Prolyl 3-Hydroxylase 2 Is a Molecular Player of Angiogenesis.

Abstract

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.

• Authors: Paola Pignata, Ivana Apicella, Valeria Cicatiello, Caterina Puglisi, Sara Magliacane Trotta, Remo Sanges, Valeria Tarallo, Sandro De Falco
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 22 Issue 8 (Apr 09 2021) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 33918807 (Publication Type: Journal Article)

Chemical References

• Collagen Type IV
• VEGFA protein, human
• Vascular Endothelial Growth Factor A
• Procollagen-Proline Dioxygenase
• P3H2 protein, human
• Vascular Endothelial Growth Factor Receptor-2
• p38 Mitogen-Activated Protein Kinases

Topics

• Animals
• Choroidal Neovascularization (diagnostic imaging, etiology, metabolism, pathology)
• Collagen Type IV (genetics, metabolism)
• Disease Models, Animal
• Fluorescent Antibody Technique
• Gene Expression Regulation
• Human Umbilical Vein Endothelial Cells (metabolism)
• Humans
• Mice
• Neovascularization, Pathologic (genetics, metabolism)
• Neovascularization, Physiologic
• Procollagen-Proline Dioxygenase (genetics, metabolism)
• Protein Binding
• Signal Transduction
• Vascular Endothelial Growth Factor A (genetics, metabolism)
• Vascular Endothelial Growth Factor Receptor-2 (metabolism)
• p38 Mitogen-Activated Protein Kinases (metabolism)