title: Direct GSK-3β inhibition enhances mesenchymal stromal cell migration by increasing expression of β-PIX and CXCR4.
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.
authors.affiliation: Department of Neurology, College of Medicine, Hanyang University, 249-1 Gyomun-dong, Guri-si, Seoul, Gyeonggi-do 471-701, Republic of Korea.
authors.forename: Young Seo
authors.initials: YS
authors.lastname: Kim
authors.forename: Min Young
authors.initials: MY
authors.lastname: Noh
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Kim
authors.forename: Hyun-Jeung
authors.initials: HJ
authors.lastname: Yu
authors.forename: Kyung Suk
authors.initials: KS
authors.lastname: Kim
authors.forename: Seung Hyun
authors.initials: SH
authors.lastname: Kim
authors.forename: Seong-Ho
authors.initials: SH
authors.lastname: Koh
mesh_terms: D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000502:physiology*; D000071679:Glycogen Synthase Kinase 3 beta; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000096:biosynthesis* / Q000502:physiology; D064067:Rho Guanine Nucleotide Exchange Factors / Q000096:biosynthesis* / Q000502:physiology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology*
chemical_list: C576606:ARHGEF7 protein, human; C527276:CXCR4 protein, human; D047428:Protein Kinase Inhibitors; D019718:Receptors, CXCR4; D064067:Rho Guanine Nucleotide Exchange Factors; C000605506:GSK3B protein, human; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3