Excess polycation mediates efficient chitosan-based gene transfer by promoting lysosomal release of the polyplexes

Thibault, Marc; Astolfi, Mélina; Tran-Khanh, Nicolas; Lavertu, Marc; Darras, Vincent; Merzouki, Abderrazzak; Buschmann, Michael D.

doi:10.1016/j.biomaterials.2011.03.010

Cited by 80 publications

(61 citation statements)

References 39 publications

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“…That amount of polycation was found to significantly rescue polyplexes from endolysosomal entrapment without causing prohibitive cytotoxicity (data not shown), similarly to previous works. 39 We resorted to that measure in order to compensate for the poor endosomal escape that had been previously associated both with polyplexes at low N/P ratios such as 6:1 39,44 and with dextran-containing nanocarriers. 45 As illustrated on Figure 8, coated polyplexes could attach to and enter cells very efficiently, up to the same performances of core polyplexes.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Tailoring the Surface of a Gene Delivery Vector with Carboxymethylated Dextran: A Systematic Analysis

et al. 2015

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Vous avez des questions? Nous pouvons vous aider. Pour communiquer directement avec un auteur, consultez la première page de la revue dans laquelle son article a été publié afin de trouver ses coordonnées. Si vous n'arrivez pas à les repérer, communiquez avec nous à PublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. Questions? Contact the NRC Publications Archive team atPublicationsArchive-ArchivesPublications@nrc-cnrc.gc.ca. If you wish to email the authors directly, please see the first page of the publication for their contact information. NRC Publications Archive Archives des publications du CNRCThis publication could be one of several versions: author's original, accepted manuscript or the publisher's version. / La version de cette publication peut être l'une des suivantes : la version prépublication de l'auteur, la version acceptée du manuscrit ou la version de l'éditeur. NRC Publications Record / Notice d'Archives des publications de CNRC:http://nparc.cisti-icist.nrc-cnrc.gc.ca/eng/view/object/?id=ab1c9b68-9dc8-474c-a7ca-0ed13201ac54 http://nparc.cisti-icist.nrc-cnrc.gc.ca/fra/voir/objet/?id=ab1c9b68-9dc8-474c-a7ca-0ed13201ac54 ABSTRACT: Polymeric nanocarriers are attractive nonviral vectors for gene delivery purposes in vivo. For such applications, numerous physiological and subcellular bottlenecks have to be overcome. In that endeavor, each structural feature of nanocarriers can be optimized with respect to its corresponding challenges. Here, we focused on the interface between a model gene delivery nanocarrier and relevant constituents of the physiological environment. We screened a library of carboxymethylated dextrans (CMD) for the electrostatic coating of positively charged nanocarriers. We evaluated the jointed influence of the CMD molecular weight and charge density upon nanocarrier coating with respect to DNase, small ions, plasma proteins, red blood cells, and target cells. A total of 4 out of 26 CMD coated nanocarriers successfully passed every screening assay, but did not yield increased reporter gene expression in target cells compared to uncoated nanocarriers. The fine-tuning of CMD for nanocarrier coating yielded a relevant shortlist of candidates that will be further tested in vivo.

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Section: ■ Results and Discussionmentioning

confidence: 99%

Tailoring the Surface of a Gene Delivery Vector with Carboxymethylated Dextran: A Systematic Analysis

et al. 2015

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“…PEI [14][15][16] and chitosan 17 Notes: among the inhibitors, cPZ was used to inhibit cMe; MβcD to inhibit lMe; Fil and gen to inhibit cvMe; and Noc to inhibit microtubule formation. The inhibitors were added to cOs-7 cells at -1 hour and removed at 2 hours posttransfection.…”

Section: Discussionmentioning

confidence: 99%

“…16 Free chitosan chains may mediate enhanced transfection efficiency by promoting the release of polyplexes from the endolysosome. 17 Moreover, excess polycationic chains increase the cytotoxicity 18 and influence the stability of polyplexes. 19,20 With regard to free polycationic chains, two aspects should be considered.…”

Section: Introductionmentioning

confidence: 99%

Elucidating the role of free polycationic chains in polycation gene carriers by free chains of polyethylenimine or N,N,N-trimethyl chitosan plus a certain polyplex

Xu¹,

Liu²,

Wang³

et al. 2014

IJN

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Polycations as gene carriers have attracted considerable attention over the past decade. Generally, polyplexes between polycations and deoxyribonucleic acid (DNA) are formed at low N/P ratios (the ratios of the numbers of nitrogen atoms in a polycation to the numbers of phosphorus atoms in DNA), but high transfection efficiency can only be obtained at much higher N/P ratios. Thus, many polycationic chains are still free in solution. In this study, we investigated the detailed functions of free polyethylenimine chains (PEI-F) and free N,N,N -trimethyl chitosan chains (TMC-F) using the same polyplex, ie, TMC polyplex (TMC-P), which has high stability in Dulbecco’s Modified Eagle’s Medium (DMEM). Meanwhile, PEI polyplex (PEI-P)/PEI-F was also evaluated rather than PEI-P/TMC-F because the stability of PEI-P is low in DMEM and, in the latter case, the TMC-F may replace the bound PEI chain in PEI-P to form TMC-P. The transfection results show that both TMC-F and PEI-F can significantly increase the transfection efficiency of TMC-P; however, PEI-F can upregulate the gene expression of TMC-P more efficiently than TMC-F. Further investigations on the endocytosis and intracellular trafficking show that PEI-P/PEI-F, TMC-P/PEI-F, and TMC-P/TMC-F exhibit similar cellular uptake efficiency. However, by shutting down the clathrin-mediated endocytosis or vacuolar proton pump, the transfection efficiency decreases in the order of PEI-P/PEI-F > TMC-P/PEI-F > TMC-P/TMC-F. These findings indicate that PEI-F and TMC-F may promote the transfection efficiency of the polyplex by affecting its cellular uptake pathway and intracellular trafficking.

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“…We recently found that some of these specific chitosan formulations could achieve a level of transfection in vitro similar to that of commercial phospholipids systems, 26 using HepG2, Caco-2, HT29, LS174T, HeLa (data not shown) and HEK293 cells. 28 We have also used these specific formulations in vivo to deliver fibroblast growth factor-2 and plateletderived growth factor-BB therapeutic proteins or to induce neutralizing antibody production in Balb/c mice depending on formulation specifics. 25 Here we show that injection of specific chitosan/GLP-1TNCs results in an increase of plasma GLP-1 concentration to fivefold higher than that in non-treated groups.…”

Section: Introductionmentioning

confidence: 99%

Effective and safe gene-based delivery of GLP-1 using chitosan/plasmid-DNA therapeutic nanocomplexes in an animal model of type 2 diabetes

et al. 2011

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Glucagon-like peptide-1 (GLP-1) is an incretin hormone that regulates blood glucose level post-prandially. It has been proposed that GLP-1 can be used in type 2 diabetes (T2D) mellitus treatment because of its insulinotropic action. Despite its remarkable advantages, GLP-1 suffers the disadvantage of an extremely short half-life owing to its degradation by the dipeptidyl peptidase IV protease. One way of overcoming this drawback is GLP-1 gene delivery. Here we show effective and safe gene-based delivery of GLP-1 using chitosan/plasmid-DNA therapeutic nanocomplexes (TNCs) in Zucker diabetic fatty (ZDF) animal model of T2D. The expression plasmid fused the GLP-1 gene to a Furin cleavage site was driven by a cytomegalovirus promoter/enhancer. TNCs were prepared by mixing this plasmid with chitosans of specific molecular weight (MW), degree of deacetylation (DDA) and ratio of chitosan amine to DNA phosphate (N:P ratio). Animals injected with the TNC chitosan 92-10-5 (DDA-MW-N:P) showed GLP-1 plasma levels of about fivefold higher than that in non-treated animals and the insulinotropic effect of recombinant GLP-1 was shown by a threefold increase in plasma insulin concentration when compared with untreated animals. Intraperitoneal glucose tolerance tests revealed an efficacious decrease of blood glucose compared with controls for up to 24 days after treatment, where injections of this formulation allowed near-normalization of blood glucose level. TNCs composed of specific chitosans and GLP-1-expressing plasmid constructs showed an impressive ability to harness the profound therapeutic potential of GLP-1 for the treatment of T2D mellitus.

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Excess polycation mediates efficient chitosan-based gene transfer by promoting lysosomal release of the polyplexes

Cited by 80 publications

References 39 publications

Tailoring the Surface of a Gene Delivery Vector with Carboxymethylated Dextran: A Systematic Analysis

Tailoring the Surface of a Gene Delivery Vector with Carboxymethylated Dextran: A Systematic Analysis

Elucidating the role of free polycationic chains in polycation gene carriers by free chains of polyethylenimine or N,N,N-trimethyl chitosan plus a certain polyplex

Effective and safe gene-based delivery of GLP-1 using chitosan/plasmid-DNA therapeutic nanocomplexes in an animal model of type 2 diabetes

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