Effect and molecular mechanism of Syndecan-1 inhibition on endometriosis

The impact of shRNA-mediated knockdown of Syndecan-1 will be assessed in the in vivo endometriosis models available to partner CONICET. WWU and KI in cooperation with CONICET will employ several in vitro assays of endometriotic stem cell phenotypic analysis, including flow cytometric side population analysis, measurement of ALDH activity, qPCR and FACS analysis of stemness-associated cell surface markers, colony formation and sphere formation assays. The impact of Syndecan-1 and the HA-CD44 assays will be studied in the endometriotic cell line 12Z, and in patient-derived primary endometriotic stroma cells by siRNA knockdown and 4-MU inhibition. TaqMan low density array analysis will be used by WWU to screen for affected pathways, and targets will be confirmed by qPCR and Western blotting.Sphere formation assays, as well as lineage-specific differentiation assays will be performed in the experimental systems subjected to Syndecan-1 shRNA treatment. Results of TaqMan low density array analysis on identified targets will be confirmed by these functional analyses.