Application of Novel Solid Lipid Nanoparticle (SLN)-Gene Vector Formulations Based on a Dimeric HIV-1 TAT-Peptide in Vitro and in Vivo
Identifieur interne : 002017 ( Main/Exploration ); précédent : 002016; suivant : 002018Application of Novel Solid Lipid Nanoparticle (SLN)-Gene Vector Formulations Based on a Dimeric HIV-1 TAT-Peptide in Vitro and in Vivo
Auteurs : Carsten Rudolph [Allemagne] ; Ulrike Schillinger [Allemagne] ; Aurora Ortiz [Allemagne] ; Kerstin Tabatt [Allemagne] ; Christian Plank [Allemagne] ; Rainer H. Müller [Allemagne] ; Joseph Rosenecker [Allemagne]Source :
- Pharmaceutical Research [ 0724-8741 ] ; 2004-09-01.
English descriptors
Abstract
Abstract: Purpose. To optimize gene delivery of SLN-based gene vectors by incorporation of a dimeric HIV-1 TAT peptide (TAT2) into SLN gene vectors. Methods. Plasmid DNA was complexed with two SLN preparations either with or without pre-compaction of DNA by TAT2, poly-l-arginine, or the mutant TAT2-M1. DNA complexed with polyethylenimine (PEI) served as a standard. Gene expression was analyzed upon transfection of bronchial epithelial cells in vitro and after intratracheal instillation or aerosol application to the lungs of mice in vivo. Stability of DNA was analyzed by agarose gel electrophoresis. Results. Incorporation of TAT2 into SLN gene vectors induced an up to 100-fold sequence-dependent increase of gene expression as compared with the mutant TAT2-M1 and was 4- to 8-times higher as compared with PEI in vitro. In vivo application of TAT2-SLN gene vectors via jet nebulization increased SLN-based gene expression but was accompanied with DNA degradation. DNA degradation was not observed when an innovative device operating on the principle of a perforated vibrating membrane was used. Conclusions. Incorporation of TAT2 into SLN gene vectors is suitable to optimize gene transfer in vitro. The use of a mild nebulization technology avoids DNA degradation and offers the opportunity for further studies in large animal models.
Url:
DOI: 10.1023/B:PHAM.0000041463.56768.ec
Affiliations:
- Allemagne
- Bavière, Berlin, District de Haute-Bavière
- Berlin, Munich
- Université Louis-et-Maximilien de Munich
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To optimize gene delivery of SLN-based gene vectors by incorporation of a dimeric HIV-1 TAT peptide (TAT2) into SLN gene vectors. Methods. Plasmid DNA was complexed with two SLN preparations either with or without pre-compaction of DNA by TAT2, poly-l-arginine, or the mutant TAT2-M1. DNA complexed with polyethylenimine (PEI) served as a standard. Gene expression was analyzed upon transfection of bronchial epithelial cells in vitro and after intratracheal instillation or aerosol application to the lungs of mice in vivo. Stability of DNA was analyzed by agarose gel electrophoresis. Results. Incorporation of TAT2 into SLN gene vectors induced an up to 100-fold sequence-dependent increase of gene expression as compared with the mutant TAT2-M1 and was 4- to 8-times higher as compared with PEI in vitro. In vivo application of TAT2-SLN gene vectors via jet nebulization increased SLN-based gene expression but was accompanied with DNA degradation. DNA degradation was not observed when an innovative device operating on the principle of a perforated vibrating membrane was used. Conclusions. Incorporation of TAT2 into SLN gene vectors is suitable to optimize gene transfer in vitro. The use of a mild nebulization technology avoids DNA degradation and offers the opportunity for further studies in large animal models.</div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>Bavière</li><li>Berlin</li><li>District de Haute-Bavière</li></region><settlement><li>Berlin</li><li>Munich</li></settlement><orgName><li>Université Louis-et-Maximilien de Munich</li></orgName></list><tree><country name="Allemagne"><region name="Bavière"><name sortKey="Rudolph, Carsten" sort="Rudolph, Carsten" uniqKey="Rudolph C" first="Carsten" last="Rudolph">Carsten Rudolph</name></region><name sortKey="Muller, Rainer H" sort="Muller, Rainer H" uniqKey="Muller R" first="Rainer H." last="Müller">Rainer H. 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