Microtopographical cues promote peripheral nerve regeneration via transient mTORC2 activation.
Microtopographical cues promote peripheral nerve regeneration via transient mTORC2 activation.
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DOI：
10.1016/j.actbio.2017.07.031
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发表时间：
2017-09-15
影响因子：
9.7
通讯作者：
Riehle MO
中科院分区：
文献类型：
--
作者：
Thomson SE;Charalambous C;Smith CA;Tsimbouri PM;Déjardin T;Kingham PJ;Hart AM;Riehle MO
尽管显微手术修复，周围神经损伤后的功能恢复是缓慢的，往往不完全。结果可以通过增加对再生分子生物学的理解和实验生物工程策略的翻译来改善。地形线索已被证明是神经突再生速度和方向性的强大调节剂，在本研究中，我们在一个器官型外植体模型中研究了线性微图案结构的下游分子效应。线性地形信号增强了神经突的生长，我们的研究结果表明mTOR通路在调节这些反应中很重要。mTOR基因表达在48 - 72 h达到峰值，与神经突快速生长和神经胶质迁移的开始时间一致，并与48 h时的神经突长度相关。mTOR蛋白位于神经胶质上，沿神经突呈点状分布。mTOR水平在72 h时达到峰值，地形图案显著增加（p < 0.05）。此外，地形线索可以超越药物抑制。下游磷酸化实验和使用雷帕霉素抑制mTORC1强调mTORC2是一个重要的介质和更特异性的治疗靶点。定量免疫组织化学证实mTORC2成分载体存在于再生前沿，它与f -肌动蛋白和血管蛋白共定位。总的来说，这些结果为地形对神经再生的作用机制提供了更深入的理解，并支持地形模式与药理mTORC2增强相结合，用于修复周围神经的生物材料结构。周围神经损伤是常见的和功能破坏性的。尽管显微外科手术修复，但愈合缓慢且不完整，并伴有持续的功能缺陷。显然，我们需要转化生物工程方法，增加我们对控制神经再生的分子过程的了解，以提高愈合的速度和成功率。地形线索是神经突生长的强大决定因素，代表了一种高度可翻译的工程策略。在这里，我们首次证明了微地形通过mTOR途径增强了神经突的生长，其中mTORC2亚型尤为重要。这些结果进一步证明了将微地形线索纳入周围神经再生导管，并表明mTORC2可能是增强神经再生的合适治疗靶点。
Despite microsurgical repair, recovery of function following peripheral nerve injury is slow and often incomplete. Outcomes could be improved by an increased understanding of the molecular biology of regeneration and by translation of experimental bioengineering strategies. Topographical cues have been shown to be powerful regulators of the rate and directionality of neurite regeneration, and in this study we investigated the downstream molecular effects of linear micropatterned structures in an organotypic explant model. Linear topographical cues enhanced neurite outgrowth and our results demonstrated that the mTOR pathway is important in regulating these responses. mTOR gene expression peaked between 48 and 72 h, coincident with the onset of rapid neurite outgrowth and glial migration, and correlated with neurite length at 48 h. mTOR protein was located to glia and in a punctate distribution along neurites. mTOR levels peaked at 72 h and were significantly increased by patterned topography (p < 0.05). Furthermore, the topographical cues could override pharmacological inhibition. Downstream phosphorylation assays and inhibition of mTORC1 using rapamycin highlighted mTORC2 as an important mediator, and more specific therapeutic target. Quantitative immunohistochemistry confirmed the presence of the mTORC2 component rictor at the regenerating front where it co-localised with F-actin and vinculin. Collectively, these results provide a deeper understanding of the mechanism of action of topography on neural regeneration, and support the incorporation of topographical patterning in combination with pharmacological mTORC2 potentiation within biomaterial constructs used to repair peripheral nerves. Peripheral nerve injury is common and functionally devastating. Despite microsurgical repair, healing is slow and incomplete, with lasting functional deficit. There is a clear need to translate bioengineering approaches and increase our knowledge of the molecular processes controlling nerve regeneration to improve the rate and success of healing. Topographical cues are powerful determinants of neurite outgrowth and represent a highly translatable engineering strategy. Here we demonstrate, for the first time, that microtopography potentiates neurite outgrowth via the mTOR pathway, with the mTORC2 subtype being of particular importance. These results give further evidence for the incorporation of microtopographical cues into peripheral nerve regeneration conduits and indicate that mTORC2 may be a suitable therapeutic target to potentiate nerve regeneration.
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影响因子：
16.6
作者：
Christie, Kimberly J.;Krishnan, Anand;Martinez, Jose A.;Purdy, Kaylynn;Singh, Bhagat;Eaton, Shane;Zochodne, Douglas
通讯作者：
Zochodne, Douglas
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通讯作者：
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