Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α

Andrea Ottria; Maili Zimmermann; Laurent M Paardekooper; Tiago Carvalheiro; Nadia Vazirpanah; Sandra Silva-Cardoso; Alsya J Affandi; Eleni Chouri; Maarten V D Kroef; Ralph G Tieland; Cornelis P J Bekker; Catharina G K Wichers; Marzia Rossato; Enric Mocholi-Gimeno; Janneke Tekstra; Evelien Ton; Jaap M van Laar; Marta Cossu; Lorenzo Beretta; Samuel Garcia Perez; Aridaman Pandit; Femke Bonte-Mineur; Kris A Reedquist; Geert van den Bogaart; Timothy R D J Radstake; Wioleta Marut

doi:10.1093/rheumatology/keab532

Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α

Andrea Ottria, Maili Zimmermann, Laurent M Paardekooper, Tiago Carvalheiro, Nadia Vazirpanah, Sandra Silva-Cardoso, Alsya J Affandi, Eleni Chouri, Maarten V D Kroef, Ralph G Tieland, Cornelis P J Bekker, Catharina G K Wichers, Marzia Rossato, Enric Mocholi-Gimeno, Janneke Tekstra, Evelien Ton, Jaap M van Laar, Marta Cossu, Lorenzo Beretta, Samuel Garcia PerezAridaman Pandit, Femke Bonte-Mineur, Kris A Reedquist, Geert van den Bogaart, Timothy R D J Radstake, Wioleta Marut

Group Nierkens

Onderzoeksoutput: Bijdrage aan tijdschrift › Artikel › peer review

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OBJECTIVE: SSc is a complex disease characterized by vascular abnormalities and inflammation culminating in hypoxia and excessive fibrosis. Previously, we identified chemokine (C-X-C motif) ligand 4 (CXCL4) as a novel predictive biomarker in SSc. Although CXCL4 is well-studied, the mechanisms driving its production are unclear. The aim of this study was to elucidate the mechanisms leading to CXCL4 production.

METHODS: Plasmacytoid dendritic cells (pDCs) from 97 healthy controls and 70 SSc patients were cultured in the presence of hypoxia or atmospheric oxygen level and/or stimulated with several toll-like receptor (TLR) agonists. Further, pro-inflammatory cytokine production, CXCL4, hypoxia-inducible factor (HIF) -1α and HIF-2α gene and protein expression were assessed using ELISA, Luminex, qPCR, FACS and western blot assays.

RESULTS: CXCL4 release was potentiated only when pDCs were simultaneously exposed to hypoxia and TLR9 agonist (P < 0.0001). Here, we demonstrated that CXCL4 production is dependent on the overproduction of mitochondrial reactive oxygen species (mtROS) (P = 0.0079) leading to stabilization of HIF-2α (P = 0.029). In addition, we show that hypoxia is fundamental for CXCL4 production by umbilical cord CD34 derived pDCs.

CONCLUSION: TLR-mediated activation of immune cells in the presence of hypoxia underpins the pathogenic production of CXCL4 in SSc. Blocking either mtROS or HIF-2α pathways may therapeutically attenuate the contribution of CXCL4 to SSc and other inflammatory diseases driven by CXCL4.

Originele taal-2	Engels
Pagina's (van-tot)	2682-2693
Aantal pagina's	12
Tijdschrift	Rheumatology (Oxford, England)
Volume	61
Nummer van het tijdschrift	6
DOI's	https://doi.org/10.1093/rheumatology/keab532
Status	Gepubliceerd - 30 mei 2022

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10.1093/rheumatology/keab532

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Ottria, A., Zimmermann, M., Paardekooper, L. M., Carvalheiro, T., Vazirpanah, N., Silva-Cardoso, S., Affandi, A. J., Chouri, E., V D Kroef, M., Tieland, R. G., Bekker, C. P. J., Wichers, C. G. K., Rossato, M., Mocholi-Gimeno, E., Tekstra, J., Ton, E., van Laar, J. M., Cossu, M., Beretta, L., ... Marut, W. (2022). Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α. Rheumatology (Oxford, England), 61(6), 2682-2693. https://doi.org/10.1093/rheumatology/keab532

@article{7e1df96907734427a2761763df1dea56,

title = "Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α",

abstract = "OBJECTIVE: SSc is a complex disease characterized by vascular abnormalities and inflammation culminating in hypoxia and excessive fibrosis. Previously, we identified chemokine (C-X-C motif) ligand 4 (CXCL4) as a novel predictive biomarker in SSc. Although CXCL4 is well-studied, the mechanisms driving its production are unclear. The aim of this study was to elucidate the mechanisms leading to CXCL4 production.METHODS: Plasmacytoid dendritic cells (pDCs) from 97 healthy controls and 70 SSc patients were cultured in the presence of hypoxia or atmospheric oxygen level and/or stimulated with several toll-like receptor (TLR) agonists. Further, pro-inflammatory cytokine production, CXCL4, hypoxia-inducible factor (HIF) -1α and HIF-2α gene and protein expression were assessed using ELISA, Luminex, qPCR, FACS and western blot assays.RESULTS: CXCL4 release was potentiated only when pDCs were simultaneously exposed to hypoxia and TLR9 agonist (P < 0.0001). Here, we demonstrated that CXCL4 production is dependent on the overproduction of mitochondrial reactive oxygen species (mtROS) (P = 0.0079) leading to stabilization of HIF-2α (P = 0.029). In addition, we show that hypoxia is fundamental for CXCL4 production by umbilical cord CD34 derived pDCs.CONCLUSION: TLR-mediated activation of immune cells in the presence of hypoxia underpins the pathogenic production of CXCL4 in SSc. Blocking either mtROS or HIF-2α pathways may therapeutically attenuate the contribution of CXCL4 to SSc and other inflammatory diseases driven by CXCL4.",

keywords = "Basic Helix-Loop-Helix Transcription Factors/metabolism, Dendritic Cells/metabolism, Humans, Hypoxia/metabolism, Hypoxia-Inducible Factor 1, alpha Subunit, Platelet Factor 4/metabolism, Reactive Oxygen Species/metabolism, Scleroderma, Systemic, Toll-Like Receptor 9",

author = "Andrea Ottria and Maili Zimmermann and Paardekooper, {Laurent M} and Tiago Carvalheiro and Nadia Vazirpanah and Sandra Silva-Cardoso and Affandi, {Alsya J} and Eleni Chouri and {V D Kroef}, Maarten and Tieland, {Ralph G} and Bekker, {Cornelis P J} and Wichers, {Catharina G K} and Marzia Rossato and Enric Mocholi-Gimeno and Janneke Tekstra and Evelien Ton and {van Laar}, {Jaap M} and Marta Cossu and Lorenzo Beretta and {Garcia Perez}, Samuel and Aridaman Pandit and Femke Bonte-Mineur and Reedquist, {Kris A} and {van den Bogaart}, Geert and Radstake, {Timothy R D J} and Wioleta Marut",

note = "{\textcopyright} The Author(s) 2021. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.com.",

year = "2022",

month = may,

day = "30",

doi = "10.1093/rheumatology/keab532",

language = "English",

volume = "61",

pages = "2682--2693",

journal = "Rheumatology (Oxford, England)",

issn = "1462-0324",

number = "6",

}

Ottria, A, Zimmermann, M, Paardekooper, LM, Carvalheiro, T, Vazirpanah, N, Silva-Cardoso, S, Affandi, AJ, Chouri, E, V D Kroef, M, Tieland, RG, Bekker, CPJ, Wichers, CGK, Rossato, M, Mocholi-Gimeno, E, Tekstra, J, Ton, E, van Laar, JM, Cossu, M, Beretta, L, Garcia Perez, S, Pandit, A, Bonte-Mineur, F, Reedquist, KA, van den Bogaart, G, Radstake, TRDJ & Marut, W 2022, 'Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α', Rheumatology (Oxford, England), vol. 61, nr. 6, blz. 2682-2693. https://doi.org/10.1093/rheumatology/keab532

TY - JOUR

T1 - Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α

AU - Ottria, Andrea

AU - Zimmermann, Maili

AU - Paardekooper, Laurent M

AU - Carvalheiro, Tiago

AU - Vazirpanah, Nadia

AU - Silva-Cardoso, Sandra

AU - Affandi, Alsya J

AU - Chouri, Eleni

AU - V D Kroef, Maarten

AU - Tieland, Ralph G

AU - Bekker, Cornelis P J

AU - Wichers, Catharina G K

AU - Rossato, Marzia

AU - Mocholi-Gimeno, Enric

AU - Tekstra, Janneke

AU - Ton, Evelien

AU - van Laar, Jaap M

AU - Cossu, Marta

AU - Beretta, Lorenzo

AU - Garcia Perez, Samuel

AU - Pandit, Aridaman

AU - Bonte-Mineur, Femke

AU - Reedquist, Kris A

AU - van den Bogaart, Geert

AU - Radstake, Timothy R D J

AU - Marut, Wioleta

PY - 2022/5/30

Y1 - 2022/5/30

N2 - OBJECTIVE: SSc is a complex disease characterized by vascular abnormalities and inflammation culminating in hypoxia and excessive fibrosis. Previously, we identified chemokine (C-X-C motif) ligand 4 (CXCL4) as a novel predictive biomarker in SSc. Although CXCL4 is well-studied, the mechanisms driving its production are unclear. The aim of this study was to elucidate the mechanisms leading to CXCL4 production.METHODS: Plasmacytoid dendritic cells (pDCs) from 97 healthy controls and 70 SSc patients were cultured in the presence of hypoxia or atmospheric oxygen level and/or stimulated with several toll-like receptor (TLR) agonists. Further, pro-inflammatory cytokine production, CXCL4, hypoxia-inducible factor (HIF) -1α and HIF-2α gene and protein expression were assessed using ELISA, Luminex, qPCR, FACS and western blot assays.RESULTS: CXCL4 release was potentiated only when pDCs were simultaneously exposed to hypoxia and TLR9 agonist (P < 0.0001). Here, we demonstrated that CXCL4 production is dependent on the overproduction of mitochondrial reactive oxygen species (mtROS) (P = 0.0079) leading to stabilization of HIF-2α (P = 0.029). In addition, we show that hypoxia is fundamental for CXCL4 production by umbilical cord CD34 derived pDCs.CONCLUSION: TLR-mediated activation of immune cells in the presence of hypoxia underpins the pathogenic production of CXCL4 in SSc. Blocking either mtROS or HIF-2α pathways may therapeutically attenuate the contribution of CXCL4 to SSc and other inflammatory diseases driven by CXCL4.

AB - OBJECTIVE: SSc is a complex disease characterized by vascular abnormalities and inflammation culminating in hypoxia and excessive fibrosis. Previously, we identified chemokine (C-X-C motif) ligand 4 (CXCL4) as a novel predictive biomarker in SSc. Although CXCL4 is well-studied, the mechanisms driving its production are unclear. The aim of this study was to elucidate the mechanisms leading to CXCL4 production.METHODS: Plasmacytoid dendritic cells (pDCs) from 97 healthy controls and 70 SSc patients were cultured in the presence of hypoxia or atmospheric oxygen level and/or stimulated with several toll-like receptor (TLR) agonists. Further, pro-inflammatory cytokine production, CXCL4, hypoxia-inducible factor (HIF) -1α and HIF-2α gene and protein expression were assessed using ELISA, Luminex, qPCR, FACS and western blot assays.RESULTS: CXCL4 release was potentiated only when pDCs were simultaneously exposed to hypoxia and TLR9 agonist (P < 0.0001). Here, we demonstrated that CXCL4 production is dependent on the overproduction of mitochondrial reactive oxygen species (mtROS) (P = 0.0079) leading to stabilization of HIF-2α (P = 0.029). In addition, we show that hypoxia is fundamental for CXCL4 production by umbilical cord CD34 derived pDCs.CONCLUSION: TLR-mediated activation of immune cells in the presence of hypoxia underpins the pathogenic production of CXCL4 in SSc. Blocking either mtROS or HIF-2α pathways may therapeutically attenuate the contribution of CXCL4 to SSc and other inflammatory diseases driven by CXCL4.

KW - Basic Helix-Loop-Helix Transcription Factors/metabolism

KW - Dendritic Cells/metabolism

KW - Humans

KW - Hypoxia/metabolism

KW - Hypoxia-Inducible Factor 1, alpha Subunit

KW - Platelet Factor 4/metabolism

KW - Reactive Oxygen Species/metabolism

KW - Scleroderma, Systemic

KW - Toll-Like Receptor 9

U2 - 10.1093/rheumatology/keab532

DO - 10.1093/rheumatology/keab532

M3 - Article

C2 - 34559222

VL - 61

SP - 2682

EP - 2693

JO - Rheumatology (Oxford, England)

JF - Rheumatology (Oxford, England)

SN - 1462-0324

IS - 6

ER -

Hypoxia and TLR9 activation drive CXCL4 production in systemic sclerosis plasmacytoid dendritic cells via mtROS and HIF-2α

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