An anti-αv-integrin antibody that blocks integrin function inhibits the development of a human melanoma in nude mice

Abstract

A series of murine monoclonal antibodies were raised against purified human alpha v beta 3 integrin and against M21 human melanoma cells. Five notable hybridomas were identified by ELISA on purified integrins, and the isolated antibodies bound the alpha v-chain. These antibodies, 17E6, 20A9, 23G5, 14D9.F8 and 10G2, recognised the extracellular domains of the integrin, and were shown to be reactive in FACS, immunoprecipitation, ELISA, and ELISA on fixed cells with M21, M21-L4, and UCLA-P3, but not with the alpha v-deficient M21-L or M21-L-IIb (M21-L transfected with GpIIb integrin). One antibody, 17E6, strongly perturbed cell attachment mediated by alpha v integrins, reacting at least with alpha v beta 3, alpha v beta 5, and alpha v beta 1, and strongly inhibiting cell attachment to alpha v-ligands vitronectin and fibronectin with an IC50 of approximately 0.1 microgram ml-1. Furthermore, 17E6 at this concentration could induce cell retraction from the substrate, while LM609 (anti-alpha v beta 3) and control antibody 14E2 (anti-200 kDa melanoma surface protein) at 1,000-fold higher concentrations had minimal effects on cell morphology. The action of 17E6 was reversible and was not due to toxic effects: in vitro 17E6 at 0.1 mg ml-1 did not affect either cell proliferation or DNA synthesis. In two nude-mouse tumour models, subcutaneous tumour development and a lung colonisation (‘experimental metastasis’) assay, injection of 17E6 strongly inhibited tumour development, while isotype-matched controls had no effect. There was no obvious mechanism of cell or of complement-mediated tumour cytotoxicity; the antibody did not mediate ADCC or AECDC, or complement fixation. The data strongly support previous studies which have indicated the importance of alpha v-integrins, and especially alpha v beta 3, in the tumour progression of human melanoma.

Tumor Targeting by Peptide-Decorated Gold Nanoparticles

“…The improvement in the uptake of peptide-decorated NPs by the B16F10 cell line is in agreement with the cytotoxicity data obtained by the MTT assay. This strongly supports that adsorption/uptake is determined by αvβ3 integrin binding mediated by the RGD sequence. , RGD-like peptides have the highest affinity for αvβ3 integrin, but they are able to bind the αvβ5 and α5β1 integrins as well, which can contribute to NP binding. , …”

“…This strongly supports that adsorption/uptake is determined by αvβ3 integrin binding mediated by the RGD sequence. 66,67 RGD-like peptides have the highest affinity for αvβ3 integrin, but they are able to bind the αvβ5 and α5β1 integrins as well, which can contribute to NP binding. 68,69 The three different NP formulations probably take advantage of different pathways to gain access to the cytoplasmic component.…”

Design, functionalization strategies and biomedical applications of targeted biodegradable/biocompatible polymer-based nanocarriers for drug delivery

“…74 It has indeed been shown that micelles slightly surface modified by antibodies accumulated in tumors, showing a better anti-tumor efficacy. 158 Antibodies-decorated nanoparticulate systems have been employed in the field of cancer (anti-CD3 Mab, 159 anti-HER2 Mab (also called trastuzumab or Herceptin), 160 EGFRMab, 161 DI17E6 Mab, 162 anti FAS Mab, 163 anti antigen rich breast cancer cells Mab, 164 AMB8LK Mab, 165 anti-CD20 mAB, 166 anti-CEA Mab 167 ), Alzheimer's disease (anti-Ab 1-42 MAb), 168 brain delivery (29B4, 169 OX26, 170 R17217, 171 anti-CD44 mAB, 172 anti-NCAM1 Mab 173 ) and autoimmune diseases (anti CD4 + ). 174 A specific target to inflamed endothelium (generally observed in various pathological settings such as cancer, cardiovascular disease and arthritis) which is characterized by overexpression of endothelial cell adhesion molecules, was achieved using anti-ICAM-1 Mab, anti VCAM-1 Mab, anti E-selectin Mab as well as anti P-selectin Mab.…”

Manipulation of the Nanoscale Presentation of Integrin Ligand Produces Cancer Cells with Enhanced Stemness and Robust Tumorigenicity

“…Cytoskeleton components (e.g., F-actin) and mechanosensitive signaling factors such as FAK and Cdc42 are known to be involved in the response of living cells to mechanical stimulations. 17,36 Immunofluorescence staining showed that the expressions of these components decreased on the substrates with increasing tether mobility (MNP2−20k−M) (Figure 2a). In contrast, cells cultured on the substrate with low/magnetically restricted ligand tether mobility (MNP2− 20k+M) showed robust expression of these mechanotransduction markers.…”