Hutchison Pilot Project Summary

The long-term goal of our work is to prioritize environmental reservoirs of pathogenic NTM, to understand NTM-mycobacteriophage relationships in drinking water treatment and distribution, and to utilize NTMspecific mycobacteriophage proteins to produce NTM-specific removal technologies for drinking water treatment. The overall objective of this proposal is to prioritize highly-pathogenic-NTM reservoirs to mine host-specific mycobacteriophages. These mycobacteriophages will be used to develop NTM-specific drinking water treatment technologies. Our focus in this proposal is on Mycobacterium abscessus (MABS). While MABS is the second most common cause of pulmonary infections in the U.S., MAB infections may have higher rates of adverse patient outcomes compared to more common NTM pathogens. The hypothesis of this study is MABS-specific mycobacteriophages sourced from MABS-rich samples can be used to identify novel, phage-based molecular targets for treatment or removal of MABS. The rationale for this project is that the development of MABS-specific mycobacteriophage tools will contribute to quantitative MABS risk assessments and promote alternative MABS treatment strategies. In order to achieve the objectives of this proposal and test our hypothesis, we will prioritize MABS-rich environments based on abundance and genetic similarity to pathogenic MABS strains using high-throughput sequencing data of environmental, human sputum, and clinically isolated samples. The basis of this comparison will use 16S rRNA for generic identification of the microbial community. To achieve a higher resolution of the Mycobacterium population, the RNA polymerase beta-subunit gene, rpoB, will be used. Once environmental MABS reservoirs have been prioritized, highly ranked environments will be mined for MABS-specific bacteriophages. Interactions between these mycobacteriophages and NTM organisms will be screened to identify novel NTM-molecular targets. The corresponding mycobacteriophage protein will be used to develop NTM-drinking water treatment technologies. Results from this study can inform immunocompromised patient behavior to avoid high-risk NTM reservoirs and determine if high-throughput sequencing of rpoB can be applied for species-level resolution of MABS in human sputum. The development of a high-throughput phage/host-protein screening platform would accelerate the identification of phage molecular targets, a potential source for the development of novel NTM antimicrobials.