长链非编码RNA000367通过调控耳蜗细胞凋亡参与顺铂致感音神经性耳聋的作用机制研究

The apoptosis of cochlea cells is the main reason of cisplatin-induced sensorineural hearing loss. Although a number of apoptotic signaling pathways have been found, the effectiveness of inhibition of single pathway is limited. So it is important to find the upstream molecular that regulates the cisplatin-induced apoptosis of cochlear cells. In our previous study, by using RNA-seq, we found a large amount of long noncoding RNA (lncRNA) that exhibited a different expression pattern between normal mice and mice with cisplatin-induced sensorineural hearing loss. The lncRNAs were chosen by CNC analysis and in vitro confirmation, and we found a lncRNA 000367 whose expression was significantly increased in cisplatin-induced cochlea apoptosis. Moreover, when the expression of lncRNA000367 was inhibited by specific siRNA, the cisplatin-induced apoptosis was also significantly weaken which indicated an important role of lncRNA000367 in cisplatin-induced apoptosis. In our project, we will firstly construct the model of lncRNA000367 knock-out mice, and confirm the role of lncRNA000367 in cisplatin-induced sensorineural hearing loss. Then we will analyse both the apopotisis-relating genes and signaling pathways regulated by lncRNA000367, and further find the target moleculars directly interacting with lncRNA000367. The finding from this work might be helpful for deciphering the mechanisms underlying the cisplatin-induced sensorineural hearing loss, which, may in turn provide a novel target for clinical treatment.

耳蜗细胞凋亡是顺铂致感音神经性耳聋的重要原因，虽已发现多种凋亡通路参与其中，但针对单一通路采取抑制措施效果有限，因此寻找调控耳蜗细胞凋亡的更上游的调控分子具有重要意义。我们前期研究利用高通量测序发现，在顺铂致感音神经性耳聋中存在长链非编码RNA的大量差异表达。经CNC分析和体外筛选验证，发现编号000367的长链非编码RNA与凋亡过程密切相关，并在顺铂损伤耳蜗中表达明显上调。利用siRNA干扰其表达后，顺铂诱导的凋亡明显被抑制，提示000367在顺铂诱导的耳蜗细胞凋亡中发挥重要作用。本课题拟通过构建基因敲除小鼠，首先在动物体内研究000367在顺铂致感音神经性耳聋中的作用。然后分析000367调控的关键凋亡基因和信号通路，并明确与之直接作用的关键靶分子，进而阐明其作用机制。本研究将从长链非编码RNA视角深入揭示顺铂致感音神经性耳聋的发病机制，为治疗提供潜在的新型有效的靶点。

耳蜗细胞凋亡是顺铂致感音神经性耳聋的重要原因，虽已发现多种凋亡通路参与其中，但针对单一通路采取抑制措施效果有限，因此寻找调控耳蜗细胞凋亡的更上游的调控分子具有重要意义。长链非编码 RNA（long noncoding RNA）是一类转录本长度超过200个核苷酸的RNA分子，是非编码RNA家族的重要成员。大量证据表明lncRNA可以通过表观遗传调控、转录调控和转录后调控等多种方式调控基因的表达，在机体的生长发育、细胞凋亡、增殖等过程中发挥重要作用。本课题利用高通量测序技术发现，在顺铂致感音神经性耳聋中存在长链非编码RNA的大量差异表达。经CNC分析和体外筛选验证，发现编号lncRNA000367的长链非编码RNA与凋亡过程密切相关，并在顺铂损伤耳蜗中表达明显上调。利用siRNA干扰其表达后，顺铂诱导的凋亡明显被抑制，说明lncRNA000367在顺铂诱导的耳蜗细胞凋亡中发挥重要作用，且lncRNA000367具有促进顺铂诱导凋亡的作用。进一步利用生物信息学技术我们发现了6个可能与目的lncRNA分子结合并发挥作用的miRNA分子。本研究从长链非编码RNA视角深入揭示顺铂致感音神经性耳聋的发病机制，并发现了一个可以促进顺铂诱导凋亡的促凋亡lncRNA分子，为临床上顺铂致感音神经性耳聋的治疗提供了新的靶点。