Abstract

During the last years the potential role of in vitro transcribed (IVT) mRNA as a vehicle to deliver genetic information has come into focus. IVT mRNA could be used for anti-cancer therapies, vaccination purposes, generation of pluripotent stem cells and also for genome engineering or protein replacement. However, the administration of IVT mRNA into the target organ is still challenging. The lung with its large surface area is not only of interest for delivery of genetic information for treatment of e.g. for cystic fibrosis or alpha-1-antitrypsin deficiency, but also for vaccination purposes. Administration of IVT mRNA to the lung can be performed by direct intratracheal instillation or by aerosol inhalation/nebulisation. The latter approach shows a non-invasive tool, although it is not known, if IVT mRNA is resistant during the process of nebulisation. Therefore, we investigated the transfection efficiency of non-nebulised and nebulised IVT mRNA polyplexes and lipoplexes in human bronchial epithelial cells (16HBE). A slight reduction in transfection efficiency was observed for lipoplexes (Lipofectamine 2000) in the nebulised part compared to the non-nebulised which can be overcome by increasing the amount of Lipofectamine. However, Lipofectamine was more than three times more efficient in transfecting 16HBE than DMRIE and linear PEI performed almost 10 times better than its branched derivative. By contrast, the nebulisation process did not affect the cationic polymer complexes. Furthermore, aerosolisation of IVT mRNA complexes did neither affect the protein duration nor the toxicity of the cationic complexes. Taken together, these data show that aerosolisation of cationic IVT mRNA complexes constitute a potentially powerful means to transfect cells in the lung with the purpose of protein replacement for genetic diseases such as cystic fibrosis or alpha-1-antitrypsin deficiency or for infectious disease vaccines, while bringing along the advantages of IVT mRNA as compared to pDNA as transfection agent.

Highlights

  • The lung with its large surface area is an attractive target organ for the delivery of drugs as it can be approached topical via the airways without bringing along the drawbacks typical for administration via the blood stream

  • Even for plasmid DNA (pDNA) complexed with lipids the aerosolisation is already used in a gene therapy trial for Cystic Fibrosis [2]

  • Our aim was to investigate the administration of in vitro transcribed (IVT) messenger RNA (mRNA) into the lung for therapeutic approaches

Read more

Summary

Introduction

The lung with its large surface area is an attractive target organ for the delivery of drugs as it can be approached topical via the airways without bringing along the drawbacks typical for administration via the blood stream. It allows higher drug concentrations to be deposited intrapulmonary at the site of action. In this context, nucleic acid delivery to this organ presents a promising therapeutic approach to substitute mutated genes causing such diseases as cystic fibrosis or alpha-1-antitrypsin deficiency [1,2,3]. Administration of gene vectors to the lung can be performed by direct intratracheal instillation or by aerosol inhalation/ nebulisation. The latter approach signifies a non-invasive and painless method of pulmonary administration. All studies published up to now employed pDNA and none investigated the potential of in vitro transcribed (IVT) messenger RNA (mRNA) delivery to the lung via aerosolisation

Objectives
Methods
Results
Conclusion
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call