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Uptake and transfection efficiency of PEGylated cationic liposome-DNA complexes with and without RGD-tagging.

Majzoub, Ramsey N; Chan, Chia-Ling; Ewert, Kai K; Silva, Bruno LU ; Liang, Keng S; Jacovetty, Erica L; Carragher, Bridget; Potter, Clinton S and Safinya, Cyrus R (2014) In Biomaterials 35(18). p.4996-5005
Abstract
Steric stabilization of cationic liposome-DNA (CL-DNA) complexes is required for in vivo applications such as gene therapy. PEGylation (PEG: poly(ethylene glycol)) of CL-DNA complexes by addition of PEG2000-lipids yields sterically stabilized nanoparticles but strongly reduces their gene delivery efficacy. PEGylation-induced weakening of the electrostatic binding of CL-DNA nanoparticles to cells (leading to reduced uptake) has been considered as a possible cause, but experimental results have been ambiguous. Using quantitative live-cell imaging in vitro, we have investigated cell attachment and uptake of PEGylated CL-DNA nanoparticles with and without a custom synthesized RGD-peptide grafted to the distal ends of PEG2000-lipids. The... (More)
Steric stabilization of cationic liposome-DNA (CL-DNA) complexes is required for in vivo applications such as gene therapy. PEGylation (PEG: poly(ethylene glycol)) of CL-DNA complexes by addition of PEG2000-lipids yields sterically stabilized nanoparticles but strongly reduces their gene delivery efficacy. PEGylation-induced weakening of the electrostatic binding of CL-DNA nanoparticles to cells (leading to reduced uptake) has been considered as a possible cause, but experimental results have been ambiguous. Using quantitative live-cell imaging in vitro, we have investigated cell attachment and uptake of PEGylated CL-DNA nanoparticles with and without a custom synthesized RGD-peptide grafted to the distal ends of PEG2000-lipids. The RGD-tagged nanoparticles exhibit strongly increased cellular attachment as well as uptake compared to nanoparticles without grafted peptide. Transfection efficiency of RGD-tagged PEGylated CL-DNA NPs increases by about an order of magnitude between NPs with low and high membrane charge density (σM; the average charge per unit area of the membrane; controlled by the molar ratio of cationic to neutral lipid), even though imaging data show that uptake of RGD-tagged particles is only slightly enhanced by high σM. This suggests that endosomal escape and, as a result, transfection efficiency of RGD-tagged NPs is facilitated by high σM. We present a model describing the interactions between PEGylated CL-DNA nanoparticles and the anionic cell membrane which shows how the PEG grafting density and membrane charge density affect adhesion of nanoparticles to the cell surface. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biomaterials
volume
35
issue
18
pages
4996 - 5005
publisher
Elsevier
external identifiers
  • pmid:24661552
  • wos:000335095900013
  • scopus:84897916261
ISSN
1878-5905
DOI
10.1016/j.biomaterials.2014.03.007
language
English
LU publication?
yes
id
3cc9d9a7-8d98-4cb0-91a9-5b5f77661b63 (old id 4379833)
date added to LUP
2014-05-07 16:56:44
date last changed
2017-11-12 03:04:14
@article{3cc9d9a7-8d98-4cb0-91a9-5b5f77661b63,
  abstract     = {Steric stabilization of cationic liposome-DNA (CL-DNA) complexes is required for in vivo applications such as gene therapy. PEGylation (PEG: poly(ethylene glycol)) of CL-DNA complexes by addition of PEG2000-lipids yields sterically stabilized nanoparticles but strongly reduces their gene delivery efficacy. PEGylation-induced weakening of the electrostatic binding of CL-DNA nanoparticles to cells (leading to reduced uptake) has been considered as a possible cause, but experimental results have been ambiguous. Using quantitative live-cell imaging in vitro, we have investigated cell attachment and uptake of PEGylated CL-DNA nanoparticles with and without a custom synthesized RGD-peptide grafted to the distal ends of PEG2000-lipids. The RGD-tagged nanoparticles exhibit strongly increased cellular attachment as well as uptake compared to nanoparticles without grafted peptide. Transfection efficiency of RGD-tagged PEGylated CL-DNA NPs increases by about an order of magnitude between NPs with low and high membrane charge density (σM; the average charge per unit area of the membrane; controlled by the molar ratio of cationic to neutral lipid), even though imaging data show that uptake of RGD-tagged particles is only slightly enhanced by high σM. This suggests that endosomal escape and, as a result, transfection efficiency of RGD-tagged NPs is facilitated by high σM. We present a model describing the interactions between PEGylated CL-DNA nanoparticles and the anionic cell membrane which shows how the PEG grafting density and membrane charge density affect adhesion of nanoparticles to the cell surface.},
  author       = {Majzoub, Ramsey N and Chan, Chia-Ling and Ewert, Kai K and Silva, Bruno and Liang, Keng S and Jacovetty, Erica L and Carragher, Bridget and Potter, Clinton S and Safinya, Cyrus R},
  issn         = {1878-5905},
  language     = {eng},
  number       = {18},
  pages        = {4996--5005},
  publisher    = {Elsevier},
  series       = {Biomaterials},
  title        = {Uptake and transfection efficiency of PEGylated cationic liposome-DNA complexes with and without RGD-tagging.},
  url          = {http://dx.doi.org/10.1016/j.biomaterials.2014.03.007},
  volume       = {35},
  year         = {2014},
}