Galectin-1-Related Modulation of Trophoblast Endothelial Interactions by Integrins α1 and ß1.

ABSTRACT

During normal trophoblast invasion, integrins α6ß4 are downregulated, and α1ß1 are upregulated in invasive cytotrophoblast cells. In preeclampsia both interstitial and endovascular invasion are shallow and cytotrophoblasts fail to upregulate α1ß1 and downregulate α6ß4. This study aims to investigate the role of integrins α1ß1 and α6ß4 on cellular pathways influencing trophoblast integration into endothelial cellular networks in vitro. Red fluorescent-labeled human uterine myometrial microvascular endothelial cells (UtMVECs) were seeded on Matrigel to form endothelial networks. Green fluorescent-labeled trophoblastic HTR-8/SVneo cells pre-incubated with 20 µg/ml of neutralizing antibodies (anti-α1, ß1, α6, ß4, α1 + ß1, or α6 + ß4) for 1 h were then co-cultured with endothelial networks with the neutralizing antibodies for 24 h. Fluorescent images were captured, and quantified utilizing Image J. Cells were retrieved to analyze mRNA expression of galectin-1, TIMP-1, and PAI-1 by quantitative PCR. MMP-2, MMP-9, free sFlt-1, and PlGF from conditioned media were measured by ELISA. The integration of trophoblast cells into endothelial cellular networks was inhibited by anti-ß1(- 28 ± 3%, p < 0.0001), and increased by anti-α6(+ 19 ± 5%, p < 0.01). Galectin-1 mRNA expression was decreased by anti-α1(- 35 ± 7%, p < 0.001), anti-ß1(- 23 ± 5%, p < 0.05), and anti-α1+ß1(- 35 ± 5%, p < 0.001). The mRNA expression of TIMP-1 was inhibited by anti-α1(- 59 ± 9%, p < 0.01) and anti-ß1(- 63 ± 7%, p < 0.001) while PAI-1 mRNA expression was increased by anti-α1 + ß1(+ 285 ± 70%, p < 0.0001). In the conditioned medium, anti-α1 reduced MMP-2(-28 ± 1%, p < 0.001), MMP-9(-27 ± 8%, p < 0.01), and sFlt-1(-27 ± 5%, p < 0.001) production. Anti-ß1 reduced MMP-2(- 15 ± 2%, p < 0.05) production. There were no changes in PlGF. Appropriate integrins α1ß1 modulate trophoblast cell integration into endothelial cellular networks in vitro through invasive pathways including galectin-1, TIMP-1, PAI-1, MMP-2, and MMP-9 production.