Differential producibility analysis reveals drug-associated carbon and nitrogen metabolite expressions in Mycobacterium tuberculosis

Mycobacterium tuberculosis (Mtb) is one of the world's successful pathogens that flexibly adapts its metabolic nature during infection of the host, and in response to drugs. Here we used genome scale metabolic modelling coupled with differential producibility analysis (DPA) to translate RNA-seq datasets into metabolite signals and identified drug-associated metabolic response profiles. We tested four tuberculosis (TB) drugs bedaquiline (BDQ), isoniazid (INH), rifampicin (RIF), and clarithromycin (CLA); conducted RNA-seq experiments of Mtb exposed to the individual drugs at subinhibitory concentrations, followed by DPA of gene expression data to map up and downregulated metabolites. Here we highlight those metabolic pathways that were flexibly used by Mtb to tolerate stress generated upon drug exposure. BDQ and INH upregulated maximum number of central carbon metabolites in glycolysis, pentose phosphate pathway and tri-carboxylic acid cycle with concomitant downregulation of lipid and amino acid metabolite classes. Oxaloacetate was significantly upregulated in all four drug-treated Mtb cells highlighting it as an important metabolite in Mtb's metabolism. Amino acid metabolism was selectively induced by different drugs. We have enhanced our knowledge on Mtb's carbon and nitrogen metabolic adaptations in the presence of drugs and identify metabolic nodes for therapeutic development against TB. Our work also provides DPA omics platform to interrogate RNA-seq datasets of any organism that can be reconstructed as a genome scale metabolic network.