Lnc-49a调控人蜕膜NK细胞的独特表型及功能

Human decidual NK (dNK) cells are functionally and phenotypically different from peripheral blood NK cells (pNK). However, little is known about the underlying mechanisms. In our preliminary data, we identified a novel lncRNA that has never been reported to-lnc-49a, overexpressed in dNK cells but down regulated in human pNK cells. In addition, we found that lnc-49a siRNA inhibited the expression of CD49a but promoted the cytotoxicity of human dNK cells. So, we hypothesized that lnc-49a plays critical role in the regulation of human NK cell phenotype and function. To test this hypothesis, we will conduct a series of experiments as follow: we plan to use the lentivirus vector systems to investigate the regulatory roles of lnc-49a in human NK cell function. Moreover, we would use transcriptomes microarray/RNA pulldown/ FACS and Western blot to investigate the possible molecular and intracellular signaling mechanisms which were related to NK cell function. We would also use ChIP-Seq/ChIP-qPCR to uncover the upstream regulation mechanisms for lnc-49a differential expressions in various human NK cell subsets. The success of this project will provide a new mechanistic explanation for regulation of NK cell function by lncRNAs and ultimately implicate a potential application of NK cells in clinical immunotherapy.

人外周血循环NK细胞（pNK）与蜕膜组织居留NK细胞（dNK）在表型及功能上存在很大差异，但机制不清。前期通过lncRNA芯片发现一个新的lncRNA（生物信息学预测其靶向CD49a，将其命名为lnc-49a），特异性高表达于dNK细胞，且干扰lnc-49a可显著下调dNK特征性表面分子 CD49a的表达，并促进dNK杀伤能力。推测lnc-49a在维持人dNK细胞特有表型及功能中起重要作用。为验证这一假设，本课题将构建lnc-49a上调和下调慢病毒载体，利用原代分离的pNK、dNK细胞及人NK细胞体外分化体系，观察lnc-49a对NK细胞表型及功能的影响；采用转录组芯片、RNA pulldown等方法阐明lnc-49a调控NK细胞功能的分子机制；通过ChIP-Seq/ChIP-qPCR等表观遗传学分析，解析 lnc-49a在NK细胞中差异表达的上游调控机制，为NK细胞的免疫治疗提供新思路。

人外周血循环NK细胞（pNK）与蜕膜组织居留NK细胞（dNK）在表型及功能上存在很大 差异，但机制不清。在本项目中，通过lncRNA芯片发现一个新的lncRNA（生物信息学预测其靶向CD 49a，将其命名为lnc-49a），特异性高表达于dNK细胞，且干扰lnc-49a可显著下调dNK特征性表面分子CD49a的表达，并促进dNK杀伤能力。此外，本项目深入研究了CD49a在dNK细胞中的功能，发现CD49a对dNK的细胞因子分泌、黏附迁移等具有重要调控作用。进一步采用RNA pulldown等方法阐明lnc-49a调控NK细胞功能的分子机制。.总之，本项目研究结果揭示了lnc49a以及CD49a在人类蜕膜NK细胞中起重要作用，lnc49a可能作为调节NK细胞效应功能的重要靶点。