Project description:We used microarrays to determine if miRNA expression in human airway smooth muscle cells is altered by a pro-inflammatory stimulus. A majority of the miRNAs on the array exhibited very low signal intensity. In ASM cells exposed to IL-1β, TNFα, and IFNγ, none of the miRNA expressed were significantly up-regulated with cytokine treatment. We did observe 11 miRNA down-regulated > 2-fold in both cultures with cytokine treatment. These miRNA include described human miRNA miR-23a, -23b, -25, -188, -320, -363, -489, as well as mouse and rat miRNA homologous to miR-140*, mouse miR-329 and a novel miRNA, abi-13268. miRNA arrays for cytokine-stimulated ASM cells were completed in duplicate from two different cultures to identify candidate miRNA for further study. Cultures were growth arrested for 48 hours and treated with 10 ng/ml IL-1β, TNFα, and IFNγ for 24 hrs. A comparison of miRNA expression under cytokine-stimulated and non-treated conditions from hybridized miRNA arrays was calculated as described.
Project description:Airway smooth muscle cells were stimulated with conditioned medium from BEAS-2B cells in the absence (Control) or Inhibition of miR-210.
Project description:We used microarrays to determine if miRNA expression in human airway smooth muscle cells is altered by a pro-inflammatory stimulus. A majority of the miRNAs on the array exhibited very low signal intensity. In ASM cells exposed to IL-1β, TNFα, and IFNγ, none of the miRNA expressed were significantly up-regulated with cytokine treatment. We did observe 11 miRNA down-regulated > 2-fold in both cultures with cytokine treatment. These miRNA include described human miRNA miR-23a, -23b, -25, -188, -320, -363, -489, as well as mouse and rat miRNA homologous to miR-140*, mouse miR-329 and a novel miRNA, abi-13268.
Project description:Human airway smooth muscle cells were co-cultured with BEAS-2B epithelial cells (or Control). Airway smooth muscle RNA was extracted and sent for Illumina HT-12 micro-array to examine gene expression.
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes