Interaction between bevacizumab and murine VEGF-A: a reassessment.

PURPOSE Bevacizumab is a humanized anti-human VEGF-A monoclonal antibody (mAb) approved by the United States Food and Drug Administration for cancer therapy and used off label to treat neovascular age-related macular degeneration. Earlier studies characterized bevacizumab as species specific and lacking the ability to neutralize murine (m) VEGF-A. However, a recent study reported that bevacizumab is a potent inhibitor of hemangiogenesis and lymphangiogenesis in murine models. The authors sought to reassess the interaction between bevacizumab and mVEGF-A. METHODS The authors performed Western blot analysis, plasmon resonance by BIAcore, and endothelial cell proliferation assays to characterize the interaction between bevacizumab and mVEGF-A. They also tested whether bevacizumab had any effects in two in vivo murine models, laser-induced choroidal neovascularization (CNV) and melanoma growth. RESULTS Western blot detected a very weak interaction, but BIAcore detected no measurable interaction between mVEGF and bevacizumab. Bevacizumab failed to inhibit mVEGF-stimulated endothelial cell proliferation. In addition, bevacizumab was indistinguishable from the control antibody in the CNV and tumor models, whereas a cross-reactive anti-VEGF-A mAb had dramatic inhibitory effects. CONCLUSIONS Bevacizumab has an extremely weak interaction with mVEGF-A, which fails to result in immunoneutralization as assessed by several bioassays.

[1]  G. Fuh,et al.  Tumor refractoriness to anti-VEGF treatment is mediated by CD11b+Gr1+ myeloid cells , 2007, Nature Biotechnology.

[2]  P. Campochiaro,et al.  VEGF‐TRAPR1R2 suppresses choroidal neovascularization and VEGF‐induced breakdown of the blood–retinal barrier , 2003 .

[3]  M. Detmar,et al.  VEGF-A produced by chronically inflamed tissue induces lymphangiogenesis in draining lymph nodes. , 2007, Blood.

[4]  Claudio Campa,et al.  Targeting VEGF-A to treat cancer and age-related macular degeneration. , 2007, Annual review of medicine.

[5]  Satoshi Hirakawa,et al.  VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis , 2005, The Journal of experimental medicine.

[6]  A. Adamis,et al.  Requirement for vascular endothelial growth factor in wound- and inflammation-related corneal neovascularization. , 1998, Investigative ophthalmology & visual science.

[7]  K. Red-Horse,et al.  Endothelium-microenvironment interactions in the developing embryo and in the adult. , 2007, Developmental cell.

[8]  B. Rouse,et al.  Contribution of Vascular Endothelial Growth Factor in the Neovascularization Process during the Pathogenesis of Herpetic Stromal Keratitis , 2001, Journal of Virology.

[9]  F. Peale,et al.  Tumor-Driven Paracrine Platelet-Derived Growth Factor Receptor α Signaling Is a Key Determinant of Stromal Cell Recruitment in a Model of Human Lung Carcinoma , 2006, Clinical Cancer Research.

[10]  R. Avery,et al.  Intravitreal bevacizumab (Avastin) for neovascular age-related macular degeneration. , 2006, Ophthalmology.

[11]  A. D. de Vos,et al.  VEGF and the Fab fragment of a humanized neutralizing antibody: crystal structure of the complex at 2.4 A resolution and mutational analysis of the interface. , 1998, Structure.

[12]  Leonard,et al.  Humanization of an anti-vascular endothelial growth factor monoclonal antibody for the therapy of solid tumors and other disorders. , 1997, Cancer research.

[13]  L. Damico,et al.  DEVELOPMENT OF RANIBIZUMAB, AN ANTI–VASCULAR ENDOTHELIAL GROWTH FACTOR ANTIGEN BINDING FRAGMENT, AS THERAPY FOR NEOVASCULAR AGE-RELATED MACULAR DEGENERATION , 2006, Retina.

[14]  D. Hinton,et al.  Transdifferentiated retinal pigment epithelial cells are immunoreactive for vascular endothelial growth factor in surgically excised age-related macular degeneration-related choroidal neovascular membranes. , 1996, Investigative ophthalmology & visual science.

[15]  N. Ferrara,et al.  The biology of VEGF and its receptors , 2003, Nature Medicine.

[16]  F. Peale,et al.  Mice expressing a humanized form of VEGF-A may provide insights into the safety and efficacy of anti-VEGF antibodies , 2007, Proceedings of the National Academy of Sciences.

[17]  J. Winer,et al.  The vascular endothelial growth factor proteins: identification of biologically relevant regions by neutralizing monoclonal antibodies. , 1992, Growth factors.

[18]  F. Peale,et al.  Cross-species Vascular Endothelial Growth Factor (VEGF)-blocking Antibodies Completely Inhibit the Growth of Human Tumor Xenografts and Measure the Contribution of Stromal VEGF* , 2006, Journal of Biological Chemistry.

[19]  Raymond M Schiffelers,et al.  Inhibition of ocular angiogenesis by siRNA targeting vascular endothelial growth factor pathway genes: therapeutic strategy for herpetic stromal keratitis. , 2004, The American journal of pathology.

[20]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[21]  Mingde Zhu,et al.  Kinetic analysis of a high-affinity antibody/antigen interaction performed by multiple Biacore users. , 2006, Analytical biochemistry.

[22]  I. Kasman,et al.  Effects of an anti-VEGF-A monoclonal antibody on laser-induced choroidal neovascularization in mice: optimizing methods to quantify vascular changes. , 2008, Investigative ophthalmology & visual science.

[23]  Kenneth J. Hillan,et al.  Discovery and development of bevacizumab, an anti-VEGF antibody for treating cancer , 2004, Nature Reviews Drug Discovery.

[24]  L. Aiello,et al.  Vascular endothelial growth factor in ocular fluid of patients with diabetic retinopathy and other retinal disorders. , 1994, The New England journal of medicine.

[25]  Napoleone Ferrara,et al.  Angiogenesis as a therapeutic target , 2005, Nature.

[26]  Björn Bachmann,et al.  Bevacizumab as a potent inhibitor of inflammatory corneal angiogenesis and lymphangiogenesis. , 2007, Investigative ophthalmology & visual science.

[27]  R. Karlsson,et al.  Experimental design for kinetic analysis of protein-protein interactions with surface plasmon resonance biosensors. , 1997, Journal of immunological methods.

[28]  D. Guyer,et al.  Pegaptanib, a targeted anti-VEGF aptamer for ocular vascular disease , 2006, Nature Reviews Drug Discovery.

[29]  P. Campochiaro,et al.  VEGF-TRAP(R1R2) suppresses choroidal neovascularization and VEGF-induced breakdown of the blood-retinal barrier. , 2003, Journal of cellular physiology.