Tissue-derived Droplet Micro-Organospheres (TDMO) As Models For High-Throughput Testing of Pathogens

Emerging infectious diseases pose grave threats to warfighters and civilians who travel to locations lacking clean food and water or are exposed to threats of bioweapons. Though many pathogens are relatively known and characterized, many others remain unknown.  This has prompted efforts aimed at developing physiologically-relevant, high-throughput assays that recapitulate host-pathogen interactions. Xilis has developed a microfluidics-based droplet technology that can successfully produce tissue-derived micro-organospheres (TDMO): a high-throughput, miniaturized version of organoids. In the current proposal, we will demonstrate that TDMO better mimic tissue response to pathogenic infections compared to 2D cell lines, show that biological responses to pathogens are maintained in TDMO after cryopreservation, and develop a high-throughput readout for detection of infection status by TDMO carrying fluorescent reporters. By identifying potential reporters capable of detecting infection status for known pathogens in Phase I, we plan to expand the TDMO assay to include reporter TDMO for detection of both gut pathogens and respiratory pathogens in Phase II. In addition, TDMO can also be used as a screening platform for analyzing the human microbiome and drugs that protect host cells from bacterial and viral pathogens.