Regulation of human feto-placental endothelial barrier integrity by vascular endothelial growth factors: competitive interplay between VEGF-A165a, VEGF-A165b, PIGF and VE-cadherin

Abstract

The human placenta nourishes and protects the developing foetus whilst influencing maternal physiology for fetal advantage. It expresses several members of the vascular endothelial growth factor (VEGF) family including the pro-angiogenic/pro-permeability VEGF-A₁₆₅a isoform, the anti-angiogenic VEGF-A₁₆₅b, placental growth factor (PIGF) and their receptors, VEGFR1 and VEGFR2. Alterations in the ratio of these factors during gestation and in complicated pregnancies have been reported; however, the impact of this on feto-placental endothelial barrier integrity is unknown. The present study investigated the interplay of these factors on junctional occupancy of VE-cadherin and macromolecular leakage in human endothelial monolayers and the perfused placental microvascular bed. Whilst VEGF-A₁₆₅a (50 ng/ml) increased endothelial monolayer albumin permeability (P0.05) or PlGF (P>0.05) did not. Moreover, VEGF-A₁₆₅b (100 ng/ml; P0.05) inhibited VEGF-A₁₆₅a-induced permeability when added singly. PlGF abolished the VEGF-A₁₆₅b-induced reduction in VEGF-A₁₆₅a-mediated permeability (P>0.05); PlGF was found to compete with VEGF-A₁₆₅b for binding to Flt-1 at equimolar affinity. Junctional occupancy of VE-cadherin matched alterations in permeability. In the perfused microvascular bed, VEGF-A₁₆₅b did not induce microvascular leakage but inhibited and reversed VEGF-A₁₆₅a-induced loss of junctional VE-cadherin and tracer leakage. These results indicate that the anti-angiogenic VEGF-A₁₆₅b isoform does not increase permeability in human placental microvessels or HUVEC primary cells and can interrupt VEGF-A₁₆₅a-induced permeability. Moreover, the interplay of these isoforms with PIGF (and s-flt1) suggests that the ratio of these three factors may be important in determining the placental and endothelial barrier in normal and complicated pregnancies.