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Comparative transcriptomic analysis of THP‑1 derived macrophages infected with Mycobacterium tuberculosis H37Rv, H37Ra and BCG

ABSTRACT: Comparative transcriptomic analysis of THP‑1 derived macrophages infected with Mycobacterium tuberculosis H37Rv, H37Ra and BCG

PROVIDER: PRJNA682755 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13202556_1.fastq.gz	Fastqsanger.gz
	SRR13202556_2.fastq.gz	Fastqsanger.gz
	SRR13202557_1.fastq.gz	Fastqsanger.gz
	SRR13202557_2.fastq.gz	Fastqsanger.gz
	SRR13202558_1.fastq.gz	Fastqsanger.gz

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Project description:Tuberculosis (TB) remains a worldwide healthcare concern, and the exploration of the host-pathogen interaction is essential to develop therapeutic modalities and strategies to control Mycobacterium tuberculosis (M.tb). In this study, RNA sequencing (transcriptome sequencing) was employed to investigate the global transcriptome changes in the macrophages during the different strains of M.tb infection. THP-1 cells derived from macrophages were exposed to the virulent M.tb strain H37Rv (Rv) or the avirulent M.tb strain H37Ra (Ra), and the M.tb BCG vaccine strain was used as a control. The cDNA libraries were prepared from M.tb-infected macrophages and then sequenced. To assess the transcriptional differences between the expressed genes, the bioinformatics analysis was performed using a standard pipeline of quality control, reference mapping, differential expression analysis, protein-protein interaction (PPI) networks, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Q-PCR and Western blot assays were also performed to validate the data. Our findings indicated that, when compared to BCG or M.tb H37Ra infection, the transcriptome analysis identified 66 differentially expressed genes in the M.tb H37Rv-infected macrophages, out of which 36 genes were up-regulated, and 30 genes were down-regulated. The up-regulated genes were associated with immune response regulation, chemokine secretion, and leucocyte chemotaxis. In contrast, the down-regulated genes were associated with amino acid biosynthetic and energy metabolism, connective tissue development and extracellular matrix organization. The Q-PCR and Western blot assays confirmed increased expression of pro-inflammatory factors, altered energy metabolic processes, enhanced activation of pro-inflammatory signalling pathways and increased pyroptosis in H37Rv-infected macrophage. Overall, our RNA sequencing-based transcriptome study successfully identified a comprehensive, in-depth gene expression/regulation profile in M.tb-infected macrophages. The results demonstrated that virulent M.tb strain H37Rv infection triggers a more severe inflammatory immune response associated with increased tissue damage, which helps in understanding the host-pathogen interaction dynamics and pathogenesis features in different strains of M.tb infection.

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Comparative transcriptomic analysis of THP‑1 derived macrophages infected with Mycobacterium tuberculosis H37Rv, H37Ra and BCG

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