<i>In Vitro </i>Gene Delivery to Hepatocytes with Galactosylated Polyethylenimine

Zanta, Maria-Antonietta; Boussif, Otmane; Adib, and Abdennaji; Behr, Jean‐Paul

doi:10.1021/bc970098f

Cited by 342 publications

(243 citation statements)

References 35 publications

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“…[3][4][5][6][7][8][9][10][11] A branched polymer containing primary, secondary, and tertiary amines, PEI is capable of complexing plasmid DNA and delivering it intracellularly to cells in vitro. The amine groups of PEI are thought to act as proton sponges, preventing endosomal acidification, which results in osmotic swelling and disruption of endosomal membranes.…”

Section: Introductionmentioning

confidence: 99%

Quantitative comparison of polyethylenimine formulations and adenoviral vectors in terms of intracellular gene delivery processes

et al. 2005

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An objective of designing molecular vehicles exhibiting viruslike transgene delivery capabilities but with low toxicity and immunogenicity continues to drive synthetic vector development. As no single step within the gene delivery pathway represents the critical limiting barrier for all vector types under all circumstances, improvements in synthetic vehicle design may be aided by quantitative analysis of the contributions of each step to the overall delivery process. To our knowledge, however, synthetic and viral gene delivery methods have not yet been explicitly compared in terms of these delivery pathway steps in a quantitative manner. As a first address of this challenge, we compare here quantitative parameters characterizing intracellular gene delivery steps for an E1/E3-deleted adenoviral vector and three polyethylenimine (PEI)-based vector formulations, as well as the liposomal transfection reagent Lipofectamine and naked DNA; the cargo is a plasmid encoding the b-galactosidase gene under a CMV promoter, and the cell host is the C3A human hepatocellular carcinoma line. The parameters were determined by applying a previously validated mathematical model to transient time-course measurements of plasmid uptake and trafficking (from whole-cell and isolated nuclei lysates, by real-time quantitative PCR), and gene expression levels, enabling discovery of those for which the adenoviral vector manifested superiority. Parameter-sensitivity analysis permitted identification of processes most critically ratelimiting for each vector. We find that the adenoviral vector advantage in delivery appears to reside partially in its import to the nuclear compartment, but that its vast superiority in transgene expression arises predominantly in our situation from postdelivery events: on the basis of per-nuclear plasmid, expression efficiency from adenovirus is superior by orders of magnitude over the PEI vectors. We find that a chemical modification of a PEI-based vector, which substantially improves its performance, appears to do so by enhancing certain trafficking rate parameters, such as binding and uptake, endosomal escape, and binding to nuclear import machinery, but leaves endosomal escape as a barrier over which transgene delivery could be most sensitively increased further for this polymer.

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Section: Introductionmentioning

confidence: 99%

Quantitative comparison of polyethylenimine formulations and adenoviral vectors in terms of intracellular gene delivery processes

et al. 2005

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“…To address the first challenge, Saravolac et al 5 incorporated amphiphilic polyethylene glycol (PEG) into cationic lipids to shield the charges of lipoplex and to provide a means of steric protection. Others have tried various types of polymers, such poly(L-lysine), [6][7] poly(ethyleneimine), [8][9] poly(methacrylate) 10 and polyamidoamine dendrimers. 11,12 To cope with this problem, we have recently described methods for the polymerization of a novel cationic acrylamide lipid and reconstitution of the resultant poly(cationic lipid) (PCL) to yield stable cationic vesicles.…”

Section: Introductionmentioning

confidence: 99%

Poly(cationic lipid)-mediated in vivo gene delivery to mouse liver

et al. 2003

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We have previously demonstrated that liposomes generated from poly(cationic lipid) (PCL) and cholesterol (Chol) have low cytotoxicity, are serum resistant, and display a transfection efficiency in vitro similar to commercially available cationic liposomes. Our in vivo experiments demonstrated that PCL-Chol liposomes bound much less avidly to serum proteins than did liposomes composed of 1,2-bis(dioleoyloxy)-3-(trimethylamonio)propane (DOTAP)-Chol or DO-TAP-L-a dioleoyl phosphatidylethanolamine (DOPE). Injection of the lipoplexes (PCL-Chol+DNA) through the portal vein after partial hepatectomy (PH) led to much higher reporter gene expression (luciferase) in the liver than did naked DNA injection. Marked green fluorescent protein expression was visualized in almost all hepatocytes in the liver of mice receiving lipoplex injection, even in the absence of PH. Subcutaneous injection of thyroid hormone triiodothyromine (T 3 ) significantly promoted hepatocyte regeneration and markedly enhanced PCL-Chol-mediated gene transfer in mouse liver when the lipoplex was administrated through either portal or tail vein. With T 3 pretreatment, PCL-Chol exerted a better gene transfer efficacy in mouse liver than DOTAP-Chol or DOTAP-DOPE. Two injections of lipoplexes through an indwelling catheter in the portal vein extended the transgene expression at a high level when T 3 injection was repeated. In conclusion, our findings demonstrate that the polymerized cationic liposomes are very stable in the blood and are effective agents for in vivo gene delivery, and that thyroid hormone administration offers a noninvasive approach to enhance liposome-mediated liver gene delivery.

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“…19 Three additional purification steps were carried out by ultrafiltration (Centricon-100 filter devices, Millipore, Bedford, MA, USA).…”

Section: Methodsmentioning

confidence: 99%