Comparison of polymeric siRNA nanocarriers in a murine LPS-activated macrophage cell line: gene silencing, toxicity and off-target gene expression
- Author
- Linda B Jensen, Joscha Griger, Broes Naeye, Amir K Varkouhi, Koen Raemdonck (UGent) , Raymond Schiffelers, Twan Lammers, Gert Storm, Stefaan De Smedt (UGent) , Brian S Sproat, Hanne M Nielsen and Camilla Foged
- Organization
- Abstract
- Tumor necrosis factor alpha (TNF-alpha) plays a key role in the progression of rheumatoid arthritis and is an important target for anti-rheumatic therapies. TNF-alpha expression can be silenced with small interfering RNA (siRNA), but efficacy is dependent on efficient and safe siRNA delivery vehicles. We aimed to identify polymeric nanocarriers for anti-TNF-alpha siRNA with optimal efficacy and minimal off-target effects in vitro. TNF-alpha silencing with polymeric siRNA nanocarriers was compared in lipopolysaccharide-activated RAW 264.7 macrophages by real-time reverse transcription (RT)-PCR. Expression of non-target genes involved in inflammation, apoptosis, and cell cycle progression was determined by RT-PCR, toxicity evaluated by propidium iodide and annexin V staining. PAMAM dendrimers (G4 and G7) and dextran nanogels mediated remarkably high concentration-dependent gene silencing and low toxicity; dioleoyltrimethylammoniumpropane-modified poly(DL-lactide-co-glycolide acid) nanoparticles, thiolated, trimethylated chitosan and poly[(2-hydroxypropyl)methacrylamide 1-methyl-2-piperidine methanol] polyplexes were less efficient transfectants. There were minor changes in the regulation of off-target genes, mainly dependent on nanocarrier and siRNA concentration. Dextran nanogels and PAMAM dendrimers mediated high gene silencing with minor toxicity and off-target transcriptional changes and are therefore expected to be suitable siRNA delivery systems in vivo.
- Keywords
- NECROSIS-FACTOR-ALPHA, RHEUMATOID-ARTHRITIS, EPITHELIAL A549 CELLS, DRUG-DELIVERY SYSTEMS, COLLAGEN-INDUCED ARTHRITIS, BIODEGRADABLE DEXTRAN NANOGELS, SMALL INTERFERING RNA, TNF-alpha, siRNA, polymer, delivery, macrophages, IN-VIVO, DNA COMPLEXES, CANCER-CELLS
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Citation
Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-2137021
- Chicago
- Jensen, Linda B, Joscha Griger, Broes Naeye, Amir K Varkouhi, Koen Raemdonck, Raymond Schiffelers, Twan Lammers, et al. 2012. “Comparison of Polymeric siRNA Nanocarriers in a Murine LPS-activated Macrophage Cell Line: Gene Silencing, Toxicity and Off-target Gene Expression.” Pharmaceutical Research 29 (3): 669–682.
- APA
- Jensen, L. B., Griger, J., Naeye, B., Varkouhi, A. K., Raemdonck, K., Schiffelers, R., Lammers, T., et al. (2012). Comparison of polymeric siRNA nanocarriers in a murine LPS-activated macrophage cell line: gene silencing, toxicity and off-target gene expression. PHARMACEUTICAL RESEARCH, 29(3), 669–682.
- Vancouver
- 1.Jensen LB, Griger J, Naeye B, Varkouhi AK, Raemdonck K, Schiffelers R, et al. Comparison of polymeric siRNA nanocarriers in a murine LPS-activated macrophage cell line: gene silencing, toxicity and off-target gene expression. PHARMACEUTICAL RESEARCH. 2012;29(3):669–82.
- MLA
- Jensen, Linda B, Joscha Griger, Broes Naeye, et al. “Comparison of Polymeric siRNA Nanocarriers in a Murine LPS-activated Macrophage Cell Line: Gene Silencing, Toxicity and Off-target Gene Expression.” PHARMACEUTICAL RESEARCH 29.3 (2012): 669–682. Print.
@article{2137021,
abstract = {Tumor necrosis factor alpha (TNF-alpha) plays a key role in the progression of rheumatoid arthritis and is an important target for anti-rheumatic therapies. TNF-alpha expression can be silenced with small interfering RNA (siRNA), but efficacy is dependent on efficient and safe siRNA delivery vehicles. We aimed to identify polymeric nanocarriers for anti-TNF-alpha siRNA with optimal efficacy and minimal off-target effects in vitro.
TNF-alpha silencing with polymeric siRNA nanocarriers was compared in lipopolysaccharide-activated RAW 264.7 macrophages by real-time reverse transcription (RT)-PCR. Expression of non-target genes involved in inflammation, apoptosis, and cell cycle progression was determined by RT-PCR, toxicity evaluated by propidium iodide and annexin V staining.
PAMAM dendrimers (G4 and G7) and dextran nanogels mediated remarkably high concentration-dependent gene silencing and low toxicity; dioleoyltrimethylammoniumpropane-modified poly(DL-lactide-co-glycolide acid) nanoparticles, thiolated, trimethylated chitosan and poly[(2-hydroxypropyl)methacrylamide 1-methyl-2-piperidine methanol] polyplexes were less efficient transfectants. There were minor changes in the regulation of off-target genes, mainly dependent on nanocarrier and siRNA concentration.
Dextran nanogels and PAMAM dendrimers mediated high gene silencing with minor toxicity and off-target transcriptional changes and are therefore expected to be suitable siRNA delivery systems in vivo.},
author = {Jensen, Linda B and Griger, Joscha and Naeye, Broes and Varkouhi, Amir K and Raemdonck, Koen and Schiffelers, Raymond and Lammers, Twan and Storm, Gert and De Smedt, Stefaan and Sproat, Brian S and Nielsen, Hanne M and Foged, Camilla},
issn = {0724-8741},
journal = {PHARMACEUTICAL RESEARCH},
keyword = {NECROSIS-FACTOR-ALPHA,RHEUMATOID-ARTHRITIS,EPITHELIAL A549 CELLS,DRUG-DELIVERY SYSTEMS,COLLAGEN-INDUCED ARTHRITIS,BIODEGRADABLE DEXTRAN NANOGELS,SMALL INTERFERING RNA,TNF-alpha,siRNA,polymer,delivery,macrophages,IN-VIVO,DNA COMPLEXES,CANCER-CELLS},
language = {eng},
number = {3},
pages = {669--682},
title = {Comparison of polymeric siRNA nanocarriers in a murine LPS-activated macrophage cell line: gene silencing, toxicity and off-target gene expression},
url = {http://dx.doi.org/10.1007/s11095-011-0589-0},
volume = {29},
year = {2012},
}
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