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Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling

AIMS: Atherosclerotic plaque hypoxia is detrimental for macrophage function. Prolyl hydroxylases (PHDs) initiate cellular hypoxic responses, possibly influencing macrophage function in plaque hypoxia. Thus, we aimed to elucidate the role of myeloid PHDs in atherosclerosis. METHODS AND RESULTS: Myelo...

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Autores principales: van Kuijk, Kim, Demandt, Jasper A F, Perales-Patón, Javier, Theelen, Thomas L, Kuppe, Christoph, Marsch, Elke, de Bruijn, Jenny, Jin, Han, Gijbels, Marion J, Matic, Ljubica, Mees, Barend M E, Reutelingsperger, Chris P M, Hedin, Ulf, Biessen, Erik A L, Carmeliet, Peter, Baker, Andrew H, Kramann, Rafael K, Schurgers, Leon J, Saez-Rodriguez, Julio, Sluimer, Judith C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8953448/
https://www.ncbi.nlm.nih.gov/pubmed/33913468
http://dx.doi.org/10.1093/cvr/cvab152
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author van Kuijk, Kim
Demandt, Jasper A F
Perales-Patón, Javier
Theelen, Thomas L
Kuppe, Christoph
Marsch, Elke
de Bruijn, Jenny
Jin, Han
Gijbels, Marion J
Matic, Ljubica
Mees, Barend M E
Reutelingsperger, Chris P M
Hedin, Ulf
Biessen, Erik A L
Carmeliet, Peter
Baker, Andrew H
Kramann, Rafael K
Schurgers, Leon J
Saez-Rodriguez, Julio
Sluimer, Judith C
author_facet van Kuijk, Kim
Demandt, Jasper A F
Perales-Patón, Javier
Theelen, Thomas L
Kuppe, Christoph
Marsch, Elke
de Bruijn, Jenny
Jin, Han
Gijbels, Marion J
Matic, Ljubica
Mees, Barend M E
Reutelingsperger, Chris P M
Hedin, Ulf
Biessen, Erik A L
Carmeliet, Peter
Baker, Andrew H
Kramann, Rafael K
Schurgers, Leon J
Saez-Rodriguez, Julio
Sluimer, Judith C
author_sort van Kuijk, Kim
collection PubMed
description AIMS: Atherosclerotic plaque hypoxia is detrimental for macrophage function. Prolyl hydroxylases (PHDs) initiate cellular hypoxic responses, possibly influencing macrophage function in plaque hypoxia. Thus, we aimed to elucidate the role of myeloid PHDs in atherosclerosis. METHODS AND RESULTS: Myeloid-specific PHD knockout (PHDko) mice were obtained via bone marrow transplantation (PHD1ko, PHD3ko) or conditional knockdown through lysozyme M-driven Cre recombinase (PHD2cko). Mice were fed high cholesterol diet for 6–12 weeks to induce atherosclerosis. Aortic root plaque size was significantly augmented 2.6-fold in PHD2cko, and 1.4-fold in PHD3ko compared to controls but was unchanged in PHD1ko mice. Macrophage apoptosis was promoted in PHD2cko and PHD3ko mice in vitro and in vivo, via the hypoxia-inducible factor (HIF) 1α/BNIP3 axis. Bulk and single-cell RNA data of PHD2cko bone marrow-derived macrophages (BMDMs) and plaque macrophages, respectively, showed enhanced HIF1α/BNIP3 signalling, which was validated in vitro by siRNA silencing. Human plaque BNIP3 mRNA was positively associated with plaque necrotic core size, suggesting similar pro-apoptotic effects in human. Furthermore, PHD2cko plaques displayed enhanced fibrosis, while macrophage collagen breakdown by matrix metalloproteinases, collagen production, and proliferation were unaltered. Instead, PHD2cko BMDMs enhanced fibroblast collagen secretion in a paracrine manner. In silico analysis of macrophage-fibroblast communication predicted SPP1 (osteopontin) signalling as regulator, which was corroborated by enhanced plaque SPP1 protein in vivo. Increased SPP1 mRNA expression upon PHD2cko was preferentially observed in foamy plaque macrophages expressing ‘triggering receptor expressed on myeloid cells-2’ (TREM2hi) evidenced by single-cell RNA, but not in neutrophils. This confirmed enhanced fibrotic signalling by PHD2cko macrophages to fibroblasts, in vitro as well as in vivo. CONCLUSION: Myeloid PHD2cko and PHD3ko enhanced atherosclerotic plaque growth and macrophage apoptosis, while PHD2cko macrophages further activated collagen secretion by fibroblasts in vitro, likely via paracrine SPP1 signalling through TREM2hi macrophages.
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spelling pubmed-89534482022-03-28 Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling van Kuijk, Kim Demandt, Jasper A F Perales-Patón, Javier Theelen, Thomas L Kuppe, Christoph Marsch, Elke de Bruijn, Jenny Jin, Han Gijbels, Marion J Matic, Ljubica Mees, Barend M E Reutelingsperger, Chris P M Hedin, Ulf Biessen, Erik A L Carmeliet, Peter Baker, Andrew H Kramann, Rafael K Schurgers, Leon J Saez-Rodriguez, Julio Sluimer, Judith C Cardiovasc Res Original Articles AIMS: Atherosclerotic plaque hypoxia is detrimental for macrophage function. Prolyl hydroxylases (PHDs) initiate cellular hypoxic responses, possibly influencing macrophage function in plaque hypoxia. Thus, we aimed to elucidate the role of myeloid PHDs in atherosclerosis. METHODS AND RESULTS: Myeloid-specific PHD knockout (PHDko) mice were obtained via bone marrow transplantation (PHD1ko, PHD3ko) or conditional knockdown through lysozyme M-driven Cre recombinase (PHD2cko). Mice were fed high cholesterol diet for 6–12 weeks to induce atherosclerosis. Aortic root plaque size was significantly augmented 2.6-fold in PHD2cko, and 1.4-fold in PHD3ko compared to controls but was unchanged in PHD1ko mice. Macrophage apoptosis was promoted in PHD2cko and PHD3ko mice in vitro and in vivo, via the hypoxia-inducible factor (HIF) 1α/BNIP3 axis. Bulk and single-cell RNA data of PHD2cko bone marrow-derived macrophages (BMDMs) and plaque macrophages, respectively, showed enhanced HIF1α/BNIP3 signalling, which was validated in vitro by siRNA silencing. Human plaque BNIP3 mRNA was positively associated with plaque necrotic core size, suggesting similar pro-apoptotic effects in human. Furthermore, PHD2cko plaques displayed enhanced fibrosis, while macrophage collagen breakdown by matrix metalloproteinases, collagen production, and proliferation were unaltered. Instead, PHD2cko BMDMs enhanced fibroblast collagen secretion in a paracrine manner. In silico analysis of macrophage-fibroblast communication predicted SPP1 (osteopontin) signalling as regulator, which was corroborated by enhanced plaque SPP1 protein in vivo. Increased SPP1 mRNA expression upon PHD2cko was preferentially observed in foamy plaque macrophages expressing ‘triggering receptor expressed on myeloid cells-2’ (TREM2hi) evidenced by single-cell RNA, but not in neutrophils. This confirmed enhanced fibrotic signalling by PHD2cko macrophages to fibroblasts, in vitro as well as in vivo. CONCLUSION: Myeloid PHD2cko and PHD3ko enhanced atherosclerotic plaque growth and macrophage apoptosis, while PHD2cko macrophages further activated collagen secretion by fibroblasts in vitro, likely via paracrine SPP1 signalling through TREM2hi macrophages. Oxford University Press 2021-04-29 /pmc/articles/PMC8953448/ /pubmed/33913468 http://dx.doi.org/10.1093/cvr/cvab152 Text en © The Author(s) 2021. Published by Oxford University Press on behalf of the European Society of Cardiology. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
van Kuijk, Kim
Demandt, Jasper A F
Perales-Patón, Javier
Theelen, Thomas L
Kuppe, Christoph
Marsch, Elke
de Bruijn, Jenny
Jin, Han
Gijbels, Marion J
Matic, Ljubica
Mees, Barend M E
Reutelingsperger, Chris P M
Hedin, Ulf
Biessen, Erik A L
Carmeliet, Peter
Baker, Andrew H
Kramann, Rafael K
Schurgers, Leon J
Saez-Rodriguez, Julio
Sluimer, Judith C
Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title_full Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title_fullStr Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title_full_unstemmed Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title_short Deficiency of myeloid PHD proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
title_sort deficiency of myeloid phd proteins aggravates atherogenesis via macrophage apoptosis and paracrine fibrotic signalling
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8953448/
https://www.ncbi.nlm.nih.gov/pubmed/33913468
http://dx.doi.org/10.1093/cvr/cvab152
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