Cooperation of Matrix Metalloproteinases with the RhoA/Rho Kinase and Mitogen‐Activated Protein Kinase Kinase‐1/Extracellular Signal‐Regulated Kinase Signaling Pathways Is Required for the Sphingosine‐1‐Phosphate‐Induced Mobilization of Marrow‐Derived Stromal Cells

Abstract

The ease of isolation and ex vivo culture of marrow-derived stromal cells (MSCs) from adult bone marrow renders them a very promising source of adult stem cells for gene transfer and cell therapy. However, little is known about the signaling pathways that control their in vivo mobilization and subsequent biodistribution. Platelet-derived sphingosine-1-phosphate (S1P), a bioactive lipid that acts via G-protein-coupled-receptors, exerts strong chemoattraction upon MSCs through yetuncharacterized signaling pathways. We show that the S1P-induced migration and morphological changes of MSCs in vitro require the activities of extracellular signal-regulated kinase (ERK), Rho kinase (ROCK), and matrix metalloproteinase (MMP) signaling molecules. Specifically, S1P-induced remodeling of the MSC cytoskeleton led to the rapid (<1 minute) formation of actin stress fibers via activation of the RhoA/ ROCK pathway and required the catalytic activity of MMPs. S1P-induced activation of the mitogen-activated protein kinase kinase-1 (MEK1)/ERK pathway also contributed to the induction of the actin stress fibers and to the redistribution of paxillin at the focal adhesions through tyrosine phosphorylation of focal adhesion kinase in an MMP-dependent manner. Moreover, MMP-and ROCK-dependent molecular events are implicated in the regulation of the S1P-induced activation of ERK. Our results demonstrate that MSC mobilization in response to S1P requires cooperation between MMP-mediated signaling events and the RhoA/ROCK and MEK1/ERK intracellular pathways. Therefore, the characterization of the cellular factors and the intracellular signaling pathways underlying MSC mobilization is crucial to achieve high efficacy in therapeutic use.

ROCK-II mediates colon cancer invasion via regulation of MMP-2 and MMP-13 at the site of invadopodia as revealed by multiphoton imaging

“…34,35 Furthermore, it identifies ROCK-II as the critical isoform of ROCK involved in activation of both MMP-2 and MMP-13. MMP-9 activity has also been linked to Rho/ROCK, 36 but our data did not show a significant decrease in MMP-9 activity with ROCK-II knockdown.…”

Epigallocatechin Gallate Targeting of Membrane Type 1 Matrix Metalloproteinase-mediated Src and Janus Kinase/Signal Transducers and Activators of Transcription 3 Signaling Inhibits Transcription of Colony-stimulating Factors 2 and 3 in Mesenchymal Stromal Cells

“…Immunoblotting Procedures-MSCs were lysed in a buffer containing 1 mM NaF and Na 3 VO 4 (14), and proteins were separated by SDS-polyacrylamide gel electrophoresis (PAGE). After electrophoresis, proteins were electrotransferred to polyvinylidene difluoride membranes, and immunoreactive material was visualized by enhanced chemiluminescence as described previously (16).…”

“…1 ated signaling in concanavalin A (ConA)-activated MSCs that could regulate the expression of the inflammation biomarker cyclooxygenase-2 (12). Interestingly, aside from its canonical role in extracellular matrix proteolysis, MT1-MMP is also involved in transducing crucial intracellular signaling that may control several processes related to MSC mobilization and cell survival (13)(14)(15). In this study, we demonstrate that, as a consequence of ConA activation, sequential Src kinase and JAK/ STAT3 signaling is required to up-regulate CSF-2 and CSF-3 transcription and that this necessitates the contribution of MT1-MMP transduction through the crucial involvement of a phosphorylatable Tyr 573 residue located within its 20-amino acid cytoplasmic domain.…”

MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines

“…32 Another inflammatory mediator, sphingosine-1-phosphate, was shown to induce chemotactic migration of hMSCs implicating MMP activity. 34 Despite those intriguing findings, detailed analysis on the role of inflammatory cytokines in hMSC invasion and identification of particular MMPs essentially involved in this process have not been performed so far.…”

“…32 Another inflammatory mediator, sphingosine-1-phosphate, was shown to induce chemotactic migration of hMSCs implicating MMP activity. 34 Despite those intriguing findings, detailed analysis on the role of inflammatory cytokines in hMSC invasion and identification of particular MMPs essentially involved in this process have not been performed so far.In this work, we have examined bone marrow-derived hMSCs for mRNA and protein expression of MMP-2, MMP-9, MT1-MMP, TIMP-1, as well as TIMP-2. RNA interference (RNAi) technology was performed to achieve selective knock-down of these genes to study their individual contribution in hMSC migration through human reconstituted basement membranes.…”