Non-degradative intracellular trafficking of highly compacted polymeric DNA nanoparticles

JOURNAL OF CONTROLLED RELEASE

卷:158

Article

Kim, Anthony J.^1,2,3  Boylan, Nicholas J.²  Suk, Jung Soo^1,3,4  Lai, Samuel K.^2,5  Hanes, Justin^1,2,3,4,5,6 

[1] Johns Hopkins Univ, Sch Med, Wilmer Eye Inst, Dept Ophthalmol, Baltimore, MD 21287 USA

[2] Johns Hopkins Univ, Dept Chem & Biomol Engn, Baltimore, MD 21218 USA

[3] Johns Hopkins Univ, Sch Med, Ctr Nanomed, Baltimore, MD 21287 USA

[4] Johns Hopkins Univ, Sch Med, Dept Biomed Engn, Baltimore, MD 21205 USA

[5] Johns Hopkins Univ, Inst NanoBioTechnol, Baltimore, MD 21218 USA

[6] Johns Hopkins Univ, Sch Med, Sidney Kimmel Comprehens Canc Ctr, Dept Oncol, Baltimore, MD 21287 USA

关键词: Gene therapy;    Nonviral;    Intracellular trafficking;    Particle tracking;    Cystic fibrosis;   

DOI  :  10.1016/j.jconrel.2011.10.031

来源: Elsevier

【 摘 要 】

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. (c) 2011 Elsevier B.V. All rights reserved.

【 授权许可】

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