Metabolite Dysregulation by Pranlukast in Mycobacterium tuberculosis

Abstract

Mycobacterium tuberculosis has been infecting millions of people worldwide over the years, causing tuberculosis. Drugs targeting distinct cellular mechanisms including synthesis of the cell wall, lipids, proteins, and nucleic acids in Mtb are currently being used for the treatment of TB. Although extensive research is being carried out at the molecular level in the infected host and pathogen, the identification of suitable drug targets and drugs remains under explored. Pranlukast, an allosteric inhibitor of MtArgJ (Mtb ornithine acetyltransferase) has previously been shown to inhibit the survival and virulence of Mtb. The main objective of this study was to identify the altered metabolic pathways and biological processes associated with the differentially expressed metabolites by PRK in Mtb. Here in this study, metabolomics was carried out using an LC-MS/MS-based approach. Collectively, 50 metabolites were identified to be differentially expressed with a significant p-value through a global metabolomic approach using a high-resolution mass spectrometer. Metabolites downstream of argJ were downregulated in the arginine biosynthetic pathway following pranlukast treatment. Predicted human protein interactors of pranlukast-treated Mtb metabolome were identified in association with autophagy, inflammation, DNA repair, and other immune-related processes. Further metabolites including N-acetylglutamate, argininosuccinate, L-arginine, succinate, ergothioneine, and L-phenylalanine were validated by multiple reaction monitoring, a targeted mass spectrometry-based metabolomic approach. This study facilitates the understanding of pranlukast-mediated metabolic changes in Mtb and holds the potential to identify novel therapeutic approaches using metabolic pathways in Mtb.

Rifampicin-Mediated Metabolic Changes in Mycobacterium tuberculosis

“…Mass spectrometry-derived data were analyzed on MZmine (Version 2.53) [43], as described previously [44]. The mzML files, derived from wiff files, were uploaded to MZmine to obtain the RT, m/z, and peak areas of the identified features.…”

Metabolomics meets systems immunology

“…Untargeted metabolomics provides a comprehensive and systematic response to the metabolome, while targeted metabolomics is more focused on the quantitation of specific metabolites. In practical applications, both approaches are often used in combination to complement each other (Asim et al , 2020 ; Thomas et al , 2020 ; Jia et al , 2022 ; Mikaeloff et al , 2022 ; Yelamanchi et al , 2022 ). For example, in the study of Xiao et al ( 2021 ), the combination of untargeted and targeted metabolomics revealed that disturbed metabolic pathways associated with hyperinflammation in severe COVID‐19 could be targeted with metabolic interventions as a possible approach to inhibit SARS‐CoV‐2‐induced cytokine release syndrome.…”

Nitrogen metabolism in mycobacteria: the key genes and targeted antimicrobials

“…Recently, it has been reported that Pranlukast (PRK) can act as an allosteric inhibitor of ArgJ during arginine biosynthesis in M. tuberculosis and inhibit its ornithine acetyltransferase activity, thereby inhibiting bacterial survival and virulence ( Mishra et al, 2019 ) (the drug-target interaction is predicted as shown in Figure 3C ). Further metabolomics studies showed that PRK caused significant differential expression of 50 metabolites ( Yelamanchi et al, 2022 ). This will contribute to a deeper understanding of PRK-mediated metabolic changes and provide a basis for the development of new therapeutic approaches.…”