Background
Methods
Cell culture
Infection assay
Exosome isolation and characterization
Exosomal RNA isolation and qualification
Small RNA library construction and miRNA sequencing
Results
Exosome characterization and exosomal RNA preparation
Infection with BCG significantly modulates the miRNA content in infected cell-derived exosomes
Infected exosomal miRNAs | Uninfected exosomal miRNA | |
---|---|---|
Common miRNAs according to expression level | mir-486, mir-21, mir-146a, mir-92a, mir-146b, let-7i, mir-423, mir-378a, let-7f, let-7g, let-7a, mir-155, mir-320a, mir-191, let-7b, mir-24-, mir-26a, mir-423, mir-140, mir-30d, mir-148a, mir-101, mir-221, mir-103a, let-7e, mir-28, mir-1307, mir-151a, mir-148b, mir-26b, mir-27a, mir-532, let-7d, mir-361, mir-99b, mir-342, mir-941, mir-511 | mir-21, mir-146b, mir-146a, let-7f, let-7i, let-7a, mir-378a, let-7g, let-7b, mir-26a, mir-191, mir-423, mir-30d, mir-155, mir-320a, mir-511, mir-423, mir-1307, mir-101, mir-24, mir-92a, mir-148a, mir-99b, mir-532, let-7d, mir-486, let-7e, mir-103a, mir-221, mir-26b, mir-140, mir-148b, mir-28, mir-151a, mir-941, mir-342, mir-27a, mir-361 |
miRNA name | Log Fc | Log CPM | p value | FDR |
---|---|---|---|---|
hsa-miR-96 | 5.89073 | 2.930204 | 0.000245 | 0.000186 |
hsa-miR-1224 | 5.217967 | 3.015217 | 0.000519 | 0.000361 |
hsa-miR-1293 | 3.171918 | 14.19242 | 0.00203 | 0.001093 |
hsa-miR-4467 | 3.174797 | 12.62878 | 0.00204 | 0.001093 |
hsa-miR-6848 | 4.143784 | 3.760948 | 0.00578 | 0.002233 |
hsa-miR-6849 | 3.118348 | 7.30397 | 0.00826 | 0.003 |
hsa-miR-4488 | 3.184877 | 5.073045 | 0.000404 | 0.010436 |
hsa-miR-425 | 3.481839 | 4.249725 | 0.000572 | 0.012611 |
hsa-miR-4732 | 2.578799 | 8.273334 | 0.000616 | 0.013178 |
hsa-miR-484 | 2.389806 | 7.714758 | 0.001566 | 0.028757 |
hsa-miR-5094 | 5.125813 | 1.680093 | 0.002864 | 0.04123 |
Differentially expressed miRNAs were associated with pathways related to pathogenesis of mycobacterial infection and intracellular survival
p value | Number of miRNAs in each pathway | |
---|---|---|
Pathways in bacterial digestion | ||
Proteasome | 0.0139 | 3 |
Ubiquitin-mediated proteolysis | 0.0057 | 7 |
Pathways in infection process | ||
Bacterial invasion of epithelial cells | 0.0310 | 7 |
Endocytosis | 0.0337 | 8 |
ECM-receptor interaction | 0.00153 | 4 |
Adherents junction | 0.00730 | 4 |
Inositol phosphate metabolism | 0.00201 | 4 |
Pathways in fatty acid metabolism | ||
Fatty acid biosynthesis | 0.00000000684 | 3 |
Fatty acid metabolism | 0.00000154 | 5 |
Fatty acid elongation | 0.000738 | 3 |
Steroid biosynthesis | 0.00631 | 3 |
2-Oxocarboxylic acid metabolism | 0.0165 | 3 |
Pathways in amino acids metabolism | ||
Phenylalanine, tyrosine and tryptophan biosynthesis | 0.000713 | 3 |
Valine, leucine and isoleucine degradation | 0.0310 | 6 |
Lysine degradation | 0.0421 | 6 |
Pathways for energy production and sugar metabolism | ||
Central carbon metabolism | 0.00435 | 5 |
2-Oxocarboxylic acid metabolism | 0.0165 | 3 |
Glycosaminoglycan biosynthesis—heparan sulfate/heparin | 0.0113 | 3 |
Proteoglycans | 0.0337 | 8 |
Cell signaling pathways | ||
Phosphatidylinositol signaling system | 0.0489 | 5 |
TGF-beta signaling pathway | 0.0489 | 4 |
Pathways | Gene | p value |
---|---|---|
Pathways in fatty acid metabolism | ||
Fatty acid biosynthesis | 2 | 0.000000000684 |
Fatty acid metabolism | 12 | 0.00000154 |
Fatty acid elongation | 7 | 0.000738 |
Steroid biosynthesis | 6 | 0.00631 |
2-Oxocarboxylic acid metabolism | 6 | 0.0165 |
Pathways in amino acids metabolism | ||
Phenylalanine, tyrosine and tryptophan biosynthesis | 4 | 0.000713 |
Valine, leucine and isoleucine degradation | 12 | 0.0310 |
Lysine degradation | 16 | 0.0421 |
Pathways of energy production and sugar metabolism | ||
Central carbon metabolism in cancer | 19 | 0.00435 |
2-Oxocarboxylic acid metabolism | 6 | 0.0165 |
Glycosaminoglycan biosynthesis—heparan sulfate/heparin | 9 | 0.0113 |
Proteoglycans | 49 | 0.0337 |
Pathways in infection | ||
Bacterial invasion of epithelial cells | 23 | 0.0310 |
Endocytosis | 51 | 0.0337 |
ECM-receptor interaction | 16 | 0.00153 |
Adheres junction | 21 | 0.00730 |
Inositol phosphate metabolism | 15 | 0.0201 |
Cell signaling pathways | ||
Phosphatidylinositol signaling system | 21 | 0.0489 |
TGF-beta signaling pathway | 18 | 0.0489 |
Pathways in bacterial digestion | ||
Proteasome | 3 | 0.0138 |
Ubiquitin mediated proteolysis | 7 | 0.00566 |
Pathways | Differentially expressed miRNA | Number target genes |
---|---|---|
Bacterial invasion pathway | hsa-miR-484 | 15 |
hsa-miR-6848- | 4 | |
hsa-miR-5094 | 1 | |
hsa-miR-425-5p | 9 | |
hsa-miR-1293 | 2 | |
hsa-miR-4488-5p | ||
hsa-miR-1224-5p | 1 | |
hsa-miR-4732- | 1 | |
Fatty acid biosynthesis pathways | hsa-miR-1293 | 1 |
hsa-miR-484 | 1 | |
hsa-miR-425-5p | 1 | |
Fatty acid metabolism | hsa-miR-5094 | 2 |
hsa-miR-425-5p | 4 | |
hsa-miR-1293 | 3 | |
hsa-miR-484 | 4 | |
hsa-miR-6849- | 1 | |
Trp/phe synthesis | hsa-miR-6848- | 1 |
hsa-miR-4467 | 1 | |
hsa-miR-484 | 2 | |
Fatty acid elongation | hsa-miR-425-5p | 4 |
hsa-miR-484 | 2 | |
hsa-miR-5094 | 1 | |
Central carbon metabolism | hsa-miR-1224-5p | 5 |
hsa-miR-484 | 10 | |
Has-mir-96 | ||
hsa-miR-425-5p | 6 | |
hsa-miR-6848- | 1 | |
hsa-miR-6849- | 1 | |
Steroid biosynthesis | hsa-miR-484 | 4 |
hsa-miR-4732- | 1 | |
hsa-miR-425-5p | 3 | |
Glucose amino glycan | hsa-miR-484 | 4 |
hsa-miR-6848- | 2 | |
hsa-miR-5094 | 1 | |
hsa-miR-1293 | 1 | |
hsa-miR-1224-5p | 1 | |
hsa-miR-6849- | 1 | |
2′-Oxycarboxyl | hsa-miR-484 | 4 |
Has-miR-96 | ||
hsa-miR-6848- | 2 | |
hsa-miR-5094 | 1 | |
hsa-miR-1293 | 1 | |
hsa-miR-1224-5p | 1 | |
hsa-miR-6849- | 1 | |
Val/leu/isoleu synthesis | hsa-miR-484 | 5 |
hsa-miR-4732- | 2 | |
hsa-miR-5094 | 3 | |
hsa-miR-425-5p | 1 | |
hsa-miR-4467 | 1 | |
hsa-miR-1293 | 1 | |
Proteoglycan | hsa-miR-5094 | 5 |
hsa-miR-484 | 27 | |
hsa-miR-425-5p | 17 | |
hsa-miR-1293 | 2 | |
hsa-miR-1224-5p | 4 | |
hsa-miR-4732- | 2 | |
hsa-miR-6849- | 3 | |
hsa-miR-6848- | 4 | |
Lysine degradation | hsa-miR-484 | 11 |
hsa-miR-5094 | 3 | |
hsa-miR-1293 | 4 | |
hsa-miR-425-5p | 4 | |
hsa-miR-6848- | 1 | |
hsa-miR-1224-5p | 1 |