ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells
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|Title:||ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells||Authors:||Ryan, Caroline M.; Brown, James A. L.; Bourke, Emer; Prendergast, Áine M.; Kavanagh, Claire; Liu, Zhonglin; Owens, Peter; Shaw, Georgina; Kolch, Walter; O'Brien, Timothy; Barry, Frank P.||Permanent link:||http://hdl.handle.net/10197/9159||Date:||24-Jul-2015||Online since:||2018-01-10T13:47:13Z||Abstract:||Bone marrow-derived stromal cells (BMSCs), also known as mesenchymal stem cells, are the focus of intensive efforts worldwide to elucidate their function and biology. Despite the importance of BMSC migration for their potential therapeutic uses, the mechanisms and signalling governing stem cell migration are still not fully elucidated. Methods: We investigated and detailed the effects of MCP-1 activation on BMSCs by using inhibitors of G protein-coupled receptor alpha beta (GPCR αβ), ROCK (Rho-associated, coiled-coil containing protein kinase), and PI3 kinase (PI3K). The effects of MCP-1 stimulation on intracellular signalling cascades were characterised by using immunoblotting and immunofluorescence. The effectors of MCP-1-mediated migration were investigated by using migration assays (both two-dimensional and three-dimensional) in combination with inhibitors. Results: We established the kinetics of the MCP-1-activated signalling cascade and show that this cascade correlates with cell surface re-localisation of chemokine (C motif) receptor 2 (CCR2) (the MCP-1 receptor) to the cell periphery following MCP-1 stimulation. We show that MCP-1-initiated signalling is dependent on the activation of βγ subunits from the GPCR αβγ complex. In addition, we characterise a novel role for PI3Kγ signalling for the activation of both PAK and ERK following MCP-1 stimulation. We present evidence that the Gβγ complex is responsible for PI3K/Akt, PAK, and ERK signalling induced by MCP-1 in BMSCs. Importantly, we found that, in BMSCs, inhibition of ROCK significantly inhibits MCP-1-induced chemotactic migration, in contrast to previous reports in other systems.Conclusions: Our results indicate differential chemotactic signalling in mouse BMSCs, which has important implications for the translation of in vivo mouse model findings into human trials. We identified novel components and interactions activated by MCP-1-mediated signalling, which are important for stem cell migration. This work has identified additional potential therapeutic targets that could be manipulated to improve BMSC delivery and homing.||Funding Details:||Science Foundation Ireland||Funding Details:||National Breast Cancer Research Institute||Type of material:||Journal Article||Publisher:||Springer Nature||Journal:||Stem Cell Research and Therapy||Volume:||6||Issue:||136||Copyright (published version):||2015 the Authors||Keywords:||ROCK; Gβγ complex; Mesenchymal stem cells; BMSCs; MCP-1 activation; MCP-1 activation; GPCR αβ; Chemotactic migration; Chemotactic signalling; Stem cell migration||DOI:||10.1186/s13287-015-0125-y||Language:||en||Status of Item:||Peer reviewed||This item is made available under a Creative Commons License:||https://creativecommons.org/licenses/by-nc-nd/3.0/ie/|
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