单核细胞lncRNA谱导致结核潜伏感染复发的分子机制及预测复发风险的临床评估

Ten percent of latent Mycobacterium tuberculosis (Mtb) infected population develops into ATBI in their lifetime, which is the core of prevention and final control of tuberculosis. However, no early diagnostic tool is available to identify latent infection with high risk of relapse. Predominant proteins expressed by Mtb are under modification during the development of disease. Monocyte derived macrophages play a crucial role in the control and elimination of invading TB, and also serve as the major residence for Mtb. The interaction between macrophages and predominant proteins of Mtb determines the LTBI outcomes. However, the macrophage presenting Mtb antigens in LTBI usually located in the inaccessible infected sites. Our primary data revealed there were 12 lncRNAs upregulated in both LTBI and ATBI. The increased lncRNAs indicated the role of lncRNA-PACER, THRIL, MacORIS in INF and TNFα pathways that implied a risk of LTBI relapse..We hypothesis that lncRNA expression profile play an important role in LTBI relapse through modulating macrophage functions, and the up-regulated lncRNA profile can serve as the biomarkers to predict the reactive risk of LTBI. In current study, to explore the modulation mechanism of lncRNA on macrophage functions, the lncRNA over-expressed or silenced macrophage will be established to explore the transcriptional modulation of lncRNA on innate immune response. The LTBI cohort will be studied in the North, West and East China. The value of lncRNA transcription profiles to predicate the LTBI reactivated risk will be valid in independent sample settings from multi-centre cooperation. This study will elucidate the molecular regulation of lncRNA and character the innate immune response in LTBI relapse, which lays a solid foundation for the development of diagnostic kit of predication of relapse risk of latent tuberculosis infection.

结核潜伏感染（LTBI）人群中约10%可发展为活动性结核（ATBI），是结核防治重点。目前缺乏有效预测LTBI复发的风险指标。我们研究发现LTBI和ATBI单核细胞共同高表达12种lncRNA，其中lncRNA-PACER、THRIL和MacORIS与干扰素和肿瘤坏死因子信号通路相关；相关报道也证实LTBI与ATBI存在共表达的基因谱。此研究结果提示：LTBI和ATBI单核巨噬细胞存在共同的lncRNA转录调控事件并参与LTBI复发，但机制不详；同时，lncRNA有望成为早期预测复发风险的标志物，但未见报道。本研究拟通过构建慢病毒介导的lncRNA沉默/过表达体系，检测lncRNA调控的单核巨噬细胞表型和功能，探讨lncRNA转录调控宿主免疫应答机制；并采用多中心独立人群联合研究，验证三种lncRNA在LTBI复发风险预测中的价值，为研发LTBI复发风险诊断试剂盒奠定理论基础。

因结核潜伏感染复发过程的隐匿性和长期性，是结核传播的主要传染源。早期发现和治疗复发高风险LTBI患者是结核防控的重点。 单核巨噬细胞lncRNA转录调控改变是LTBI复发的关键环节；高表达的lncRNA-MacORIS在转录水平调控单核巨噬细胞的分化及功能是导致LTBI复发的主要因素。本研究利用慢病毒介导的lncRNA过表达/沉默技术，转染单核巨噬细胞，结合流式细胞术和Luminex技术，鉴定单核巨噬细胞的分化和功能，探讨了MacORIS介导LTBI临床转归的分子机制。同时，采用不同人群的联合研究，进一步阐明ATBI和LTBI人群不同的免疫特征，以及IL-10在卡介苗临床应用中的作用。