Efficient and non-toxic gene delivery by anionic lipoplexes based on polyprenyl ammonium salts and their effects on cell physiology

Rak, Monika; Ochałek, Anna; Bielecka, Ewa; Latasiewicz, Joanna; Gawarecka, Katarzyna; Sroka, Jolanta; Czyż, Jarosław; Piwowarczyk, Katarzyna; Masnyk, Marek; Chmielewski, Marek; Chojnacki, Tadeusz; Świeżewska, Ewa; Madeja, Zbigniew

doi:10.1002/jgm.2930

Cited by 10 publications

(11 citation statements)

References 64 publications

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“…Recently, PTAI were demonstrated by us to allow effective transfection of plasmid DNA complexes into cells [ 10 , 11 ]. The aim of this study was evaluation of PTAI as an efficient carrier of DNA vaccines.…”

Section: Discussionmentioning

confidence: 99%

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Effective usage of cationic derivatives of polyprenols as carriers of DNA vaccines against influenza virus

Stachyra

Rak

Redkiewicz

et al. 2017

Virol J

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BackgroundCationic derivatives of polyprenols (trimethylpolyprenylammonium iodides – PTAI) with variable chain length between 6 and 15 isoprene units prepared from naturally occurring poly-cis-prenols were tested as DNA vaccine carriers in chickens and mice. This study aimed to investigate if PTAI could be used as an efficient carrier of a DNA vaccine.MethodsSeveral vaccine mixtures were prepared by combining different proportions of the vaccine plasmid (carrying cDNA encoding a vaccine antigen, hemagglutinin from H5N1 influenza virus) and various compositions of PTAI. The vaccines were delivered by intramuscular injection to either chickens or mice. The presence of specific antibodies in sera collected from the immunized animals was analyzed by enzyme-linked immunosorbent assay (ELISA) and hemagglutination inhibition (HI) test.ResultsThe mixtures of PTAI with helper lipids, such as DOPE (1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine), DC-cholesterol [{3ß-[N-(N′,N′-dimethylaminoethane)-carbamoyl] cholesterol} hydrochloride] or DOPC (1,2-dioleoyl-sn-glycero-3-phosphatidylcholine) induced strong humoral response to the antigen encoded by the DNA vaccine plasmid.ConclusionThe animal immunization results confirmed that PTAI compositions, especially mixtures of PTAI with DOPE and DC-cholesterol, do work as effective carriers of DNA vaccines, comparable to the commercially available lipid transfection reagent.

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Section: Discussionmentioning

confidence: 99%

“…1 ). The PTAI were obtained as previously described [ 10 , 11 ], (patent PL 2012, B1 211824, Patent Office of the Republic of Poland). PTAI and helper lipids (DOPE, DC-cholesterol and DOPC were dissolved in 99.8% ethanol).…”

Section: Methodsmentioning

confidence: 99%

Effective usage of cationic derivatives of polyprenols as carriers of DNA vaccines against influenza virus

Stachyra

Rak

Redkiewicz

et al. 2017

Virol J

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“…Among factors regulating the activity of transfection complexes, the size of the lipoplexes has been reported to be a major determinant of transfection efficiency [ 42 , 43 , 44 ]. The morphology and size of the lipoplexes formed by 1,4-DHP amphiphiles C12-Man-Q and D19 were analysed by the transmission electron microscopy (TEM) technique.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro

et al. 2018

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New amphiphilic 1,4-DHP derivative C12-Man-Q with remoted cationic moieties at positions 2 and 6 was synthesised to study DNA delivery activity. The results were compared with data obtained for cationic 1,4-DHP derivative D19, which is known to be the most efficient one among the previously tested 1,4-DHP amphiphiles. We analysed the effects of C12-Man-Q concentration, complexation media, and complex/cell contact time on the gene delivery effectiveness and cell viability. Transmission electron microscopy data confirms that lipoplexes formed by the compound C12-Man-Q were quite uniform, vesicular-like structures with sizes of about 50 nm, and lipoplexes produced by compound D19 were of irregular shapes, varied in size in the range of 25–80 nm. Additionally, confocal microscopy results revealed that both amphiphiles effectively delivered green fluorescent protein expression plasmid into BHK-21 cells and produced a fluorescent signal with satisfactory efficiency, although compound C12-Man-Q was more cytotoxic to the BHK-21 cells with an increase of concentration. It can be concluded that optimal conditions for C12-Man-Q lipoplexes delivery in BHK-21 cells were the serum free media without 0.15 M NaCl, at an N/P ratio of 0.9. Compound D19 showed higher transfection efficiency to transfect BHK-21 and Cos-7 cell lines, when transfecting active proliferating cells. Although D19 was not able to transfect all studied cell lines we propose that it could be cell type specific. The compound C12-Man-Q showed modest delivery activity in all used cell lines, and higher activity was obtained in the case of H2-35 and B16 cells. The transfection efficiency in cell lines MCF-7, HeLa, and Huh-7 appears to be comparable to the reference compound D19 and minimal in the HepG2 cell line.

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“…In our previous studies we have shown that cationic polyisoprenoids are effective components of lipofecting mixtures (Rak et al 2016) and DNA vaccines vehicles (Stachyra et al 2017). In the present study, we have tested PTAI-6-8 and PTAI-10-14 in mixtures with helper lipids in the in vitro model of XC rat Rous sarcoma cell line before using these formulations in vivo in rats.…”

Section: In Vitromentioning

confidence: 92%

“…They are involved in cell response to environmental stress, glycosylation, and prenylation of proteins, and also they intensify the fusion and increase the permeability of model membranes (Swiezewska and Danikiewicz 2005). Recent studies (Gawrys et al 2014b(Gawrys et al , 2018Grecka et al 2016;Rak et al 2016;Stachyra et al 2017) show that cationic derivatives of polyisoprenoid alcohols, called PTAI, are non-toxic and may serve as lipofectants. In the present study, the effectiveness of two PTAI mixtures was investigated for plasmid DNA delivery into rats' cells and to rats and compared to the commercially available reagents for transfection: in vivo-jetPEI® (Polyplus-transfection) and Avalanche®-in vivo Transfection Reagent (EZ Biosystems LLC).…”

Section: Introductionmentioning

confidence: 99%

Polyprenol-Based Lipofecting Agents for In Vivo Delivery of Therapeutic DNA to Treat Hypertensive Rats

et al. 2020

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Development of efficient vectors for transfection is one of the major challenges in genetic engineering. Previous research demonstrated that cationic derivatives of polyisoprenoids (PTAI) may serve as carriers of nucleic acids. In the present study, the effectiveness of two PTAI-based formulations (PTAI-6-8 and 10-14) was investigated and compared to the commercial reagents. The purpose of applied gene therapy was to enhance the expression of vascular endothelial growth factor (VEGF-A) in the renal medulla of spontaneously hypertensive rats (SHR) and to test its potential as a novel antihypertensive intervention. In the first part of the study (in vitro), we confirmed that PTAI-based lipoplexes efficiently transfect XC rat sarcoma cells and are stable in 37 °C for 7 days. In the in vivo experiments, we administered selected lipoplexes directly to the kidneys of conscious SHR (via osmotic pumps). There were no blood pressure changes and VEGF-A level in renal medulla was significantly higher only for PTAI-10-14-based formulation. In conclusion, despite the promising results, we were not able to achieve VEGF-A expression level high enough to verify VEGF-A gene therapy usefulness in SHR. However, results of our study give important indications for the future development of PTAI-based DNA carriers and kidney-targeted gene delivery.

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Efficient and non-toxic gene delivery by anionic lipoplexes based on polyprenyl ammonium salts and their effects on cell physiology

Abstract: The developed carriers constitute a group of promising lipofecting agents of a new type that can be utilized as effective lipofecting agents in vitro and they are also an encouraging basis for in vivo applications.

Cited by 10 publications

References 64 publications

Effective usage of cationic derivatives of polyprenols as carriers of DNA vaccines against influenza virus

Effective usage of cationic derivatives of polyprenols as carriers of DNA vaccines against influenza virus

Synthesis and Comparative Evaluation of Novel Cationic Amphiphile C12-Man-Q as an Efficient DNA Delivery Agent In Vitro

Polyprenol-Based Lipofecting Agents for In Vivo Delivery of Therapeutic DNA to Treat Hypertensive Rats

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