Disulfiram and Copper Combination Therapy Targets NPL4, Cancer Stem Cells and Prolongs Survival in Group 3 Medulloblastoma. (16th November 2020)

• Record Type: Journal Article
• Authors: Serra, Riccardo; Zhao, Tianna; Huq, Sakibul; Gorelick, Noah L; Casaos, Joshua; Cecia, Arba; Mangraviti, Antonella; Bai, Renyuan; Olivi, Alessandro; Brem, Henry; Jackson, Eric M; Tyler, Betty
• Is Part Of: Neurosurgery. Volume 67(2010)Supplement 1
• Publication Date: 2020-11-16
• Subjects: Nervous system -- Surgery -- Periodicals; 617.48005
• DOI: 10.1093/neuros/nyaa447_908
• Languages: English
• ISSNs: 0148-396X

Abstract

Abstract: INTRODUCTION: Medulloblastoma (MB) is the most common brain malignancy in children. Among its molecular subtypes, Shh-driven with mutated Tp53 and Group 3 are still responsible for significant mortality and morbidity in the pediatric population. METHODS: The cytotoxic and anti-cancer stem-cell (CSC) effects of DSF and DSF/Cu ++ were evaluated with clonogenic assays, flow cytometry with AnnV/PI, PI/Ki67, Aldefluor ®, anti-Nestin and anti-CD133 antibody, immunofluorescence, and western-blotting. Human cell lines - ONS76, UW228 (SHH-driven with Tp53m), D425med, D283 and D341 (Group 3) were used. In vivo survival and nuclear protein localization protein-4 (NPL4), Ki67, Cleaved-Caspase-3, GFAP and NeuN expression were assessed in two xenografts of Group 3 MB with immunohistochemistry and western-blotting. RESULTS: Significant in vitro cytotoxicity was demonstrated at nanomolar concentrations of DSF. DSF/Cu ++ induced cell death (increased AnnV/PI, cleaved-Poly(ADP-ribose)polymerase fraction, and Apoptosis Inducing Factor) through NPL4 accumulation in cell nucleus and intracellular buildup of poly-ubiquitylated proteins. DNA damage was detected with western-blotting and H2AX foci on immunofluorescence. Flow cytometry analysis demonstrated a significant reduction in ALDH, Nestin-positive and CD133-positive cells following DSF/Cu ++ treatment. Anti-CSC effect was confirmed with western-blotting in vitro and in vivo . DSF/Cu ++ prolonged survival, and increased nuclear NPL4 expression in vivo in two models of Group 3 MB. IHC showed reduced expression of Ki67, and an increase in Cleaved-Caspase-3 + cells. GFAP and NeuN expression was not affected by treatment. CONCLUSION: DSF/Cu++ demonstrated a potent therapeutic effect on Shh-driven and Group 3 MB by inducing apoptosis and targeting CSCs. Our data suggest that this combination may serve as a novel treatment, alone or with existing therapies, for aggressive subtypes of pediatric MB.