Abstract
Bcl-2 family proteins regulate mitochondrial apoptosis downstream of diverse stressors. Cancer cells frequently deregulate Bcl-2 proteins leading to chemoresistance. We have optimized a platform for solid tumors in which Bcl-2 family resistance patterns are inferred. Functional mitochondria were isolated from neuroblastoma (NB) cell lines, exposed to distinct BH3-domain peptides, and assayed for cytochrome c release. Such BH3 profiles revealed three patterns of cytochrome c response. A subset had a dominant NoxaBH3 response implying Mcl1 dependence. These cells were more sensitive to small molecules that antagonize Mcl1 (AT-101) than those that antagonize Bcl-2, Bcl-xL and Bcl-w (ABT-737). A second subset had a dominant BikBH3 response, implying a Bcl-xL/-w dependence, and was exquisitely sensitive to ABT-737 (IC50 <200 nM). Finally, most NB cell lines derived at relapse were relatively resistant to pro-death BH3 peptides and Bcl-2 antagonists. Our findings define heterogeneity for apoptosis resistance in NB, help triage emerging Bcl-2 antagonists for clinical use, and provide a platform for studies to characterize post-therapy resistance mechanisms for NB and other solid tumors.
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Abbreviations
- NB:
-
neuroblastoma
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Acknowledgements
ABT-737 and AT-101 were generously provided by Abbott Laboratories and Ascenta Therapeutics, respectively. We thank Steven Elmore from Abbott Laboratories and Lance Leopold from Ascenta Therapeutics for their direction and assistance with drug usage. We thank David Teachey for provision of T cells, Josh Courtright for xenograft assistance, and Anthony Letai for help in optimizing mitochondrial isolation assays. This work was supported by NIH CA97323, the King Family, the Richard and Sheila Sanford Chair in Pediatric Oncology, and the Alex's Lemonade Stand Foundation (to MDH); and NIH K08-CA128925, The Caitlin Robb Foundation, and Hope Street Kids Foundation (to KCG).
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Goldsmith, K., Lestini, B., Gross, M. et al. BH3 response profiles from neuroblastoma mitochondria predict activity of small molecule Bcl-2 family antagonists. Cell Death Differ 17, 872–882 (2010). https://doi.org/10.1038/cdd.2009.171
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DOI: https://doi.org/10.1038/cdd.2009.171
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