PMID- 32153563 OWN - NLM STAT- MEDLINE DCOM- 20210308 LR - 20210308 IS - 1664-3224 (Electronic) IS - 1664-3224 (Linking) VI - 11 DP - 2020 TI - Estrogen Receptor, Inflammatory, and FOXO Transcription Factors Regulate Expression of Myasthenia Gravis-Associated Circulating microRNAs. PG - 151 LID - 10.3389/fimmu.2020.00151 [doi] LID - 151 AB - MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate important intracellular biological processes. In myasthenia gravis (MG), a disease-specific pattern of elevated circulating miRNAs has been found, and proposed as potential biomarkers. These elevated miRNAs include miR-150-5p, miR-21-5p, and miR-30e-5p in acetylcholine receptor antibody seropositive (AChR+) MG and miR-151a-3p, miR-423-5p, let-7a-5p, and let-7f-5p in muscle-specific tyrosine kinase antibody seropositive (MuSK+) MG. In this study, we examined the regulation of each of these miRNAs using chromatin immunoprecipitation sequencing (ChIP-seq) data from the Encyclopedia of DNA Elements (ENCODE) to gain insight into the transcription factor pathways that drive their expression in MG. Our aim was to look at the transcription factors that regulate miRNAs and then validate some of those in vivo with cell lines that have sufficient expression of these transcription factors This analysis revealed several transcription factor families that regulate MG-specific miRNAs including the Forkhead box or the FOXO proteins (FoxA1, FoxA2, FoxM1, FoxP2), AP-1, interferon regulatory factors (IRF1, IRF3, IRF4), and signal transducer and activator of transcription proteins (Stat1, Stat3, Stat5a). We also found binding sites for nuclear factor of activated T-cells (NFATC1), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), early growth response factor (EGR1), and the estrogen receptor 1 (ESR1). AChR+ MG miRNAs showed a stronger overall regulation by the FOXO transcription factors, and of this group, miR-21-5p, let-7a, and let 7f were found to possess ESR1 binding sites. Using a murine macrophage cell line, we found activation of NF-kappaB -mediated inflammation by LPS induced expression of miR-21-5p, miR-30e-5p, miR-423-5p, let-7a, and let-7f. Pre-treatment of cells with the anti-inflammatory drugs prednisone or deflazacort attenuated induction of inflammation-induced miRNAs. Interestingly, the activation of inflammation induced packaging of the AChR+-specific miRNAs miR-21-5p and miR-30e-5p into exosomes, suggesting a possible mechanism for the elevation of these miRNAs in MG patient serum. In conclusion, our study summarizes the regulatory transcription factors that drive expression of AChR+ and MuSK+ MG-associated miRNAs. Our findings of elevated miR-21-5p and miR-30e-5p expression in immune cells upon inflammatory stimulation and the suppressive effect of corticosteroids strengthens the putative role of these miRNAs in the MG autoimmune response. CI - Copyright (c) 2020 Fiorillo, Heier, Huang, Tully, Punga and Punga. FAU - Fiorillo, Alyson A AU - Fiorillo AA AD - Center for Genetic Medicine Research, Children's Research Institute, Washington, DC, United States. AD - Genomics & Precision Medicine, The George Washington University, Washington, DC, United States. FAU - Heier, Christopher R AU - Heier CR AD - Center for Genetic Medicine Research, Children's Research Institute, Washington, DC, United States. AD - Genomics & Precision Medicine, The George Washington University, Washington, DC, United States. FAU - Huang, Yu-Fang AU - Huang YF AD - Department of Neuroscience, Clinical Neurophysiology, Uppsala University, Uppsala, Sweden. FAU - Tully, Christopher B AU - Tully CB AD - Center for Genetic Medicine Research, Children's Research Institute, Washington, DC, United States. FAU - Punga, Tanel AU - Punga T AD - Department of Medical Biochemistry and Microbiology, Uppsala University, Uppsala, Sweden. FAU - Punga, Anna Rostedt AU - Punga AR AD - Department of Neuroscience, Clinical Neurophysiology, Uppsala University, Uppsala, Sweden. LA - eng GR - K99 HL130035/HL/NHLBI NIH HHS/United States GR - L40 AR068727/AR/NIAMS NIH HHS/United States GR - L40 AR070539/AR/NIAMS NIH HHS/United States GR - R00 HL130035/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20200221 PL - Switzerland TA - Front Immunol JT - Frontiers in immunology JID - 101560960 RN - 0 (Antibodies) RN - 0 (Circulating MicroRNA) RN - 0 (Forkhead Transcription Factors) RN - 0 (Interferon Regulatory Factors) RN - 0 (RNA, Messenger) RN - 0 (Receptors, Cholinergic) RN - 0 (Receptors, Estrogen) RN - 0 (STAT Transcription Factors) RN - EC 2.7.10.1 (MUSK protein, human) RN - EC 2.7.10.1 (Receptor Protein-Tyrosine Kinases) SB - IM MH - Adult MH - Aged MH - Aged, 80 and over MH - Animals MH - Antibodies/immunology MH - Circulating MicroRNA/*genetics/*metabolism MH - Cohort Studies MH - Female MH - Forkhead Transcription Factors/*metabolism MH - *Gene Expression Regulation MH - Humans MH - Interferon Regulatory Factors/*metabolism MH - Male MH - Mice MH - Middle Aged MH - Myasthenia Gravis/*metabolism MH - RAW 264.7 Cells MH - RNA, Messenger/genetics MH - Receptor Protein-Tyrosine Kinases/immunology MH - Receptors, Cholinergic/immunology MH - Receptors, Estrogen/*metabolism MH - STAT Transcription Factors/*metabolism MH - Signal Transduction/genetics MH - T-Lymphocytes/metabolism PMC - PMC7046803 OTO - NOTNLM OT - FOXO OT - NF-kappaB OT - estradiol OT - miR-21-5p OT - microRNA OT - myasthenia gravis EDAT- 2020/03/11 06:00 MHDA- 2021/03/09 06:00 PMCR- 2020/01/01 CRDT- 2020/03/11 06:00 PHST- 2019/11/09 00:00 [received] PHST- 2020/01/20 00:00 [accepted] PHST- 2020/03/11 06:00 [entrez] PHST- 2020/03/11 06:00 [pubmed] PHST- 2021/03/09 06:00 [medline] PHST- 2020/01/01 00:00 [pmc-release] AID - 10.3389/fimmu.2020.00151 [doi] PST - epublish SO - Front Immunol. 2020 Feb 21;11:151. doi: 10.3389/fimmu.2020.00151. eCollection 2020.