In Vitro Investigations on Optimizing and Nebulization of IVT-mRNA Formulations for Potential Pulmonary-Based Alpha-1-Antitrypsin Deficiency Treatment

In vitro-transcribed (IVT) mRNA has come into focus in recent years as a potential therapeutic approach for the treatment of genetic diseases. The nebulized formulations of IVT-mRNA-encoding alpha-1-antitrypsin (A1AT-mRNA) would be a highly acceptable and tolerable remedy for the protein replacement therapy for alpha-1-antitrypsin deficiency in the future. Here we show that lipoplexes containing A1AT-mRNA prepared in optimum conditions could successfully transfect human bronchial epithelial cells without significant toxicity. A reduction in transfection efficiency was observed for aerosolized lipoplexes that can be partially overcome by increasing the initial number of components. A1AT produced from cells transfected by nebulized A1AT-mRNA lipoplexes is functional and could successfully inhibit the enzyme activity of trypsin as well as elastase. Our data indicate that aerosolization of A1AT-mRNA therapy constitutes a potentially powerful means to transfect airway epithelial cells with the purpose of producing functional A1AT, while bringing along the unique advantages of IVT-mRNA.

Automated summary

IVT mRNA has emerged as a potential therapeutic approach for genetic diseases, with nebulized A1AT-mRNA formulations showing promise as a protein replacement therapy for alpha-1-antitrypsin deficiency. Lipoplexes containing A1AT-mRNA can successfully transfect human bronchial epithelial cells without significant toxicity, and increasing the initial number of components can help overcome a reduction in transfection efficiency. The functional A1AT produced from nebulized A1AT-mRNA lipoplexes is capable of inhibiting the enzyme activity of trypsin and elastase, highlighting the potential of aerosolization of A1AT-mRNA therapy for transfecting airway epithelial cells.