In vivo gene silencing in solid tumors by targeted electrically mediated siRNA delivery

Golzio, Muriel; Mazzolini, Laurent; Ledoux, Adeline; Paganin, A; Izard, M. E.; Hellaudais, Laëtitia; Bieth, Anne; Pillaire, Marie-Jeanne; Cazaux, Christophe; Hoffmann, Jean-Sébastien; Couderc, Bettina; Teissié, Justin

doi:10.1038/sj.gt.3302920

Cited by 86 publications

(66 citation statements)

References 45 publications

(69 reference statements)

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“…Moreover, EP has proven to be an efficient method for in vitro and in vivo delivery of siRNA/shRNA molecules into tumor cells resulting in successful silencing of reporter or therapeutic genes, including K-ras. 11,[33][34][35][36] Previous in vivo study performed on pancreatic tumors showed that electrotransfection of subcutaneous or orthotopic pancreatic tumors (Panc-1) with synthetic siRNA or plasmid DNA encoding shRNA directed against K-ras dramatically reduced the tumor growth. 11 Results obtained from pancreatic tumor model and our results are consistent with the observation that a single electrotransfection of colorectal tumors with pmiRNA-Kras resulted in growth delay of 8 days, whereas in the study by Rejiba et al 11 the growth delay of 15 days was achieved after two consecutive electrotransfections of pancreatic tumors with pshRNA-K-ras (time interval: 7 days).…”

Section: Discussionmentioning

confidence: 99%

“…Results of previous in vitro and in vivo studies targeting reporter gene GFP with siRNA/shRNA showed that the highest gene silencing is obtained between the second and the fifth day after electrotransfection. [33][34][35] In our in vitro and in vivo experiments, it was demonstrated that not only the miRNA-K-ras contributed to the reduction in cell and tumor growth, but also the apparent toxicity of plasmid DNA. This was observed from the reduced cell survival and from growth of tumors after electrotransfection with control plasmid DNA (pmiRNA-ctrl).…”

Section: Discussionmentioning

confidence: 99%

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MicroRNAs targeting mutant K-ras by electrotransfer inhibit human colorectal adenocarcinoma cell growth in vitro and in vivo

et al. 2010

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Mutations of K-ras have been found in 30-60% of colorectal carcinomas and are believed to be associated with tumor initiation, tumor progression and metastasis formation. Therefore, silencing of mutant K-ras expression has become an attractive therapeutic strategy for colorectal cancer treatment. The aim of our study was to investigate the effect of microRNA (miRNA) molecules directed against K-ras (miRNA-K-ras) on K-ras expression level and the growth of colorectal carcinoma cell line LoVo in vitro and in vivo. In addition, we evaluated electroporation as a gene delivery method for transfection of LoVo cells and tumors with plasmid DNA encoding miRNA-K-ras (pmiRNA-K-ras). Results of our study indicated that miRNAs targeting K-ras efficiently reduced K-ras expression and cell survival after in vitro electrotransfection of LoVo cells with pmiRNA-K-ras. In vivo, electroporation has proven to be a simple and efficient delivery method for local administration of pmiRNA-K-ras molecules into LoVo tumors. This therapy shows pronounced antitumor effectiveness and has no side effects. The obtained results demonstrate that electrogene therapy with miRNA-K-ras molecules can be potential therapeutic strategy for treatment of colorectal cancers harboring K-ras mutations.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

MicroRNAs targeting mutant K-ras by electrotransfer inhibit human colorectal adenocarcinoma cell growth in vitro and in vivo

et al. 2010

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“…Strategies that have been considered for in vivo delivery of synthetic siRNA in laboratory animals are hydrodynamic injection of naked siRNA [3] or siRNA conjugates [4], electroporation [5][6][7][8] and the use of cationic carriers [9][10][11][12][13][14]. However, several aspects limit the applicability of these methods in humans.…”

Section: Introductionmentioning

confidence: 99%

Ultrasound assisted siRNA delivery using PEG-siPlex loaded microbubbles

Vandenbroucke

Lentacker

Demeester

et al. 2008

Journal of Controlled Release

109

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Short interfering RNA (siRNA) attracts much attention for the treatment of various diseases. However, its delivery, especially via systemic routes, remains a challenge. Indeed, naked siRNAs are rapidly degraded, while complexed siRNAs massively aggregate in the blood or are captured by macrophages. Although this can be circumvented by PEGylation, we found that PEGylation had a strong negative effect on the gene silencing efficiency of siRNA-liposome complexes (siPlexes). Recently, ultrasound combined with microbubbles has been used to deliver naked siRNA but the gene silencing efficiency is rather low and very high amounts of siRNA are required. To overcome the negative effects of PEGylation and to enhance the efficiency of ultrasound assisted siRNA delivery, we coupled PEGylated siPlexes (PEG-siPlexes) to microbubbles. Ultrasound radiation of these microbubbles resulted in massive release of unaltered PEG-siPlexes. Interestingly, PEG-siPlexes loaded on microbubbles were able to enter cells after exposure to ultrasound, in contrast to free PEG-siPlexes, which were not able to enter cells rapidly. Furthermore, these PEG-siPlex loaded microbubbles induced, in the presence of ultrasound, much higher gene silencing than free PEG-siPlexes. Additionally, the PEG-siPlex loaded microbubbles only silenced the expression of genes in the presence of ultrasound, which allows space and time controlled gene silencing.

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“…6 It has been used in the laboratory for gene delivery to cells in vitro or in vivo, and is recently receiving much attention in cancer gene therapy as a clinically applicable method for enhancing gene delivery, based on it being carried out safely and easily with low cost. [7][8][9] In this method, plasmid DNA is injected into target tissue after which a series of electric pulses that induce temporary and reversible breakdown of cell membrane and pore formation is conducted to augment DNA transport into the cells. It has been applied to not only accessible surface tissue but also to deep tissues.…”

Section: Introductionmentioning

confidence: 99%

Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging

Aung¹,

Hasegawa²,

Koshikawa-Yano³

et al. 2009

Gene Ther

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In vivo electroporation (EP) is an efficient method for effective gene transfer and is highly expected for application in anticancer gene therapy. Non-invasive monitoring of gene transfer/expression is critical for optimal gene therapy. Here we report in vivo optical and high-field magnetic resonance imaging (MRI) of EP-mediated transgene expression in a tumor model. Initially, we observed spatio-temporal change in in vivo EP-mediated transgene expression by optical imaging using red fluorescence protein (RFP) as a reporter gene. Next, we constructed a dual-reporter plasmid carrying a gene-encoding MRI reporter ferritin heavy chain and RFP gene to visualize the intratumoral transgene expression by dual modality. Cells transfected with this plasmid showed lower signal intensity on in vitro T 2 -weighted cellular MRI and quantitatively increased the transverse relaxation rate (1/T 2 ) compared with control cells. After conducting in vivo EP in an experimental tumor, the plasmidinjected region showed both fluorescent emissions in optical imaging and detectably lowered signal on T 2 -weighted MRI. The correlative immunohistological findings confirmed that both the reporter transgenes were co-expressed in this region. Thus, our strategy provides a platform for evaluating EP-mediated cancer gene therapy easily and safely without administering contrast agent or substrate.

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In vivo gene silencing in solid tumors by targeted electrically mediated siRNA delivery

Cited by 86 publications

References 45 publications

MicroRNAs targeting mutant K-ras by electrotransfer inhibit human colorectal adenocarcinoma cell growth in vitro and in vivo

MicroRNAs targeting mutant K-ras by electrotransfer inhibit human colorectal adenocarcinoma cell growth in vitro and in vivo

Ultrasound assisted siRNA delivery using PEG-siPlex loaded microbubbles

Visualization of in vivo electroporation-mediated transgene expression in experimental tumors by optical and magnetic resonance imaging

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