Nanoconjugation modulates the trafficking and mechanism of antibody induced receptor endocytosis.

Publication Type:

Journal Article

Source:

Proceedings of the National Academy of Sciences of the United States of America, Volume 107, Issue 33, p.14541-6 (2010)

Keywords:

Antibodies, Monoclonaldigestive disease, digestive deseases Antineoplastic Agentsdigestive disease, digestive deseases Biological Transportdigestive disease, digestive deseases Carbocyaninesdigestive disease, digestive deseases Cell Line, Tumordigestive disease, digestive deseases Dynamin IIdigestive disease, digestive deseases Endocytosisdigestive disease, digestive deseases Endosomesdigestive disease, digestive deseases Golddigestive disease, digestive deseases Golgi Apparatusdigestive disease, digestive deseases Humansdigestive disease, digestive deseases Lysosomesdigestive disease, digestive deseases Metal Nanoparticlesdigestive disease, digestive deseases Microscopy, Confocaldigestive disease, digestive deseases Microscopy, Electron, Transmissiondigestive disease, digestive deseases Models, Biologicaldigestive disease, digestive deseases Mutationdigestive disease, digestive deseases Pancreatic Neoplasmsdigestive disease, digestive deseases Receptor, Epidermal Growth Factor

Abstract:

Treatment with monoclonal antibody (mAbs) is a viable therapeutic option in cancer. Recently, these mAbs such as cetuximab, herceptin, etc., have been used as targeting agents to selectively deliver chemotherapeutics to cancerous cells. However, mechanisms of nanoparticles-mAbs interactions with the target cells and its effect on intracellular trafficking and mechanism are currently unknown. In this paper, we demonstrate that the distinct patterning and dynamics of anti-EGFR (epidermal growth factor receptor) antibody cetuximab (C225)- induced EGFR internalization in pancreatic cancer cells with variable receptor expression is altered upon nanoconjugation. Nanoconjugation uniformly enhanced C225-induced EGFR endocytosis in PANC-1, AsPC-1, and MiaPaca-2 cells, influenced its compartmentalization and regulated the involvement of dynamin-2 in the endocytic processes. Receptor endocytosis and its intracellular trafficking were monitored by confocal microscopy and transmission electron microscopy. The role of dynamin-2 in EGFR endocytosis was determined after overexpressing either wild-type dynamin-2 or mutant dynamin-2 in pancreatic cancer cells followed by tracking the receptor-antibody complex internalization by confocal microscopy. Significantly, these findings demonstrate that the nanoconjugation cannot be construed as an innocuous reaction involved in attaching the targeting agent to the nanoparticle, instead it may distinctly alter the cellular processes at the molecular level, at least antibody induced receptor endocytosis. This information is critical for successful design of a nanoparticle-based targeted drug delivery system for future clinical translation.