Nanoscale Integrin Ligand Patterns Determine Melanoma Cell Behavior

Cells use integrin receptors to adhere onto surfaces by binding to ligands such as the arginine-glycine-aspartic acid (RGD) motif. Cancer cells make use of this adhesion process, which has motivated the development of integrin-directed drugs. However, those drugs may exert paradoxical effects on tumor progression, which raises the question of how integrin function is governed in tumor cells on the nanoscale. We have utilized precisely defined and tunable RGD ligand site densities spanning 1 order of magnitude, i.e., 103 to 1145 ligand sites/μm(2), by using RGD-functionalized gold nanoparticle patterns immobilized on glass by block copolymer (micellar) nanolithography. In an αVβ3 integrin-dependent fashion, human melanoma cells spread, formed focal contacts, and reorganized cytoskeletal fibers on a physiologically relevant RGD density of 349 sites/μm(2). Intriguingly, low doses of solute RGD "shifted" the optimal densities of immobilized ligand along with corresponding melanoma cell integrin clusters and cytoskeletal changes toward those typical for "intermediate" ligand presentation. Consequently, melanoma cells were forced into a "permissive" state, optimizing interactions with suboptimal nanostructured biomimetic surfaces, thus providing an explanation for the seemingly paradoxical effects on tumor progression and a potential clue for individualized antitumoral therapies.