Gene Delivery to Lung Epithelial Cells Using a Cell Penetrating Peptide

Author:

Gene Delivery to Lung Epithelial Cells Using a Cell Penetrating Peptide

Larissa Gomes dos Reis1, Maree Svolos1, Lyn M Moir 1, Rima Jaber2, David Fecher2, Norbert Windhab2, Paul M Young1 & Daniela Traini1

1 Respiratory Technology, Woolcock Institute of Medical Research and Discipline of Pharmacology, Faculty of Medicine and Health, The University of Sydney, NSW, 2037, Australia

2 Evonik Nutrition and Care GmbH, Kirschenallee, 64293, Darmstadt, Germany

Summary

In this study, plasmid DNA (pDNA)-encapsulated nanoparticles (NPs) were manufactured (<200 nm). Addition of a novel proprietary cell penetrating peptide (CPP) to the formulation was essential for efficient internalisation. Cellular uptake in Beas-2B occurred rapidly (less than 3h) and efficiently (83.9% of the cell), with NP-DNA-CPP localised in the nucleus. Low level of protein expression was observed after 96 h of incubation. The NP-DNA-CPP did not affect mitochondrial respiration or membrane integrity. Reactive Oxygen Species (ROS) and Interleukin (IL-8) concentrations were reduced for formulations containing CPP. The increase in necrosis observed for NP-DNA-CPP is likely to be related to the endosomal-escape mechanism that occurs when exposed to these particles. Although further investigation with other lung cells is needed, this system appears to be a safe delivery method for pDNA to the lungs.