Molecular mechanism in noise-induced hearing loss

摘要 这项研究的长期目标是阐明噪音诱导的细胞凋亡的分子机制。 听力损失（NIHL），并提出合理的治疗干预措施，以预防。基于我们之前 研究结果，并与其他系统的研究结果一致，我们将测试的假设，延长激活 AMPKα（T172）通过钙/钙调蛋白依赖性激酶激酶2（CaMKK 2）和肝激酶B1（LKB 1）或 活性氧（ROS）触发外毛细胞（OHC）死亡。我们进一步提出，激活的CaMKK 2 促进活化T细胞核因子（NFAT）易位到OHC核中，导致细胞因子释放 基因.最后，我们将使用siRNA和药物化合物来阻断多种死亡途径， 针对NIHL的协同保护作用。我们将采用全面的实验方法，包括体内 成年小鼠的研究，以确定潜在的分子机制，使用短发夹RNA（shRNA），通过 病毒转染，单细胞RNA-seq（scRNA-seq）与基因集富集分析（GSEA），和 用RNAscope进行荧光原位杂交。这些研究探索创新的治疗策略 旨在确定预防NIHL的新途径，使受影响者的生活质量受益 个人和降低医疗成本。此外，该提案所产生的数据将产生重大影响， 这有助于我们理解广泛的内耳疾病，因为相似性已经显示在 与噪声、药物和年龄相关性听力损失相关的分子事件。

ABSTRACT The long-term goals of this research are to elucidate the molecular mechanisms underlying noise-induced hearing loss (NIHL) and to propose rational therapeutic interventions for prevention. Based on our previous findings and consistent with findings in other systems, we will test the hypothesis that prolonged activation of AMPKα (T172) by calcium/calmodulin-dependent kinase kinase 2 (CaMKK2) and liver kinase B1 (LKB1) or reactive oxygen species (ROS) triggers outer hair cell (OHC) death. We further propose that activated CaMKK2 facilitates translocation of nuclear factor of activated T-cells (NFAT) into OHC nuclei, causing release of cytokine genes. Finally, we will use siRNA and pharmacological compounds to block multiple death pathways for potential synergistic protection against NIHL. We will employ a comprehensive experimental approach including in-vivo studies with adult mice to define the underlying molecular mechanisms using short hairpin RNA (shRNA) via virus transfection, single cell RNA-seq (scRNA-seq) with gene set enrichment analysis (GSEA), and fluorescence in-situ hybridization with RNAscope. These studies investigating innovative therapeutic strategies are designed to identify novel avenues for the prevention of NIHL, benefiting the quality of life of affected individuals and reducing healthcare costs. In addition, the data generated by this proposal will make a significant contribution to our understanding of a broad range of inner ear disorders since similarities have been shown in the molecular events associated with noise-induced, drug-induced, and age-related hearing loss.