title: Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3.
abstract: BACKGROUND
  Latent tuberculosis infection (LTBI) relies on a homeostasis of macrophages and Mycobacterium tuberculosis (Mtb). The small heat shock protein, Mtb Hsp16.3 (also known as latency-associated antigen), plays an important role in Mtb persistence within macrophages. However, the mechanism of LTBI remains elusive. The aim of this study was to delineate LTBI-related miRNA expression in U937 macrophages expressing Mtb Hsp16.3 protein. U937 macrophages were infected with an integrase-deficient Lentivirus vector to transiently express Mtb Hsp16.3, and green fluorescent protein (GFP) as a control. We used a microRNA (miRNA) microarray chip containing more than 1000 probes to identify the significant differentially expressed miRNAs in the infected U937 cells, and employed real-time quantitative polymerase chain reaction (qRT-PCR) for validation. Furthermore, we confirmed these candidate LTBI-related miRNAs in peripheral blood mononuclear cells from subjects with LTBI and in healthy control individuals. Functional annotation prediction of miRNA target genes and pathway enrichment analyses were used to explore the putative links between these miRNAs and LTBI.
  
  
  RESULTS
  Analysis of the miRNA expression profile identified 149 miRNAs that were differentially expressed in U937 macrophages expressing Mtb Hsp16.3 compared with the control expressing GFP. The expression level of seven miRNAs (miR-424-5p, miR-493-5p, miR-296-5p, miR-27b-3p, miR-377-5p, miR-3680-5p, miR-191-5p) were validated by qRT-PCR. The expression level of four miRNAs (miR-424-5p, miR-27b-3p, miR-377-5p, miR-3680-5p) in the peripheral blood mononuclear cells samples from LTBI and healthy participants reflected the altered patterns observed in the microarray profile. The bioinformatic analyses suggest that the miRNAs may regulate Mtb latent infection by affecting the development of macrophage cells.
  
  
  CONCLUSIONS
  The results suggest that miRNA expression may play a considerable role in the pathogenesis of LTBI, and this would increase our understanding of the molecular basis of Hsp16.3-facilitated Mtb survival in macrophages.
authors: Qing-Lin
authors: QL
authors: Meng
authors: Fei
authors: F
authors: Liu
authors: Xing-Yuan
authors: XY
authors: Yang
authors: Xiao-Mei
authors: XM
authors: Liu
authors: Xia
authors: X
authors: Zhang
authors: Suzhou Municipal Key Laboratory of Molecular Diagnostics and Therapeutics, Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou 215163, Jiangsu, China. chunzhang@sibet.ac.cn.
authors: Chun
authors: C
authors: Zhang
authors: Zong-De
authors: ZD
authors: Zhang
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001769:Blood / Q000276:immunology; D002478:Cells, Cultured; D018833:Chaperonins / Q000096:biosynthesis* / Q000235:genetics; D020869:Gene Expression Profiling; D054884:Host-Pathogen Interactions; D006801:Humans; D055985:Latent Tuberculosis / Q000276:immunology* / Q000382:microbiology*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology*; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000276:immunology*
chemical_list: D001426:Bacterial Proteins; C431133:Hsp16.3 protein, Mycobacterium tuberculosis; D035683:MicroRNAs; D018833:Chaperonins