Next-generation sequencing of wild type and 5XFAD transcriptomes for Alzheimer's disease study. Next-generation sequencing of wild type and 5XFAD transcriptomes for Alzheimer's disease study

We found some novel miRNA that targets SIRT1 may contribute to the pathogenesis in AD mouse model. We first identified that SIRT1 is significantly reduced in Alzheimer patient`s precentral gyrus and 5XFAD mice. To determine whether the strategy of inhibiting some miRNA affects AD pathology, we synthesized antisense oligonucleotides of miRNA (miRNA ASO) and studied this sequence in the brain of 5XFAD through direct intracerebral ventricular injection using stereotaxic instrument. We identified miRNA ASO significantly reduced amyloid beta plaque and amyloid production enzyme. Importantly, the attenuation of amyloid beta plaques through miRNA ASO was caused by a phagocytic activity of glial cells, by which it can directly target CD36. miRNA ASO also decreases inflammatory responses. These results inhibit neuronal loss caused by amyloid beta and leading to improvement of cognitive impairment. These insights of miRNA ASO suggests as therapeutic scope against AD pathology. Overall design: Frontal cortex mRNA profiles of 6- and 11-month-old wild type (WT) and 5XFAD mice were generated by next-generation sequencing, using Illumina NovaSeq 6000 system.

本研究发现若干靶向沉默信息调节因子1（SIRT1）的新型微小RNA（miRNA）可能参与阿尔茨海默病（AD）小鼠模型的发病机制。本研究首先证实，AD患者中央前回及5XFAD小鼠脑组织中的SIRT1表达水平显著降低。为探究靶向抑制miRNA的策略能否改善AD病理，我们合成了miRNA反义寡核苷酸（miRNA ASO），并通过立体定位仪介导的直接脑室内注射方式，将该序列注入5XFAD小鼠脑中开展相关研究。研究发现，miRNA ASO可显著降低β淀粉样蛋白斑块及淀粉样蛋白生成酶的水平。值得注意的是，miRNA ASO介导的β淀粉样蛋白斑块消减作用，依赖于胶质细胞的吞噬活性，该过程可直接靶向CD36。此外，miRNA ASO还可抑制炎症反应。上述结果表明，该疗法可抑制β淀粉样蛋白诱导的神经元丢失，进而改善认知功能障碍。上述关于miRNA ASO的研究结果，为AD病理的靶向治疗提供了新的应用前景。实验整体设计：采用Illumina NovaSeq 6000测序平台，通过二代测序技术获取6月龄及11月龄野生型（WT）与5XFAD小鼠的额叶皮层信使RNA（mRNA）表达谱。