On the nanotoxicity of PAMAM dendrimers: Superfect® stimulates the EGFR–ERK1/2 signal transduction pathway via an oxidative stress-dependent mechanism in HEK 293 cells

• Published in: International journal of pharmaceutics Vol. 448; no. 1; pp. 239 - 246
• Main Authors: Akhtar, Saghir, Chandrasekhar, Bindu, Attur, Sreeja, Yousif, Mariam H.M., Benter, Ibrahim F.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.05.2013
• Subjects: adverse effects; antagonists; antioxidants; Antioxidants - pharmacology; apoptosis; catalase; cell growth; Cell Survival - drug effects; Dendrimer delivery system; Dendrimers - toxicity; drug delivery systems; drugs; EGFR; epidermal growth factor; Epidermal Growth Factor - pharmacology; epidermal growth factor receptors; ERK1/2; HEK293 Cells; Humans; Hydrogen Peroxide - pharmacology; kidneys; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinase 3 - metabolism; oxidative stress; Oxidative Stress - drug effects; PAMAM; phosphorylation; Receptor, Epidermal Growth Factor - metabolism; RNA interference; RNAi; signal transduction; Signal Transduction - drug effects; Toxicity; transfection; tyrosine; ERK1/2; RNAi; EGF; Toxicity; Dendrimer delivery system; SOD; EGFR; PAMAM
• ISSN: 0378-5173; 1873-3476
• DOI: 10.1016/j.ijpharm.2013.03.039

Polyamidoamine (PAMAM) dendrimers are cationic branch-like macromolecules that may serve as drug delivery systems for gene-based therapies such as RNA interference. For their safe use in the clinic, they should ideally only enhance drug delivery to target tissues and exhibit no adverse effects. However, little is known about their toxicological profiles in terms of their interactions with cellular signal transduction pathways such as the epidermal growth factor receptor (EGFR). The EGFR is an important signaling cascade that regulates cell growth, differentiation, migration, survival and apoptosis. Here, we investigated the impact of naked, unmodified Superfect (SF), a commercially available generation 6 PAMAM dendrimer, on the epidermal growth factor receptor (EGFR) tyrosine kinase–extracellular-regulated kinase 1/2 (ERK1/2) signaling pathway in human embryonic kidney (HEK 293) cells. At concentrations routinely used for transfection, SF exhibited time and dose-dependent stimulation of EGFR and ERK1/2 phosphorylation whereas AG1478, a selective EGFR tyrosine kinase antagonist, inhibited EGFR–ERK1/2 signaling. SF-induced phosphorylation of EGFR for 1h was partly reversible upon removal of the dendrimer and examination of cells 24 later. Co-treatment of SF with epidermal growth factor (EGF) ligand resulted in greater EGFR stimulation than either agent alone implying that the stimulatory effects of SF and the ligand are synergistic. Dendrimer-induced stimulation of EGFR–ERK1/2 signaling could be attenuated by the antioxidants apocynin, catalase and tempol implying that an oxidative stress dependent mechanism was involved. These results show for the first time that PAMAM dendrimers, aside from their ability to improve drug delivery, can modulate the important EGFR–ERK1/2 cellular signal transduction pathway – a novel finding that may have a bearing on their safe application as drug delivery systems.