title: The Glycolytic Enzyme PFKFB3 Is Involved in Estrogen-Mediated Angiogenesis via GPER1.
abstract: The endogenous estrogen 17β-estradiol (E2) is a key factor in promoting endothelial healing and angiogenesis. Recently, proangiogenic signals including vascular endothelial growth factor and others have been shown to converge in endothelial cell metabolism. Because inhibition of the glycolytic enzyme activator phosphofructokinase-2/fructose-2,6-bisphosphatase 3 (PFKFB3) reduces pathologic angiogenesis and estrogen receptor (ER) signaling stimulates glucose uptake and glycolysis by inducing PFKFB3 in breast cancer, we hypothesized that E2 triggers angiogenesis in endothelial cells via rapid ER signaling that requires PFKFB3 as a downstream effector. We report that treatment with the selective G protein-coupled estrogen receptor (GPER1) agonist G-1 (10-10to 10-7M) mimicked the chemotactic and proangiogenic effect of E2 as measured in a number of short-term angiogenesis assays in human umbilical vein endothelial cells (HUVECs); in addition, E2 treatment upregulated PFKFB3 expression in a time- and concentration-dependent manner. Such an effect peaked at 3 hours and was also induced by G-1 and abolished by pretreatment with the GPER1 antagonist G-15 or GPER1 siRNA, consistent with engagement of membrane ER. Experiments with the PFKFB3 inhibitor 3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one showed that PFKFB3 activity was required for estrogen-mediated HUVEC migration via GPER1. In conclusion, E2-induced angiogenesis was mediated at least in part by the membrane GPER1 and required upregulation of the glycolytic activator PFKFB3 in HUVECs. These findings unravel a previously unrecognized mechanism of estrogen-dependent endocrine-metabolic crosstalk in HUVECs and may have implications in angiogenesis occurring in ischemic or hypoxic tissues.
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Trenti
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Tedesco
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Carlotta
authors.initials: C
authors.lastname: Boscaro
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Ferri
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy andrea.cignarella@unipd.it.
authors.forename: Andrea
authors.initials: A
authors.lastname: Cignarella
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Trevisi
authors.affiliation: Department of Pharmaceutical and Pharmacological Sciences (A.T., S.T., Ca.B., N.F., L.T., Ch.B) and Department of Medicine (A.C.), University of Padova, Padova, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Bolego
mesh_terms: D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects* / Q000502:physiology; D025481:Phosphofructokinase-2 / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D034741:RNA, Small Interfering / Q000494:pharmacology; D011960:Receptors, Estrogen / Q000037:antagonists & inhibitors / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: C109584:GPER1 protein, human; D034741:RNA, Small Interfering; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D004958:Estradiol; C107345:PFKFB3 protein, human; D025481:Phosphofructokinase-2