title: Small-molecule Bcl-2 inhibitors sensitise tumour cells to immune-mediated destruction.
abstract: The cytotoxic effects of anticancer immune cells are mediated by perforin/granzyme-B, Fas ligand and tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), and therefore depend on intact apoptotic responses in target tumour cells. As killing by all three of these mechanisms is blocked by the frequently overexpressed antiapoptotic oncoprotein Bcl-2, we hypothesised that coexposure to a Bcl-2 inhibitor might enhance anticancer immune responses. We evaluated this in U937 lymphoma cells, and A02 melanoma cells, which both show strong Bcl-2 expression. Valpha24(+) Vbeta11(+) natural killer T (NKT) cells expanded from peripheral blood of normal donors (n=3) were coincubated with PKH26-labelled U937 cells, and cytotoxicity was determined by flow cytometry after annexin-V-FITC and 7-AAD staining. In all cases, addition of the HA14-1 small-molecule Bcl-2 inhibitor to the cocultures significantly increased apoptosis in the target U937 cells. Using a similar assay, killing of A02 cells by the cytotoxic T-lymphocyte clone 1H3 was shown to be amplified by coexposure to the potent small-molecule Bcl-2 inhibitor ABT-737. Experiments with immune effectors preincubated with concanamycin-A suggested that sensitisation to perforin/granzyme-B may underlie enhanced target-cell killing observed in the presence of Bcl-2 inhibitors. We conclude that immune destruction of malignant cells can be amplified by molecular interventions that overcome Bcl-2-mediated resistance to apoptosis.
authors.affiliation: Clive Berghofer Cancer Research Centre, Queensland Institute of Medical Research, Herston, Queensland 4029, Australia. Jason_Lickliter@health.qld.gov.au
authors.forename: J D
authors.initials: JD
authors.lastname: Lickliter
authors.forename: J
authors.initials: J
authors.lastname: Cox
authors.forename: J
authors.initials: J
authors.lastname: McCarron
authors.forename: N R
authors.initials: NR
authors.lastname: Martinez
authors.forename: C W
authors.initials: CW
authors.lastname: Schmidt
authors.forename: H
authors.initials: H
authors.lastname: Lin
authors.forename: M
authors.initials: M
authors.lastname: Nieda
authors.forename: A J
authors.initials: AJ
authors.lastname: Nicol
mesh_terms: D000328:Adult; D000970:Antineoplastic Agents / Q000494:pharmacology; D001578:Benzopyrans / Q000494:pharmacology*; D001713:Biphenyl Compounds / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D053804:Granzymes / Q000187:drug effects / Q000276:immunology; D006801:Humans; D008223:Lymphoma / Q000188:drug therapy* / Q000276:immunology; D008545:Melanoma / Q000188:drug therapy* / Q000276:immunology; D008562:Membrane Glycoproteins / Q000187:drug effects / Q000276:immunology; D009570:Nitriles / Q000494:pharmacology*; D009596:Nitrophenols / Q000494:pharmacology*; D054353:Perforin; D010879:Piperazines / Q000494:pharmacology; D052899:Pore Forming Cytotoxic Proteins / Q000187:drug effects / Q000276:immunology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors* / Q000276:immunology; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship; D013449:Sulfonamides / Q000494:pharmacology*
chemical_list: C501332:ABT-737; D000970:Antineoplastic Agents; D001578:Benzopyrans; D001713:Biphenyl Compounds; D004791:Enzyme Inhibitors; D008562:Membrane Glycoproteins; D009570:Nitriles; D009596:Nitrophenols; D010879:Piperazines; D052899:Pore Forming Cytotoxic Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D013449:Sulfonamides; C456800:ethyl 2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate; D054353:Perforin; D053804:Granzymes