Patient-derived glioblastoma cells show significant heterogeneity in treatment responses to the inhibitor-of-apoptosis-protein antagonist birinapant.
BACKGROUND: Resistance to temozolomide (TMZ) greatly limits chemotherapeutic effectiveness in glioblastoma (GBM). Here we analysed the ability of the Inhibitor-of-apoptosis-protein (IAP) antagonist birinapant to enhance treatment responses to TMZ in both commercially available and patient-derived GBM cells.
METHODS: Responses to TMZ and birinapant were analysed in a panel of commercial and patient-derived GBM cell lines using colorimetric viability assays, flow cytometry, morphological analysis and protein expression profiling of pro- and antiapoptotic proteins. Responses in vivo were analysed in an orthotopic xenograft GBM model.
RESULTS: Single-agent treatment experiments categorised GBM cells into TMZ-sensitive cells, birinapant-sensitive cells, and cells that were insensitive to either treatment. Combination treatment allowed sensitisation to therapy in only a subset of resistant GBM cells. Cell death analysis identified three principal response patterns: Type A cells that readily activated caspase-8 and cell death in response to TMZ while addition of birinapant further sensitised the cells to TMZ-induced cell death; Type B cells that readily activated caspase-8 and cell death in response to birinapant but did not show further sensitisation with TMZ; and Type C cells that showed no significant cell death or moderately enhanced cell death in the combined treatment paradigm. Furthermore, in vivo, a Type C patient-derived cell line that was TMZ-insensitive in vitro and showed a strong sensitivity to TMZ and TMZ plus birinapant treatments.
CONCLUSIONS: Our results demonstrate remarkable differences in responses of patient-derived GBM cells to birinapant single and combination treatments, and suggest that therapeutic responses in vivo may be greatly affected by the tumour microenvironment.
Funding is acknowledged from Science Foundation Ireland (13/IA/1881 and 14/IA/2582) and the European Union’s Seventh Frame-work Programme for research, technological development, anddemonstration under grant agreement 306021 (APO-DECIDE) toJHMP and MR. ATB is funded under the European Union’sSeventh Framework Programme under grant agreement 278981(AngioPredict). AT is funded by the Irish Cancer Society(CRF13TIV) and supported by Tesco Charity of the Year.
CommentsThis article is available at http://www.nature.com/bjc/journal/v114/n2/pdf/bjc2015420a.pdf
Published CitationZakaria Z, Tivnan A, Flanagan L, Murray DW, Salvucci M, Stringer BW, Day BW, Boyd AW, Ko ̈gel D, Rehm M, O’Brien DF, Byrne AT, Prehn JHM. Patient-derived glioblastoma cells showsignificant heterogeneity in treatmentresponses to the inhibitor-of-apoptosis-protein antagonist birinapant. British Journal of Cancer. 2016;114(2):188-98.
- Beaumont Hospital
- Centre for Systems Medicine
- Physiology and Medical Physics