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MicroRNA-132 targets HB-EGF upon IgE-mediated activation in murine and human mast cells

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Abstract

MicroRNAs provide an additional layer in the regulation of gene expression acting as repressors with several targets at the posttranscriptional level. This study describes microRNA expression patterns during differentiation and activation of mast cells. The expression levels of 567 different mouse miRNAs were compared by microarray between c-Kit+ committed progenitors, mucosal mast cells, resting and IgE-crosslinked BMMCs in vitro. The strongest upregulation of miR-132 upon IgE-mediated activation was validated in human cord blood-derived mast cells as well. HB-EGF growth factor also upregulated upon activation and was ranked high by more prediction algorithms. Co-transfection of miR-132 mimicking precursor and the 3′UTR of human Hbegf-containing luciferase vector proves that the predicted binding site is functional. In line with this, neutralization of miR-132 by anti-miR inhibitor leads to sustained production of HB-EGF protein in activated mast cells. Our data provide a novel example for negative regulation of a growth factor by an upregulated miRNA.

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Abbreviations

3′-UTR:

3′-Untranslated region

BMMC:

Bone marrow-derived mast cell

CREB:

Cyclic AMP-response-element-binding protein

CTMC:

Connective tissue-type mast cells

DNP-HSA:

Dinitrophenyl-human serum albumin

FAM:

Fluorescein amidite

FBS:

Fetal bovine serum

FGF2:

Fibroblast growth factor 2

FYN:

Oncogene related to SRC, FGR, YES

HB-EGF:

Heparin-binding EGF-like growth factor

HER2:

Human epidermal growth factor receptor 2

IL-3:

Interleukin-3

LYN:

v-yes-1 Yamaguchi sarcoma viral related oncogene homolog

MeCP2:

Methyl CpG-binding domain protein-2

MMC:

Mucosal mast cells

mMcpt1:

Mouse Mast Cell Protease-1

NGF:

Nerve growth factor

PLC-γ:

Phospholipase C-γ

SCF:

Stem cell factor

SYK:

Spleen tyrosine kinase

TGF-β:

Transforming growth factor beta

VEGF:

Vascular endothelial growth factor

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Acknowledgments

This work is supported by the National Research Foundation of the Hungarian Academy of Sciences (grant OTKA 67955).

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Authors and Affiliations

  1. Department of Genetics, Cell and Immunobiology, Semmelweis University, Budapest, Hungary

    Viktor Molnár, Barbara Érsek, Zoltán Wiener & András Falus

  2. 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary

    Zsófia Tömböl, Péter M. Szabó & Péter Igaz

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  1. Viktor Molnár
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  2. Barbara Érsek
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Correspondence to Viktor Molnár.

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Molnár, V., Érsek, B., Wiener, Z. et al. MicroRNA-132 targets HB-EGF upon IgE-mediated activation in murine and human mast cells. Cell. Mol. Life Sci. 69, 793–808 (2012). https://doi.org/10.1007/s00018-011-0786-3

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