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尿沉渣miR-150-5p在IgA肾病无创诊断中的价值

Value of urinary sediment miR-150-5p in noninvasive diagnosis of IgA nephropathy

    摘要
  • 摘要:
      背景  IgA肾病(IgA nephropathy，IgAN)是一种发病率高且呈进展性的疾病，现主要确诊手段为肾穿刺活检，临床诊疗中需要研究无创且具有特异性的IgAN诊断标志物。
      目的  检测IgAN患者尿沉渣miR-150-5p表达水平并评估其诊断IgAN的价值。
      方法  根据已经发表的3项有关IgAN尿沉渣miRNA芯片的研究数据，筛选IgAN患者与正常人群之间差异性表达的miRNA，最终确定miR-150-5p在IgAN患者与正常人群中表达差异显著且趋势一致，是正常人群的(30.22 ~ 598.34) 倍。为了验证芯片筛选的准确性，选取解放军总医院第一医学中心2018年10月- 2019年2月的30例IgAN患者和30例正常人进行了小样本RT-PCR验证。为进一步明确其是否具有疾病特异性，选取解放军总医院第一医学中心2019年3月- 2022年5月的144例IgAN患者、84例其他肾病患者60例膜性肾病(membranous nephropathy，MN)、8例微小病变型肾病(minimal change disease，MCD)、16例局灶性节段性肾小球硬化(focal segmental glomerular sclerosis，FSGS)、67例正常人进行了较大样本的验证，比较miR-150-5p在IgAN组与不同疾病对照组之间的差异表达。绘制受试者工作特征(receiver operating characteristic，ROC)曲线评估差异性表达miRNA在无创诊断IgAN中的价值。
      结果  在小样本验证研究中，RT-PCR实验显示IgAN患者尿沉渣miR-150-5p与正常对照组相比显著升高Md(IQR)：25.632(5.868 ~ 61.523) vs 0.557(0.218 ~ 1.258)，P<0.0001，与文献中芯片测量结果基本一致。在加入疾病对照组的较大样本研究中，IgAN组miR-150-5p表达水平显著高于疾病对照组Md(IQR)：12.606(3.939 ~ 34.431) vs 2.808(1.420 ~ 9.616)，P<0.0001。miR-150-5p在IgAN组中表达水平较MN组和 FSGS组高(P<0.001)，与MCD组之间差异无统计学意义(P=0.613)。miR-150-5p诊断IgAN的ROC曲线下面积(area under curve，AUC)为0.844，95% CI为0.804 ~ 0.885，最佳截断值为2.505，敏感度71.51%，特异度84.48%。
      结论  miR-150-5p或可作为无创诊断IgAN的潜在标志物。

     

    Abstract:
      Background  IgA nephropathy (IgAN) is a progressive disease with high incidence. The main diagnostic method is renal puncture biopsy. Non-invasive and specific diagnostic markers of IgAN need to be studied in clinical practice.
      Objective  To detect the expression level of miR-150-5p in urine sediment of IgAN patients and evaluate its value in the diagnosis of IgAN.
      Methods  According to the data of three published studies on IgAN urinary sediment miRNAs profiles, the commonnly and differentially expressed miRNA between IgAN and healthy people were screened. miR-150-5p was selected because its expression in urine sediment was significantly higher in the IgAN patients than that in the normal controls (30.22-598.34 fold-changes). In order to verify the accuracy of chip screening results, 30 IgAN patients and 30 normal controls in the First Medical Center of Chinese PLA General Hospital from October 2018 to February 2019 were selected for RT-PCR verification. In order to further clarify whether it was disease-specific, 144 patients with IgAN, 84 patients with other nephropathy (60 patients with membranous nephropathy MN, 8 patients with minimal change disease MCD, 16 patients with focal segmental glomerular sclerosis FSGS) and 67 normal controls in the First Medical Center of Chinese PLA General Hospital from March 2019 to May 2022 were selected for experimental verification with larger samples, and the differential expression of miRNA between IgAN and control groups with different diseases was compared. Receiver operating characteristic curve (ROC curve) was plotted to evaluate the value of miR-150-5p in non-invasive diagnosis of IgAN.
      Results  In the small sample validation study, RT-PCR assay showed that miR-150-5p in urine sediment of the IgAN patients was significantly higher than that of the normal controls (Md IQR: 25.632 5.868-61.523 vs 0.557 0.218-1.258, P<0.0001), which was consistent with the results of the profiles in the literature. In the larger sample study including disease control group, the expression level of miR-150-5p in the IgAN group was significantly higher than that in the disease control group (Md IQR: 12.606 3.939-34.431 vs 2.808 1.420-9.616, P<0.0001). The expression level of miR-150-5p in IgAN was higher than that in MN and FSGS (P <0.001), and there was no significant difference between IgAN and MCD (P=0.613). The area under the ROC curve (AUC) for the diagnosis of IgAN by miR-150-5p was 0.844 (95%CI: 0.804-0.885), the optimal cut-off value was 2.505, with the sensitivity of 71.51%, and the specificity of 84.48%.
      Conclusion  miR-150-5p may be a potential marker for non-invasive diagnosis of IgAN.

     

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