METHODS: We conducted a cross-sectional study recruiting subjects diagnosed with active-TB (drug-sensitive and drug-resistant) and healthy controls. Sera samples were collected and utilized for preparing small RNA libraries. Quantitative patterns of circulating sncRNAs (miRNAs, piRNAs and tRFs) were identified via high-throughput sequencing and DeSeq2 analysis and validated in independent active-TB cohorts. Functional knockdown for two selected miRNAs were also performed.
RESULTS: A diagnostic signature of four sncRNAs for both drug-sensitive and drug-resistant active-TB cases was validated, exhibiting an AUC of 0.96 (95% CI: 0.937-0.996, p < 0.001) with 86.7% sensitivity (95% CI: 0.775-0.932) and 91.7% specificity (95% CI: 0.730-0.990) in ROC analysis. Functional knockdown demonstrated regulatory roles of hsa-miR-223-5p and hsa-miR-10b-5p in Mycobacterium tuberculosis (Mtb) growth and pro-inflammatory cytokine expression (IL-6 and IL-8).
CONCLUSIONS: The study identified a diagnostic tool utilizing a signature of four sncRNAs with high specificity and sensitivity, enhancing our understanding of sncRNAs as ATB diagnostic biomarker. Additionally, hsa-miR-223-5p and hsa-miR-10b-5p demonstrated potential roles in Mtb pathogenesis and host-response to infection.
方法:我们进行了一项横断面研究,招募了被诊断为活动性TB(药物敏感性和耐药性)和健康对照的受试者。收集血清样品并用于制备小RNA文库。循环sncRNAs的定量模式(miRNAs,piRNA和tRF)通过高通量测序和DeSeq2分析进行鉴定,并在独立的活动性TB队列中进行验证。还进行了两种选择的miRNA的功能敲低。
结果:对药物敏感和耐药的活动性TB病例的4种sncRNA的诊断特征进行了验证,在ROC分析中表现出0.96的AUC(95%CI:0.937-0.996,p<0.001),86.7%的灵敏度(95%CI:0.775-0.932)和91.7%的特异性(95%CI:0.730-0.990)。功能性敲除证明了hsa-miR-223-5p和hsa-miR-10b-5p在结核分枝杆菌(Mtb)生长和促炎细胞因子表达(IL-6和IL-8)中的调节作用。
结论:本研究确定了一种诊断工具,该工具利用了4种具有高特异性和敏感性的sncRNAs。增强我们对sncRNAs作为ATB诊断生物标志物的理解。此外,hsa-miR-223-5p和hsa-miR-10b-5p在Mtb发病机制和宿主对感染的反应中显示出潜在的作用。