Department of Microbiology and Molecular Biology, ICMR-National JALMA Institute for Leprosy and Other Mycobacterial Diseases, Agra, Uttar Pradesh, India.
Department of Tuberculosis and Chest, Sarojini Naidu Medical College, Agra, Uttar Pradesh, India.
Indian J Med Res. 2024 Jun;159(6):653-662. doi: 10.25259/IJMR_1281_23.
Background & objectives Despite the evidence of population differences in miRNA expression, limited information is available about the expression profile of miRNAs in Indian tuberculosis (TB) patients. The present study aimed to investigate the expression profile of candidate serum exosomal microRNAs in Indian patients with and without HIV-TB coinfection. Methods The pool samples of serum exosomes of study participants (HIV-TB coinfection, extra-pulmonary TB, HIV mono-infection, pulmonary TB) and healthy humans were processed for the isolation of total RNA followed by miRNA analysis using miRCURY LNA human focus PCR panel by real-time PCR. The significantly altered miRNAs were identified using differential expression analysis. The target genes prediction and potential functional analysis of exclusively differentially expressed miRNAs were performed using bioinformatics tools. Results The expression profile of 57, 58, 49 and 11 miRNAs was significantly altered in exosome samples of HIV-TB coinfected, extra-pulmonary TB, HIV mono-infected and pulmonary TB patients compared to healthy controls, respectively. The set of three (hsa-let-7i-5p, hsa-miR-24-3p, hsa-miR-92a-3p), three (hsa-miR-20a-5p, hsa-let-7e-5p, hsa-miR-26a-5p) and four (hsa-miR-21-5p, hsa-miR-19a-3p, hsa-miR-19b-3p, hsa-miR-146a-5p) miRNAs were exclusively significantly differentially expressed in study participants with HIV-TB coinfection, extra-pulmonary TB and pulmonary TB, respectively. Most of the target genes of exclusively differentially expressed miRNAs were enriched in pathways in cancer, MAPK signalling pathway and Ras signalling pathway. Interpretation & conclusions The present study demonstrates a distinct expression profile of miRNAs in serum exosomes of the study participants and identified crucial miRNAs which may have a significant impact on the biomarker analysis and pathogenesis of TB in Indian patients.
背景与目的 尽管有证据表明 miRNA 表达存在人群差异，但有关印度结核病（TB）患者中 miRNA 表达谱的信息有限。本研究旨在探讨印度 HIV-TB 合并感染、肺外结核、HIV 单一感染和肺结核患者血清外泌体候选 miRNA 的表达谱。
方法 对研究参与者（HIV-TB 合并感染、肺外结核、HIV 单一感染、肺结核）和健康人的血清外泌体混合样本进行处理，以分离总 RNA，然后使用实时 PCR 通过 miRCURY LNA 人类焦点 PCR 面板进行 miRNA 分析。使用差异表达分析鉴定显著改变的 miRNA。使用生物信息学工具对唯一差异表达 miRNA 的靶基因预测和潜在功能分析。
结果 与健康对照组相比，HIV-TB 合并感染、肺外结核、HIV 单一感染和肺结核患者的血清外泌体样本中，分别有 57、58、49 和 11 个 miRNA 的表达谱显著改变。在 HIV-TB 合并感染、肺外结核和肺结核患者中，有三个（hsa-let-7i-5p、hsa-miR-24-3p、hsa-miR-92a-3p）、三个（hsa-miR-20a-5p、hsa-let-7e-5p、hsa-miR-26a-5p）和四个（hsa-miR-21-5p、hsa-miR-19a-3p、hsa-miR-19b-3p、hsa-miR-146a-5p）miRNA 是唯一显著差异表达的。大多数唯一差异表达 miRNA 的靶基因富集在癌症途径、MAPK 信号通路和 Ras 信号通路中。
结论 本研究表明，印度研究参与者血清外泌体中的 miRNA 表达谱存在明显差异，并鉴定了可能对印度 TB 患者的生物标志物分析和发病机制有重大影响的关键 miRNA。
