协助朊病毒感染、致病的lncRNA鉴定及其功能分析

lncRNA is a group of important long non-coding RNA discovered in recent years. It plays an important role in life biological process. It is probably an important co-factor in prion disease pathogenesis. Further study for lncRNA will give us a new hope to understand the mechanism of prion pathogenesis. Therefore, there is a great need to study the relationship between lncRNA and prion disease. By using mouse lncRNA microarray and whole-genome microarray, we studied the lncRNA and mRNA expression in mouse brains infected with prion. Our preliminary study suggested that lncRNA played a role in host and virus interaction. In this study, we will focus on analysis of the differentially expressed lncRNA, then select the highest difference of lncRNA. The expression profiles of significant lncRNA will be analysed. Based on the march of lncRNA and mRNA expression pattern, we can identify the specific target effect of lncRNA. We will analyse the interaction of lncRNAs inside the cells during host-infected with prion. Therefore, we can obtain specific lncRNAs during prion infection and pathogenesis, and analyse their functions, especially the change of brain molecules level after those specific lncRNA changed during prion infection. This study will help to understand the molecular mechanism of prion infection and pathogenesis, which contributes to the further clarification of relation, molecular mechanisms of lncRNA and host genome involved prion infection. It will certainly help to introduce new therapeutic and preventive methods in prion infection.

lncRNA是近年来发现的一类重要长链非编码RNA分子，参与生命过程中的一系列重要进程，可能是朊病毒致病的重要协助因子，它给朊病毒病发病机理研究带来新的希望，因此，迫切需要人们开展lncRNA与朊病毒病相关性的研究。本课题前期对朊病毒病小鼠脑lncRNA、mRNA表达谱进行分析，初步实验结果表明朊病毒与lncRNA水平上存在着相互作用。本研究以从lncRNA表达谱结果筛选出表达差异显著的lncRNA为研究对象，基于lncRNA与mRNA的配对表达谱可高精度分辨出功能性lncRNA的靶向关系，研究朊病毒与宿主在lncRNA调控体系水平上的相互作用，获得朊病毒感染、致病相关的重要lncRNA分子，并深入研究其功能。本研究有助于认识朊病毒感染和致病的分子机理及朊病毒与宿主基因功能关系的一般规律，对研究和引入新型的朊病毒防治措施具有推动作用。

LncRNA是近些年发现的一种重要的长链非编码RNA，参与生命过程中一系列重要进程，它可能是朊病毒致病的重要协助因子，它的研究给朊病毒病的发病机理带来了希望。本课题建立朊病毒病模型小鼠脑部LncRNA和mRNA表达谱，并通过荧光定量PCR进行了验证，最终检测出6条LncRNA表达量相比对照组下调，13条LncRNA表达量相比对照组上调；建立了小鼠海马组织中mRNA芯片，共检测出170条差异表达的mRNA分子，其中25条mRNA表达下调，145条miRNA表达上调。通过生物信息学分析预测，得到差异性表达LncRNA作用的靶基因以及靶基因的功能和参与细胞信号转导通路。荧光定量PCR分析得知过表达LncRNA-2对Prnp基因并没有显著影响，Western Blotting检测ScN2a细胞内PrPSc含量说明，过表达LnRNA-2后能够抑制PrPSc蛋白的复制。为后续研究长链非编码RNA以及朊病毒相关分子伴侣提供了一个理论的支持，奠定了基础。