On the mechanism whereby cationic lipids promote intracellular delivery of polynucleic acids

Hafez, IM; Maurer, Norbert; Cullis, P. R.

doi:10.1038/sj.gt.3301506

Cited by 523 publications

(381 citation statements)

References 67 publications

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“…4, DOTAP: Chol/DNA lipoplexes were the most efficient to transfect C6 cells, and both primary hippocampal and cortical neurons. This enhancing effect can be attributed to the inclusion of Chol as a helper lipid in liposomes, as already reported by others (Crook et al, 1998;Hafez et al, 2001). Besides improving the stability of liposomes, the presence of Chol reduces the binding of serum proteins to the lipoplexes, thus improving their biological stability (Faneca et al, 2002).…”

Section: Influence Of Liposome Composition On Transfection Activitysupporting

confidence: 60%

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

Cruz

Simões

Lima

2004

Experimental Neurology

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The development of methodologies for gene transfer into the central nervous system is crucial for gene therapy of neurological disorders. In this study, different cationic liposome formulations were used to transfer DNA into C6 glioma cells and primary hippocampal and cortical neurons by varying the nature of the helper lipid (DOPE, Chol) or a mixture of DOPE and cholesterol (Chol) associated to DOTAP. In addition, the effect of the lipid/DNA (+/À) charge ratio, the association of the ligand transferrin to the lipoplexes, and the stage of differentiation of the primary cells on the levels of transfection activity, transfection efficiency, and duration of gene expression were evaluated. Mechanistic studies were also performed to investigate the route of delivery of the complexes into neurons. Our results indicate that DOTAP:Chol (1:1 mol ratio) was the best formulation to transfer a reporter gene into C6 glioma cells, primary hippocampal neurons, and primary cortical neurons. The use of transferrin-associated lipoplexes resulted in a significant enhancement of transfection activity, as compared to plain lipoplexes, which can be partially attributed to the promotion of their internalization mediated by transferrin. While for hippocampal neurons the levels of luciferase gene expression are very low, for primary cortical neurons the levels of transgene expression are high and relatively stable, although only 4% of the cells has been transfected. The stage of cell differentiation revealed to be critical to the levels of gene expression. Consistent with previous findings on the mechanisms of cell internalization, the experiments with inhibitors of the endocytotic pathway clearly indicate that transferrin-associated lipoplexes are internalized into primary neurons by endocytosis. Promising results were obtained in terms of the levels and duration of gene expression, particularly in cortical neurons when transfected with the Tfassociated lipoplexes, this finding suggesting the usefulness of these lipid-based carriers to deliver genes within the CNS. D 2004 Elsevier Inc. All rights reserved.

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Section: Influence Of Liposome Composition On Transfection Activitysupporting

confidence: 60%

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

Cruz

Simões

Lima

2004

Experimental Neurology

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“…Once in the endosome, some lipids may destabilize the endosomal membrane and encourage the release of DNA into the cytosol, thus avoiding the lysosomal degradation pathway. [55][56][57] Such lipoplexes can effectively deliver transgenes to myoblasts in culture and many different types of cells in vivo, including epithelial cells and hepatocytes. [58][59][60][61][62] However, the effectiveness of lipoplexes on gene transfer in muscle in vivo has been disappointing, both as reported by other groups 61 and in our laboratory.…”

Section: Lipids Liposomes and Polymersmentioning

confidence: 99%

Non-viral gene delivery in skeletal muscle: a protein factory

Lu¹,

Bou‐Gharios²,

Partridge³

2003

Gene Ther

170

109

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Ever since the publication of the first reports in 1990 using skeletal muscle as a direct target for expressing foreign transgenes, an avalanche of papers has identified a variety of proteins that can be synthesized and correctly processed by skeletal muscle. The impetus to the development of such applications is not only amelioration of muscle diseases, but also a range of therapeutic applications, from immunization to delivery of therapeutic proteins, such as clotting factors and hormones. Although the most efficient way of introducing transgenes into muscle fibres has been by a variety of recombinant viral vectors, there are potential benefits in the use of non-viral vectors. In this review we assess the recent advances in construction and delivery of naked plasmid DNA to skeletal muscle and highlight the options available for further improvements to raise efficiency to therapeutic levels.

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“…Endosomal escape of lipoplexes is mediated by membrane fusion or disruption of endosomes. 95,96) PEGylation confers steric stability on complexes and inhibits fusion with the endosomal membrane. Therefore, valid strategies for removing the PEG moiety from complexes at the target site in response to the local environment are expected to improve cellular uptake and the subsequent endosomal escape of carriers.…”

Section: -28)mentioning

confidence: 99%

The Polyethyleneglycol Dilemma: Advantage and Disadvantage of PEGylation of Liposomes for Systemic Genes and Nucleic Acids Delivery to Tumors

Hatakeyama

Akita

Harashima

2013

Biological & Pharmaceutical Bulletin

425

276

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Gene and nucleic acid therapy is expected to play a major role in the next generation of agents for cancer treatment. We have recently developed a multifunctional envelope-type nano device (MEND) for use as a novel nonviral gene delivery system. The modification of polyethyleneglycol (PEG), i.e., PEGylation, is a useful method for achieving a longer circulation time for the delivery of MEND to a tumor via the enhanced permeability and retention (EPR) effect. However, PEGylation strongly inhibits cellular uptake and endosomal escape, which results in significant loss of activity of the delivery system. For successful nucleic acid delivery for cancer treatment, the crucial problem associated with the use of PEG, i.e., the "PEG dilemma" must be resolved. In this review, we describe the development and applications of MEND and discuss various strategies for overcoming the PEG dilemma based on the manipulation of both pharmacokinetics and intracellular trafficking of cellular uptake and endosomal release. To increase cellular uptake, target ligands including proteins, peptides, antibodies and aptamers that recognize molecules specifically expressed on tumors are first introduced. Second, cleavable PEG systems are described. The cleavage of PEG from carriers was achieved in response to the intracellular environment as well as the tumor microenvironment, which improvs cellular uptake and endosomal escape. Then, endosomal fusogenic peptides are discussed. Finally, pH-sensitive liposomes using pH-sensitive lipids are described.

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On the mechanism whereby cationic lipids promote intracellular delivery of polynucleic acids

Cited by 523 publications

References 67 publications

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

Improving lipoplex-mediated gene transfer into C6 glioma cells and primary neurons

Non-viral gene delivery in skeletal muscle: a protein factory

The Polyethyleneglycol Dilemma: Advantage and Disadvantage of PEGylation of Liposomes for Systemic Genes and Nucleic Acids Delivery to Tumors

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