TY - JOUR
T1 - Cytokine-mediated modulation of the hepatic miRNome
T2 - miR-146b-5p is an IL-6-inducible miRNA with multiple targets
AU - Kirchmeyer, Mélanie
AU - Servais, Florence A.
AU - Hamdorf, Matthias
AU - Nazarov, Petr V.
AU - Ginolhac, Aurélien
AU - Halder, Rashi
AU - Vallar, Laurent
AU - Glanemann, Matthias
AU - Rubie, Claudia
AU - Lammert, Frank
AU - Kreis, Stephanie
AU - Behrmann, Iris
N1 - Funding Information:
This work was funded by Fonds National de la Recherche Luxembourg and the Deutsche Forschungsgemeinschaft (C12/BM/3975937, FL/997/7-1, Inter project “HepmiRSTAT”) and by the Internal Research Project «IL6LongLiv» of the University of Luxembourg. We thank Prof. Stefan Rose-John (Christian-Albrechts-University, Kiel, Germany) for providing hyper-IL-6 and we are grateful to Prof. Nobuyuki Kato, Okayama University, Japan, for providing PH5CH8 cells. We obtained expert advice as well as TRAF6 and IRAK1 expression plasmids from Prof. Michael Martin (Giessen University, Germany), and we also thank Prof. Henning Walczak and Dr. Eva Rieser (UCL Cancer Institute, London, UK) for helpful advice. We thank Nathalie Nicot for performing the microarray experiments. The RNA-Seq analysis was carried out using the HPC facilities of the University of Luxembourg.36
Funding Information:
I.B. and S.K. conceived the study, I.B., F.A.S., M.K., M.H., and S.K. designed the research. F.A.S., M.K., M.H. performed experiments. F.A.S. and M.K. analyzed the data. R.H. prepared the library for sequencing. P.V.N. and A.G. did bioinformatics analysis of the microarray and RNA-Seq data, respectively. S.K. and L.V. gave conceptual advice. F.L., M.G., and C.R. provided access to patient samples. F.A.S., M.K., and I.B. wrote the manuscript. All authors revised and approved the final draft of the manuscript. M.K. and F.A.S. contributed equally to this work. This work was funded by Fonds National de la Recherche Luxembourg and the Deutsche Forschungsgemeinschaft (C12/BM/3975937, FL/997/7-1, Inter project ?HepmiRSTAT?) and by the Internal Research Project ?IL6LongLiv? of the University of Luxembourg. We thank Prof. Stefan Rose-John (Christian-Albrechts-University, Kiel, Germany) for providing hyper-IL-6 and we are grateful to Prof. Nobuyuki Kato, Okayama University, Japan, for providing PH5CH8 cells. We obtained expert advice as well as TRAF6 and IRAK1 expression plasmids from Prof. Michael Martin (Giessen University, Germany), and we also thank Prof. Henning Walczak and Dr. Eva Rieser (UCL Cancer Institute, London, UK) for helpful advice. We thank Nathalie Nicot for performing the microarray experiments. The RNA-Seq analysis was carried out using the HPC facilities of the University of Luxembourg. The authors declare no conflicts of interest.
Publisher Copyright:
©2018 The Authors. Society for Leukocyte Biology Published by Wiley Periodicals, Inc.
PY - 2018/11
Y1 - 2018/11
N2 - Interleukin-6 (IL-6)-type cytokines play important roles in liver (patho-)biology. For instance, they regulate the acute phase response to inflammatory signals and are involved in hepatocarcinogenesis. Much is known about the regulation of protein-coding genes by cytokines whereas their effects on the miRNome is less well understood. We performed a microarray screen to identify microRNAs (miRNAs) in human hepatocytes which are modulated by IL-6-type cytokines. Using samples of 2 donors, 27 and 68 miRNAs (out of 1,733) were found to be differentially expressed upon stimulation with hyper-IL-6 (HIL-6) for up to 72 h, with an overlap of 15 commonly regulated miRNAs. qPCR validation revealed that miR-146b-5p was also consistently up-regulated in hepatocytes derived from 2 other donors. Interestingly, miR-146b-5p (but not miR-146a-5p) was induced by IL-6-type cytokines (HIL-6 and OSM) in non-transformed liver-derived PH5CH8 and THLE2 cells and in Huh-7 hepatoma cells, but not in HepG2 or Hep3B hepatoma cells. We did not find evidence for a differential regulation of miR-146b-5p expression by promoter methylation, also when analyzing the TCGA data set on liver cancer samples. Inducible overexpression of miR-146b-5p in PH5CH8 cells followed by RNA-Seq analysis revealed effects on multiple mRNAs, including those encoding IRAK1 and TRAF6 crucial for Toll-like receptor signaling. Indeed, LPS-mediated signaling was attenuated upon overexpression of miR-146b-5p, suggesting a regulatory loop to modulate inflammatory signaling in hepatocytes. Further validation experiments suggest DNAJC6, MAGEE1, MPHOSPH6, PPP2R1B, SLC10A3, SNRNP27, and TIMM17B to be novel targets for miR-146b-5p (and miR-146a-5p).
AB - Interleukin-6 (IL-6)-type cytokines play important roles in liver (patho-)biology. For instance, they regulate the acute phase response to inflammatory signals and are involved in hepatocarcinogenesis. Much is known about the regulation of protein-coding genes by cytokines whereas their effects on the miRNome is less well understood. We performed a microarray screen to identify microRNAs (miRNAs) in human hepatocytes which are modulated by IL-6-type cytokines. Using samples of 2 donors, 27 and 68 miRNAs (out of 1,733) were found to be differentially expressed upon stimulation with hyper-IL-6 (HIL-6) for up to 72 h, with an overlap of 15 commonly regulated miRNAs. qPCR validation revealed that miR-146b-5p was also consistently up-regulated in hepatocytes derived from 2 other donors. Interestingly, miR-146b-5p (but not miR-146a-5p) was induced by IL-6-type cytokines (HIL-6 and OSM) in non-transformed liver-derived PH5CH8 and THLE2 cells and in Huh-7 hepatoma cells, but not in HepG2 or Hep3B hepatoma cells. We did not find evidence for a differential regulation of miR-146b-5p expression by promoter methylation, also when analyzing the TCGA data set on liver cancer samples. Inducible overexpression of miR-146b-5p in PH5CH8 cells followed by RNA-Seq analysis revealed effects on multiple mRNAs, including those encoding IRAK1 and TRAF6 crucial for Toll-like receptor signaling. Indeed, LPS-mediated signaling was attenuated upon overexpression of miR-146b-5p, suggesting a regulatory loop to modulate inflammatory signaling in hepatocytes. Further validation experiments suggest DNAJC6, MAGEE1, MPHOSPH6, PPP2R1B, SLC10A3, SNRNP27, and TIMM17B to be novel targets for miR-146b-5p (and miR-146a-5p).
KW - IL-6-type cytokines
KW - hepatocarcinogenesis
KW - liver
KW - miR-146a-5p
KW - microRNAs
UR - http://www.scopus.com/inward/record.url?scp=85052476544&partnerID=8YFLogxK
U2 - 10.1002/JLB.MA1217-499RR
DO - 10.1002/JLB.MA1217-499RR
M3 - Article
C2 - 30145833
AN - SCOPUS:85052476544
SN - 0741-5400
VL - 104
SP - 987
EP - 1002
JO - Journal of Leukocyte Biology
JF - Journal of Leukocyte Biology
IS - 5
ER -