CXCL4 drives fibrosis by promoting several key cellular and molecular processes

Alsya J. Affandi; Tiago Carvalheiro; Andrea Ottria; Judith J. de Haan; Maike A.D. Brans; Maarten M. Brandt; Ralph G. Tieland; Ana P. Lopes; Beatriz Malvar Fernández; Cornelis P.J. Bekker; Maarten van der Linden; Maili Zimmermann; Barbara Giovannone; Catharina G.K. Wichers; Samuel Garcia; Michael de Kok; Giuseppina Stifano; Yan Juan Xu; M. Anna Kowalska; Maaike Waasdorp; Caroline Cheng; Susan Gibbs; Saskia C.A. de Jager; Joel A.G. van Roon; Timothy R.D.J. Radstake; Wioleta Marut

doi:10.1016/j.celrep.2021.110189

CXCL4 drives fibrosis by promoting several key cellular and molecular processes

Alsya J. Affandi, Tiago Carvalheiro, Andrea Ottria, Judith J. de Haan, Maike A.D. Brans, Maarten M. Brandt, Ralph G. Tieland, Ana P. Lopes, Beatriz Malvar Fernández, Cornelis P.J. Bekker, Maarten van der Linden, Maili Zimmermann, Barbara Giovannone, Catharina G.K. Wichers, Samuel Garcia, Michael de Kok, Giuseppina Stifano, Yan Juan Xu, M. Anna Kowalska, Maaike WaasdorpCaroline Cheng, Susan Gibbs, Saskia C.A. de Jager, Joel A.G. van Roon, Timothy R.D.J. Radstake, Wioleta Marut^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Fibrosis is a major cause of mortality worldwide, characterized by myofibroblast activation and excessive extracellular matrix deposition. Systemic sclerosis is a prototypic fibrotic disease in which CXCL4 is increased and strongly correlates with skin and lung fibrosis. Here we aim to elucidate the role of CXCL4 in fibrosis development. CXCL4 levels are increased in multiple inflammatory and fibrotic mouse models, and, using CXCL4-deficient mice, we demonstrate the essential role of CXCL4 in promoting fibrotic events in the skin, lungs, and heart. Overexpressing human CXCL4 in mice aggravates, whereas blocking CXCL4 reduces, bleomycin-induced fibrosis. Single-cell ligand-receptor analysis predicts CXCL4 to affect endothelial cells and fibroblasts. In vitro, we confirm that CXCL4 directly induces myofibroblast differentiation and collagen synthesis in different precursor cells, including endothelial cells, by stimulating endothelial-to-mesenchymal transition. Our findings identify a pivotal role of CXCL4 in fibrosis, further substantiating the potential role of neutralizing CXCL4 as a therapeutic strategy.

Original language	English
Article number	110189
Journal	Cell Reports
Volume	38
Issue number	1
DOIs	https://doi.org/10.1016/j.celrep.2021.110189
Publication status	Published - 4 Jan 2022
Externally published	Yes

Bibliographical note

Funding Information:
A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology: Funda??o para a Ci?ncia e a Tecnologia [SFRH/BD/93526/2013]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant (ERC-2011-StG, Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript. A.J.A. T.C. T.R.D.J.R. and W.M. conceived the experiments and drafted and revised the manuscript. A.J.A. T.C. C.C. M.A.K. S. Gibbs, S.C.A.d.J. J.A.G.v.R. T.R.D.J.R. and W.M. designed the experiments. A.J.A. T.C. A.O. J.J.d.H. M.A.D.B. M.M.B. R.G.T. A.P.L. B.M.F. C.P.J.B. M.v.d.L. M.Z. B.G. C.G.K.W. S. Garcia, M.d.K. M.W. G.S. and Y.J.X. carried out the experiments and analyses. C.P.J.B. M.A.K. J.A.G.v.R. S. Garcia, M.W. and S.C.A.d.J. revised the manuscript. J.A.G.v.R. T.R.D.J.R. and W.M. supervised the study. All authors made substantial, direct, and intellectual contributions to the work and approved the final version of the manuscript. The authors declare no competing interests.

Funding Information:
A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301 ) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014 ). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology : Fundação para a Ciência e a Tecnologia [ SFRH/BD/93526/2013 ]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant ( ERC-2011-StG , Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149 ). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript.

Publisher Copyright:
© 2021 The Authors

Funding

A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301 ) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014 ). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology : Fundação para a Ciência e a Tecnologia [ SFRH/BD/93526/2013 ]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant ( ERC-2011-StG , Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149 ). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript.

Keywords

bleomycin
CXCL4
endothelial-to-mesenchymal transition
fibrosis
inflammation
myofibroblast
systemic sclerosis

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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Cite this

Affandi, A. J., Carvalheiro, T., Ottria, A., de Haan, J. J., Brans, M. A. D., Brandt, M. M., Tieland, R. G., Lopes, A. P., Fernández, B. M., Bekker, C. P. J., van der Linden, M., Zimmermann, M., Giovannone, B., Wichers, C. G. K., Garcia, S., de Kok, M., Stifano, G., Xu, Y. J., Kowalska, M. A., ... Marut, W. (2022). CXCL4 drives fibrosis by promoting several key cellular and molecular processes. Cell Reports, 38(1), Article 110189. https://doi.org/10.1016/j.celrep.2021.110189

@article{8280bb1630ff4a4ea08fc9f370b4a39f,

title = "CXCL4 drives fibrosis by promoting several key cellular and molecular processes",

abstract = "Fibrosis is a major cause of mortality worldwide, characterized by myofibroblast activation and excessive extracellular matrix deposition. Systemic sclerosis is a prototypic fibrotic disease in which CXCL4 is increased and strongly correlates with skin and lung fibrosis. Here we aim to elucidate the role of CXCL4 in fibrosis development. CXCL4 levels are increased in multiple inflammatory and fibrotic mouse models, and, using CXCL4-deficient mice, we demonstrate the essential role of CXCL4 in promoting fibrotic events in the skin, lungs, and heart. Overexpressing human CXCL4 in mice aggravates, whereas blocking CXCL4 reduces, bleomycin-induced fibrosis. Single-cell ligand-receptor analysis predicts CXCL4 to affect endothelial cells and fibroblasts. In vitro, we confirm that CXCL4 directly induces myofibroblast differentiation and collagen synthesis in different precursor cells, including endothelial cells, by stimulating endothelial-to-mesenchymal transition. Our findings identify a pivotal role of CXCL4 in fibrosis, further substantiating the potential role of neutralizing CXCL4 as a therapeutic strategy.",

keywords = "bleomycin, CXCL4, endothelial-to-mesenchymal transition, fibrosis, inflammation, myofibroblast, systemic sclerosis",

author = "Affandi, {Alsya J.} and Tiago Carvalheiro and Andrea Ottria and {de Haan}, {Judith J.} and Brans, {Maike A.D.} and Brandt, {Maarten M.} and Tieland, {Ralph G.} and Lopes, {Ana P.} and Fern{\'a}ndez, {Beatriz Malvar} and Bekker, {Cornelis P.J.} and {van der Linden}, Maarten and Maili Zimmermann and Barbara Giovannone and Wichers, {Catharina G.K.} and Samuel Garcia and {de Kok}, Michael and Giuseppina Stifano and Xu, {Yan Juan} and Kowalska, {M. Anna} and Maaike Waasdorp and Caroline Cheng and Susan Gibbs and {de Jager}, {Saskia C.A.} and {van Roon}, {Joel A.G.} and Radstake, {Timothy R.D.J.} and Wioleta Marut",

note = "Funding Information: A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology: Funda??o para a Ci?ncia e a Tecnologia [SFRH/BD/93526/2013]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant (ERC-2011-StG, Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript. A.J.A. T.C. T.R.D.J.R. and W.M. conceived the experiments and drafted and revised the manuscript. A.J.A. T.C. C.C. M.A.K. S. Gibbs, S.C.A.d.J. J.A.G.v.R. T.R.D.J.R. and W.M. designed the experiments. A.J.A. T.C. A.O. J.J.d.H. M.A.D.B. M.M.B. R.G.T. A.P.L. B.M.F. C.P.J.B. M.v.d.L. M.Z. B.G. C.G.K.W. S. Garcia, M.d.K. M.W. G.S. and Y.J.X. carried out the experiments and analyses. C.P.J.B. M.A.K. J.A.G.v.R. S. Garcia, M.W. and S.C.A.d.J. revised the manuscript. J.A.G.v.R. T.R.D.J.R. and W.M. supervised the study. All authors made substantial, direct, and intellectual contributions to the work and approved the final version of the manuscript. The authors declare no competing interests. Funding Information: A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301 ) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014 ). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology : Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia [ SFRH/BD/93526/2013 ]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant ( ERC-2011-StG , Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149 ). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript. Publisher Copyright: {\textcopyright} 2021 The Authors",

year = "2022",

month = jan,

day = "4",

doi = "10.1016/j.celrep.2021.110189",

language = "English",

volume = "38",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Elsevier Saunders",

number = "1",

}

Affandi, AJ, Carvalheiro, T, Ottria, A, de Haan, JJ, Brans, MAD, Brandt, MM, Tieland, RG, Lopes, AP, Fernández, BM, Bekker, CPJ, van der Linden, M, Zimmermann, M, Giovannone, B, Wichers, CGK, Garcia, S, de Kok, M, Stifano, G, Xu, YJ, Kowalska, MA, Waasdorp, M, Cheng, C, Gibbs, S, de Jager, SCA, van Roon, JAG, Radstake, TRDJ & Marut, W 2022, 'CXCL4 drives fibrosis by promoting several key cellular and molecular processes', Cell Reports, vol. 38, no. 1, 110189. https://doi.org/10.1016/j.celrep.2021.110189

TY - JOUR

T1 - CXCL4 drives fibrosis by promoting several key cellular and molecular processes

AU - Affandi, Alsya J.

AU - Carvalheiro, Tiago

AU - Ottria, Andrea

AU - de Haan, Judith J.

AU - Brans, Maike A.D.

AU - Brandt, Maarten M.

AU - Tieland, Ralph G.

AU - Lopes, Ana P.

AU - Fernández, Beatriz Malvar

AU - Bekker, Cornelis P.J.

AU - van der Linden, Maarten

AU - Zimmermann, Maili

AU - Giovannone, Barbara

AU - Wichers, Catharina G.K.

AU - Garcia, Samuel

AU - de Kok, Michael

AU - Stifano, Giuseppina

AU - Xu, Yan Juan

AU - Kowalska, M. Anna

AU - Waasdorp, Maaike

AU - Cheng, Caroline

AU - Gibbs, Susan

AU - de Jager, Saskia C.A.

AU - van Roon, Joel A.G.

AU - Radstake, Timothy R.D.J.

AU - Marut, Wioleta

N1 - Funding Information: A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology: Funda??o para a Ci?ncia e a Tecnologia [SFRH/BD/93526/2013]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant (ERC-2011-StG, Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript. A.J.A. T.C. T.R.D.J.R. and W.M. conceived the experiments and drafted and revised the manuscript. A.J.A. T.C. C.C. M.A.K. S. Gibbs, S.C.A.d.J. J.A.G.v.R. T.R.D.J.R. and W.M. designed the experiments. A.J.A. T.C. A.O. J.J.d.H. M.A.D.B. M.M.B. R.G.T. A.P.L. B.M.F. C.P.J.B. M.v.d.L. M.Z. B.G. C.G.K.W. S. Garcia, M.d.K. M.W. G.S. and Y.J.X. carried out the experiments and analyses. C.P.J.B. M.A.K. J.A.G.v.R. S. Garcia, M.W. and S.C.A.d.J. revised the manuscript. J.A.G.v.R. T.R.D.J.R. and W.M. supervised the study. All authors made substantial, direct, and intellectual contributions to the work and approved the final version of the manuscript. The authors declare no competing interests. Funding Information: A.J.A. was supported by the Dutch Arthritis Association (Reumafonds grant NR-10-1-301 ) and the Netherlands Organisation for Scientific Research (NWO, Mosaic grant 017.008.014 ). T.C. was supported by a grant from the Portuguese National Funding Agency for Science, Research and Technology : Fundação para a Ciência e a Tecnologia [ SFRH/BD/93526/2013 ]. T.R.D.J.R and J.A.G.v.R. received funding from an European Research Council (ERC) Starting Grant ( ERC-2011-StG , Circumvent). W.M. obtained funding from the Marie Curie Intra-European Fellowship (Proposal number: 624871) and from the NWO VENI (grant number 91919149 ). We thank Mike DiMarzio, Giuseppina A. Farina (Boston University School of Medicine), Sarita Hartgring, Aniek Meijers, Karin de Cortie, Wouter van Leeuwen, Dirk-Jan Heijnen, Enric Mocholi-Gimeno, and Christian van Dijk (University Medical Center Utrecht) for technical advice, support, and expertise. We also thank Kris A. Reedquist for critical review of the manuscript. Publisher Copyright: © 2021 The Authors

PY - 2022/1/4

Y1 - 2022/1/4

N2 - Fibrosis is a major cause of mortality worldwide, characterized by myofibroblast activation and excessive extracellular matrix deposition. Systemic sclerosis is a prototypic fibrotic disease in which CXCL4 is increased and strongly correlates with skin and lung fibrosis. Here we aim to elucidate the role of CXCL4 in fibrosis development. CXCL4 levels are increased in multiple inflammatory and fibrotic mouse models, and, using CXCL4-deficient mice, we demonstrate the essential role of CXCL4 in promoting fibrotic events in the skin, lungs, and heart. Overexpressing human CXCL4 in mice aggravates, whereas blocking CXCL4 reduces, bleomycin-induced fibrosis. Single-cell ligand-receptor analysis predicts CXCL4 to affect endothelial cells and fibroblasts. In vitro, we confirm that CXCL4 directly induces myofibroblast differentiation and collagen synthesis in different precursor cells, including endothelial cells, by stimulating endothelial-to-mesenchymal transition. Our findings identify a pivotal role of CXCL4 in fibrosis, further substantiating the potential role of neutralizing CXCL4 as a therapeutic strategy.

AB - Fibrosis is a major cause of mortality worldwide, characterized by myofibroblast activation and excessive extracellular matrix deposition. Systemic sclerosis is a prototypic fibrotic disease in which CXCL4 is increased and strongly correlates with skin and lung fibrosis. Here we aim to elucidate the role of CXCL4 in fibrosis development. CXCL4 levels are increased in multiple inflammatory and fibrotic mouse models, and, using CXCL4-deficient mice, we demonstrate the essential role of CXCL4 in promoting fibrotic events in the skin, lungs, and heart. Overexpressing human CXCL4 in mice aggravates, whereas blocking CXCL4 reduces, bleomycin-induced fibrosis. Single-cell ligand-receptor analysis predicts CXCL4 to affect endothelial cells and fibroblasts. In vitro, we confirm that CXCL4 directly induces myofibroblast differentiation and collagen synthesis in different precursor cells, including endothelial cells, by stimulating endothelial-to-mesenchymal transition. Our findings identify a pivotal role of CXCL4 in fibrosis, further substantiating the potential role of neutralizing CXCL4 as a therapeutic strategy.

KW - bleomycin

KW - CXCL4

KW - endothelial-to-mesenchymal transition

KW - fibrosis

KW - inflammation

KW - myofibroblast

KW - systemic sclerosis

UR - http://www.scopus.com/inward/record.url?scp=85122040732&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2021.110189

DO - 10.1016/j.celrep.2021.110189

M3 - Article

AN - SCOPUS:85122040732

SN - 2211-1247

VL - 38

JO - Cell Reports

JF - Cell Reports

IS - 1

M1 - 110189

ER -

CXCL4 drives fibrosis by promoting several key cellular and molecular processes

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

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