Inhibition of Glycogen Synthase Kinase 3 (GSK3) Increases the Cytotoxicity of Enzastaurin

Cutaneous T cell lymphomas (CTCL) represent a spectrum of several distinct non-Hodgkin's lymphomas that are characterized by an invasion of the skin by malignant, clonal lymphocytes. Our lab has previously demonstrated that the Protein Kinase C (PKC) β inhibitor Enzastaurin increases apoptosis in malignant lymphocytes of CTCL. These results directly led to a clinical trial for Enzastaurin in CTCL where it was well tolerated and showed modest activity. To ascertain a means of improving the efficacy of Enzastaurin, we investigated complimentary signaling pathways and identified Glycogen Synthase Kinase 3 (GSK3) as important in survival signaling in CTCL. Enzastaurin combined with GSK3 inhibitors demonstrated anenhancement of cytotoxicity. Treatment with a combination of Enzastaurin and the GSK3 inhibitor AR-A014418 resulted in up-regulation of β catenin total protein and β catenin-mediated transcription. Inhibition of β catenin-mediated transcription or shRNA knockdown of β catenin decreased the cytotoxic effects of Enzastaurin plus AR-A014418. In addition, treatment with Enzastaurin and AR-A014418 decreased the mRNA levels and surface expression of CD44. shRNA knockdown of β catenin also restored CD44 surface expression. Our observations provide a rationale for the combined targeting of PKC and GSK3 signaling pathways in CTCL to enhance the therapeutic outcome.