ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells.
ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells.
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DOI：
10.1186/s13287-015-0125-y
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发表时间：
2015-07-24
影响因子：
7.5
通讯作者：
Barry FP
中科院分区：
文献类型：
--
作者：
Ryan CM;Brown JA;Bourke E;Prendergast ÁM;Kavanagh C;Liu Z;Owens P;Shaw G;Kolch W;O'Brien T;Barry FP
骨髓源性基质细胞（BMSCs），也称为间充质干细胞，是全世界致力于阐明其功能和生物学的焦点。尽管BMSC迁移对于其潜在的治疗用途的重要性，但支配干细胞迁移的机制和信号传导仍然没有完全阐明。我们通过使用G蛋白偶联受体α β（GPCR αβ）、ROCK（Rho相关的卷曲螺旋蛋白激酶）和PI 3激酶（PI 3 K）的抑制剂，研究并详细描述了MCP-1活化对BMSCs的影响。MCP-1刺激对细胞内信号级联的影响通过免疫印迹和免疫荧光进行表征。MCP-1介导的迁移的效应器进行了研究，通过使用迁移试验（二维和三维）结合抑制剂。我们建立了MCP-1激活的信号级联反应的动力学，并表明该级联反应与MCP-1刺激后趋化因子（C基序）受体2（CCR 2）（MCP-1受体）向细胞外周的细胞表面再定位相关。我们发现MCP-1启动的信号传导依赖于GPCR αβγ复合物中βγ亚基的激活。此外，我们还发现了PI 3 K γ信号在MCP-1刺激后PAK和ERK激活中的新作用。我们提出的证据表明，Gβγ复合物是负责PI 3 K/Akt，PAK，ERK信号转导由MCP-1诱导的BMSCs。重要的是，我们发现，在BMSC中，ROCK的抑制显着抑制MCP-1诱导的趋化迁移，在其他系统中的先前报告相反。我们的研究结果表明，小鼠骨髓间充质干细胞的差异趋化性信号，这有重要意义的翻译在体内小鼠模型的研究结果到人体试验。我们确定了新的组件和MCP-1介导的信号激活的相互作用，这是重要的干细胞迁移。这项工作已经确定了其他潜在的治疗靶点，可以操纵这些靶点来改善BMSC的递送和归巢。本文的在线版本（doi：10.1186/s13287-015-0125-y）包含补充材料，可供授权用户使用。
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. 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. 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. 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. The online version of this article (doi:10.1186/s13287-015-0125-y) contains supplementary material, which is available to authorized users.