Macropinocytosis of Bevacizumab by Glioblastoma Cells in the Perivascular Niche Affects their Survival.

Abstract

Purpose: Bevacizumab, a humanized monoclonal antibody to VEGF, is used routinely in the treatment of patients with recurrent glioblastoma (GBM). However, very little is known regarding the effects of bevacizumab on the cells in the perivascular space in tumors.Experimental Design: Established orthotopic xenograft and syngeneic models of GBM were used to determine entry of monoclonal anti-VEGF-A into, and uptake by cells in, the perivascular space. Based on the results, we examined CD133+ cells derived from GBM tumors in vitro Bevacizumab internalization, trafficking, and effects on cell survival were analyzed using multilabel confocal microscopy, immunoblotting, and cytotoxicity assays in the presence/absence of inhibitors.Results: In the GBM mouse models, administered anti-mouse-VEGF-A entered the perivascular tumor niche and was internalized by Sox2+/CD44+ tumor cells. In the perivascular tumor cells, bevacizumab was detected in the recycling compartment or the lysosomes, and increased autophagy was found. Bevacizumab was internalized rapidly by CD133+/Sox2+-GBM cells in vitro through macropinocytosis with a fraction being trafficked to a recycling compartment, independent of FcRn, and a fraction to lysosomes. Bevacizumab treatment of CD133+ GBM cells depleted VEGF-A and induced autophagy thereby improving cell survival. An inhibitor of lysosomal acidification decreased bevacizumab-induced autophagy and increased cell death. Inhibition of macropinocytosis increased cell death, suggesting macropinocytosis of bevacizumab promotes CD133+ cell survival.Conclusions: We demonstrate that bevacizumab is internalized by Sox2+/CD44+-GBM tumor cells residing in the perivascular tumor niche. Macropinocytosis of bevacizumab and trafficking to the lysosomes promotes CD133+ cell survival, as does the autophagy induced by bevacizumab depletion of VEGF-A. Clin Cancer Res; 23(22); 7059-71. ©2017 AACR.

• Authors: Gaëlle Müller-Greven, Cathleen R Carlin, Monica E Burgett, Manmeet S Ahluwalia, Adam Lauko, Amy S Nowacki, Cameron J Herting, Maha A Qadan, Markus Bredel, Steven A Toms, Justin D Lathia, Dolores Hambardzumyan, Jann N Sarkaria, Petra Hamerlik, Candece L Gladson
• Journal: Clinical cancer research : an official journal of the American Association for Cancer Research (Clin Cancer Res) Vol. 23 Issue 22 Pg. 7059-7071 (Nov 15 2017) ISSN: 1557-3265 [Electronic] United States
• PMID: 28912141 (Publication Type: Journal Article)
• Copyright: ©2017 American Association for Cancer Research.

Chemical References

• Actins
• Antineoplastic Agents, Immunological
• Biomarkers
• Bevacizumab

Topics

• Actins (metabolism)
• Animals
• Antineoplastic Agents, Immunological (immunology, pharmacology)
• Autophagy (drug effects)
• Bevacizumab (immunology, pharmacology)
• Biomarkers
• Cell Line, Tumor
• Cell Membrane (metabolism)
• Disease Models, Animal
• Endosomes (metabolism)
• Glioblastoma (immunology, metabolism, mortality, pathology)
• Humans
• Immunophenotyping
• Lysosomes (metabolism)
• Mice
• Neovascularization, Pathologic (immunology, metabolism)
• Pinocytosis (immunology)
• Tumor Microenvironment (drug effects, immunology)
• Xenograft Model Antitumor Assays