VIP INDUCES THE DECIDUALIZATION PROGRAM ON HUMAN ENDOMETRIAL STROMAL CELLS AND CONDITIONS DENDRITIC CELLS PROFILE

GRASSO E; GORI S; PAPARINI D; SALAMONE G; MARTINEZ G; IRIGOYEN M; RUHLMANN C; PÉREZ LEIRÓS C; RAMHORST R

Lisboa

Congreso; Annual Meeting ESRHE 2015; 2015

ESHRE

Study question: To investigate VIP (vasoactive intestinal peptide) contributioninto the decidualization program, from phenotypic and functional aspects andits effect on dendritic cells (DC) immune-profi le. Here, we used an in vitroimplantation model based in the co-culture of blastocyst-like spheroids fromtrophoblast cells cultured on hESC monolayer decidualized with VIP.Summary answer: Our results suggest that VIP contributes to the decidualizationprocess on hESC cells inducing phenotypic markers and chemokinesexpression. From the functional aspect, VIP-decidualizated hESC cells allowthe blastocyst-like spheroides invasion and condition Dendritic cells to a tolerogenicprofi le preventing the increase of costimualtory molecules and inducingIL-10 secretion.What is known already: The decidualization program involves phenotype andfunctional changes on endometrial cells that facilitate the attachment and invasionof the blastocyst. Although progesterone is a key hormone, the trigger ofthis process is an intracellular cAMP increase. The decidualization involves themodulation of different mediators such as cytokines, chemokines and growthfactors. Particularly, vasoactive intestinal peptide (VIP) is a neuropeptide producedby endometrial stromal cells among others, and displays multiple targetcircuits that allow immunotolerance.Study design, size, duration: We used an in vitro implantation model based inthe co-culture of blastocyst-like spheroids from trophoblast cells line Swan71cultured on hESC monolayer decidualized with VIP.Participants/materials, setting, methods: Human endometrial stromal cellline (hESC) was cultured with/without VIP or MPA + dbcAMP. VIP/VPACsystem and decidualization markers were evaluated by RTPCR and ELISA.Blastocyst-like spheroids (BLS) were obtained from Swan71cells and culturedon hESC monolayer. Monocytes were isolated from PBMCs, differentiated toDC and studied by FACS.Main results and the role of chance: The MPA + dbcAMP decidualizationincreased VIP expression and secretion (p < 0.05). When decidualization wasinduced by VIP, we observed an increase of markers IGFBP1, PRL, KLF13/KLF9 ratio, IL-8 and SDF1 expression in a concentration-dependent manner(p < 0.05). To evaluate functional aspects of VIP-decidualization, an invasionassay was performed. BLS were able to invade hESC monolayer decidualizatedwith VIP or MPA + dbcAMP. When these assays were performed with conditionedmedia obtained from human blastocyst we found an increase in BLSinvasion on VIP-decidualizated hESC (p < 0.05). Finally, we evaluate the immunomodulatoryeffects of decidualized cells on DC and monocytes. DecidualizedhESC conditioned media induced a semi-mature profi le on DC preventingthe induction of CD83 and CD86 expression, and increasing IL-10 secretion(p < 0.05). Monocytes profi le was not modulated by conditioned media. Limitations, reason for caution: The present results were studied using immortalizedcell lines. Further studies are necessary to elucidate the precisemechanisms involve and the role of VIP in decidualization.Wider implications of the fi ndings: Our results suggest that VIP may have animportant role during the decidualization process as it is able to induce the differentiationon the stromal cells, not only from the phenotypic aspect but also froma functional one, by allowing blastocyst invasion while contributing to controlthe immune micro-environment by inducing a tolerogenic profi le on DC. Wepropose that VIP may be a new participant in this differentiation program.Study funding/competing interest(s): Funding by University(ies) ? Fundingby national/international organization(s). Universidad de Buenos Aires, Argentina;Consejo Nacional de Investigaciones Científi cas y Tecnológicas.Trial registration number: NA.