NIX-mediated mitophagy regulate metabolic reprogramming in phagocytic cells during mycobacterial infection.

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Analysis of publicly available transcriptomic data from the NCBI-GEO database indicated that M. bovis (BCG) infection also induces similar rewiring of metabolic and mitophagy pathways. Mtb chronic infection and BCG in-vitro infection both downregulated oxidative phosphorylation and upregulated glycolysis and mitophagy; therefore, we used non-pathogenic mycobacterial species BCG as a model for Mtb infection to gain molecular insights and outcomes of this phenomenon. BCG infection in PBMCs and THP-1 macrophages induce mitophagy and glycolysis, leading to differentiation of naïve macrophage to M1 phenotype. Glucose consumption and lactate production were quantified by NMR, while the mitochondrial mass assessment was performed by mitotracker red uptake assay. Infected macrophages predominantly exhibit M1-phenotype, which is indicated by an increase in M1 specific cytokines (IL-6, TNF-α, and IL-1β) and increased NOS2/ARG1, CD86/CD206 ratio. NIX knockdown abrogates this upregulation of glycolysis, mitophagy, and secretion of pro-inflammatory cytokines in BCG infected cells, indicating that mycobacterial infection-induced immunometabolic changes are executed via NIX mediated mitophagy and are essential for macrophage differentiation and resolution of infection.", "claims": [{"id": "claim-method", "statement": "A two-tailed unpaired t-test was performed on data involving two independent experimental groups, while two-way ANOVA was performed on data involving two or more related experimental groups.", "kind": "methodological", "target": null, "evidence": [{"asset": "source:pmid:33421909", "locator": "sections[14] \"Statistical Analysis\" paragraph 2", "quote": "A two-tailed unpaired t-test was performed on data involving two independent experimental groups, while two-way ANOVA was performed on data involving two or more related experimental groups."}], "state": "complete"}, {"id": "claim-data-access", "statement": "Raw and processed RNA-Seq data files have been submitted to NCBI-GEO. 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