Elevated pulmonary tuberculosis biomarker miR-423-5p plays critical role in the occurrence of active TB by inhibiting autophagosome-lysosome fusion
Elevated pulmonary tuberculosis biomarker miR-423-5p plays critical role in the occurrence of active TB by inhibiting autophagosome-lysosome fusion
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肺结核生物标志物miR-423-5p升高，通过抑制自噬体-溶酶体融合，在活动性结核病的发生中发挥关键作用
DOI：
10.1080/22221751.2019.1590129
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发表时间：
2019-03-22
影响因子：
13.2
通讯作者：
Li, Jicheng
中科院分区：
文献类型：
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作者：
Tu, Huihui;Yang, Su;Li, Jicheng
摘要快速诊断肺结核是预防肺结核传播的有效措施。然而，严峻的事实是缺乏新的、快速的、安全的临床诊断方法。此外，尽管自身溶酶体在清除结核分枝杆菌中起着至关重要的作用，但作为结核病生物标志物的microrna在自噬体成熟中的病理意义尚不清楚。本文通过Solexa测序和qPCR验证对这些microrna进行了研究，并通过逻辑回归建立了潜在的诊断模型。此外，还研究了其中一个参与结核病发生的microrna的机制。结果显示，结核患者血清中miR-423-5p、miR-17-5p、miR-20b-5p的表达明显升高。这三种microrna联合建立的模型诊断结核病的准确率为78.18%，曲线下面积值为0.908。生物信息学分析揭示了miR-423-5p是调控自噬体成熟的最有可能的候选者。上调miR-423-5p可通过抑制巨噬细胞中自噬体-溶酶体融合来抑制自噬体成熟。进一步研究发现，VPS33A是miR-423-5p的直接靶点，VPS33A 3′-UTR中的两个CUGCCCCUC结构域是miR-423-5p的直接调控位点。此外，在结核病患者外周血单个核细胞中发现VPS33A与miR-423-5p呈负相关。由于抑制自噬酶体的形成在结核病的发生中起着至关重要的作用，我们的研究结果表明，miR-423-5p可以通过转录后调控VPS33A抑制自噬体-溶酶体的融合，这可能是活动性结核病发生的重要原因。
ABSTRACT Rapid diagnosis of pulmonary tuberculosis is an effective measure to prevent the spread of tuberculosis. However, the grim fact is that the new, rapid, and safe methods for clinical diagnosis are lacking. Moreover, although auto-lysosome is critical in clearing Mycobacterium tuberculosis, the pathological significance of microRNAs, as biomarkers of tuberculosis, in autophagosome maturation is unclear. Here, these microRNAs were investigated by Solexa sequencing and qPCR validation, and a potential diagnostic model was established by logistic regression. Besides that, the mechanism of one of the microRNAs involved in the occurrence of tuberculosis was studied. The results showed that the expression of miR-423-5p, miR-17-5p, and miR-20b-5p were significantly increased in the serum of patients with tuberculosis. The combination of these three microRNAs established a model to diagnose tuberculosis with an accuracy of 78.18%, and an area under the curve value of 0.908. Bioinformatics analysis unveiled miR-423-5p as the most likely candidate in regulating autophagosome maturation. The up-regulation of miR-423-5p could inhibit autophagosome maturation through suppressing autophagosome–lysosome fusion in macrophages. Further investigations showed that VPS33A was the direct target of miR-423-5p, and the two CUGCCCCUC domains in VPS33A 3’-UTR were the direct regulatory sites for miR-423-5p. In addition, an inverse correlation between VPS33A and miR-423-5p was found in peripheral blood mononuclear cells of patients with tuberculosis. Since the inhibition of autolysosome formation plays a critical role in tuberculosis occurrence, our findings suggests that miR-423-5p could suppress autophagosome–lysosome fusion by post-transcriptional regulation of VPS33A, which might be important for the occurrence of active tuberculosis.