RNA Sequencing of Mycobacterium tuberculosis-infected macrophages treated with meclizine.

The ability of Mycobacterium tuberculosis (Mtb) to tolerate antibiotics is a major impediment in the treatment of tuberculosis. This antibiotic tolerance is influenced by the host cells in which the bacteria reside, as Mtb senses host cues, and the bacterial response can provide an enhanced ability to withstand anti-TB drugs. Previouslyt, we have shown that Mtb exhibits redox heterogeneity in terms of its mycothiol redox potential (EMSH) giving rise to three broad subpopulations EMSH-reduced, EMSH-basal, and EMSH-oxidized Mtb specifically inside the macrophages. Among these, the EMSH-reduced Mtb exhibit enahnced tolerance to several anti-TB drugs while the EMSH-basal and EMSH-oxidized subpopulations are more susceptible. In this study, we have discovered a novel host-directed therapy molecule- meclizine, which enhances the drug susceptible population and diminishes the tolerant population. Overall design: Bone marrow-derived macrophages were infected with Mtb expressing Mrx1-roGFP2 and were treated with 20uM meclizine or equivalent amount of DMSO (0.2%). At 24h post-infection, infected macrophages were flow-sorted using GFP as a marker for infection and RNA-Seq was performed for these sorted macrophages.