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

Olga Gawrys,Monika Rak,Ewa Swiezewska,Marek Masnyk,Sylwia Bobis-Wozowicz,Elzbieta Kompanowska-Jezierska,Karolina Szaro,Elzbieta Karnas,Iwona Baranowska,Zbigniew Madeja,Marek Chmielewski

doi:10.1007/s10528-020-09992-9

Abstract

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.

Highlights

Development of efficient and safe vectors for transfection is one of the major challenges in gene therapy (Ramamoorth and Narvekar 2015)
We aimed at testing if efficiency of PTAI lipofecting mixtures is the same after preparation of lipoplexes in culture medium and 5% glucose as planned for in vivo experiments
The results showed no statistically significant differences between lipoplexes prepared in these two different solutions (Fig. 2a and b). Both compositions showed the same pattern of slightly lower lipofection efficiency in the presence of serum compared to serum-free conditions

Summary

Introduction

Development of efficient and safe vectors for transfection is one of the major challenges in gene therapy (Ramamoorth and Narvekar 2015). Virus-based vectors are considered more effective, but plagued by safety concerns, due to their immunogenicity and cytotoxicity (Jafari et al 2012). Several studies indicate that cationic lipids enhance the efficiency of nucleic acids and drug delivery to the cells (Namiki et al 2011; Jafari et al 2012). Addition of neutral lipids ( called helper or co‐lipids), such as l-α-phosphatidylethanolamine dioleyl (DOPE) or cholesterol, change the properties of lipoplexes making them more suitable for some applications. Despite the fact that numerous cationic and helper lipids have been synthesized (many of them being the subjects of patent applications (Koynova and Tenchov 2011), none of the developed lipofecting agents appear to be perfect. Intensive research has been carried out aiming to elaborate an optimal lipofecting formulation

Methods

Results

Discussion

Conclusion