HuD及其靶标lncRNA对阿尔茨海默病关键相关基因的调控机制

The expression or dysfunction of related gene is closely associated with the occurrence and development of Alzheimer's disease (AD). However, the regulatory mechanism of these genes is far from being clarified. Our preliminary study showed: 1) the expression of HuD(RNA binding protein) in superior temporal gyrus (STG) from patients with AD was significantly increased. 2) the expression of APP,BACE1,and BACE1AS (lncRNA) was also elevated accompanied by obviously Aβ deposition in the cortex and hippocampus of HuD transgenic mouse. 3) HuD combined with 57 lncRNA including BACE1AS. 4) HuD stabilized the BACE1AS RNA. Based on the above results, we hypothesize that HuD may affect the expression of key AD-related genes by interacting with AD-related lncRNA, then affect the deposition of Aβ. This project will further investigate the regulatory effect of HuD and AD-related lncRNA on the expression of key AD-related gene, discuss the interaction between HuD and AD-related lncRNA, explore whether HuD mediates the regulation of lncRNA on key AD-related gene. This study will not only help to resolve the pathogenesis of AD, but also provide experimental evidence for clinical treatment of AD.

相关基因表达或功能失调与阿尔茨海默病(AD)的发生发展密切相关，但这些基因的调控机制远未得到阐明。申请者前期研究发现：1) RNA结合蛋白HuD水平在AD病人脑的颞上回区显著上调；2) HuD转基因小鼠的皮层和海马区APP，BACE1以及lncRNA BACE1AS表达升高并伴有明显的Aβ沉积；3) HuD结合包括BACE1AS在内的57个lncRNA；4）HuD稳定BACE1AS RNA。基于上述结果，我们假设：HuD可能通过与AD相关lncRNA互作影响关键AD相关基因的表达，进而影响AD过程中Aβ的沉积。本课题将在前期工作的基础上，进一步探讨HuD及其相关lncRNA对关键AD相关基因的表达的影响，探讨HuD对与之结合的lncRNA 水平及功能的影响，探讨HuD是否介导lncRNA对关键AD相关基因的表达的调控。这些研究不仅有助于解析AD的发病机制，也将为AD的临床治疗提供实验依据

阿尔茨海默病(AD)患者激增，但其发病机制尚未完全阐明，近来研究表明AD 疾病可能和基因转录后的调控有关，鉴于实验室前期研究提示HuD（RNA结合蛋白）有调控编码和非编码RNA的作用。为了揭示HuD调控的分子机制，本课题结合了体内外实验进行了深入研究。首先，我们采用RIP法在神经细胞SH-SY5Y中筛选了与HuD相关的57个长链非编码基因（lncRNA）和7个编码基因，接着用实时荧光定量PCR和RNA pull-down实验进一步验证。用分子生物学方法建立了siHuD和过表达lncRNA JPX的细胞模型，并运用QRT-PCR\免疫组化\WB\双荧光素酶等方法研究HuD调控的相关通路。结果表明HuD调控并稳定lncRNA JPX和重要的剪接调控因子NOVA1的表达，而JPX 也对NOVA1的表达有调控作用。同时，联合利用AD病人的脑部组织及衰老大鼠动物模型进行体内研究，结果发现，HuD和NOVA1在AD病人的脑部皮层区高表达，在大鼠的DG 区高表达。本研究表明HuD具有主调节器的功能，其分子机制可能通过调控lncRNA JPX和NOVA1发挥作用。上述结果证实了我们的科学假说，这将对AD疾病新机制有所揭示，为其防治提供新靶点。