前庭毛细胞(HC)损失导致平衡障碍。目前多基因诱导等可获得HC再生，然而再生HC未成熟，转分化消耗了支持细胞(SC)。精确的平面极性(PCP)及突触连接等均为平衡功能恢复的必要条件，目前尚无法实现。申请人前期利用体细胞编程使小鼠前庭上皮产生多能干细胞(iPSC)，并鉴定了Atoh1过表达诱导的HC类型和部分电生理功能。拟利用此模型结合前庭发育规律，研究前庭结构和功能再生的可行性。包括①利用Oct4、Sox2、Klf4a、c-Myc（OSKM）重编程联合调控Atoh1、Notch、Wnt、BMP、FGF信号通路，促进前庭上皮iPSC增殖并分化为HC，诱导原位HC和SC匹配再生；②完善前庭发育机制，指导再生上皮极性建立、突触形成及定向分化；③通过单细胞测序技术筛选HC发育成熟关键基因，促进再生HC成熟，利用膜片钳技术鉴定再生HC类型和功能

Vestibular hair cell (HCs) loss leads to balance disorder. Previous studies showed genetic reprogramming technique could induce the vestibular hair cells regeneration, but the regeneration process consumes support cells and resulted in the structural imbalance. Precise planar cell polarity (PCP) and synaptic connections are necessary for the balance of functional recovery, which are hard to achieve. In the pilot study, we have generated induced pluripotent stem cells (iPSC) in the vestibular epithelium by genetic reprogramming technique in mouse. We identified HCs types and electrical-physiological function by patch clamp in newly formed hair cells induced by Atoh1 overexpression and found most of newly formed hair cells were type II HCs. Base on this, our plan includes (1) achieve SCs proliferation as well as vestibular HCs regeneration using iPSC technology combined with Atoh1 over-expression, inhibition of the Notch signal, activation of the Wnt, BMP, FGF signaling pathways; (2) explore the developmental law of vestibular epithelium and guide the directional differentiation and planar cell polarity(PCP) of regenerated HCs and SCs, facilitate vestibular hair cells afferent and efferent nerve synapse formation, fulfill vestibular organ structure regeneration; (3) facilitate HCs maturation through screening the key genes for HC maturation by single-cell sequencing technology and identify the function and subtype of HCs by patch clamp technique