ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China; NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai, 200031, China.
ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China; NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai, 200031, China.
Exp Cell Res. 2021 Jan 1;398(1):112395. doi: 10.1016/j.yexcr.2020.112395. Epub 2020 Dec 3.
Loss of hair cells from vestibular epithelium results in balance dysfunction. The current therapeutic regimen for vestibular diseases is limited. Upon injury or Atoh1 overexpression, hair cell replacement occurs rapidly in the mammalian utricle, suggesting a promising approach to induce vestibular hair cell regeneration. In this study, we applied simultaneous gentamicin-mediated hair cell ablation and Atoh1 overexpression to induce neonatal utricular hair cell formation in vitro. We confirmed that type I hair cells were the primary targets of gentamicin. Furthermore, injury and Atoh1 overexpression promoted hair cell regeneration in a timely and efficient manner through robust viral transfection. Hair cells regenerated with type II characteristics in the striola and type I/II characteristics in non-sensory regions. Rare EdU/myosin7a cells in sensory regions and robust EdU/myosin7a signals in ectopic regions indicate that transdifferentiation of supporting cells in situ, and mitosis and differentiation of non-sensory epithelial cells in ectopic regions, are sources of regenerative hair cells. Distinct regeneration patterns in in situ and ectopic regions suggested robust plasticity of vestibular non-sensory epithelium, generating more developed hair cell subtypes and thus providing a promising stem cell-like source of hair cells. These findings suggest that simultaneously causing injury and overexpressing Atoh1 promotes hair cell regeneration efficacy and maturity, thus expanding the understanding of ectopic plasticity in neonatal vestibular organs.
前庭上皮细胞毛细胞的丧失会导致平衡功能障碍。目前，前庭疾病的治疗方案有限。在损伤或 Atoh1 过表达时，哺乳动物的前庭囊中毛细胞会迅速替代，这表明一种有前途的诱导前庭毛细胞再生的方法。在这项研究中，我们应用同时给予庆大霉素介导的毛细胞消融和 Atoh1 过表达，以在体外诱导新生前庭毛细胞的形成。我们证实 I 型毛细胞是庆大霉素的主要靶标。此外，损伤和 Atoh1 过表达通过强大的病毒转染以及时和有效的方式促进毛细胞再生。在条纹区再生具有 II 型特征的毛细胞，在非感觉区具有 I/II 型特征的毛细胞。感觉区中罕见的 EdU/myosin7a 细胞和异位区中丰富的 EdU/myosin7a 信号表明，原位支持细胞的转分化，以及异位区非感觉上皮细胞的有丝分裂和分化，是再生毛细胞的来源。原位和异位区的不同再生模式表明前庭非感觉上皮具有强大的可塑性，产生更发达的毛细胞亚型，从而为毛细胞提供了有前途的干细胞样来源。这些发现表明，同时造成损伤和过表达 Atoh1 可提高毛细胞再生的效果和成熟度，从而扩展了对新生前庭器官异位可塑性的理解。
