miRNA expression in moncytes in response to estradiol

This profiling experiment evaluates the global changes in miRNA expression in monocytic cells in response to 10 nM Estradiol treatment. Levels of circulating 17β-estradiol (E2) may influence the progression of many human diseases. We hypothesize that E2 modulates the inflammatory response of circulating innate immune cells through microRNA (miRNA)-based modulation of secretory leucoprotease inhibitor (SLPI), a multifunctional antiprotease with immunomodulatory functions. Monocytes were treated with E2 or control, and differentially expressed miRNAs in monocytes were identified using PCR profiling. Cells were transfected with miRNA mimics or antimiRs and SLPI mRNA and protein levels were quantified by qPCR and ELISA, respectively. Luciferase activity assay was performed using SLPI-3’UTR reporter constructs. ChIP was carried out on monocytes treated with E2. SLPI expression is downregulated, and miR-19 is upregulated in response to E2 in monocytes, via increased MIR17HG promoter activity mediated by c-MYC. Transfection of monocytic cells with premiR-19b reduced SLPI expression. This was abrogated using antimiRs against miR-19b. miR-19b directly binds the SLPI 3’UTR as determined by luciferase activity assay. The data show that E2 decreases expression of SLPI in human monocytic cells, via changes in miRNA expression and further highlights the potential for E2 to modulate key components of innate immunity. Modulation of these miRNAs may offer a new therapeutic approach in the treatment of chronic inflammatory diseases. This profiling experiment evaluates the global changes in miRNA expression in monocytic cells in response to 10 nM Estradiol treatment.