Abstract
In spite of the extensive potential of human mesenchymal stem cells (hMSCs) in cell therapy, little is known about the molecular mechanisms that regulate their therapeutic properties. We aimed to identify microRNAs (miRNAs) involved in controlling the transition between the resting and reparative phenotypes of hMSCs, hypothesizing that these miRNAs must be present in the undifferentiated cells and downregulated to allow initiation of distinct activation/differentiation programs. Differential miRNA expression analyses revealed that miR-335 is significantly downregulated upon hMSC differentiation. In addition, hMSCs derived from a variety of tissues express miR-335 at a higher level than human skin fibroblasts, and overexpression of miR-335 in hMSCs inhibited their proliferation and migration, as well as their osteogenic and adipogenic potential. Expression of miR-335 in hMSCs was upregulated by the canonical Wnt signaling pathway, a positive regulator of MSC self-renewal, and downregulated by interferon-γ (IFN-γ), a pro-inflammatory cytokine that has an important role in activating the immunomodulatory properties of hMSCs. Differential gene expression analyses, in combination with computational searches, defined a cluster of 62 putative target genes for miR-335 in hMSCs. Western blot and 3′UTR reporter assays confirmed RUNX2 as a direct target of miR-335 in hMSCs. These results strongly suggest that miR-335 downregulation is critical for the acquisition of reparative MSC phenotypes.
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Accession codes
Abbreviations
- hMSC:
-
human mesenchymal stem cells
- fdr:
-
false discovery rate
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Acknowledgements
We thank S Calleja and R Alvarez (CNIC) for technical help with microarrays and real-time RT-PCR; P Fernández and JC Ramírez (CNIC) for lentivirus production; F Cabo (CNIC) for bioinformatics and statistical support; C Carreiro and A Esteban (CNIC) for preparation and maintenance of plasmid stocks; M García-Arranz (Hospital Univesitario La Paz, Madrid, Spain), C Trigueros, A Izeta and A García (F Inbiomed, San Sebastián, Spain) for biological samples; S Bartlett (CNIC) for English revision and M Ramón for secretarial support. This work was supported by grants to AB from the Spanish Plan Nacional de Salud y Farmacia/CICYT (SAF 2008-02099), Comunidad Autónoma de Madrid (P-BIO-0306-2006) and Red de Terapia Celular del Instituto de Salud Carlos III (TerCel); MAG is supported by the ‘Miguel Servet’ Program (CP07/00306) of the Instituto de Salud Carlos III (Ministerio de Ciencia e Innovación, Spain); JCS is supported by the ACI-PLAN-E Program (Ministerio de Ciencia e Innovación, Spain). The CNIC is supported by the Spanish Ministry of Science and Innovation and the Pro-CNIC Foundation.
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Tomé, M., López-Romero, P., Albo, C. et al. miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells. Cell Death Differ 18, 985–995 (2011). https://doi.org/10.1038/cdd.2010.167
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DOI: https://doi.org/10.1038/cdd.2010.167
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