Novel anti-denatured collagen humanized antibody D93 inhibits angiogenesis and tumor growth: An extracellular matrix-based therapeutic approach.
Matrix metalloproteases (MMPs) secreted by both tumor and endothelial cells proteolytically degrade collagen during tumor growth and neo-vascularization. This exposes cryptic binding sites on collagen with functional relevance for angiogenesis. In this report, we characterized a novel humanized monoclonal IgG1 antibody, D93. After humanization, the antibody retained the binding specificity of the parental murine IgM antibody for denatured (dn) collagen. D93 recognized dn-collagen but not native (nat) collagen of different species, including mouse, chicken, and human, indicating that its cryptic binding site(s) is conserved across species. In immunohistochemistry (IHC) studies, D93 stained the basement membrane of blood vessels in several xenograft human tumors or in surgically removed tumor tissues from patients with different types of malignancies. D93 staining was rarely or not present in normal blood vessels of healthy tissues. In in vivo experiments, D93 significantly inhibited basic fibroblast growth factor (bFGF)-induced angiogenesis in chick embryo chorioallantoic membrane (CAM) and the tumor growth of pre-established orthotopic human breast (MDA-MB-435) tumors in mice. D93 i.v. administered in mice was subsequently detected in the subendothelial basement membrane of tumor blood vessels but not blood vessels of normal tissues. Inhibition of growth of pre-established orthotopic human breast MDA-MB-435 tumors was more effective when D93 was combined with Taxol, than either treatment alone. In addition, tumors from animals treated with D93 and/or Taxol showed significantly reduced levels of the endothelial cell-marker CD31. Our data suggest that blockade of cryptic epitopes exposed on collagen IV during angiogenesis and tumor growth by a monoclonal antibody may provide a novel therapeutic modality for treatment of cancer and pathogenic neo-vascularization.
Pernasetti
Flavia
F
Cell-Matrix, Inc., a subsidiary of Micromet Corporation, Carlsbad, CA 92008, USA. fpernase@hotmail.com
Nickel
Jeffrey
J
Clark
Derek
D
Baeuerle
Patrick A
PA
Van Epps
Dennis
D
Freimark
Bruce
B
D000328:Adult; D000368:Aged; D020533:Angiogenesis Inhibitors / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology*; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D001485:Basement Membrane / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D001943:Breast Neoplasms / Q000098:blood supply / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D048708:Cell Growth Processes / Q000187:drug effects; D002642:Chick Embryo; D024141:Collagen Type IV / Q000276:immunology* / Q000378:metabolism; D004306:Dose-Response Relationship, Immunologic; D004357:Drug Synergism; D000939:Epitopes; D005109:Extracellular Matrix / Q000378:metabolism*; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007075:Immunoglobulin M / Q000276:immunology; D008297:Male; D008545:Melanoma / Q000098:blood supply / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008875:Middle Aged; D009389:Neovascularization, Pathologic / Q000276:immunology / Q000378:metabolism / Q000473:pathology / Q000628:therapy; D017239:Paclitaxel / Q000008:administration & dosage; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology
D020533:Angiogenesis Inhibitors; D000911:Antibodies, Monoclonal; D024141:Collagen Type IV; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D017239:Paclitaxel