P26 - Reproducción

Extracellular vesicles from trophoblast cells overexpressing VIPR2 induce angiogenesis

Avalos, Julieta¹ - Aguilera, Franco¹ -Hauk, Vanesa¹ - Grasso, Daniel² - García,M. Noé² - Lara, Brenda¹ - Merech, Fátima¹ - Charó, Nancy³ - Petrelli, Valentina⁴ - Aiello, Horacio⁴ - Meller, Cesar⁴ - Ramhorst, Rosanna¹ - Grasso Esteban¹ - Pérez Leirós, Claudia¹ - Vota, Daiana M¹* - Paparini, Daniel E¹*

1) Laboratorio de Inmunofarmacologia, Instituto de Química Biologica de la Facultad de Ciencias Exactas y Naturales (IQUIBICEN-CONICET),Universidad de Buenos Aires (UBA).
2)Instituto de Estudios de la Inmunidad Humoral (IDEHU), CONICET-Universidad de Buenos Aires, Junín 956, Piso4, Buenos Aires CP 1113, Argentina.
3) Instituto de medicina experimental (IMEX)-CONICET/Academia Nacional de Medicina de Buenos Aires, Buenos Aires, Argentina.
4) Hospital Italiano de Buenos Aires, Servicio de Obstetricia, Buenos Aires, Argentina. * Equal contribution
Contacto: julieta.a.avalos@gmail.com

Vascular remodeling is a critical step in correct placentation. Along with this process, the expression of vasoactive intestinal peptide receptor 2 (VIPR2) increases in human placental villi- it can exert proangiogenic effects on the murine vasculature. During pregnancy, the number of extracellular vesicles (EVs) increases, and those derived from trophoblast cells EVs (Tb-EVs) are known to play a key role in regulating placentation.

However, whether VIPR2 expression by Tb cells contributes to the pro-angiogenic effects of EVs during placentation remains unknown. In this study, we examined the effect of human trophoblast cell line overexpressing VIPR2 (Tb-VIPR2) EVs on endothelial cells function. EVs were obtained from the human trophoblast cell line Swan 71, transfected either with a VIPR2 plasmid (Tb-VIPR2) or an empty vector (control, Tb-C) using differential centrifugation. Tb-EVs were characterized using transmission electron microscopy, nanoparticle tracking analysis and imaging cytometry analysis. Endothelial cells, EA.hy926 and HUVEC, were used for wound healing, reactive oxygen species (ROS) detection and tube formation assays. Tb-VIPR2 EVs carried VIPR2 on the membrane along with CD9 and CD81 tetraspanins. Compared to Tb-C EVs, they exhibited a smaller size range and slightly lower particles/mL. Tb-VPR2 EVs enhanced endothelial cell migration (*P< 0.05), showed a
trend toward reduced ROS production and increased the number of meshes, nodes, pieces and segments in tubulogenic assays respect to Tb-C EVs (*P< 0.05).

These findings support the modulatory effect of extracellular vesicles produced by human trophoblast cells expressing VIPR2 on endothelial cell function. These suggest a regulatory role for VIPR2 in human placental-associated adaptations.

Palabras clave: PREGNANCY, TROPHOBLAST CELLS, ENDOTHELIAL CELLS