POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS

M. A. Nielsen; D. Køster; K. Stengaard-Pedersen; P. Busson; P. Junker; K. Hørslev-Petersen; M. L. Hetland; M. Østergaard; M. Hvid; T. W. Kragstrup; B. Deleuran

doi:10.1136/annrheumdis-2022-eular.1213

POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS

M. A. Nielsen, D. Køster, K. Stengaard-Pedersen, P. Busson, P. Junker, K. Hørslev-Petersen, M. L. Hetland, M. Østergaard, M. Hvid, T. W. Kragstrup, B. Deleuran

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Konferenceabstrakt i tidsskrift › Forskning › peer review

Abstract

Background Fibroblast-like synoviocytes (FLS) are pivotal mediators of rheumatoid synovitis expansive growth and invasiveness which respond insufficiently to disease-modifying antirheumatic drugs. Galectin-9 (Gal-9) is a lectin with well-conserved carbohydrate-recognition domains. Gal-9 has been reported to have both pro- and anti-inflammatory properties based on its ability to modulate RA FLS. Gal-9 levels are elevated in serum of rheumatoid arthritis (RA) patients, and high levels of Gal-9 have been identified in both the synovial fluid and in the inflamed synovial tissue of patients with RA.Objectives To investigate the role of Gal-9 as a biomarker for disease severity in treatment naive patients with early RA and to study aspects of Gal-9 effects on inflammatory RA FLS.Methods Soluble plasma Gal-9 was measured in patients with newly diagnosed, treatment-naive RA (n = 98) and in Healthy (HC) (n = 48) (Table 1). Over a 2-year period patients were randomized to either MTX alone or MTX and anti-TNF antibody treatments. Serial measurements of disease activity (DAS28CRP) were collected to evaluate the disease course. In another cohort of patients with established RA, plasma and synovial fluid samples were also examined for Gal-9 (n = 18) (Table 1). Synovial fluid mononuclear cells (SFMC) from established RA patients were used to harvest RA FLS (n=7). Osteoarthritis FLS were used as disease controls and obtained from patients with knee OA undergoing joint replacement surgery (n = 5). Monocultures of synovial fluid derived FLS (SF-FLS) (n= 6) and autologous co-cultures of SF-FLS and peripheral blood mononuclear cells (PBMC) were established (n=7) and subsequently analyzed by flow cytometry, MTT assay, and ELISA. In vitro, cultures were treated with a neutralizing anti-Gal-9 antibody.View this table:Table 1. Results Patients with early and established RA had increased plasma levels of Gal-9 compared with HCs (P lt; 0.05) (Table 1) and levels of Gal-9 correlated positively with swollen joint counts at baseline (rho = 0.344, P lt; 0.05). The levels remained unaffected by treatment with MTX alone or by a combination of MTX and anti-TNF antibodies. Gal-9 levels were markedly elevated in the synovial fluid of chronic RA patients compared with the corresponding plasma samples (P lt; 0.05) (Table 1). In vitro, a neutralizing Gal-9 antibody mediated a 401 secretion (P lt; 0.05) and a 306 secretion (P lt; 0.05) in RA FLS monocultures. In OA FLS, addition of anti-Gal-9 antibodies comparably decreased the production of both MCP-1 (P lt; 0.05) and IL-6 (P lt; 0.05). The changes in cytokine production were not attributable to reduction in the fraction of inflammatory FLS (CD34-PDPN+THY1+), decreased viability or proliferation. We further investigated if the effect of neutralizing Gal-9 persisted in co-cultures between FLS and autologous PBMC. Also in this setting, neutralization of Gal-9 mediated a 401 and IL-6 (P lt; 0.05, P lt; 0.05).Conclusion Pretreatment plasma Gal-9 levels in patients with newly diagnosed RA were increased and correlated with baseline clinical disease activity but remained elevated during goal directed synovitis suppressive therapy. In addition, in vitro neutralization of Gal-9 decreased MCP-1 and IL-6 production in a subset of FLS linked to RA pathology. Collectively these findings indicate that Gal-9 overexpression is a co-player in the causation of acute and persistent RA synovitis by enhancing pro-inflammatory FLS pathways.Acknowledgements We thank Karin Skovgr ard Sørensen (Dept. of Biomedicine, Aarhus University) for technical assistance concerning the ELISA data and the FACS Core Facility (Aarhus University, Denmark) for technical assistant regarding Flow cytometry. We thank medical doctors and nurses at the Department of Rheumatology, Aarhus University Hospital for helping to collect the patient samples. We kindly acknowledge the generous grants from Aarhus University Research Foundation and the Danish Rheumatism Association.Disclosure of Interests None declared.

Originalsprog	Udefineret/Ukendt
Tidsskrift	Annals of the Rheumatic Diseases
Vol/bind	81
Nummer	Suppl 1
Sider (fra-til)	462-463
Antal sider	2
ISSN	0003-4967
DOI	https://doi.org/10.1136/annrheumdis-2022-eular.1213
Status	Udgivet - 2022

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10.1136/annrheumdis-2022-eular.1213

https://ard.bmj.com/content/81/Suppl_1/462.1

Citationsformater

Nielsen, M. A., Køster, D., Stengaard-Pedersen, K., Busson, P., Junker, P., Hørslev-Petersen, K., Hetland, M. L., Østergaard, M., Hvid, M., Kragstrup, T. W., & Deleuran, B. (2022). POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS. Annals of the Rheumatic Diseases, 81(Suppl 1), 462-463. https://doi.org/10.1136/annrheumdis-2022-eular.1213

Nielsen, M. A. ; Køster, D. ; Stengaard-Pedersen, K. et al. / POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS. I: Annals of the Rheumatic Diseases. 2022 ; Bind 81, Nr. Suppl 1. s. 462-463.

@article{846c9c25de8144fd838e315768c01276,

title = "POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NA{\"I}VE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS",

abstract = "Background Fibroblast-like synoviocytes (FLS) are pivotal mediators of rheumatoid synovitis expansive growth and invasiveness which respond insufficiently to disease-modifying antirheumatic drugs. Galectin-9 (Gal-9) is a lectin with well-conserved carbohydrate-recognition domains. Gal-9 has been reported to have both pro- and anti-inflammatory properties based on its ability to modulate RA FLS. Gal-9 levels are elevated in serum of rheumatoid arthritis (RA) patients, and high levels of Gal-9 have been identified in both the synovial fluid and in the inflamed synovial tissue of patients with RA.Objectives To investigate the role of Gal-9 as a biomarker for disease severity in treatment naive patients with early RA and to study aspects of Gal-9 effects on inflammatory RA FLS.Methods Soluble plasma Gal-9 was measured in patients with newly diagnosed, treatment-naive RA (n = 98) and in Healthy (HC) (n = 48) (Table 1). Over a 2-year period patients were randomized to either MTX alone or MTX and anti-TNF antibody treatments. Serial measurements of disease activity (DAS28CRP) were collected to evaluate the disease course. In another cohort of patients with established RA, plasma and synovial fluid samples were also examined for Gal-9 (n = 18) (Table 1). Synovial fluid mononuclear cells (SFMC) from established RA patients were used to harvest RA FLS (n=7). Osteoarthritis FLS were used as disease controls and obtained from patients with knee OA undergoing joint replacement surgery (n = 5). Monocultures of synovial fluid derived FLS (SF-FLS) (n= 6) and autologous co-cultures of SF-FLS and peripheral blood mononuclear cells (PBMC) were established (n=7) and subsequently analyzed by flow cytometry, MTT assay, and ELISA. In vitro, cultures were treated with a neutralizing anti-Gal-9 antibody.View this table:Table 1. Results Patients with early and established RA had increased plasma levels of Gal-9 compared with HCs (P lt; 0.05) (Table 1) and levels of Gal-9 correlated positively with swollen joint counts at baseline (rho = 0.344, P lt; 0.05). The levels remained unaffected by treatment with MTX alone or by a combination of MTX and anti-TNF antibodies. Gal-9 levels were markedly elevated in the synovial fluid of chronic RA patients compared with the corresponding plasma samples (P lt; 0.05) (Table 1). In vitro, a neutralizing Gal-9 antibody mediated a 401 secretion (P lt; 0.05) and a 306 secretion (P lt; 0.05) in RA FLS monocultures. In OA FLS, addition of anti-Gal-9 antibodies comparably decreased the production of both MCP-1 (P lt; 0.05) and IL-6 (P lt; 0.05). The changes in cytokine production were not attributable to reduction in the fraction of inflammatory FLS (CD34-PDPN+THY1+), decreased viability or proliferation. We further investigated if the effect of neutralizing Gal-9 persisted in co-cultures between FLS and autologous PBMC. Also in this setting, neutralization of Gal-9 mediated a 401 and IL-6 (P lt; 0.05, P lt; 0.05).Conclusion Pretreatment plasma Gal-9 levels in patients with newly diagnosed RA were increased and correlated with baseline clinical disease activity but remained elevated during goal directed synovitis suppressive therapy. In addition, in vitro neutralization of Gal-9 decreased MCP-1 and IL-6 production in a subset of FLS linked to RA pathology. Collectively these findings indicate that Gal-9 overexpression is a co-player in the causation of acute and persistent RA synovitis by enhancing pro-inflammatory FLS pathways.Acknowledgements We thank Karin Skovgr ard S{\o}rensen (Dept. of Biomedicine, Aarhus University) for technical assistance concerning the ELISA data and the FACS Core Facility (Aarhus University, Denmark) for technical assistant regarding Flow cytometry. We thank medical doctors and nurses at the Department of Rheumatology, Aarhus University Hospital for helping to collect the patient samples. We kindly acknowledge the generous grants from Aarhus University Research Foundation and the Danish Rheumatism Association.Disclosure of Interests None declared.",

author = "Nielsen, {M. A.} and D. K{\o}ster and K. Stengaard-Pedersen and P. Busson and P. Junker and K. H{\o}rslev-Petersen and Hetland, {M. L.} and M. {\O}stergaard and M. Hvid and Kragstrup, {T. W.} and B. Deleuran",

year = "2022",

doi = "10.1136/annrheumdis-2022-eular.1213",

language = "Udefineret/Ukendt",

volume = "81",

pages = "462--463",

journal = "Annals of the Rheumatic Diseases",

issn = "0003-4967",

publisher = "BMJ Publishing Group",

number = "Suppl 1",

}

Nielsen, MA, Køster, D, Stengaard-Pedersen, K, Busson, P, Junker, P, Hørslev-Petersen, K, Hetland, ML, Østergaard, M, Hvid, M , Kragstrup, TW & Deleuran, B 2022, 'POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS', Annals of the Rheumatic Diseases, bind 81, nr. Suppl 1, s. 462-463. https://doi.org/10.1136/annrheumdis-2022-eular.1213

POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS. / Nielsen, M. A.; Køster, D.; Stengaard-Pedersen, K. et al.
I: Annals of the Rheumatic Diseases, Bind 81, Nr. Suppl 1, 2022, s. 462-463.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Konferenceabstrakt i tidsskrift › Forskning › peer review

TY - ABST

T1 - POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS

AU - Nielsen, M. A.

AU - Køster, D.

AU - Stengaard-Pedersen, K.

AU - Busson, P.

AU - Junker, P.

AU - Hørslev-Petersen, K.

AU - Hetland, M. L.

AU - Østergaard, M.

AU - Hvid, M.

AU - Kragstrup, T. W.

AU - Deleuran, B.

PY - 2022

Y1 - 2022

N2 - Background Fibroblast-like synoviocytes (FLS) are pivotal mediators of rheumatoid synovitis expansive growth and invasiveness which respond insufficiently to disease-modifying antirheumatic drugs. Galectin-9 (Gal-9) is a lectin with well-conserved carbohydrate-recognition domains. Gal-9 has been reported to have both pro- and anti-inflammatory properties based on its ability to modulate RA FLS. Gal-9 levels are elevated in serum of rheumatoid arthritis (RA) patients, and high levels of Gal-9 have been identified in both the synovial fluid and in the inflamed synovial tissue of patients with RA.Objectives To investigate the role of Gal-9 as a biomarker for disease severity in treatment naive patients with early RA and to study aspects of Gal-9 effects on inflammatory RA FLS.Methods Soluble plasma Gal-9 was measured in patients with newly diagnosed, treatment-naive RA (n = 98) and in Healthy (HC) (n = 48) (Table 1). Over a 2-year period patients were randomized to either MTX alone or MTX and anti-TNF antibody treatments. Serial measurements of disease activity (DAS28CRP) were collected to evaluate the disease course. In another cohort of patients with established RA, plasma and synovial fluid samples were also examined for Gal-9 (n = 18) (Table 1). Synovial fluid mononuclear cells (SFMC) from established RA patients were used to harvest RA FLS (n=7). Osteoarthritis FLS were used as disease controls and obtained from patients with knee OA undergoing joint replacement surgery (n = 5). Monocultures of synovial fluid derived FLS (SF-FLS) (n= 6) and autologous co-cultures of SF-FLS and peripheral blood mononuclear cells (PBMC) were established (n=7) and subsequently analyzed by flow cytometry, MTT assay, and ELISA. In vitro, cultures were treated with a neutralizing anti-Gal-9 antibody.View this table:Table 1. Results Patients with early and established RA had increased plasma levels of Gal-9 compared with HCs (P lt; 0.05) (Table 1) and levels of Gal-9 correlated positively with swollen joint counts at baseline (rho = 0.344, P lt; 0.05). The levels remained unaffected by treatment with MTX alone or by a combination of MTX and anti-TNF antibodies. Gal-9 levels were markedly elevated in the synovial fluid of chronic RA patients compared with the corresponding plasma samples (P lt; 0.05) (Table 1). In vitro, a neutralizing Gal-9 antibody mediated a 401 secretion (P lt; 0.05) and a 306 secretion (P lt; 0.05) in RA FLS monocultures. In OA FLS, addition of anti-Gal-9 antibodies comparably decreased the production of both MCP-1 (P lt; 0.05) and IL-6 (P lt; 0.05). The changes in cytokine production were not attributable to reduction in the fraction of inflammatory FLS (CD34-PDPN+THY1+), decreased viability or proliferation. We further investigated if the effect of neutralizing Gal-9 persisted in co-cultures between FLS and autologous PBMC. Also in this setting, neutralization of Gal-9 mediated a 401 and IL-6 (P lt; 0.05, P lt; 0.05).Conclusion Pretreatment plasma Gal-9 levels in patients with newly diagnosed RA were increased and correlated with baseline clinical disease activity but remained elevated during goal directed synovitis suppressive therapy. In addition, in vitro neutralization of Gal-9 decreased MCP-1 and IL-6 production in a subset of FLS linked to RA pathology. Collectively these findings indicate that Gal-9 overexpression is a co-player in the causation of acute and persistent RA synovitis by enhancing pro-inflammatory FLS pathways.Acknowledgements We thank Karin Skovgr ard Sørensen (Dept. of Biomedicine, Aarhus University) for technical assistance concerning the ELISA data and the FACS Core Facility (Aarhus University, Denmark) for technical assistant regarding Flow cytometry. We thank medical doctors and nurses at the Department of Rheumatology, Aarhus University Hospital for helping to collect the patient samples. We kindly acknowledge the generous grants from Aarhus University Research Foundation and the Danish Rheumatism Association.Disclosure of Interests None declared.

AB - Background Fibroblast-like synoviocytes (FLS) are pivotal mediators of rheumatoid synovitis expansive growth and invasiveness which respond insufficiently to disease-modifying antirheumatic drugs. Galectin-9 (Gal-9) is a lectin with well-conserved carbohydrate-recognition domains. Gal-9 has been reported to have both pro- and anti-inflammatory properties based on its ability to modulate RA FLS. Gal-9 levels are elevated in serum of rheumatoid arthritis (RA) patients, and high levels of Gal-9 have been identified in both the synovial fluid and in the inflamed synovial tissue of patients with RA.Objectives To investigate the role of Gal-9 as a biomarker for disease severity in treatment naive patients with early RA and to study aspects of Gal-9 effects on inflammatory RA FLS.Methods Soluble plasma Gal-9 was measured in patients with newly diagnosed, treatment-naive RA (n = 98) and in Healthy (HC) (n = 48) (Table 1). Over a 2-year period patients were randomized to either MTX alone or MTX and anti-TNF antibody treatments. Serial measurements of disease activity (DAS28CRP) were collected to evaluate the disease course. In another cohort of patients with established RA, plasma and synovial fluid samples were also examined for Gal-9 (n = 18) (Table 1). Synovial fluid mononuclear cells (SFMC) from established RA patients were used to harvest RA FLS (n=7). Osteoarthritis FLS were used as disease controls and obtained from patients with knee OA undergoing joint replacement surgery (n = 5). Monocultures of synovial fluid derived FLS (SF-FLS) (n= 6) and autologous co-cultures of SF-FLS and peripheral blood mononuclear cells (PBMC) were established (n=7) and subsequently analyzed by flow cytometry, MTT assay, and ELISA. In vitro, cultures were treated with a neutralizing anti-Gal-9 antibody.View this table:Table 1. Results Patients with early and established RA had increased plasma levels of Gal-9 compared with HCs (P lt; 0.05) (Table 1) and levels of Gal-9 correlated positively with swollen joint counts at baseline (rho = 0.344, P lt; 0.05). The levels remained unaffected by treatment with MTX alone or by a combination of MTX and anti-TNF antibodies. Gal-9 levels were markedly elevated in the synovial fluid of chronic RA patients compared with the corresponding plasma samples (P lt; 0.05) (Table 1). In vitro, a neutralizing Gal-9 antibody mediated a 401 secretion (P lt; 0.05) and a 306 secretion (P lt; 0.05) in RA FLS monocultures. In OA FLS, addition of anti-Gal-9 antibodies comparably decreased the production of both MCP-1 (P lt; 0.05) and IL-6 (P lt; 0.05). The changes in cytokine production were not attributable to reduction in the fraction of inflammatory FLS (CD34-PDPN+THY1+), decreased viability or proliferation. We further investigated if the effect of neutralizing Gal-9 persisted in co-cultures between FLS and autologous PBMC. Also in this setting, neutralization of Gal-9 mediated a 401 and IL-6 (P lt; 0.05, P lt; 0.05).Conclusion Pretreatment plasma Gal-9 levels in patients with newly diagnosed RA were increased and correlated with baseline clinical disease activity but remained elevated during goal directed synovitis suppressive therapy. In addition, in vitro neutralization of Gal-9 decreased MCP-1 and IL-6 production in a subset of FLS linked to RA pathology. Collectively these findings indicate that Gal-9 overexpression is a co-player in the causation of acute and persistent RA synovitis by enhancing pro-inflammatory FLS pathways.Acknowledgements We thank Karin Skovgr ard Sørensen (Dept. of Biomedicine, Aarhus University) for technical assistance concerning the ELISA data and the FACS Core Facility (Aarhus University, Denmark) for technical assistant regarding Flow cytometry. We thank medical doctors and nurses at the Department of Rheumatology, Aarhus University Hospital for helping to collect the patient samples. We kindly acknowledge the generous grants from Aarhus University Research Foundation and the Danish Rheumatism Association.Disclosure of Interests None declared.

U2 - 10.1136/annrheumdis-2022-eular.1213

DO - 10.1136/annrheumdis-2022-eular.1213

M3 - Konferenceabstrakt i tidsskrift

SN - 0003-4967

VL - 81

SP - 462

EP - 463

JO - Annals of the Rheumatic Diseases

JF - Annals of the Rheumatic Diseases

IS - Suppl 1

ER -

Nielsen MA, Køster D, Stengaard-Pedersen K, Busson P, Junker P, Hørslev-Petersen K et al. POS0415 PLASMA GALECTIN-9 IS ASSOCIATED WITH CLINICAL DISEASE ACTIVITY IN TREATMENT NAÏVE EARLY RHEUMATOID ARTHRITIS, UNAFFECTED BY DMARD TREATMENT, AND IN VITRO MODULATES THE SECRETION OF MCP-1 AND IL-6 IN RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLASTS. Annals of the Rheumatic Diseases. 2022;81(Suppl 1):462-463. doi: 10.1136/annrheumdis-2022-eular.1213