Organotypic lung tissue culture as a preclinical model to study host-influenza A viral infection: A case for repurposing of nafamostat mesylate

Abstract

Reliable and effective models for recapitulation of host-pathogen interactions are imperative for the discovery of potential therapeutics. Ex vivo models can fulfill these requirements as the multicellular native environment in the tissue is preserved and be utilized for toxicology vaccine infection and drug efficacy studies due to the presence of immune cells. Drug repurposing involves the identification of new applications for already approved drugs that are not related to the prime medical indication and emerged as a strategy to cope with slow pace of drug discovery due to high costs and necessary phases to reach the patients. Within the scope of the study broadspectrum serine protease inhibitor nafamostat mesylate was repurposed to inhibit influenza A infection and evaluated by a translational ex vivo organotypic model in which human organ-level responses can be achieved in preclinical safety studies of potential antiviral agents along with in in vitro lung airway culture. The safe doses were determined as 10 mu M for in vitro whereas 22 mu M for ex vivo to be applied for evaluation of host-pathogen interactions which reduced virus infectivity increased cell/tissue viability and protected total protein content by reducing cell death with the inflammatory response. When the gene expression levels of specific proinflammatory anti-inflammatory and cell surface markers involved in antiviral responses were examined the significant inflammatory response represented by highly elevated mRNA gene expression levels of cytokines and chemokines combined with CDH5 downregulated by 5.1-fold supported the antiviral efficacy of NM and usability of ex vivo model as a preclinical infection model.