An HGF–MSP chimera disassociates the trophic properties of scatter factors from their pro-invasive activity

Abstract

Hepatocyte growth factor (HGF) and macrophage-stimulating protein (MSP) have an intrinsic dual nature: they are trophic cytokines preventing apoptosis on one side and scatter factors promoting invasion on the other. For therapeutic use, their anti-apoptotic activity must be separated from their pro-invasive activity. To this end, we engineered chimeric factors containing selected functional domains of HGF and/or MSP in different combinations, and tested their biological activity. Here we present a chimeric cytokine derived from the alpha-chains of HGF and MSP, named Metron factor 1 for its ability to concomitantly activate the HGF receptor (Met) and the MSP receptor (Ron). We provide evidence that Metron factor 1 prevents apoptosis and stimulates cell proliferation at nanomolar concentrations, but is devoid of any pro-invasive activity. In an in vivo murine model of drug-induced nephrotoxicity, intravenous injection of recombinant Metron factor 1 prevented renal damage and preserved tubular integrity.

Monovalency Unleashes the Full Therapeutic Potential of the DN-30 Anti-Met Antibody

“…The recombinant antibodies were purified from conditioned medium by immobilized metal affinity chromatography as described (20).…”

“…1D). (15,20). Cells were stimulated for 15 min with increasing amounts of DN-30 mAb or DN-30 Fab, and Met activation was determined by immunoblotting with anti-phosphotyrosine antibodies.…”

“Active” Cancer Immunotherapy by Anti-Met Antibody Gene Transfer

“…After saturation with 0.5% bovine serum albumin for 1 h at 37jC, different dilutions of the sample and increasing concentrations (0-100 ng/mL) of DN-30RF purified by immobilized-metal affinity chromatography, as described in ref. 24, were bound overnight at 4jC. Bound antibodies were revealed by a first incubation with anti-Flag M2 biotin conjugates (Sigma) followed by incubation with streptavidinhorseradish peroxidase (HRP; Amersham).…”

Depleting MET-Expressing Tumor Cells by ADCC Provides a Therapeutic Advantage over Inhibiting HGF/MET Signaling

“…4E). Finally, in an HGF-induced branching morphogenesis assay using SV40 T-antigen-transformed LOC human kidney epithelial cell spheroids (23) and increasing antibody concentrations, ARGX-111 significantly inhibited HGF-induced branched tubule formation already at a 1:1 molar ratio with recombinant HGF (Fig. 4F).…”

“…All cells were cultured according to the protocols provided by the supplier and used within 6 months after receipt or validated by short tandem repeat profiling using Cell ID System (Promega). LOC cells (23), immortalized human mammary fibroblasts (24) and CRCM tumors (25) have been described previously.…”