Top

Published in:

Open Access 01-12-2021 | Ankylosing Spondylitis | Research article

Monocyte transcriptomes from patients with axial spondyloarthritis reveal dysregulated monocytopoiesis and a distinct inflammatory imprint

Authors: Fabian Karow, Biljana Smiljanovic, Joachim R. Grün, Denis Poddubnyy, Fabian Proft, Alice Talpin, Christophe Hue, Anne Boland, Jean-François Deleuze, Henri-Jean Garchon, Ilkay Ergenç, Ann-Sophie De Craemer, Ulrike Erben, Thomas Häupl, Dirk Elewaut, Maxime Breban, Andreas Grützkau, Uta Syrbe

Published in: Arthritis Research & Therapy | Issue 1/2021

Abstract

Background

In patients with axial spondyloarthritis (axSpA), monocytes show a pre-activated phenotype. Gut inflammation is a trigger of monocyte activation and may also affect their development in the bone marrow (BM). As gut inflammation is commonly observed in axSpA patients, we performed a detailed analysis of monocyte transcriptomes of axSpA patients in two cohorts and searched for signs of activation and developmental adaptations as putative imprints of gut inflammation.

Methods

Transcriptomes of blood CD14⁺ monocytes of HLA-B27+ axSpA patients and healthy controls (HC) were generated by microarrays from cohort 1 and by RNA-sequencing from cohort 2. Differentially expressed genes from both analyses were subjected to gene set enrichment analysis (GSEA) and to co-expression analysis in reference transcriptomes from BM cells, blood cells and activated monocytes. As serological markers of translocation, 1,3 beta-glycan, intestinal fatty acid binding protein, and lipopolysaccharide binding protein (LBP) were determined by LAL and ELISA.

Results

Transcriptome analysis identified axSpA-specific monocyte signatures showing an imprint of LPS/cytokine-activated monocytes, late granulopoietic BM cells, blood neutrophils, and G-CSF-mobilized blood cells, which suggests LPS/TNF activation and more prominent BM adaptation promoting a neutrophil-like phenotype. GSEA mapped axSpA upregulated genes to inflammatory responses and TNFα signaling and downregulated probe-sets to metabolic pathways. Among translocation markers, LBP levels were significantly increased in axSpA patients vs. HC (p < 0.001).

Stratified analysis by disease activity and stage identified an “active disease signature” (BASDAI ≥ 4) with an imprint of LPS/cytokine-activated monocytes and CD16⁺ monocyte subsets. The “AS signature” (vs. non-radiographic axSpA) showed a reinforced neutrophil-like phenotype due to deprivation of dendritic cell transcripts.

Conclusions

The neutrophil-like phenotype of axSpA monocytes points towards a biased monocytopoiesis from granulocyte-monocyte progenitors. This shift in monocytopoiesis and the LPS/cytokine imprint as well as the elevated LBP levels are indicators of systemic inflammation, which may result from bacterial translocation. The BM adaptation is most prominent in AS patients while disease activity appears to be linked to activation and trafficking of monocytes.

Available only for authorised users

Brewerton DA, Hart FD, Nicholls A, Caffrey M, James DC, Sturrock RD. Ankylosing spondylitis and HL-A 27. Lancet. 1973;1(7809):904–7. https://doi.org/10.1016/S0140-6736(73)91360-3.CrossRefPubMed

Cortes A, Hadler J, Pointon JP, Robinson PC, Karaderi T, Leo P, et al. Identification of multiple risk variants for ankylosing spondylitis through high-density genotyping of immune-related loci. Nat Genet. 2013;45(7):730–8. https://doi.org/10.1038/ng.2667.CrossRefPubMedPubMedCentral

Ellinghaus D, Jostins L, Spain SL, Cortes A, Bethune J, Han B, et al. Analysis of five chronic inflammatory diseases identifies 27 new associations and highlights disease-specific patterns at shared loci. Nat Genet. 2016;48(5):510–8. https://doi.org/10.1038/ng.3528.CrossRefPubMedPubMedCentral

Gracey E, Vereecke L, McGovern D, Frohling M, Schett G, Danese S, et al. Revisiting the gut-joint axis: links between gut inflammation and spondyloarthritis. Nat Rev Rheumatol. 2020;16(8):415–33. https://doi.org/10.1038/s41584-020-0454-9.CrossRefPubMed

Menegatti S, Guillemot V, Latis E, Yahia-Cherbal H, Mittermuller D, Rouilly V, et al. Immune response profiling of patients with spondyloarthritis reveals signalling networks mediating TNF-blocker function in vivo. Ann Rheum Dis. 2020;80(4):475–86. https://doi.org/10.1136/annrheumdis-2020-218304.CrossRef

Wright C. Edelmann M, diGleria K, Kollnberger S, Kramer H, McGowan S, McHugh K, Taylor S, Kessler B, Bowness P: Ankylosing spondylitis monocytes show upregulation of proteins involved in inflammation and the ubiquitin proteasome pathway. Ann Rheum Dis. 2009;68(10):1626–32. https://doi.org/10.1136/ard.2008.097204.CrossRefPubMed

Conrad K, Wu P, Sieper J, Syrbe U. In vivo pre-activation of monocytes in patients with axial spondyloarthritis. Arthritis Res Ther. 2015;17(1):179. https://doi.org/10.1186/s13075-015-0694-2.CrossRefPubMedPubMedCentral

Ziegler-Heitbrock L, Ancuta P, Crowe S, Dalod M, Grau V, Hart DN, et al. Nomenclature of monocytes and dendritic cells in blood. Blood. 2010;116(16):e74–80. https://doi.org/10.1182/blood-2010-02-258558.CrossRefPubMed

Patel AA, Zhang Y, Fullerton JN, Boelen L, Rongvaux A, Maini AA, et al. The fate and lifespan of human monocyte subsets in steady state and systemic inflammation. J Exp Med. 2017;214(7):1913–23. https://doi.org/10.1084/jem.20170355.CrossRefPubMedPubMedCentral

10.

Tak T, van Groenendael R, Pickkers P, Koenderman L. Monocyte subsets are differentially lost from the circulation during acute inflammation induced by human experimental endotoxemia. J Innate Immun. 2017;9(5):464–74. https://doi.org/10.1159/000475665.CrossRefPubMedPubMedCentral

11.

Stolwijk C, van Tubergen A, Castillo-Ortiz JD, Boonen A. Prevalence of extra-articular manifestations in patients with ankylosing spondylitis: a systematic review and meta-analysis. Ann Rheum Dis. 2013;72(Suppl 3):A520.2–A521. https://doi.org/10.1136/annrheumdis-2013-eular.1558.CrossRef

12.

Mielants H, Veys EM, Cuvelier C, de Vos M. Ileocolonoscopic findings in seronegative spondylarthropathies. Br J Rheumatol. 1988;27(Suppl 2):95–105.CrossRefPubMed

13.

Van Praet L, Van den Bosch FE, Jacques P, Carron P, Jans L, Colman R, et al. Microscopic gut inflammation in axial spondyloarthritis: a multiparametric predictive model. Ann Rheum Dis. 2013;72(3):414–7. https://doi.org/10.1136/annrheumdis-2012-202135.CrossRefPubMed

14.

Chavakis T, Mitroulis I, Hajishengallis G. Hematopoietic progenitor cells as integrative hubs for adaptation to and fine-tuning of inflammation. Nat Immunol. 2019;20(7):802–11. https://doi.org/10.1038/s41590-019-0402-5.CrossRefPubMedPubMedCentral

15.

Yanez A, Coetzee SG, Olsson A, Muench DE, Berman BP, Hazelett DJ, et al. Granulocyte-monocyte progenitors and monocyte-dendritic cell progenitors ndependently produce eunctionally distinct monocytes. Immunity. 2017;47(5):890–902 e894. https://doi.org/10.1016/j.immuni.2017.10.021.CrossRefPubMedPubMedCentral

16.

Rudwaleit M, van der Heijde D, Landewe R, Listing J, Akkoc N, Brandt J, et al. The development of Assessment of SpondyloArthritis International Society classification criteria for axial spondyloarthritis (part II): validation and final selection. Ann Rheum Dis. 2009;68(6):777–83. https://doi.org/10.1136/ard.2009.108233.CrossRefPubMed

17.

van der Linden S, Valkenburg HA, Cats A. Evaluation of diagnostic criteria for ankylosing spondylitis. A proposal for modification of the New York criteria. Arthritis Rheum. 1984;27(4):361–8. https://doi.org/10.1002/art.1780270401.CrossRefPubMed

18.

Garrett S, Jenkinson T, Kennedy LG, Whitelock H, Gaisford P, Calin A. A new approach to defining disease status in ankylosing spondylitis: the Bath Ankylosing Spondylitis Disease Activity Index. J Rheumatol. 1994;21(12):2286–91.PubMed

19.

Lukas C, Landewe R, Sieper J, Dougados M, Davis J, Braun J, et al. Assessment of SpondyloArthritis International S: Development of an ASAS-endorsed disease activity score (ASDAS) in patients with ankylosing spondylitis. Ann Rheum Dis. 2009;68(1):18–24. https://doi.org/10.1136/ard.2008.094870.CrossRefPubMed

20.

Cypers H, Varkas G, Beeckman S, Debusschere K, Vogl T, Roth J, et al. Elevated calprotectin levels reveal bowel inflammation in spondyloarthritis. Ann Rheum Dis. 2016;75(7):1357–62. https://doi.org/10.1136/annrheumdis-2015-208025.CrossRefPubMed

21.

Smiljanovic B, Grun JR, Steinbrich-Zollner M, Stuhlmuller B, Haupl T, Burmester GR, et al. Defining TNF-alpha- and LPS-induced gene signatures in monocytes to unravel the complexity of peripheral blood transcriptomes in health and disease. J Mol Med (Berl). 2010;88(10):1065–79. https://doi.org/10.1007/s00109-010-0648-8.CrossRef

22.

Lepre J, Rice JJ, Tu Y, Stolovitzky G. Genes@Work: an efficient algorithm for pattern discovery and multivariate feature selection in gene expression data. Bioinformatics. 2004;20(7):1033–44. https://doi.org/10.1093/bioinformatics/bth035.CrossRefPubMed

23.

Subramanian A, Tamayo P, Mootha VK, Mukherjee S, Ebert BL, Gillette MA, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A. 2005;102(43):15545–50. https://doi.org/10.1073/pnas.0506580102.CrossRefPubMedPubMedCentral

24.

Mootha VK, Lindgren CM, Eriksson KF, Subramanian A, Sihag S, Lehar J, et al. PGC-1alpha-responsive genes involved in oxidative phosphorylation are coordinately downregulated in human diabetes. Nat Genet. 2003;34(3):267–73. https://doi.org/10.1038/ng1180.CrossRefPubMed

25.

Smiljanovic B, Grutzkau A, Sorensen T, Grun JR, Vogl T, Bonin M, et al. Synovial tissue transcriptomes of long-standing rheumatoid arthritis are dominated by activated macrophages that reflect microbial stimulation. Sci Rep. 2020;10(1):7907. https://doi.org/10.1038/s41598-020-64431-4.CrossRefPubMedPubMedCentral

26.

Grützkau A, Radbruch A. Separation of whole blood cells and its impact on gene expression. In: Bosio A, Gerstmayer B, editors. Birkhäuser Basel Microarrays in inflammation progress in inflammation research; 2008.

27.

Wesemann DR, Benveniste EN. STAT-1 alpha and IFN-gamma as modulators of TNF-alpha signaling in macrophages: regulation and functional implications of the TNF receptor 1:STAT-1 alpha complex. J Immunol. 2003;171(10):5313–9. https://doi.org/10.4049/jimmunol.171.10.5313.CrossRefPubMed

28.

Ma X, Jiang Z, Li N, Jiang W, Gao P, Yang M, et al. Ets2 suppresses inflammatory cytokines through MAPK/NF-kappaB signaling and directly binds to the IL-6 promoter in macrophages. Aging (Albany NY). 2019;11(22):10610–25. https://doi.org/10.18632/aging.102480.CrossRef

29.

Rapin N, Bagger FO, Jendholm J, Mora-Jensen H, Krogh A, Kohlmann A, et al. Comparing cancer vs normal gene expression profiles identifies new disease entities and common transcriptional programs in AML patients. Blood. 2014;123(6):894–904. https://doi.org/10.1182/blood-2013-02-485771.CrossRefPubMed

30.

Buzzeo MP, Yang J, Casella G, Reddy V. Hematopoietic stem cell mobilization with G-CSF induces innate inflammation yet suppresses adaptive immune gene expression as revealed by microarray analysis. Exp Hematol. 2007;35(9):1456–65. https://doi.org/10.1016/j.exphem.2007.06.001.CrossRefPubMed

31.

Smiljanovic B, Grun JR, Biesen R, Schulte-Wrede U, Baumgrass R, Stuhlmuller B, et al. The multifaceted balance of TNF-alpha and type I/II interferon responses in SLE and RA: how monocytes manage the impact of cytokines. J Mol Med (Berl). 2012;90(11):1295–309. https://doi.org/10.1007/s00109-012-0907-y.CrossRef

32.

Evrard M, Kwok IWH, Chong SZ, Teng KWW, Becht E, Chen J, et al. Developmental analysis of bone marrow neutrophils reveals populations specialized in expansion, trafficking, and effector functions. Immunity. 2018;48(2):364–79 e368. https://doi.org/10.1016/j.immuni.2018.02.002.CrossRefPubMed

33.

Adriaanse MP, Leffler DA, Kelly CP, Schuppan D, Najarian RM, Goldsmith JD, et al. Serum I-FABP detects gluten responsiveness in adult celiac disease patients on a short-term gluten challenge. Am J Gastroenterol. 2016;111(7):1014–22. https://doi.org/10.1038/ajg.2016.162.CrossRefPubMed

34.

Lakatos PL, Kiss LS, Palatka K, Altorjay I, Antal-Szalmas P, Palyu E, et al. Serum lipopolysaccharide-binding protein and soluble CD14 are markers of disease activity in patients with Crohn's disease. Inflamm Bowel Dis. 2011;17(3):767–77. https://doi.org/10.1002/ibd.21402.CrossRefPubMed

35.

Ramendra R, Isnard S, Mehraj V, Chen J, Zhang Y, Finkelman M, et al. Circulating LPS and (1-->3)-beta-D-Glucan: a folie a deux contributing to HIV-associated immune activation. Front Immunol. 2019;10:465. https://doi.org/10.3389/fimmu.2019.00465.CrossRefPubMedPubMedCentral

36.

Ramiro S, van der Heijde D, van Tubergen A, Stolwijk C, Dougados M, van den Bosch F, et al. Higher disease activity leads to more structural damage in the spine in ankylosing spondylitis: 12-year longitudinal data from the OASIS cohort. Ann Rheum Dis. 2014;73(8):1455–61. https://doi.org/10.1136/annrheumdis-2014-205178.CrossRefPubMed

37.

Bleil J, Maier R, Hempfing A, Schlichting U, Appel H, Sieper J, et al. Histomorphological and histomorphometric characteristics of zygapophyseal joint remodelling in ankylosing spondylitis. Arthritis Rheum. 2014;66(7):1745–54. https://doi.org/10.1002/art.38404.CrossRef

38.

Regan-Komito D, Swann JW, Demetriou P, Cohen ES, Horwood NJ, Sansom SN, et al. GM-CSF drives dysregulated hematopoietic stem cell activity and pathogenic extramedullary myelopoiesis in experimental spondyloarthritis. Nat Commun. 2020;11(1):155. https://doi.org/10.1038/s41467-019-13853-4.CrossRefPubMedPubMedCentral

39.

Molinaro A, Koh A, Wu H, Schoeler M, Faggi MI, Carreras A, et al. Hepatic expression of lipopolysaccharide-binding protein (Lbp) is induced by the gut microbiota through Myd88 and impairs glucose tolerance in mice independent of obesity. Mol Metab. 2020;37:100997. https://doi.org/10.1016/j.molmet.2020.100997.CrossRefPubMedPubMedCentral

40.

Blairon L, Wittebole X, Laterre PF. Lipopolysaccharide-binding protein serum levels in patients with severe sepsis due to gram-positive and fungal infections. J Infect Dis. 2003;187(2):287–91. https://doi.org/10.1086/346046.CrossRefPubMed

41.

Zhao S, Fung-Leung WP, Bittner A, Ngo K, Liu X. Comparison of RNA-Seq and microarray in transcriptome profiling of activated T cells. PLoS One. 2014;9(1):e78644. https://doi.org/10.1371/journal.pone.0078644.CrossRefPubMedPubMedCentral

42.

Deodhar A, Strand V, Kay J, Braun J. The term 'non-radiographic axial spondyloarthritis' is much more important to classify than to diagnose patients with axial spondyloarthritis. Ann Rheum Dis. 2016;75(5):791–4. https://doi.org/10.1136/annrheumdis-2015-208852.CrossRefPubMed

43.

Pearce EJ, Everts B. Dendritic cell metabolism. Nat Rev Immunol. 2015;15(1):18–29. https://doi.org/10.1038/nri3771.CrossRefPubMedPubMedCentral

44.

Injarabian L, Devin A, Ransac S, Marteyn BS. Neutrophil metabolic shift during their lifecycle: impact on their survival and activation. Int J Mol Sci. 2019;21(1). https://doi.org/10.3390/ijms21010287.

45.

Im CH, Kang EH, Ki JY, Shin DW, Choi HJ, Chang EJ, et al. Receptor activator of nuclear factor kappa B ligand-mediated osteoclastogenesis is elevated in ankylosing spondylitis. Clin Exp Rheumatol. 2009;27(4):620–5.PubMed

46.

Lemma S, Sboarina M, Porporato PE, Zini N, Sonveaux P, Di Pompo G, et al. Energy metabolism in osteoclast formation and activity. Int J Biochem Cell Biol. 2016;79:168–80. https://doi.org/10.1016/j.biocel.2016.08.034.CrossRefPubMed

Title: Monocyte transcriptomes from patients with axial spondyloarthritis reveal dysregulated monocytopoiesis and a distinct inflammatory imprint
Authors: Fabian Karow
Biljana Smiljanovic
Joachim R. Grün
Denis Poddubnyy
Fabian Proft
Alice Talpin
Christophe Hue
Anne Boland
Jean-François Deleuze
Henri-Jean Garchon
Ilkay Ergenç
Ann-Sophie De Craemer
Ulrike Erben
Thomas Häupl
Dirk Elewaut
Maxime Breban
Andreas Grützkau
Uta Syrbe
Publication date: 01-12-2021
Publisher: BioMed Central
Keywords: Ankylosing Spondylitis
Spondyloarthropathy
Published in: Arthritis Research & Therapy / Issue 1/2021
Electronic ISSN: 1478-6362
DOI: https://doi.org/10.1186/s13075-021-02623-7

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Pediatric Cardiology Video

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

Cardiopulmonary Comorbidity Video

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Valvular Heart Disease Video

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Heart Failure Video

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.

Watch now

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

16-04-2024 Type 2 Diabetes News

Incentivised to exercise

11-04-2024 Lifestyle Modifications News

New intervention for STEMI-related cardiogenic shock

10-04-2024 Myocardial Infarction News

» View more highlights on the ACC 2024 congress hub page

Image Credits

STEP AAG HeroTeaserCollection image/© Tarek / Generated with AI / Stock.adobe.com, ACC 2024 Pediatric and Congenital Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Pulmonary Vascular Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 Valvular Heart Disease: Year in Review/© 2024 American College of Cardiology, ACC 2024 HF and Cardiomyopathies: Year in Review/© 2024 American College of Cardiology, Woman out walking/© Seventyfour / stock.adobe.com (symbolic image with model), Woman checking smartwatch /© saltdium / stock.adobe.com (symbolic image with model), Cardiac catheterization/© Damian / stock.adobe.com

At a glance: The STEP trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2021

Diagnosis of early stage knee osteoarthritis based on early clinical course: data from the CHECK cohort

Serial Lipocalin 2 and Oncostatin M levels reflect inflammation status and treatment response in axial spondyloarthritis

Sine scleroderma, limited cutaneous, and diffused cutaneous systemic sclerosis survival and predictors of mortality

The role of lung ultrasound B-lines and serum KL-6 in the screening and follow-up of rheumatoid arthritis patients for an identification of interstitial lung disease: review of the literature, proposal for a preliminary algorithm, and clinical application to cases

Involvement of lncRNA IL21-AS1 in interleukin-2 and T follicular regulatory cell activation in systemic lupus erythematosus

Increase of aerobic glycolysis mediated by activated T helper cells drives synovial fibroblasts towards an inflammatory phenotype: new targets for therapy?

Highlights from the ACC 2024 Congress

ACC 2024 Congress Coverage