Tf-lipoplexes for neuronal siRNA delivery: A promising system to mediate gene silencing in the CNS

被引:68
作者
Cardoso, A. L. C. [1 ,2 ]
Simoes, S. [2 ,3 ]
de Almeida, L. P. [2 ,3 ]
Plesnila, N. [4 ,5 ]
Pedroso de Lima, M. C. [1 ,2 ]
Wagner, E. [6 ]
Culmsee, C. [6 ]
机构
[1] Univ Coimbra, Fac Sci & Technol, Dept Biochem, P-3001401 Coimbra, Portugal
[2] Univ Coimbra, Ctr Neurosci & Cell Biol, P-3004517 Coimbra, Portugal
[3] Univ Coimbra, Fac Pharm, Pharmaceut Technol Lab, P-3000295 Coimbra, Portugal
[4] Univ Munich, Dept Neurosurg, Med Ctr Grosshadern, Munich, Germany
[5] Univ Munich, Inst Surg Res, Med Ctr Grosshadern, D-8000 Munich, Germany
[6] Univ Munich, Dept Pharm, Munich, Germany
关键词
Cationic liposomes; Transferring; siRNA; Gene silencing; Brain; In vivo;
D O I
10.1016/j.jconrel.2008.08.014
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Although RNAi-based gene silencing holds a great potential for treatment of neurological disorders, its application to the CNS has been restricted by low levels of tissue distribution and cellular uptake. In this work we report that cationic lipid-based vectors can enhance siRNA delivery to neurons both in vitro and in vivo. DOTAP:Chol liposomes associated with transferrin (Tf) and complexed with siRNAs (Tf-lipoplexes) were delivered to primary cultures of luciferase-expressing cortical neurons. Confocal microscopy studies revealed efficient cellular uptake of Cy3-labelled siRNAs after Tf-lipoplex delivery, which was reduced but not completely inhibited by blocking the Tf-receptor with excess Tf. Gene silencing was also evaluated after delivery of anti-luciferase or anti-c-Jun siRNAs. Our results demonstrate that Tf-lipoplexes achieve up to 50% luciferase and c-Jun knockdown, 48 h after transfection, without significant cytotoxicity. Similar results were observed in vivo, where a 40% reduction of luciferase activity was found in the striatum of luciferase mice. In addition, fluorescence microscopy studies showed extensive local distribution and internalization of Tf-lipoplex-associated Cy3-siRNAs without tissue toxicity. Overall, our results demonstrate that Tf-lipoplexes can mediate efficient gene silencing in neuronal cells, both in vitro an in vivo, which may prove useful in therapeutic approaches to neuronal protection and repair. (C) 2008 Elsevier B.V. All rights reserved.
引用
收藏
页码:113 / 123
页数:11
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