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RIP1 is required for IAP inhibitor-mediated sensitization for TRAIL-induced apoptosis via a RIP1/FADD/caspase-8 cell death complex

Oncogene volume 32, pages 3263–3273 (2013)Cite this article

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Abstract

Inhibitor of apoptosis (IAP) proteins represent promising therapeutic targets due to their high expression in many cancers. Here, we report that small-molecule IAP inhibitors at subtoxic concentrations cooperate with monoclonal antibodies against TRAIL receptor 1 (Mapatumumab) or TRAIL-R2 (Lexatumumab) to induce apoptosis in neuroblastoma cells in a highly synergistic manner (combination index <0.1). Importantly, we identify receptor-activating protein 1 (RIP1) as a critical mediator of this synergism. RIP1 is required for the formation of a RIP1/FADD/caspase-8 complex that drives caspase-8 activation, cleavage of Bid into tBid, mitochondrial outer membrane permeabilization, full activation of caspase-3 and caspase-dependent apoptosis. Indeed, knockdown of RIP1 abolishes formation of the RIP1/FADD/caspase-8 complex, caspase activation and apoptosis upon combination treatment. Similarly, inhibition of RIP1 kinase activity by Necrostatin-1 inhibits IAP inhibitor- and TRAIL receptor-triggered apoptosis. In contrast, overexpression of the dominant-negative superrepressor IκBα-SR or addition of the tumor necrosis factor (TNF)α-blocking antibody Enbrel do not interfere with cotreatment-induced apoptosis, pointing to a nuclear factor-κB- and TNFα-independent mechanism. Of note, IAP inhibitor also sensitizes primary cultured neuroblastoma cells for TRAIL receptor-mediated loss of viability, underscoring the clinical relevance. By identifying RIP1 as a critical mediator of IAP inhibitor-mediated sensitization for Mapatumumab- or Lexatumumab-induced apoptosis, our findings provide new insights into the synergistic interaction of IAP inhibitors together with TRAIL receptor agonists.

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Acknowledgements

We thank X Wang for providing RIP3 antibody and C Hugenberg for expert secretarial assistance. This work has been partially supported by grants from the Deutsche Forschungsgemeinschaft, European Community (ApopTrain, APO-SYS) and IAP6/18 (to SF).

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Authors and Affiliations

  1. Institute for Experimental Cancer Research in Pediatrics, Goethe-University, Frankfurt, Germany

    B A Abhari, S Cristofanon & S Fulda

  2. Department of Pediatric Surgery, Dr von Haunerschen Children’s Hospital, Ludwig-Maximilians-University Munich, Germany

    R Kappler & D von Schweinitz

  3. Oncology Research Department, Human Genome Sciences, Inc., Rockville, MD, USA

    R Humphreys

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  1. B A Abhari
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Correspondence to S Fulda.

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Abhari, B., Cristofanon, S., Kappler, R. et al. RIP1 is required for IAP inhibitor-mediated sensitization for TRAIL-induced apoptosis via a RIP1/FADD/caspase-8 cell death complex. Oncogene 32, 3263–3273 (2013). https://doi.org/10.1038/onc.2012.337

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