Abstract
Abstract
Background: Resistance to antiangiogenic therapy (AT) in patients with glioblastoma (GBM) treated with bevacizumab (BEV) is characterized by local recurrence and distant dissemination of gliomas associated with remodeling of tumor vessels and pronounced hypoxia known to promote glioma cell invasion. Expression of the chemokine receptor CXCR4 and its ligand stromal cell-derived factor-1α (SDF-1α) is enhanced in invading tumor cells (CXCR4) and neurons and blood vessels (SDF-1α) in GBM and associated with tumor hypoxia, proliferation, invasion and angiogenesis. Using Protein Epitope Mimetics (PEM) technology (Robinson JA et al., 2008), Polyphor Ltd. has developed selective, highly potent CXCR4 antagonists (De Marco SJ et al., 2006) (U.S. Patent no. 8,716,242), such as POL5551. To address the problem of resistance to AT, we sought to determine whether combined therapy (CTx) with POL5551 and the murine equivalent of BEV (antibody B20-4.1.1) could inhibit the invasion and associated pathologic characteristics of gliomas in vivo.
Methods: Adult C57BL/6 mice implanted orthotopically with syngeneic CT-2A or GL261 glioma cells were randomized on day 14 into 4 groups: 1) control, 2) POL5551 (5 mg/kg s.c.), 3) anti-murine vascular endothelial growth factor (VEGF) monoclonal antibody B20-4.1.1 (5 mg/kg i.p.; Genentech Inc.) (Bagri A et al., 2010) and 4) combined POL5551 and B20-4.1.1 (CTx). On day 28, brain tissues were processed, sections analyzed for tumor volume and invasiveness (Sottoriva A et al., 2010) (H&E), hypoxia (Hypoxyprobe), proliferation (pHH3, phosphohistone H3), vascular density (CD105), apoptosis (cleaved caspase 3) and microglial (IBA1, ionized calcium binding adaptor molecule 1) and stem (nestin) cellular fractions and the results analyzed by analysis of variance (ANOVA). The tissue concentrations of POL5551 were measured by mass spectrometry.
Results: Following B20-4.1.1, both glioma models recapitulated the increased invasive phenotype seen in human GBM associated with increased tumor hypoxia and reduced cellular proliferation and vascular density. Following POL5551 alone, tumor proliferation was slightly diminished. Nestin immunopositivity in CT-2A glioma was markedly reduced following POL5551, and to an intermediate level with CTx, consistent with a previous report (Barone A et al., 2014). In both models, CTx reduced tumor invasiveness (by up to 39.3% compared with B20-4.1.1 alone) and vascular density together with increased apoptosis. In CT-2A glioma, CTx also diminished IBA1 immunopositivity. The concentration of POL5551 was higher in GL261 glioma (45.8 nM) than in brain adjacent to tumor (23.7 nM) or normal brain (11.1 nM) and sufficient to account for its biological effects given the high binding affinity (0.5 nM) of POL5551 for CXCR4. In preliminary work, POL5551 reduced the rate of migration of GL261 glioma cells in vitro.
Conclusions: Following anti-VEGF antibody treatment, both murine glioma models recapitulated the increased invasive phenotype seen in human GBM associated with a more hypoxic tumor microenvironment. In both tumor models, CTx with POL5551 and B20-4.1.1 may be beneficial in overcoming AT-induced glioma dissemination. Importantly, POL5551 crossed the blood-brain barrier and accumulated to biologically active levels in glioma tissue. Our results suggest the potential clinical utility of PEM CXCR4 antagonists as adjunct therapy for patients with GBM.
Citation Format: Jean-Pierre Gagner, Yasmeen Sarfraz, Fawaz M. Alotaibi, Valerio Ortenzi, Awab T. Tayyib, Luis A. Chiriboga, Garry J. Douglas, Eric Chevalier, Barbara Romagnoli, Gerald Tuffin, Klaus Dembowsky, David Zagzag. A novel CXCR4 antagonist interferes with antivascular endothelial growth factor therapy-induced glioma dissemination. [abstract]. In: Proceedings of the AACR Special Conference: Advances in Brain Cancer Research; May 27-30, 2015; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2015;75(23 Suppl):Abstract nr A20.