
After intranasal instillation of LNPs encapsulating luciferase reporter mRNA, they primarily observed luciferase expression around the nasal cavity of tested mice, with minor expression in the lungs. investigated in vivo transfection of LNPs to the nasal respiratory epithelia. Specifically, they observed that intravenously administered LNPs with a molar percentage of 50% 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) achieved selective targeting to the lung. showed that adding varying concentrations of permanently cationic or anionic lipids to conventional LNPs resulted in nanoparticles with selective organ targeting (SORT). In particular, lipid nanoparticles (LNPs) for delivery of nucleic acids such as messenger RNA (mRNA) are promising gene therapies, and recent studies have demonstrated that LNPs could be used for pulmonary delivery of mRNA. Polymer and lipid non-viral delivery platforms are promising carriers to protect and encapsulate these larger sized nucleic acid payloads with less immunogenicity compared to viral vectors such as adeno-associated virus. Pulmonary delivery of therapeutics is attractive due to local deposition at high concentrations of medicines directly within the lungs. However, it is a challenge to systemically deliver gene therapies at sufficient therapeutic concentrations at the site of action. Successful delivery of nucleic acids for gene replacement therapy and editing offers the promise of long-term correction and a cure for genetic diseases affecting the lung, such as cystic fibrosis (CF) and alpha-1 antitrypsin deficiency. This study demonstrated that LNPs hold promise to be applied for aerosolization-mediated pulmonary mRNA delivery. It was found that luciferase protein was predominately expressed in the mouse lung for the four lead formulations before and after nebulization. We identified four formulations possessing higher intracellular protein expression ability in vitro even after aerosolization which were then assessed in in vivo studies. An increasing molar ratio of poly-(ethylene) glycol (PEG)-lipid significantly decreased size but also intracellular protein expression of mRNA.

Generally, upon nebulization, LNP formulations showed increased particle size and decreased encapsulation efficiencies.

Lipid nanoparticles (LNPs) showed stable physicochemical properties for at least 14 days of storage at 4 ☌, and most formulations exhibited high encapsulation efficiencies greater than 80%. In this study, we employed a Design of Experiments (DOE) strategy to screen a library of lipid nanoparticle compositions to identify formulations possessing high potency both before and after aerosolization. The development of a stable and efficacious mRNA pulmonary delivery system would enable high therapeutic concentrations locally in the lungs to improve efficacy and limit potential toxicities. Messenger RNA is a class of promising nucleic acid therapeutics to treat a variety of diseases, including genetic diseases.
