LSD1抑制剂调控组蛋白H3K4甲基化对耳毒性药物损伤中耳蜗的保护作用及其机制研究

药物、噪声、感染等因素可致耳蜗损伤和永久听力损失。研究耳蜗细胞保护及其作用机制对防治感音神经性聋至关重要。我们既往发现传统LSD1抑制剂在体外可通过上调H3K4me2表达水平发挥耳蜗抗凋亡保护作用。前期我们设计出一类易溶于水、保护性更优的新型LSD1抑制剂（芳香环丙基胺化合物）。但这类抑制剂（传统和新型）发挥保护作用的内在机制仍未知。本项目拟在前期基础上采用斑马鱼及小鼠模型进一步检测LSD1抑制剂生物安全性；建立小鼠体内外耳毒性药物损伤模型进一步验证、比较传统和新型抑制剂对耳蜗毛细胞、螺旋神经元及听觉功能保护效应；通过流式细胞分选毛细胞，应用染色质免疫共沉淀测序筛选H3K4甲基化修饰差异区域，联合RNA-Seq分析鉴定靶基因及调控网络，揭示其发挥保护作用的分子机制。该成果不仅有助于揭示这类抑制剂调控H3K4甲基化修饰对损伤中耳蜗的保护机制，且可从表观遗传调控角度为耳聋防治提供新靶点。

Drugs, noise, infection and other factors can cause damage to cochlear cells and permanent hearing loss. The study of effective protection of cochlear cells and investigation of the mechanism are particularly important for prevention of sensorineural hearing loss. We have previously found that traditional lysine specific demethylase 1 (LSD1) inhibitor can upregulate H3K4 dimethylation expression mediated inhibition of apoptosis to achieve cochlear protective effect in vitro. In our preliminary works, we designed a novel kind of LSD1 inhibitor named aromatic cyclopropyl amine compound which is soluble in water and found it has better effect than the traditional one on the protection of cochlear cells through the regulation of histone H3K4 methylation by inhibiting the expression of LSD1. But the internal protective mechanism of this kind of inhibitors (traditional and novel) is still unknown. Based on our preliminary works, we will use zebrafish and mice models in vivo to detect the biosafety of these inhibitors in this study; ototoxic drug injury mice models will be established to verify and compare their protective effects on hair cells, spiral ganglion neurons and auditory functions in vitro and in vivo; hair cells are going to be purified by flow cytometry and then chromatin immunoprecipitation sequencing (ChIP-Seq) combined with transcriptome sequencing (RNA-Seq) will be applied for screening the different regions of H3K4 methylation modification and the target genes among the regulatory networks for the purpose to reveal the molecular mechanism of their protective effect. The results not only help to reveal the protective mechanism of H3K4 methylation modification on damaged cochlea through the inhibitors, but also provide new targets for the prevention and treatment of sensorineural hearing loss from the perspective of epigenetic regulation.