Bortezomib Primes Glioblastoma, Including Glioblastoma Stem Cells, for TRAIL by Increasing tBid Stability and Mitochondrial Apoptosis

• Published in: Clinical cancer research 2011-06, Vol.17 (12), p.4019-4030
• Main Authors: UNTERKIRCHER, Thomas, CRISTOFANON, Silvia, KRISHNA VELLANKI, Sri Hari, NONNENMACHER, Lisa, KARPEL-MASSLER, Georg, WIRTZ, Christian Rainer, DEBATIN, Klaus-Michael, FULDA, Simone
• Format: Article
• Language: English
• Subjects: Antineoplastic agents; Antineoplastic Agents - pharmacology; Apoptosis; Apoptosis - drug effects; BAK protein; Bax protein; Bcl-2 protein; BH3 Interacting Domain Death Agonist Protein - metabolism; BID protein; Biological and medical sciences; Boronic Acids - pharmacology; Bortezomib; Cancer; Cell culture; Cell Death - drug effects; Cell Line, Tumor; Cell survival; Colonies; Death Domain Receptor Signaling Adaptor Proteins - metabolism; Glioblastoma; Glioblastoma - pathology; glioblastoma cells; Humans; Medical sciences; Membrane potential; Mitochondria; Mitochondria - drug effects; Molecular modelling; Neoplastic Stem Cells - drug effects; Neurology; Pharmacology. Drug treatments; proteasome inhibitors; Protein Stability - drug effects; Proto-Oncogene Proteins c-bcl-2 - metabolism; Pyrazines - pharmacology; Receptors, TNF-Related Apoptosis-Inducing Ligand - metabolism; Signal transduction; Signal Transduction - drug effects; Stem cells; TNF-Related Apoptosis-Inducing Ligand - metabolism; TRAIL protein; Tumors of the nervous system. Phacomatoses
• ISSN: 1078-0432; 1557-3265
• DOI: 10.1158/1078-0432.CCR-11-0075

Searching for novel approaches to sensitize glioblastoma for cell death, we investigated the proteasome inhibitor bortezomib. The effect of bortezomib on tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis signaling pathways was analyzed in glioblastoma cell lines, primary glioblastoma cultures, and in an in vivo model. Bortezomib and TRAIL synergistically trigger cell death and reduce colony formation of glioblastoma cells (combination index < 0.1). Investigations into the underlying molecular mechanisms reveal that bortezomib and TRAIL act in concert to cause accumulation of tBid, the active cleavage product of Bid. Also, the stability of TRAIL-derived tBid markedly increases on proteasome inhibition. Notably, knockdown of Bid significantly decreases bortezomib- and TRAIL-mediated cell death. By comparison, silencing of Noxa, which is also upregulated by bortezomib, does not confer protection. Coinciding with tBid accumulation, the activation of Bax/Bak and loss of mitochondrial membrane potential are strongly increased in cotreated cells. Overexpression of Bcl-2 significantly reduces mitochondrial perturbations and cell death, underscoring the functional relevance of the mitochondrial pathway. In addition, bortezomib cooperates with TRAIL to reduce colony formation of glioblastoma cells, showing an effect on long-term survival. Of note, bortezomib profoundly enhances TRAIL-triggered cell death in primary cultured glioblastoma cells and in patient-derived glioblastoma stem cells, underlining the clinical relevance. Importantly, bortezomib cooperates with TRAIL to suppress tumor growth in an in vivo glioblastoma model. These findings provide compelling evidence that the combination of bortezomib and TRAIL presents a promising novel strategy to trigger cell death in glioblastoma, including glioblastoma stem cells, which warrants further investigation.