Modeling synergistic drug inhibition of Mycobacterium tuberculosisgrowth in murine macrophages

Abstract

We developed a metabolism-based systems biology framework to model drug-induced growth inhibition of Mycobacterium tuberculosis in murine macrophage cells. We used it to simulate ex vivo bacterial growth inhibition due to 3-nitropropionate (3-NP) and calculated the corresponding time- and drug concentration-dependent dose-response curves. 3-NP targets the isocitrate lyase 1 (ICL1) and ICL2 enzymes in the glyoxylate shunt, an essential component in carbon metabolism of many important prokaryotic organisms. We used the framework to in silico mimic drugging additional enzymes in combination with 3-NP to understand how synergy can arise among metabolic enzyme targets. In particular, we focused on exploring additional targets among the central carbon metabolism pathways and ascertaining the impact of jointly inhibiting these targets and the ICL1/ICL2 enzymes. Thus, additionally inhibiting the malate synthase (MS) enzyme in the glyoxylate shunt did not produce synergistic effects, whereas additional inhibition of the glycerol-3-phosphate dehydrogenase (G3PD) enzyme showed a reduction in bacterial growth beyond what each single inhibition could achieve. Whereas the ICL1/ICL2-MS pair essentially works on the same branch of the metabolic pathway processing lipids as carbon sources (the glyoxylate shunt), the ICL1/ICL2-G3PD pair inhibition targets different branches among the lipid utilization pathways. This allowed the ICL1/ICL2-G3PD drug combination to synergistically inhibit carbon processing and ultimately affect cellular growth. Our previously developed model for in vitro conditions failed to capture these effects, highlighting the importance of constructing accurate representations of the experimental ex vivo macrophage system.