Non-degradative intracellular trafficking of highly compacted polymeric DNA nanoparticles

作者：Kim Anthony J; Boylan Nicholas J; Suk Jung Soo; Lai Samuel K; Hanes Justin^*

来源：Journal of Controlled Release, 2012, 158(1): 102-107.

DOI：10.1016/j.jconrel.2011.10.031

• Gene therapy
• Nonviral
• Intracellular trafficking
• Particle tracking
• Cystic fibrosis

Highly compacted DNA nanoparticles (DNPs) composed of polyethylene glycol linked to a 30-mer of poly-L-lysine via a single cysteine residue (CK(30)PEG) have previously been shown to provide efficient gene delivery to the brain, eyes and lungs. In this study, we used a combination of flow cytometry, high-resolution live-cell confocal microscopy, and multiple particle tracking ( MPT) to investigate the intracellular trafficking of highly compacted CK(30)PEG DNPs made using two different molecular weights of PEG, CK(30)PEG(10k) and CK(30)PEG(5k). We found that PEG MW did not have a major effect on particle morphology nor nanoparticle intracellular transport. CK(30)PEG(10k) and CK(30)PEG(5k) DNPs both entered human bronchial epithelial (BEAS-2B) cells via a caveolae-mediated pathway, bypassing degradative endolysosomal trafficking. Both nanoparticle formulations were found to rapidly accumulate in the perinuclear region of cells within 2 h, 37 +/- 19% and 47 +/- 8% for CK(30)PEG(10k) and CK(30)PEG(5k), respectively. CK(30)PEG(10k) and CK(30)PEG(5k) DNPs moved within live cells at average velocities of 0.09 +/- 0.04 mu m/s and 0.11 +/- 0.04 mu m/s, respectively, in good agreement with reported values for caveolae. These findings show that highly compacted DNPs employ highly regulated trafficking mechanisms similar to biological pathogens to target specific intracellular compartments.

• 出版日期2012-2-28