Identifying quantitative sncRNAs signature using global sequencing as a potential biomarker for tuberculosis diagnosis and their role in regulating host response

Objectives: The study aimed to identify a quantitative signature of circulating small non -coding RNAs (sncRNAs) as a biomarker for pulmonary tuberculosis disease (active-TB/ATB) and explore their regulatory roles in hostpathogen interactions and disease progression. 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). Conclusion: 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.