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IKK-2 is required for TNF-α-induced invasion and proliferation of human mesenchymal stem cells

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Abstract

Mesenchymal stem cells (MSCs) can contribute to tissue repair by actively migrating to sites of tissue injury. However, the cellular and molecular mechanisms of MSC recruitment are largely unknown. The nuclear factor (NF)-κB pathway plays a pivotal role in regulating genes that influence cell migration, cell differentiation, inflammation, and proliferation. One of the major cytokines released at sites of injury is tumor necrosis factor-α (TNF-α), which is known to be a key regulator of the NF-κB pathway. Therefore, we hypothesized that TNF-α may lead to MSC invasion and proliferation by activation of the NF-κB pathway. TNF-receptor 1 and 2, NF-κB (p65), and IκB kinase 2 (IKK-2) are expressed in human MSCs (hMSCs). Stimulation of hMSCs with TNF-α caused a p65 translocation from the cytoplasm to nucleoplasm but did not change the expression profile of MSC markers. TNF-α strongly augmented the migration of hMSCs through the human extracellular matrix. Using lentiviral gene transfer, overexpressing a dominant-negative mutant of IKK-2 (dn-IKK-2) significantly blocked this effect. NF-κB target genes associated with migration (vascular cell adhesion molecule-1, CD44, and matrix metalloproteinase 9) were upregulated by TNF-α stimulation and blocked by dn-IKK-2. Moreover, using the bromodeoxyuridine assay, we showed that the inhibition of the NF-κB pathway caused a significant reduction in the basal proliferation rate. TNF-α stimulated the proliferation of hMSCs, whereas overexpression of dn-IKK-2 significantly blocked this effect. TNF-α led to the upregulated expression of the proliferation-associated gene cyclin D1. In conclusion, we demonstrated that the NF-κB pathway components, p65 and IKK-2, are expressed in hMSCs. Our data provide evidence that this signal transduction pathway is implicated in TNF-α-mediated invasion and proliferation of hMSCs. Therefore, hMSC recruitment to sites of tissue injury may, at least in part, be regulated by the NF-κB signal transduction pathway.

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Acknowledgment

We would like to thank Prof. Dr. Med. Ulrich Koszinowski and Priv. Doz. Dr. med. Dr. rer. nat. Jürgen Haas (Max-von-Pettenkofer-Institute, Virology Section, University of Munich) for providing us the S2 laboratory and the intense discussion on lentiviral gene transfer and Martina Burggraf for PCR technical help and Dr. Eduard Fellows for assistance in the apoptosis assay. This project was supported by the AO Research Fund of the AO Foundation, Davos-Platz, Switzerland (AO grants: 05-B42 and 05-D83). We also appreciate the financial support of the Friedrich-Baur-Foundation (University of Munich) on lentiviral gene transfer in hMSCs, and Denitsa Docheva and Cvetan Popov acknowledge the financial support of the Bavarian Research Foundation (grant DPA-51/05).

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Authors and Affiliations

  1. Experimental Surgery and Regenerative Medicine, Department of Surgery, University of Munich (LMU), Nussbaumstrasse 20, 80336, Munich, Germany

    Wolfgang Böcker, Denitsa Docheva, Wolf Christian Prall, Emmanouil Pappou, Oliver Roßmann, Cvetan Popov, Wolf Mutschler & Matthias Schieker

  2. Division of Clinical Chemistry and Biochemistry, Department of Surgery, University of Munich (LMU), Munich, Germany

    Virginia Egea & Christian Ries

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  1. Wolfgang Böcker
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Correspondence to Matthias Schieker.

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Böcker, W., Docheva, D., Prall, W.C. et al. IKK-2 is required for TNF-α-induced invasion and proliferation of human mesenchymal stem cells. J Mol Med 86, 1183–1192 (2008). https://doi.org/10.1007/s00109-008-0378-3

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