title: TRAIL-R2-specific antibodies and recombinant TRAIL can synergise to kill cancer cells.
abstract: Tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptosis in cancer cells while sparing normal tissues. Despite promising preclinical results, few patients responded to treatment with recombinant TRAIL (Apo2L/Dulanermin) or TRAIL-R2-specific antibodies, such as conatumumab (AMG655). It is unknown whether this was due to intrinsic TRAIL resistance within primary human cancers or insufficient agonistic activity of the TRAIL-receptor (TRAIL-R)-targeting drugs. Fcγ receptors (FcγR)-mediated crosslinking increases the cancer-cell-killing activity of TRAIL-R2-specific antibodies in vivo. We tested this phenomenon using FcγR-expressing immune cells from patients with ovarian cancer. However, even in the presence of high numbers of FcγR-expressing immune cells, as found in ovarian cancer ascites, AMG655-induced apoptosis was not enabled to any significant degree, indicating that this concept may not translate into clinical use. On the basis of these results, we next set out to determine whether AMG655 possibly interferes with apoptosis induction by endogenous TRAIL, which could be expressed by immune cells. To do so, we tested how AMG655 affected apoptosis induction by recombinant TRAIL. This, however, resulted in the surprising discovery of a striking synergy between AMG655 and non-tagged TRAIL (Apo2L/TRAIL) in killing cancer cells. This combination was as effective in killing cancer cells as highly active recombinant isoleucine-zipper-tagged TRAIL (iz-TRAIL). The increased killing efficiency was due to enhanced formation of the TRAIL death-inducing signalling complex, enabled by concomitant binding of Apo2L/TRAIL and AMG655 to TRAIL-R2. The synergy of AMG655 with Apo2L/TRAIL extended to primary ovarian cancer cells and was further enhanced by combination with the proteasome inhibitor bortezomib or a second mitochondrial-derived activator of caspases (SMAC) mimetic. Importantly, primary human hepatocytes were not killed by the AMG655-Apo2L/TRAIL combination, also not when further combined with bortezomib or a SMAC mimetic. We therefore propose that clinical-grade non-tagged recombinant forms of TRAIL, such as dulanermin, could be combined with antibodies such as AMG655 to introduce a highly active TRAIL-R2-agonistic therapy into the cancer clinic.
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.|MRC Clinical Sciences Centre, Imperial College London, London, UK.
authors.forename: Mark H
authors.initials: MH
authors.lastname: Tuthill
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Antonella
authors.initials: A
authors.lastname: Montinaro
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Julia
authors.initials: J
authors.lastname: Zinngrebe
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Katharina
authors.initials: K
authors.lastname: Prieske
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Draber
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Stefan
authors.initials: S
authors.lastname: Prieske
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Tom
authors.initials: T
authors.lastname: Newsom-Davis
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Silvia
authors.initials: S
authors.lastname: von Karstedt
authors.affiliation: Amgen Inc., Seattle, USA.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Graves
authors.affiliation: Tumour Immunology Unit, Imperial College London, London, UK.
authors.forename: Henning
authors.initials: H
authors.lastname: Walczak
mesh_terms: D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001897:Boronic Acids / Q000627:therapeutic use; D000069286:Bortezomib; D002470:Cell Survival / Q000187:drug effects; D004357:Drug Synergism; D005260:Female; D022781:Hepatocytes / Q000187:drug effects; D006801:Humans; D010051:Ovarian Neoplasms / Q000188:drug therapy* / Q000276:immunology / Q000473:pathology; D011485:Protein Binding; D011719:Pyrazines / Q000627:therapeutic use; D017452:Receptors, IgG / Q000037:antagonists & inhibitors; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000037:antagonists & inhibitors / Q000276:immunology / Q000627:therapeutic use*; D011994:Recombinant Proteins / Q000627:therapeutic use*
chemical_list: D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D001897:Boronic Acids; C489017:FCGR1A protein, human; D011719:Pyrazines; D017452:Receptors, IgG; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D011994:Recombinant Proteins; C505639:TNFRSF10B protein, human; C554537:conatumumab; D000069286:Bortezomib