Efficient in vivo transfection and safety profile of a CpG-free and codon optimized luciferase plasmid using a cationic lipophosphoramidate in a multiple intravenous administration procedure.
As any drug, the success of gene therapy is largely dependent on the vehicle that has to selectively and efficiently deliver therapeutic nucleic acids into targeted cells with minimal side-effects. In the case of chronic diseases that require a life-long treatment, non-viral gene delivery vehicles are less likely to induce an immune response, thereby allowing for repeated administration. Beyond the gene delivery efficiency of a given vector, the nature of nucleic acid constructs also has a central importance in gene therapy protocols. Herein, we investigated the impact of two firefly luciferase encoding plasmids on the transgene expression profile following systemic delivery of lipoplexes in mice, as well as their potential to be safely and efficiently readministered. Whereas pTG11033 plasmid is driven by a strong ubiquitous cytomegalovirus promoter, pGM144 plasmid, which has been designed to avoid inflammation and provide sustained transgene expression in lungs, is CpG-free and is under control of the human elongation factor-1 alpha promoter. Combined to the efficient cationic lipophosphoramidate BSV4, bioluminescence data showed that both plasmids were mostly expressed in the lungs of mice following a primary injection of lipoplexes. However, mice transfected with pGM144 exhibited a higher and more sustained transgene expression than those treated with pTG11033. Repeated administration studies revealed that several injections of lipoplexes could lead to similar transgene expression profiles if an interval of several weeks between subsequent injections was respected. A transient hepatotoxicity and a partial inflammatory response were caused by lipoplex injection, irrespective of the plasmid used. Altogether, these results indicate that repeated systemic administration of lipophosphoramidate-based lipoplexes in mice conducts to an effective lung transfection without serious side effects, and highlight the need to use long-lasting expressing and well tolerated plasmids in order to efficiently renew transgene expression by the successive doses.
Unité INSERM 1078, SFR 148 ScInBioS, Faculté de Médecine, Université de Bretagne Occidentale, Université Européenne de Bretagne, 22 Avenue Camille Desmoulins, 29238 Brest Cedex 2, France. Electronic address: mattiasf.lindberg@gmail.com.
Mattias F
MF
Lindberg
Unité INSERM 1078, SFR 148 ScInBioS, Faculté de Médecine, Université de Bretagne Occidentale, Université Européenne de Bretagne, 22 Avenue Camille Desmoulins, 29238 Brest Cedex 2, France.
Tony
T
Le Gall
Unité INSERM 1078, SFR 148 ScInBioS, Faculté de Médecine, Université de Bretagne Occidentale, Université Européenne de Bretagne, 22 Avenue Camille Desmoulins, 29238 Brest Cedex 2, France; Plateforme SynNanoVect, SFR 148 ScInBioS, Biogenouest, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France.
Nathalie
N
Carmoy
Plateforme SynNanoVect, SFR 148 ScInBioS, Biogenouest, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France; UMR CNRS 6521, SFR 148 ScInBioS, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France.
Mathieu
M
Berchel
Gene Medicine Group, Nuffield Division of Clinical Laboratory Sciences, University of Oxford John Radcliffe Hospital, Oxford, UK.
Stephen C
SC
Hyde
Gene Medicine Group, Nuffield Division of Clinical Laboratory Sciences, University of Oxford John Radcliffe Hospital, Oxford, UK.
Deborah R
DR
Gill
Plateforme SynNanoVect, SFR 148 ScInBioS, Biogenouest, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France; UMR CNRS 6521, SFR 148 ScInBioS, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France.
Paul-Alain
PA
Jaffrès
Unité INSERM 1078, SFR 148 ScInBioS, Faculté de Médecine, Université de Bretagne Occidentale, Université Européenne de Bretagne, 22 Avenue Camille Desmoulins, 29238 Brest Cedex 2, France; Laboratoire de génétique moléculaire et d'histocompatibilité, Hôpital Morvan, CHRU de Brest, 2 Avenue du maréchal Foch, 29609 Brest Cedex, France; DUMG, Faculté de Médecine et des Sciences de la Santé, 22 Avenue Camille Desmoulins, 29238 Brest, France.
Pierre
P
Lehn
Unité INSERM 1078, SFR 148 ScInBioS, Faculté de Médecine, Université de Bretagne Occidentale, Université Européenne de Bretagne, 22 Avenue Camille Desmoulins, 29238 Brest Cedex 2, France; Plateforme SynNanoVect, SFR 148 ScInBioS, Biogenouest, Université de Bretagne Occidentale, Université Européenne de Bretagne, Brest, France; Laboratoire de génétique moléculaire et d'histocompatibilité, Hôpital Morvan, CHRU de Brest, 2 Avenue du maréchal Foch, 29609 Brest Cedex, France; DUMG, Faculté de Médecine et des Sciences de la Santé, 22 Avenue Camille Desmoulins, 29238 Brest, France. Electronic address: tristan.montier@univ-brest.fr.
Tristan
T
Montier
D061605:Administration, Intravenous; D000577:Amides / Q000737:chemistry*; D000818:Animals; D002412:Cations; D002460:Cell Line; D003062:Codon; D018899:CpG Islands; D004247:DNA / Q000008:administration & dosage; D005260:Female; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D010756:Phosphoric Acids / Q000737:chemistry*; D010957:Plasmids; D014162:Transfection
D000577:Amides; D002412:Cations; D003062:Codon; D010756:Phosphoric Acids; D004247:DNA; C011067:phosphoramidic acid; D008156:Luciferases
Cationic lipids; CpG; Inflammation; In vivo bioluminescence; Non-viral gene transfer; Repeated administration; Transfection