title: Selective IAP inhibition results in sensitization of unstimulated but not CD40-stimulated chronic lymphocytic leukaemia cells to TRAIL-induced apoptosis.
abstract: Despite recent advances in therapy, chronic lymphocytic leukaemia (CLL) remains incurable and new treatment strategies are therefore urgently required. Inhibitor of apoptosis proteins (IAPs) are over-expressed in CLL, suggesting both a role in disease pathogenesis and the potential for therapeutic targeting. To explore these questions, we evaluated the effects on primary CLL cells of AZD5582, a novel potent and selective inhibitor of IAPs. AZD5582 at nanomolar concentrations induced extensive degradation of cIAP-1 and cIAP-2, but minimally of X chromosome-linked IAP (XIAP). However, these effects of AZD5582 produced little or no direct cytotoxicity, nor did they sensitize CLL cells to p53-dependent killing by fludarabine or p53-independent killing by dexamethasone. In contrast, AZD5582 significantly enhanced apoptosis induced by the death receptor (DR) agonist tumour necrosis factor-related apoptosis-inducing ligand (TRAIL). Importantly, killing by TRAIL plus AZD5582 was independent of adverse prognostic features including TP53 deletion which is strongly associated with chemoresistance in CLL. Coculture experiments involving transfected mouse fibroblasts expressing human CD40L (CD154) to mimic the effect of T cells at sites of tissue involvement showed that CD40 stimulation almost completely prevented the killing of CLL cells by TRAIL plus AZD5582 despite up-regulating TRAIL receptors 1 and 2. In conclusion, our findings confirm the rate-limiting, upstream involvement of IAPs in the extrinsic but not intrinsic apoptotic pathway of CLL cells and suggest that drug combinations that simultaneously activate DRs and inhibit IAPs may have therapeutic potential in patients with CLL who have failed T-cell-depleting chemotherapy.
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, University of Liverpool Liverpool, United Kingdom.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Zhuang
authors.affiliation: AstraZeneca R&D Boston Waltham, Massachusetts.
authors.forename: Naomi
authors.initials: N
authors.lastname: Laing
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, University of Liverpool Liverpool, United Kingdom.
authors.forename: Melanie
authors.initials: M
authors.lastname: Oates
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, University of Liverpool Liverpool, United Kingdom ; Royal Liverpool & Broadgreen University Hospitals NHS Trust Liverpool, United Kingdom.
authors.forename: Ke
authors.initials: K
authors.lastname: Lin
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, University of Liverpool Liverpool, United Kingdom ; Royal Liverpool & Broadgreen University Hospitals NHS Trust Liverpool, United Kingdom.
authors.forename: Gillian
authors.initials: G
authors.lastname: Johnson
authors.affiliation: Department of Molecular and Clinical Cancer Medicine, University of Liverpool Liverpool, United Kingdom ; Royal Liverpool & Broadgreen University Hospitals NHS Trust Liverpool, United Kingdom.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Pettitt
keywords: Apoptosis; CD40-stimulation; CLL; IAPs; TRAIL