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Direct GSK-3β Inhibition Enhances Mesenchymal Stromal Cell Migration by Increasing Expression of Beta-PIX and CXCR4

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Abstract

Mesenchymal stromal cells (MSCs) are emerging as candidate cells for the treatment of neurological diseases because of their neural replacement, neuroprotective, and neurotrophic effects. However, the majority of MSCs transplanted by various routes fail to reach the site of injury, and they have demonstrated only minimal therapeutic benefit in clinical trials. Therefore, enhancing the migration of MSCs to target sites is essential for this therapeutic strategy to be effective. In this study, we assessed whether inhibition of glycogen synthase kinase-3β (GSK-3β) increases the migration capacity of MSCs during ex vivo expansion. Human bone marrow MSCs (hBM-MSCs) were cultured with various GSK-3β inhibitors (LiCl, SB-415286, and AR-A014418). Using a migration assay kit, we found that the motility of hBM-MSCs was significantly enhanced by GSK-3β inhibition. Western blot analysis revealed increased levels of migration-related signaling proteins such as phospho-GSK-3β, β-catenin, phospho-c-Raf, phospho-extracellular signal-regulated kinase (ERK), phospho-β-PAK-interacting exchange factor (PIX), and CXC chemokine receptor 4 (CXCR4). In addition, real-time polymerase chain reaction demonstrated increased expression of matrix metalloproteinase-2 (MMP-2), membrane-type MMP-1 (MT1-MMP), and β-PIX. In the reverse approach, treatment with β-PIX shRNA or CXCR4 inhibitor (AMD 3100) reduced hBM-MSC migration. These findings suggest that inhibition of GSK-3β during ex vivo expansion of hBM-MSCs may enhance their migration capacity by increasing expression of β-catenin, phospho-c-Raf, phospho-ERK, and β-PIX and the subsequent up-regulation of CXCR4. Enhancing the migration capacity of hBM-MSCs by treating these cells with GSK-3β inhibitors may increase their therapeutic potential.

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Acknowledgments

This study was supported by grants from the Korea Healthcare Technology R&D Project, Ministry for Health & Welfare Affairs, Republic of Korea (A101712). The funding organization had no role in the design, conduct, analysis, or preparation of this report.

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No competing financial interests exist.

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Authors and Affiliations

  1. Department of Neurology, College of Medicine, Hanyang University, 249-1 Gyomun-dong, Guri-si, Seoul, Gyeonggi-do, 471-701, Republic of Korea

    Young Seo Kim, Min Young Noh, Seung Hyun Kim & Seong-Ho Koh

  2. Department of Nuclear Medicine, College of Medicine, Hanyang University, Seoul, Republic of Korea

    Ji Young Kim

  3. Department of Neurology, Bundang Jesaeng General Hospital, Seongnam, Republic of Korea

    Hyun-Jeung Yu

  4. Bioengineering Institute, CoreStem Inc., Seoul, Republic of Korea

    Kyung Suk Kim

  5. Department of Translational Medicine, Graduate School of Biomedical Science and Engineering, Hanyang University, Seoul, Republic of Korea

    Seung Hyun Kim & Seong-Ho Koh

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  1. Young Seo Kim
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Correspondence to Seong-Ho Koh.

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Young Seo Kim and Min Young Noh contributed equally to this work.

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Kim, Y.S., Noh, M.Y., Kim, J.Y. et al. Direct GSK-3β Inhibition Enhances Mesenchymal Stromal Cell Migration by Increasing Expression of Beta-PIX and CXCR4. Mol Neurobiol 47, 811–820 (2013). https://doi.org/10.1007/s12035-012-8393-3

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  • DOI: https://doi.org/10.1007/s12035-012-8393-3

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