The hyaluronan synthesis inhibitor 4-methylumbelliferone inhibits cell proliferation and wound healing on in vitro endometriosis models.

CN OLIVARES; D MADANES; RI BARAÑAO; B MCCORMACK; A RANDI; F CHIAPPINI ; CM IBAÑEZ PADILLA ; GF MERESMAN

Congreso; 35th Annual Meeting of the European Society of Human Reproduction and Embryology; 2019

ESHRE

Study question (30 words): What is the effect of 4-methylumbelliferone (4MU) on cellproliferation, wound healing process and gelatinase activity on human endometrial stromal (t-HESC) or epithelial (ECC-1) cell lines?Summary answer (25 words): Cell proliferation and wound healing were significantly inhibited by4MU, whereas MMP-2 and MMP-9 activity would not be modulated by this treatment.What is known already (100 words): Endometriosis is a benign gynecological disease affecting10% of women of reproductive age, characterized by the presence of endometriotic foci outsidethe uterine cavity. We have already demonstrated that 4MU has antiangiogenic properties in twoangiogenesis models of endometriosis, both in vitro and in vivo, and it has been reported thatbinding of hyaluronan to its CD44 receptor is involved in proliferation, migration and invasion incancer cells.Study design, size and duration (75 words): We evaluated the effect of 0.1, 0.5, 1, 2 and 4mM4MU on in vitro endometriosis models comparing control versus treatment. Cell proliferation wasassessed on ECC-1 (n=10) and t-HESC (n=5), and gelatinase activity (n=2) and wound healing (n=6)process were evaluated on t-HESC. For cell proliferation and gelatinase activity cells were treatedfor 24hs and for the wound healing assay cells were treated during 20hs.Participants/materials, setting, methods (75 words): t-HESC and ECC-1 endometrial cell lineswere stimulated with different concentrations of 4MU. Cell proliferation was evaluated after 24hswith the cell proliferation reagent WST-1. The area of a scratch closed by the t-HESC wascalculated in a wound healing assay, for this purpose photomicrographs were taken at time 0h andtime 20hs; and the gelatinase activity in conditioned media of t-HESC was evaluated byzimography. Only p<0.05 was considered as statistically significant.Main results and the role of chance (200 words): After 24hs of treatment the cell proliferation ofECC-1 was significantly inhibited by 0.5, 1 and 2 mM 4MU (p<0.01, p<0.001 and p<0.01,respectively versus control), whereas cell proliferation of t-HESC was significantly inhibited by 2and 4 mM 4MU (p<0.01 versus control). Nevertheless, when evaluated at 20hs of treatment,already at 0.5 mM 4MU, the migration of t-HESC cells was significantly inhibited and the scratchwas closed in a low percentage (p<0.01 for 0.5 and 1 mM 4MU versus control, p<0.001 for 2 and 4mM 4MU). Preliminary results of MMP-2 and MMP-9 activities revealed that these gelatinaseswould not be modulated after 24hs of 4MU treatment.Limitations, reasons for caution: This study has only focused on in vitro results. More studies areneeded to understand the implication of hyaluronan synthesis inhibition on the migratory capacityof the cells and which molecules are involved in this matter.Wider implications of the findings (50 words): Previous studies from our laboratory demonstrateda strong antiangiogenic activity of 4MU. The present results are, in part, in agreement with thosereported by other authors. Given these and our background on targeting hyaluronan onendometriosis models we are encouraged to continue investigating on this path.