Target phenotypic variation to enhance tuberculosis therapy – TREATABLE

TREATABLE project focuses on tuberculosis that remains a major global-health threat, also causing a quarter of the global antimicrobial resistance emergency, and is further exacerbated by the COVID-19 pandemic. The ability of Mycobacterium tuberculosis to persist despite the pressure exerted by the host and antimicrobials is associated with its intrinsic phenotypic variation and its enhanced diversification potential under stressful conditions. Here we propose to target persistence-associated biomarkers, formerly identified at the subpopulation level, aiming to counter this phenomenon, and to potentiate and accelerate the anti-tubercular therapy. To achieve this goal, we established an interdisciplinary consortium, where both academia and private sector will work in synergy. Together we will implement a state-of-the-art research program, merging complementary expertise: bulk- and single-cell microbiology; molecular and cell biology; advanced microfluidic culture-systems engineering; control of microflow dynamics; computer-aided drug design and drug discovery. In particular, we plan to: 1) develop a robust and automated microfluidic system for single-cell imaging, based on an existing prototype; 2) carry out in silico drug discovery using a chemically diverse compound library and focusing on a shortlist of persistence biomarkers; 3) characterize the single-cell activity of our putative anti-persistence compounds, pre-selected at the population level; 4) validate our compound hits and look for synergies with standard anti-tubercular drugs. With TREATABLE, we will increase our fundamental knowledge of the single-cell bases of M. tuberculosis persistence, contribute to a technological breakthrough, which will prove useful to probe single-cell biology in different fields, we will generate new weapons for healthcare intervention and improve tuberculosis therapeutics.