Controlling integrin specificity and stem cell differentiation in 2D and 3D environments through regulation of fibronectin domain stability.
Controlling integrin specificity and stem cell differentiation in 2D and 3D environments through regulation of fibronectin domain stability.
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DOI：
10.1016/j.biomaterials.2008.10.047
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发表时间：
2009-02
期刊：
影响因子：
14
通讯作者：
Barker, Thomas H.
中科院分区：
文献类型：
--
作者：
Martino, Mikael M.;Mochizuki, Mayumi;Rothenfluh, Dominique A.;Rempel, Sandra A.;Hubbell, Jeffrey A.;Barker, Thomas H.
细胞外基质（ECM）对许多细胞现象（包括干细胞分化）发挥强大的控制作用。因此，设计和调节ECM类似物以连接特异性整联蛋白是用于再生医学策略的体外和体内控制细胞过程的有前景的方法。虽然纤连蛋白（FN），在组织发育和修复中的一个重要的ECM蛋白，其RGD肽被广泛用于细胞粘附，混杂与他们从事整合素导致难以控制的受体特异性相互作用。最近的模拟力介导的FN结构域的展开和人与小鼠FN的序列分析表明，FN的中央细胞结合结构域（FN III 9 -10）的结构稳定性影响其整合素的特异性。通过生产具有可变稳定性的FN III 9 -10变体，我们获得了对整合素α5β1具有不同特异性并且可以共价连接到纤维蛋白基质中的配体。在这里，我们证明了α5β1整合素特异性参与影响人类间充质干细胞（MSC）在2D和3D环境中的行为的能力。我们的数据表明α5β1在MSC成骨分化的控制中具有重要作用。与αvβ3相比，对α5β1特异性增加的FN片段导致MSC在2D和临床相关的3D纤维蛋白基质系统中的成骨分化显著增强，尽管附着/扩散和增殖与全长FN相当。这项工作显示了整合素依赖性细胞与ECM的相互作用如何在适合于3D细胞培养和组织工程的系统内被工程化以控制干细胞的命运。
The extracellular matrix (ECM) exerts powerful control over many cellular phenomena, including stem cell differentiation. As such, design and modulation of ECM analogs to ligate specific integrin is a promising approach to control cellular processes in vitro and in vivo for regenerative medicine strategies. Although fibronectin (FN), a crucial ECM protein in tissue development and repair, and its RGD peptide are widely used for cell adhesion, the promiscuity with which they engage integrins leads to difficulty in control of receptor-specific interactions. Recent simulations of force-mediated unfolding of FN domains and sequences analysis of human versus mouse FN suggest that the structural stability of the FN’s central cell-binding domains (FN III9-10) affects its integrin specificity. Through production of FN III9-10 variants with variable stabilities, we obtained ligands that present different specificities for the integrin α5β1 and that can be covalently linked into fibrin matrices. Here, we demonstrate the capacity of α5β1 integrin-specific engagement to influence human mesenchymal stem cell (MSC) behavior in 2D and 3D environments. Our data indicate that α5β1 has an important role in the control of MSC osteogenic differentiation. FN fragments with increased specificity for α5β1 versus αvβ3 results in significantly enhanced osteogenic differentiation of MSCs in 2D and in a clinically relevant 3D fibrin matrix system, although attachment/spreading and proliferation were comparable with that on full-length FN. This work shows how integrin-dependant cellular interactions with the ECM can be engineered to control stem cell fate, within a system appropriate for both 3D cell culture and tissue engineering.
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域间倾斜角度决定了人纤连蛋白的第九和第十FIII结构域的整联蛋白依赖性功能。
影响因子：
4.8
作者：
Altroff, H;Schlinkert, R;van der Walle, CF;Bernini, A;Campbell, ID;Werner, JM;Mardon, HJ
通讯作者：
Mardon, HJ
影响因子：
10.8
作者：
Zisch, AH;Schenk, U;Hubbell, JA
通讯作者：
Hubbell, JA
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DOI：
10.1093/protein/15.12.1021
发表时间：
2002-12
期刊：
Protein engineering
影响因子：
--
作者：
van der Walle CF;Altroff H;Mardon HJ
通讯作者：
Mardon HJ
影响因子：
14
作者：
Stephansson, SN;Byers, BA;García, AJ
通讯作者：
García, AJ
影响因子：
4.8
作者：
Altroff, H;van der Walle, CF;Mardon, HJ
通讯作者：
Mardon, HJ