PROJECT SUMMARY Despite overwhelming evidence demonstrating the importance of microbes to maintain human health, efforts to develop them as therapeutics are impeded because most are currently impossible to manufacture. The long- term goal of this work is to develop self-assembling nanomaterials as a universal “shield” to protect any microbe from processing, transportation, and storage stresses. The proposed coatings have been shown to protect both Gram-positive and Gram-negative strains during freeze-drying and subsequent storage in non-ideal conditions. These results beg the question: how do the coatings protect microbes from environmental stressors, including freeze-drying, UV-light exposure, extreme temperatures, and high humidity? The overall aim of this application is to elucidate the mechanism of microbial protection afforded by these coatings. To accomplish this goal, the physical and chemical properties of coatings with diverse compositions will be established. The ability of these coatings to protect therapeutically-relevant microbial strains from processing, storage, and transport stresses will then be studied. Finally, by combining findings on microbial survival upon exposure to stressors with physicochemical characterization of the nanoscale coatings, an algorithm will be developed to predict optimal coatings for any novel strain of interest without necessitating screening to identify coating compositions with the desired protective properties. This work will provide fundamental insights into the physical properties of these nanoscale coatings and the impact of these properties on microbial viability. This knowledge will enable the production of important but challenging microbial therapeutics with unprecedented ease. Even partial success in this work will yield unprecedented information on how best to protect microbes during manufacture by elucidating the mechanism of protection. This knowledge can directly enable and improve the development of therapeutics. Microbial medicines are especially attractive as treatments for hard-to-treat diseases such as multidrug-resistant infections and chemotherapy-resistant cancers, making any progress in their development highly important. Not only will this technology enable the development of new therapeutics, it will also provide equitable global access to such biologics by enabling their delivery and storage in non-ideal environments.

PROJECT NARRATIVE The proposed work is relevant to public health because it will enable an understanding of the mechanism protection of beneficial microbes with self-assembled coatings during processing and storage. The findings will further facilitate the prediction of optimal coatings for new microbial therapeutics. In turn, this work will support the development of microbes as novel therapeutics, especially for the treatment of antibiotic-resistant infections and chemotherapy-resistance cancers, which will improve patient outcomes and quality of life.