Background: In humans, the endocannabinoid system (ECS) plays an important role in several physiological processes, including reproductive system functions (Ligresti et al., 2009; Maccarrone et al., 2010). The components of ECS have been extensively characterized and comprise two main receptors, CB1 and CB2, two main ligands, anandamide (AEA) and 2-arachidonoylglycerol (2-AG) and their metabolic enzymes involved in their synthesis and degradation (Ligresti et al., 2009; Maccarrone et al., 2010). The ECS has been shown to regulate pathways involved in the process of implantation and its dysregulation might be a determinant of infertility (Maia et al.;2009). In particular, it has been demonstrated that sustained levels of anandamide (AEA) impair decidualization, a process known to be necessary for endometrial receptivity. We are currently investigating whether the adverse effect of AEA on decidualization might be a consequence of its negative regulation of the EGFL7/NOTCH axis. The role of the NOTCH pathway in the regulation of endometrial receptivity has been well established (Afshar et al, 2012; Su et al, 2015). We recently demonstrated that EGFL7 can drive the differentiation of human primary endometrial stromal cells towards the decidual phenotype by activating the NOTCH pathway (Lacconi et al., 2023). Results and discussion: HESC were decidualized in vitro (DESC) with medroxyprogesterone acetate (MPA), estradiol and 8-Bromo-cyclic AMP (cAMP). At 4 e 6 day decidualized cells were treated with AEA (10µM) for 24 and 72h. qRT-PCR showed that HESC and DESC express the CB1 receptor and the major enzymes involved in AEA metabolism (FAAH). In agreement with data from the literature, AEA culture supplementation downregulates the expression of the decidualization markers IGFBP1 and Prolactin (Almada et al.,2016). Concomitantly we observed downregulation of the NOTCH target gene HEY1, whose expression is normally increased upon decidualization (Lacconi et al., 2023). As we previously demonstrated decidualization upregulates the expression of EGFL7 in stromal cells, however in contrast with what expected AEA treatment further increased its expression. We hypotheses that AEA-induced EGFL7 excessive increase may negatively impact on the activation of NOTCH signalling, as demonstrated in other biological systems (Nichol and Stuhlmann 2012), this in turn negatively impact on decidualization.
The endocannabinoid-EGFL7/Notch axis in endometrial preparation to implantation
Gina La Sala;
2023
Abstract
Background: In humans, the endocannabinoid system (ECS) plays an important role in several physiological processes, including reproductive system functions (Ligresti et al., 2009; Maccarrone et al., 2010). The components of ECS have been extensively characterized and comprise two main receptors, CB1 and CB2, two main ligands, anandamide (AEA) and 2-arachidonoylglycerol (2-AG) and their metabolic enzymes involved in their synthesis and degradation (Ligresti et al., 2009; Maccarrone et al., 2010). The ECS has been shown to regulate pathways involved in the process of implantation and its dysregulation might be a determinant of infertility (Maia et al.;2009). In particular, it has been demonstrated that sustained levels of anandamide (AEA) impair decidualization, a process known to be necessary for endometrial receptivity. We are currently investigating whether the adverse effect of AEA on decidualization might be a consequence of its negative regulation of the EGFL7/NOTCH axis. The role of the NOTCH pathway in the regulation of endometrial receptivity has been well established (Afshar et al, 2012; Su et al, 2015). We recently demonstrated that EGFL7 can drive the differentiation of human primary endometrial stromal cells towards the decidual phenotype by activating the NOTCH pathway (Lacconi et al., 2023). Results and discussion: HESC were decidualized in vitro (DESC) with medroxyprogesterone acetate (MPA), estradiol and 8-Bromo-cyclic AMP (cAMP). At 4 e 6 day decidualized cells were treated with AEA (10µM) for 24 and 72h. qRT-PCR showed that HESC and DESC express the CB1 receptor and the major enzymes involved in AEA metabolism (FAAH). In agreement with data from the literature, AEA culture supplementation downregulates the expression of the decidualization markers IGFBP1 and Prolactin (Almada et al.,2016). Concomitantly we observed downregulation of the NOTCH target gene HEY1, whose expression is normally increased upon decidualization (Lacconi et al., 2023). As we previously demonstrated decidualization upregulates the expression of EGFL7 in stromal cells, however in contrast with what expected AEA treatment further increased its expression. We hypotheses that AEA-induced EGFL7 excessive increase may negatively impact on the activation of NOTCH signalling, as demonstrated in other biological systems (Nichol and Stuhlmann 2012), this in turn negatively impact on decidualization.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
