Comparison of the Gene Transfer Efficiency of mRNA/GL67 and pDNA/GL67 Complexes in Respiratory Cells

被引：24

作者：

Andries, Oliwia ^{[1
]}

De Filette, Marina ^{[1
]}

Rejman, Joanna ^{[2
]}

De Smedt, Stefaan C. ^{[2
]}

Demeester, Jo ^{[2
]}

Van Poucke, Mario ^{[3
]}

Peelman, Luc ^{[3
]}

Peleman, Cindy ^{[4
]}

Lahoutte, Tony ^{[4
]}

Sanders, Niek N. ^{[1
]}

机构：

[1] Univ Ghent, Fac Vet Med, Dept Nutr Genet & Ethol, Lab Gene Therapy, B-9820 Merelbeke, Belgium

[2] Univ Ghent, Fac Pharmaceut Sci, Ghent Res Grp Nanomed, Lab Gen Biochem & Phys Pharm, B-9000 Ghent, Belgium

[3] Univ Ghent, Fac Vet Med, Dept Nutr Genet & Ethol, Lab Anim Genet, B-9820 Merelbeke, Belgium

[4] UZ Brussel, Lab Vivo Cellular & Mol Imaging ICMI, B-1090 Brussels, Belgium

来源：

MOLECULAR PHARMACEUTICS | 2012年 / 9卷 / 08期

关键词：

mRNA; gene therapy; GL67; lung; nonviral vectors; gene delivery; MESSENGER-RNA; IN-VIVO; MAMMALIAN-CELLS; DELIVERY; DNA; EXPRESSION; STABILITY; RECEPTOR; ACID; LUNG;

D O I：

10.1021/mp200604h

中图分类号：

R-3 [医学研究方法]; R3 [基础医学];

学科分类号：

100103 [病原生物学]; 100218 [急诊医学];

摘要：

Complexes between mRNA and GL67:DOPE:DMPE-PEGS000 (GL67) liposomes were formulated and characterized. Subsequently, the in vitro and in vivo expression characteristics of mRNA/GL67 complexes and pDNA/GL67 complexes, each produced at their optimal ratio, were compared in respiratory cells. Transfection of A549 cells with mRNA/GL67 complexes resulted in a much faster expression than after transfection with pDNA/GL67 complexes. The percentage of GFP-positive cells after mRNA and pDNA transfection peaked after 8 and 24 h, respectively. At these time points the percentage of GFP-positive cells was two times higher after mRNA transfection than after pDNA transfection. Furthermore, the efficacy of mRNA/GL67 complexes was independent of the cell cycle. This was in sharp contrast with pDNA/GL67 complexes that caused only a weak expression in nondividing cells. This confirms that the nuclear barrier is a crucial obstacle for pDNA but not for mRNA. Finally, mRNA/GL67 and pDNA/GL67 complexes encoding luciferase were administered intranasally to the lungs of mice. The mRNA/GL67 complexes did not give rise to a measurable luciferase expression in the murine lungs. In contrast, a detectable bioluminescent signal was present in the lungs of mice that received the pDNA/GL67 complexes. We showed that mRNA/GL67 complexes have a lower stability in biological fluids. Consequently, this may be an explanation for their lower performance in vivo.

引用

页码：2136 / 2145

页数：10

共 45 条

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Stability of mRNA/cationic lipid lipoplexes in human and rat cerebrospinal fluid: Methods and evidence for nonviral mRNA gene delivery to the central nervous system [J].