Delivery and efficacy of synthetic oligonucleotide vectors to an ARDS model by vibrating mesh nebulisation

Acute respiratory distress syndrome (ARDS) continues to have a high mortality with no effective specific therapy. Anti-inflammatory and anti-oxidant therapeutic proteins have previously shown promise in in vivo ARDS models. Similarly, targeting cell permeability (‘leakiness’) associated with ARDS has shown potential. A synthetic mRNA vector for rapid production of protein, coupled with direct delivery to the injured lung via nebulisation, could overcome these limitations. mRNA complexed with a novel transfection reagent (Factor Bioscience Ltd.) encoding green fluorescent protein (GFP), anti-inflammatory and anti-oxidant therapeutic proteins was administered to aseptic or septic rat lungs through a vibrating mesh nebuliser (Aerogen), via the flexiVent (SCIREQ). Animals were harvested and physiological and histological parameters were assessed. An in vitro model of cell permeability employing Calu-3 epithelial cells was also examined using TIE2 and ANGPT1 mRNA targets. A system incorporating Ussing chambers facilitating the measurement of specific ionic transport mechanisms, and by extension fluid dynamics across cell monolayers, was enrolled. GFP mRNA expressed rapidly in lung epithelial cells (