Project description:The peritoneal macrophages were infected with Mtb H37Rv for 4 hours, and the miRNA expression profile were analyzed with deep sequencing.
Project description:The goal of this study is to perform transcriptome profiling of both infected and uninfected WT and HIF-1ɑ-/- peritoneal macrophages using RNA sequencing techniques. For that, WT and HIF-1ɑ-/- peritoneal macrophages were infeced for 6 hours with L. donovani promastigotes (or left uninfected as a control) and RNA samples were stored in TRI reagent (Sigma), for further analysis.
Project description:Noncoding RNAs regulate the process of Mycobacterium tuberculosis (M. tb) infecting the host, but there is no simultaneous transcriptional information of long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) and the global regulatory networks of non-coding RNA. Rv1759c, a virulence factor, is a member of protein family containing the proline-glutamic acid (PE) in M. tb, which can increase M. tb survival. To reveal the noncoding RNA regulatory networks and the effect of Rv1759c on non-coding RNA expression during M. tb infection, we collected samples of H37Rv- and H37Rv△1759c-infected macrophages and explored the full transcriptome expression profile. We found 356 mRNAs, 433 lncRNAs, 168 circRNAs, and 12 miRNAs differentially expressed during H37Rv infection, 356 mRNAs, 433 lncRNAs, 168 circRNAs, and 12 miRNAs differentially expressed during H37Rv△1759c infection. We constructed lncRNA/circRNA-miRNA-mRNA regulatory networks during H37Rv and H37Rv△1759c infection. We demonstrated the role of one of the hubs of the networks, hsa-miR-181b-3p, for H37Rv survival in macrophages. We discovered that the expression changes of 68 mRNAs, 92 lncRNAs, 26 circRNAs, and 3 miRNAs were only related to the deletion of Rv1759c by comparing the transcription profiles of H37Rv and H37Rv△1759c. Here, our study comprehensively characterizes the transcriptional profiles in THP1-derived-macrophages infected with H37Rv and H37Rv△1759c, which provides support and new directions for in-depth exploration of noncoding RNA and PE/PPE family functions during the infection process.
Project description:To explore the regulatory network of noncoding RNAs after M.tb infection and the role of Rv1759c in the infection process, we collected samples of H37Rv- and H37Rv△1759c-infected macrophages and explored the full transcriptome expression profile. We constructed DE-lncRNA/DE-circRNA-DE-miRNA-DE-mRNA regulatory networks during H37Rv and H37Rv△1759c infection. In addition, we first discovered the close relationship between Rv1759c and chemokines during M.tb infection by comparing the transcription profiles of H37Rv and H37Rv△1759c and bioinformatics analysis. Here, our study comprehensively characterizes the ncRNA and mRNA profiles in macrophages infected with H37Rv and H37Rv△1759c, which provides support and new directions for in-depth exploration of ncRNA and PE/PPE family functions during the infection process.
Project description:The C57B/6 mice were infected with Mtb H37Rv (CFU=200) for 28 days, and the miRNA expression profile from lung tissues were analyzed with deep sequencing.
Project description:Purpose: The goals of this study are to screen differentially expressed genes in mouse peritoneal macrophages treated by vehicle or R-2-HG and to explore potential mechanisms of how R-2-HG regulating the process of macrophage activation. Methods: After RNA extraction, purification, and library construction, the samples were paired-end (PE) sequenced by Next-Generation Sequencing using an Illumina sequencing platform (Illumina HiSeq 2500). After sequencing, the filtered high-quality sequences (Clean Data) filtered from the original raw data were mapped to the Mus_musculus GRCm38.p4 (mm10) genome. Based on the mapped results, the expression level of each gene was calculated, and the samples of the different groups were subjected to further analysis for differences in expression, enrichment, and cluster analysis. Results: The mRNA expression levels of only a few genes were changed after R-2-HG treatment in mouse peritoneal macrophages, but HIF1A was not included. Conclusions:Regulating the macrophage activation by R-2-HG may not be due to its effect on gene transcription, and R-2-HG doses not affect the mRNA expression of HIF1A.
