A dynamic shift of VEGF isoforms with a transient and selective progesterone-induced expression of VEGF₁₈₉regulates angiogenesis and vascular permeability in human uterus

Abstract

A key mechanism underlying physiological angiogenesis of the human endometrium is its ability to regenerate the vascular capillary network and to perform vascular remodeling (i.e., development of spiral arteries). Vascular endothelial growth factor (VEGF) is associated with angiogenesis and capillary permeability in this tissue. VEGF is expressed as several spliced variants, its main human isoforms contain 121 and 165 aa; 17␤-estradiol (E2) increases endometrial VEGF, possibly in all isoforms. Here we show that progesterone (P) selectively increases the expression of the VEGF189 (V189) isoform in the human uterus. V189 is identified in the conditioned medium of stromal cells treated with E2 ؉ P; its presence in this in vitro model of decidual stromal cells is detected after 6 -8 days, using ELISA, and after 8 -10 days, using Western blot analysis with different antibodies, including one specific for V189. The secretion pattern of V189 parallels that of the decidual protein IGFBP-1. V189 is secreted as a native isoform, as compared with the migration of recombinant V189 by SDS͞PAGE. In situ hybridization and immunocytochemistry, performed on the same biopsies, suggest that decidual cells express V189 during the midlate secretory phase of the menstrual cycle and early gestation. Finally, using an in vivo permeability assay, we show that native V189 increases capillary permeability. These observations demonstrate that P regulates V189 expression in decidual cells, which could have important implications for understanding uterine vascular remodeling and implantation, and may be relevant in a range of disease states such as edema and irregular bleeding.A s a tissue that exhibits rapid cyclic changes (1), the human endometrium is a good model for the study of physiological angiogenesis, which is under the control of 17␤ estradiol (E 2 ), progesterone (P), and vascular endothelial growth factor (VEGF) (2-7).VEGF is a protein with angiogenic activity and is a potent stimulator of microvascular permeability (8, 9); it plays an important role in physiological and pathological neovascularization, via its receptors Flt-1͞VEGFR1 and Flk-1͞VEGFR2 (10-13). Molecular cloning of the cDNA for VEGF revealed that differential exon splicing generates several isoforms containing 121, 165, 189, and 206 aa (V 121, V 165, V 189, and V 206 ) (14, 15). In most systems V 121 and V 165 are the major species expressed, whereas V 189 is only minimally present and V 206 is limited to embryonic tissue (9, 15). The different isoforms appear to have similar functions, but differ in their binding affinity for extracellular matrix (ECM). In contrast to VEGF 121 , which is secreted and found freely soluble in the culture medium, the bioavailability of VEGF 165 and to a greater extent V 189 appears to be regulated by binding to heparan sulfate proteoglycans (HSPG) in the ECM (9, 16). Bound VEGF isoforms could provide a reserve of growth factors available after cleavage by heparinase or urokinase-type plasminogen activator (uPA) (17, 18).Previous stud...

Spatio-temporal analysis of vascular endothelial growth factor expression and blood vessel remodelling in pig ovarian follicles during the periovulatory period

“…The production of VEGF isoforms with high molecular weight (i.e. VEGF 189 ), characterized by a lower solubility, could explain these results, as demonstrated in the endometrium when it is stimulated by a modified steroid milieu (Ancelin et al 2002). In addition, it cannot be excluded that a change in the local heparin concentration or in the composition of cell surface heparan sulphate may condition VEGF bioavailability in the follicle (Robinson & Stringer 2001).…”

Transforming Growth Factor-β1 Stimulates Vascular Endothelial Growth Factor 164 via Mitogen-activated Protein Kinase Kinase 3-p38α and p38δ Mitogen-activated Protein Kinase-dependent Pathway in Murine Mesangial Cells

“…VEGF 188 mRNA transcription occurs as a late event during lipogenesis distinct from earlier induction of VEGF 120 and VEGF 164 mRNAs during differentiation (45). In the human uterus, 17␤-estradiol increases endometrial expression of all VEGF isoforms, whereas progesterone selectively increases the expression of the VEGF 189 isoform (46). Studies in hyperoxic acute lung injury by Corne et al (47) show that interleukin-13 selectively stimulates VEGF 164 , whereas interleukin-13 plus hyperoxia stimulate VEGF 120 and VEGF 188 .…”

Regulation of endometrial vascular remodelling: role of the vascular endothelial growth factor family and the angiopoietin–TIE signalling system

“…Various studies have now shown that the different VEGFA isoforms are present in the endometrium (Charnock-Jones et al 1993, Halder et al 2000, Sharkey et al 2000, Ancelin et al 2002, Niklaus et al 2002, Sugino et al 2002. As in other tissues, VEGFA 164 is believed to be the dominant isoform in human endometrium.…”