Hair cell stereociliary bundle regeneration by espin gene transduction after aminoglycoside damage and hair cell induction by Notch inhibition.
Hair cell stereociliary bundle regeneration by espin gene transduction after aminoglycoside damage and hair cell induction by Notch inhibition.
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DOI：
10.1038/gt.2016.12
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发表时间：
2016-05
期刊：
影响因子：
5.1
通讯作者：
Ryan AF
中科院分区：
文献类型：
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作者：
Taura A;Taura K;Koyama Y;Yamamoto N;Nakagawa T;Ito J;Ryan AF
一旦内耳毛细胞 (HC) 因药物、噪音或老化而受损，其顶端结构（包括立体纤毛阵列）通常是最先丧失的细胞特征。虽然这可能导致 HC 体细胞逐渐丧失，但即使在静纤毛丧失后，仍有大量 HC 体仍然存在。然而，在没有静纤毛的情况下，它们就没有功能。 HC 有时可以通过 Atoh1 转导或 Notch 抑制来再生，但它们也可能缺乏静纤毛束。因此，开发静纤毛再生方法以实现 HC 功能恢复非常重要。 Espin 是一种肌动蛋白捆绑蛋白，已知参与发育过程中的立体纤毛伸长。我们使用两种 espin 同工型的病毒载体转导，评估了组织培养中受损前庭感觉上皮的立体纤毛阵列再生。氨基糖甙类药物会损伤椭圆囊 HC。然后用γ-分泌酶抑制剂处理椭圆囊，然后进行espin或对照转导和组织化学处理。虽然 γ-分泌酶抑制增加了 HC 的数量，但很少有 HC 具有立体纤毛阵列。相反，espin1 转导 46 小时后，观察到发束样结构显着增加。通过扫描电子显微镜证实这些是未成熟的立体纤毛阵列。 FM1-43 摄取的增加提供了立体纤毛功能的证据。 Espin4 转导没有效果。结果表明，espin1 基因治疗可以恢复受损或再生的 HC 上的静纤毛。
Once inner ear hair cells (HCs) are damaged by drugs, noise or aging, their apical structures including the stereociliary arrays are frequently the first cellular feature to be lost. While this can be followed by progressive loss of HC somata, a significant number of HC bodies often remain even after stereociliary loss. However, in the absence of stereocilia they are nonfunctional. HCs can sometimes be regenerated by Atoh1 transduction or Notch inhibition, but they also may lack stereociliary bundles. It is therefore important to develop methods for the regeneration of stereocilia, in order to achieve HC functional recovery. Espin is an actin bundling protein known to participate in sterociliary elongation during development. We evaluated stereociliary array regeneration in damaged vestibular sensory epithelia in tissue culture, using viral vector transduction of two espin isoforms. Utricular HCs were damaged with aminoglycosides. The utricles were then treated with a γ-secretase inhibitor, followed by espin or control transduction and histochemistry. While γ-secretase inhibition increased the number of HCs, few had stereociliary arrays. In contrast, 46 hrs after espin1 transduction, a significant increase in hair-bundle-like structures was observed. These were confirmed to be immature stereociliary arrays by scanning electron microscopy. Increased uptake of FM1–43 uptake provided evidence of stereociliary function. Espin4 transduction had no effect. The results demonstrate that espin1 gene therapy can restore stereocilia on damaged or regenerated HCs.