Fig. 8: Eliglustat inhibits OC formation from MM patients while exogenous GSLs reverse eliglustat inhibition.
a BM mononuclear cells from MM patients were seeded at 1.5 × 105 cells/well into 48-well-plate and differentiated to OCs with 50 ng/ml human M-CSF and 75 ng/ml human RANKL in αMEM media. BM mononuclear cells were treated with increasing doses of eliglustat (0.1, 1, 10, 25 μM) and OCs were identified by TRAP staining on day 11. b–d Number of OCs per filed of view, number of nuclei per TRAP + cell and % OC area over total area were quantified. n = 9 patients. e Eliglustat was applied to the BM mononuclear cells from MM patients with or without C16 LacCer, C16 GlcCer, and C24 LacCer during in vitro OC formation process and mature OCs were recognized by TRAP staining. f–h Number of OCs per filed of view, number of nuclei per TRAP + cell and % OC area over total area were quantified. Scale bar represents 200 μm. n = 9 patients, each with 5–7 replicates. i Schematic graph illustrating the overall proposed mechanism. Binding of RANKL to RANK receptor induces the degradation of TRAF3 via autophagy, which allows NF-kB signaling to occur; this is required for OC formation. Eliglustat reduces the amount of GlcCer and LacCer, preventing autophagic flux, thereby reducing TRAF3 degradation and OC formation. Results from a and e are representative of 3 independent experiments with 9 patient samples. Data are presented as mean values ± SEM. Exact p-values are depicted in the figure. Statistical analysis was performed using one-way ANOVA. Source data are provided as a Source Data file.
