Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis
Prolyl 3-hydroxylase 2 (P3H2) catalyzes the post-translational formation of 3-hydroxyproline on collagens, mainly on type IV. Its activity has never been directly associated to angiogenesis. Here, we identified P3H2 gene through a deep-sequencing transcriptome analysis of human umbilical vein endoth...
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doaj-af7aface4a1f4b6ba1f25f9b96b1dcf62021-04-09T23:04:41ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-04-01223896389610.3390/ijms22083896Prolyl 3-Hydroxylase 2 Is a Molecular Player of AngiogenesisPaola Pignata0Ivana Apicella1Valeria Cicatiello2Caterina Puglisi3Sara Magliacane Magliacane Trotta4Remo Sanges5Valeria Tarallo6Sandro De De Falco7Institute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyInstitute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyInstitute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyIOM Ricerca s.r.l., 95029 Viagrande, ItalyInstitute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyComputational Genomics Laboratory, International School for Advanced Studies (SISSA), 34136 Trieste, ItalyInstitute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyInstitute of Genetics and Biophysics ‘Adriano Buzzati-Traverso’—CNR, Angiogenesis LAB, 80131 Naples, ItalyProlyl 3-hydroxylase 2 (P3H2) catalyzes the post-translational formation of 3-hydroxyproline on collagens, mainly on type IV. Its activity has never been directly associated to angiogenesis. Here, we identified P3H2 gene through a deep-sequencing transcriptome analysis of human umbilical vein endothelial cells (HUVECs) stimulated with vascular endothelial growth factor A (VEGF-A). Differently from many previous studies we carried out the stimulation not on starved HUVECs, but on cells grown to maintain the best condition for their in vitro survival and propagation. We showed that P3H2 is induced by VEGF-A in two primary human endothelial cell lines and that its transcription is modulated by VEGF-A/VEGF receptor 2 (VEGFR-2) signaling pathway through p38 mitogen-activated protein kinase (MAPK). Then, we demonstrated that P3H2, through its activity on type IV Collagen, is essential for angiogenesis properties of endothelial cells in vitro by performing experiments of gain- and loss-of-function. Immunofluorescence studies showed that the overexpression of P3H2 induced a more condensed status of Collagen IV, accompanied by an alignment of the cells along the Collagen IV bundles, so towards an evident pro-angiogenic status. Finally, we found that P3h2 knockdown prevents pathological angiogenesis in vivo, in the model of laser-induced choroid neovascularization. Together these findings reveal that P3H2 is a new molecular player involved in new vessels formation and could be considered as a potential target for anti-angiogenesis therapy.https://www.mdpi.com/1422-0067/22/8/3896angiogenesisProlyl 3-hydroxylase 2Collagen IVvascular endothelial growth factor Achoroidal neovascularizationage-related macular degeneration |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Paola Pignata Ivana Apicella Valeria Cicatiello Caterina Puglisi Sara Magliacane Magliacane Trotta Remo Sanges Valeria Tarallo Sandro De De Falco |
spellingShingle |
Paola Pignata Ivana Apicella Valeria Cicatiello Caterina Puglisi Sara Magliacane Magliacane Trotta Remo Sanges Valeria Tarallo Sandro De De Falco Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis International Journal of Molecular Sciences angiogenesis Prolyl 3-hydroxylase 2 Collagen IV vascular endothelial growth factor A choroidal neovascularization age-related macular degeneration |
author_facet |
Paola Pignata Ivana Apicella Valeria Cicatiello Caterina Puglisi Sara Magliacane Magliacane Trotta Remo Sanges Valeria Tarallo Sandro De De Falco |
author_sort |
Paola Pignata |
title |
Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis |
title_short |
Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis |
title_full |
Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis |
title_fullStr |
Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis |
title_full_unstemmed |
Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis |
title_sort |
prolyl 3-hydroxylase 2 is a molecular player of angiogenesis |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-04-01 |
description |
Prolyl 3-hydroxylase 2 (P3H2) catalyzes the post-translational formation of 3-hydroxyproline on collagens, mainly on type IV. Its activity has never been directly associated to angiogenesis. Here, we identified P3H2 gene through a deep-sequencing transcriptome analysis of human umbilical vein endothelial cells (HUVECs) stimulated with vascular endothelial growth factor A (VEGF-A). Differently from many previous studies we carried out the stimulation not on starved HUVECs, but on cells grown to maintain the best condition for their in vitro survival and propagation. We showed that P3H2 is induced by VEGF-A in two primary human endothelial cell lines and that its transcription is modulated by VEGF-A/VEGF receptor 2 (VEGFR-2) signaling pathway through p38 mitogen-activated protein kinase (MAPK). Then, we demonstrated that P3H2, through its activity on type IV Collagen, is essential for angiogenesis properties of endothelial cells in vitro by performing experiments of gain- and loss-of-function. Immunofluorescence studies showed that the overexpression of P3H2 induced a more condensed status of Collagen IV, accompanied by an alignment of the cells along the Collagen IV bundles, so towards an evident pro-angiogenic status. Finally, we found that P3h2 knockdown prevents pathological angiogenesis in vivo, in the model of laser-induced choroid neovascularization. Together these findings reveal that P3H2 is a new molecular player involved in new vessels formation and could be considered as a potential target for anti-angiogenesis therapy. |
topic |
angiogenesis Prolyl 3-hydroxylase 2 Collagen IV vascular endothelial growth factor A choroidal neovascularization age-related macular degeneration |
url |
https://www.mdpi.com/1422-0067/22/8/3896 |
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