∗ Claire Vidon a Wasim Abbas b Xavier Guillot a Eric Toussirot a,b Georges Herbein b,c,∗ a Department of Rheumatology, CHRU de Besanc¸on, boulevard Fleming, 25030 Besanc¸on, France b University of Franche-Comté, UPRES EA 4266 Pathogens & Inflammation Laboratory, SFR FED4234, Besanc¸on, France c Department of Virology, CHRU de Besanc¸on, place Saint-Jacques, 25030 Besanc¸on, France ∗ Corresponding
authors. Department of Rheumatology, CHRU de Besanc¸on, boulevard Fleming, 25030 Besanc¸on, France. E-mail addresses: [email protected] (D. Wendling), [email protected] (G. Herbein) Accepted 13 February 2014 Available online xxx
Please cite this article in press as: Wendling D, et al. Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis. Joint Bone Spine (2014), doi:10.1016/j.jbspin.2014.02.006
ARTICLE IN PRESS Joint Bone Spine xxx (2014) xxx–xxx
Available online at www.sciencedirect.com
Letter to the Editor Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis
a r t i c l e
i n f o
(Fig. 1c), and nuclear activity was higher in patients treated with corticosteroids (P = 0.03). After resveratrol treatment [6], no significant changes in TNF, IL-6 or IL-8 levels were found (data not shown).
Keywords: Epigenetics Rheumatoid arthritis Sirtuin 1 HDAC IL-6
Sirtuin 1 (Sirt1) is mostly a nuclear enzyme from the class III histone deacetylases (HDACs) modulating gene expression, involved in the regulation of various biological processes (cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis), local and systemic inflammation, as well as in bone and cartilage remodeling [1,2]. In RA synovial cells, TNF induced over-expression of Sirt1 contributes to chronic inflammation by promoting inflammatory cytokine production and inhibiting apoptosis [3,4]. The main objective of the study was to evaluate Sirt1 activity in peripheral blood mononuclear cells (PBMCs) by venous blood aspiration in rheumatoid arthritis (RA) patients, compared to those of control patients, and to analyze the relationship between Sirt1 activity, disease activity and production of mediators of inflammation and cytokines (TNF alpha, IL-6, IL-8) by the cells, before and after ex vivo treatment with a Sirtuin activator, resveratrol. A prospective and comparative monocentric study was performed in order to compare the activity of Sirt1 in patients with RA (according to ACR criteria) and controls. Disease activity was assessed by DAS28 (ESR) and CRP. PBMCs were isolated from venous blood, and Sirt1 activity was evaluated from cytoplasmic and nuclear compartments using a fluorometric assay (SIRT1 fluorimetric kit, BML-AK-555, Enzo Life Sciences, Villeurbanne, France) at the 15 min point. Culture supernatant levels of TNF alpha, IL-6, IL8 were quantified before and after resveratrol (1 mol and 5 mol) ex vivo treatment, with commercial kits, as previously described [5]. Statistical analysis used Wilcoxon and t tests; significance: P less than 0.05. Twenty-two patients with RA (age 55 ± 12 years, mean DAS 28: 4.27; disease duration 15 ± 10 years; 75% ACPA + mean CRP: 12.7 mg/l) and 18 controls (age 54 ± 13 years) were included. No differences were found in cytoplasmic or nuclear Sirt1 activity between patients and controls. Cytoplasmic and nuclear Sirt1 activities were correlated in patients and controls (Fig. 1a). Sirt1 activity (nuclear and cytoplasmic) was not correlated with DAS28, CRP or ESR, but cytoplasmic Sirt1 activity was correlated to baseline IL-6 (P = 0.02) (Fig. 1b) and baseline TNF (P = 0.04) in RA and controls, but not with IL-8. Sirt1 activity (cytoplasmic and nuclear) was lower in ACPA-positive RA compared to ACPA-negative (P = 0.01)
Fig. 1. a: correlation in rheumatoid arthritis (RA) patients between nuclear and cytoplasmic Sirt1 activity; b: correlation between cytoplasmic Sirt1 activity and IL6 levels in RA patients; c: comparison of cytoplasmic Sirt1 activity according to ACPA positivity (1: ACPA+, 0: ACPA–).
1297-319X/$ – see front matter © 2014 Société franc¸aise de rhumatologie. Published by Elsevier Masson SAS. All rights reserved. doi:10.1016/j.jbspin.2014.02.006
Please cite this article in press as: Wendling D, et al. Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis. Joint Bone Spine (2014), doi:10.1016/j.jbspin.2014.02.006
G Model BONSOI-3994; No. of Pages 2
ARTICLE IN PRESS Letter to the Editor / Joint Bone Spine xxx (2014) xxx–xxx
2
Sirt1 activity (cytoplasmic and nuclear) from PBMCs was not different between RA patients and controls in this study, whereas HDAC activity was found increased in PBMCs from RA patients compared to controls in another study [7]. Nevertheless, the correlation in RA between Sirt1 activity and TNF and IL-6 levels, as well as reduced Sirt1 activity in ACPA-positive patients may suggest an implication of Sirt1-mediated epigenetic regulation in RA. Epigenetics, and particularly Sirtuins represent a matter of investigation in osteoarticular diseases [8–10]. Disclosure of interest The authors declare that they have no conflicts of interest concerning this article. References [1] Herbein G, Wendling D. Histone deacetylases in viral infections. Clin Epigenetics 2010;1:13–24. [2] Viatte S, Plant D, Raychaudhuri S. Genetics and epigenetics of rheumatoid arthritis. Nat Rev Rheumatol 2013;9:141–53. [3] Niederer F, Ospelt C, Brentano F, et al. SIRT1 over-expression in the rheumatoid arthritis synovium contributes to proinflammatory cytokine production and apoptosis resistance. Ann Rheum Dis 2011;70:1866–73. [4] Grabiec AM, Krausz S, de Jager W, et al. Histone deacetylase inhibitors suppress inflammatory activation of rheumatoid arthritis patient synovial macrophages and tissue. J Immunol 2010;184:2718–28. [5] Wendling D, Abbas W, Godfrin-Valnet M, et al. Resveratrol, a sirtuin 1 activator, increases IL-6 production by peripheral blood mononuclear cells of patients with knee osteoarthritis. Clin Epigenetics 2013;5:10, doi:10.1186/1868-7083-5-10. [6] Nakayama H, Yaguchi T, Yoshiya S, et al. Resveratrol induces apoptosis MH7A human rheumatoid arthritis synovial cells in a sirtuin 1-dependent manner. Rheumatol Int 2012;32:151–7. [7] Gillespie J, Savic S, Wong C, et al. Histone deacetylases are dysregulated in rheumatoid arthritis and a novel histone deacetylase 3-selective inhibitor
reduces interleukin-6 production by peripheral blood mononuclear cells from rheumatoid arthritis patients. Arthritis Rheum 2012;64:418–22. [8] Weitzman J. Epigenetics in health and disease. Joint Bone Spine 2012; 79:565. [9] Gabay O, Sanchez C. Epigenetics, sirtuins and osteoarthritis. Joint Bone Spine 2012;79:570–3. [10] Wendling D, Herbein G. Sirt1 and osteoarthritis. Comments on the paper by Gabay et al.: “Sirt1-deficient mice exhibit an altered cartilage phenotype”. Joint Bone Spine 2013. Joint Bone Spine 2014;81:104–5.
Daniel Wendling a,b,∗ Claire Vidon a Wasim Abbas b Xavier Guillot a Eric Toussirot a,b Georges Herbein b,c,∗ a Department of Rheumatology, CHRU de Besanc¸on, boulevard Fleming, 25030 Besanc¸on, France b University of Franche-Comté, UPRES EA 4266 Pathogens & Inflammation Laboratory, SFR FED4234, Besanc¸on, France c Department of Virology, CHRU de Besanc¸on, place Saint-Jacques, 25030 Besanc¸on, France ∗ Corresponding
authors. Department of Rheumatology, CHRU de Besanc¸on, boulevard Fleming, 25030 Besanc¸on, France. E-mail addresses: [email protected] (D. Wendling), [email protected] (G. Herbein) Accepted 13 February 2014 Available online xxx
Please cite this article in press as: Wendling D, et al. Sirt1 activity in peripheral blood mononuclear cells from patients with rheumatoid arthritis. Joint Bone Spine (2014), doi:10.1016/j.jbspin.2014.02.006
