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Smac mimetic Birinapant induces apoptosis and enhances TRAIL potency in inflammatory breast cancer cells in an IAP-dependent and TNF-α-independent mechanism

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Abstract

X-linked inhibitor of apoptosis protein (XIAP), the most potent mammalian caspase inhibitor, has been associated with acquired therapeutic resistance in inflammatory breast cancer (IBC), an aggressive subset of breast cancer with an extremely poor survival rate. The second mitochondria-derived activator of caspases (Smac) protein is a potent antagonist of IAP proteins and the basis for the development of Smac mimetic drugs. Here, we report for the first time that bivalent Smac mimetic Birinapant induces cell death as a single agent in TRAIL-insensitive SUM190 (ErbB2-overexpressing) cells and significantly increases potency of TRAIL-induced apoptosis in TRAIL-sensitive SUM149 (triple-negative, EGFR-activated) cells, two patient tumor-derived IBC models. Birinapant has high binding affinity (nM range) for cIAP1/2 and XIAP. Using isogenic SUM149- and SUM190-derived cells with differential XIAP expression (SUM149 wtXIAP, SUM190 shXIAP) and another bivalent Smac mimetic (GT13402) with high cIAP1/2 but low XIAP binding affinity (K d > 1 μM), we show that XIAP inhibition is necessary for increasing TRAIL potency. In contrast, single agent efficacy of Birinapant is due to pan-IAP antagonism. Birinapant caused rapid cIAP1 degradation, caspase activation, PARP cleavage, and NF-κB activation. A modest increase in TNF-α production was seen in SUM190 cells following Birinapant treatment, but no increase occurred in SUM149 cells. Exogenous TNF-α addition did not increase Birinapant efficacy. Neutralizing antibodies against TNF-α or TNFR1 knockdown did not reverse cell death. However, pan-caspase inhibitor Q-VD-OPh reversed Birinapant-mediated cell death. In addition, Birinapant in combination or as a single agent decreased colony formation and anchorage-independent growth potential of IBC cells. By demonstrating that Birinapant primes cancer cells for death in an IAP-dependent manner, these findings support the development of Smac mimetics for IBC treatment.

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Acknowledgments

The authors thank Amy Aldrich, Jess Hendin, and Katherine Aird for technical support. Grant support was received from American Cancer Society-RSG-08-290-01-CCE (GRD) and Duke University Chancellor’s Scholarship (JLA).

Conflict of interest

GRD has previously received a philanthropic gift for breast cancer research from TetraLogic Pharmaceuticals.

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This manuscript complies with the current laws of the United States of America.

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Authors and Affiliations

  1. Department of Surgery, Duke University Medical Center, Durham, NC, 27710, USA

    Jennifer L. Allensworth, Scott J. Sauer, H. Kim Lyerly & Gayathri R. Devi

  2. Department of Medicine, Duke University Medical Center, Durham, NC, 27710, USA

    Michael A. Morse

  3. Department of Pathology, Duke University Medical Center, Durham, NC, 27710, USA

    Jennifer L. Allensworth & Gayathri R. Devi

  4. Duke Cancer Institute, DUMC Duke University Medical Center, Box 2606, Durham, NC, 27710, USA

    H. Kim Lyerly, Michael A. Morse & Gayathri R. Devi

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  1. Jennifer L. Allensworth
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  2. Scott J. Sauer
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  5. Gayathri R. Devi
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Correspondence to Gayathri R. Devi.

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Jennifer L. Allensworth and Scott J. Sauer contributed equally and share co-first authorship.

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Allensworth, J.L., Sauer, S.J., Lyerly, H.K. et al. Smac mimetic Birinapant induces apoptosis and enhances TRAIL potency in inflammatory breast cancer cells in an IAP-dependent and TNF-α-independent mechanism. Breast Cancer Res Treat 137, 359–371 (2013). https://doi.org/10.1007/s10549-012-2352-6

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