RGD-modified dihydrolipoamide dehydrogenase conjugated to titanium dioxide nanoparticles – switchable integrin-targeted photodynamic treatment of melanoma cells

Abstract

The photocytotoxic effect of UVA-excited titanium dioxide (TiO₂), which is caused by the generation of reactive oxygen species (ROS), is often used in medical applications, such as cancer treatment. Photodynamic-therapy (PDT) is applied in several cancer models including cutaneous melanoma (CM), however the lack of selectivity causing damage to surrounding healthy tissues limits its applicability and novel targeted-delivery approaches are required. As cancer cells often overexpress integrin receptors (e.g. αvβ₃) on their cell surface, targeted delivery of TiO₂ nanoparticles (NPs) via an Arg-Gly-Asp (RGD) motif would make PDT more selective. We have recently reported that the mitochondrial enzyme dihydrolipoamide dehydrogenase (DLDH) strongly and specifically conjugates TiO₂ via coordinative bonds. In this work we have modified DLDH with RGD moieties (DLDH^RGD), creating a molecular bridge between the integrin-expressing cancer cells and the photo-excitable TiO₂ nanoparticles. Physicochemical assays have indicated that the hybrid-conjugated nanobiocomplex, TiO₂–DLDH^RGD, is producing controlled-release ROS under UVA illumination, with anatase NPs being the most photoreactive TiO₂ form. This drug delivery system exhibited a cytotoxic effect in αvβ₃ integrin-expressing mice melanoma cells (B16F10), but not in normal cells lacking this integrin (HEK293). No cytotoxic effect was observed in the absence of UV illumination. Our results demonstrate the feasibility of combining the high efficiency of TiO₂-based PDT, with an integrin-mediated tumor-targeted drug delivery for nanomedicine.