Targeted depletion of BTF3a in macrophages activates autophagic pathway to eliminate Mycobacterium tuberculosis

• Published in: Life sciences (1973) 2019-03, Vol.220, p.21-31
• Main Authors: Rawat, Kavita, Das, Swetarka, Vivek Vinod, Budai S., Vekariya, Umeshkumar, Garg, Tanu, Dasgupta, Arunava, Tripathi, Raj Kamal
• Format: Article
• Language: English
• Subjects: Autophagy; Bacillus Calmette-Guerin vaccine; BCG; Biodegradation; BTF3a; Casein; Confocal microscopy; CRISPR; Depletion; gRNA; Immune clearance; Immunoblotting; Intracellular; Lysosomes; Macrophage; Macrophages; Microscopy; Mycobacteria; Mycobacterium tuberculosis; Pathogenesis; Phagocytosis; Phagosomes; Survival; Therapeutic applications; Tuberculosis
• ISSN: 0024-3205; 1879-0631
• DOI: 10.1016/j.lfs.2019.01.035

β casein fragment peptide (54–59) downregulates Basic Transcription factor 3a (BTF3a) in macrophages and exhibits enhanced clearance of M. bovis BCG and several other intracellular pathogens. However, the direct effect of BTF3a downregulation on Mycobacterium tuberculosis (Mtb) survival and the probable pathways involved have not yet been studied. Therefore, the present study was undertaken to deduce the antimycobacterial significance of BTF3a in human macrophages. CRISPR/Cas 9 gRNA was designed to downregulate BTF3a in THP1 derived macrophages. Fold change in BTF3a, p62 and Lamp 1 expression was evaluated through immune blot analysis. CFU assay was done to enumerate the intracellular burden of Mtb H37Rv. LC3B-II turnover and Lamp 1 expression was checked through immunoblotting and also visualized through confocal microscopy. Colocalization of Mtb H37Rv with LC3B, Lysotracker and Rab 7 was visualized through confocal microscopy. The current study identifies BTF3a as a critical host factor assisting intracellular survival of Mtb. In THP1 derived macrophages, infection with Mtb H37Rv resulted in upregulation of BTF3a and targeted depletion of BTF3a resulted in augmented Mtb clearance. Furthermore, BTF3a knockdown demonstrated increased autophagy flux and ameliorated the lysosomal targeting of Mtb containing autophagosomes for lysosomal degradation. Deep understanding of macrophage-Mtb interactions and their roles in the pathogenesis can offer exciting new therapeutic targets for alternative host-specific adjunct therapies in tuberculosis treatment. The present study highlights a novel and significant role of BTF3a in curbing the intracellular survival of Mtb through modulation of autophagy and lysosome biogenesis.