Abstract
Our goal was to develop a potent humanized antibody against mouse/human CXCL12. This report summarized its in vitro and in vivo activities. Cell surface binding and cell migration assays were used to select neutralizing hamster antibodies, followed by testing in several animal models. Monoclonal antibody (mAb) 30D8 was selected for humanization based on its in vitro and in vivo activities. 30D8, a hamster antibody against mouse and human CXCL12α, CXCL12β, and CXCL12γ, was shown to dose-dependently block CXCL12α binding to CXCR4 and CXCR7, and CXCL12α-induced Jurkat cell migration in vitro. Inhibition of primary tumor growth and/or metastasis was observed in several models. 30D8 alone significantly ameliorated arthritis in a mouse collagen-induced arthritis model (CIA). Combination with a TNF-α antagonist was additive. In addition, 30D8 inhibited 50% of laser-induced choroidal neovascularization (CNV) in mice. Humanized 30D8 (hu30D8) showed similar in vitro and in vivo activities as the parental hamster antibody. A crystal structure of the hu30D8 Fab/CXCL12α complex in combination with mutational analysis revealed a "hot spot" around residues Asn(44)/Asn(45) of CXCL12α and part of the RFFESH region required for CXCL12α binding to CXCR4 and CXCR7. Finally, hu30D8 exhibited fast clearance in cynomolgus monkeys but not in rats. CXCL12 is an attractive target for treatment of cancer and inflammation-related diseases; hu30D8 is suitable for testing this hypothesis in humans.
Highlights
CXCL12, known as stromal-derived factor 1 (SDF1), is a CXC chemokine that binds to two G-protein–coupled receptors, CXCR4 and CXCR7
30D8, a hamster antibody against mouse and human CXCL12a, CXCL12b, and CXCL12g, was shown to dose-dependently block CXCL12a binding to CXCR4 and CXCR7, and CXCL12a-induced Jurkat cell migration in vitro
Inhibition of primary tumor growth and/or metastasis was observed in several models. 30D8 alone significantly ameliorated arthritis in a mouse collagen-induced arthritis model (CIA)
Summary
CXCL12, known as stromal-derived factor 1 (SDF1), is a CXC chemokine that binds to two G-protein–coupled receptors, CXCR4 and CXCR7 (reviewed in ref. 1). CXCL12, known as stromal-derived factor 1 (SDF1), is a CXC chemokine that binds to two G-protein–coupled receptors, CXCR4 and CXCR7 It participates in many developmental and physiologic processes, including hematopoiesis and angiogenesis. Gene deletion of CXCL12 or CXCR4 results in embryonic lethality at E18.5, with severe developmental defects affecting central nervous system, bone marrow, heart, and vascular system [2, 3]. CXCR7 inactivation, on the other hand, seems to affect primarily cardiac development, defects in neuronal migration have been noted [4]. Note: Supplementary data for this article are available at Clinical Cancer Research Online (http://clincancerres.aacrjournals.org/).
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