RNA sequencing identified specific circulating miRNA biomarkers for early detection of diabetes retinopathy
RNA sequencing identified specific circulating miRNA biomarkers for early detection of diabetes retinopathy
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DOI：
10.1152/ajpendo.00021.2018
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发表时间：
2018-09-01
影响因子：
5.1
通讯作者：
Zhai, Ri H.
中科院分区：
文献类型：
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作者：
Liang, Zhen;Gao, Kai P.;Zhai, Ri H.
糖尿病视网膜病变(DR)是糖尿病患者致盲的主要原因。然而，用于早期检测DR的生物标记物仍然缺乏。MicroRNAs(MiRNAs)调节多种生物学功能，在糖尿病患者中常被解除调节。我们的目的是探讨循环中的miRNAs是否可以作为早期糖尿病患者的生物标志物。我们用RNA-seq和qRT-PCR方法鉴定了2型糖尿病合并DR患者(T2 DM-DR)、T2 DM非DR患者(T2 DM-NO-DR)和健康对照组血清中不同的miRNAs。我们使用qRT-PCR方法验证了两个阶段的差异循环miRNAs。RNA-seq分析鉴定出7个T2 DM-DR和T2 DM-NO-DR之间的差异循环miRNAs，以及47个T2 DM-DR和健康人之间的差异miRNAs。两阶段分析证实5个血清miRNAs(hsa-let-7a-5p、hsa-miR-new-chr5_15976、hsa-miR-28-3p、has-miR-151a-5p、has-miR-148a-3p)与T2 DM-DR显著相关。受试者-操作者特征分析显示，hsa-let-7a-5p、hsa-miR-new-chr5_15976和hsa-miR-28-3p对T2 DM-DR和T2 DM-no-DR的鉴别敏感性和特异性分别为0.92和0.94，对早期T2 DM-DR和晚期DR的敏感性和特异性分别为0.93和0.86。慢病毒介导的hsa-let-7a-5p在人视网膜微血管内皮细胞(HRMECs)中过表达可显著提高HRMECs的增殖率。综上所述，血清中的3-miRNA特征可作为DR的非侵入性诊断生物标志物。此外，我们还表明DR相关的miRNAs可能至少部分地通过改变HRMEC的增殖参与了DR的发病。
Diabetic retinopathy (DR) is the leading cause of blindness in patients with diabetes. However, biomarkers for early detection of DR are still lacking. MicroRNAs (miRNAs) regulate multiple biological functions and are often deregulated in DR. We aimed to investigate whether circulating miRNAs can be used as biomarkers of early-stage DR. We used RNA-seq and qRT-PCR to identify differential serum miRNAs in patients with type 2 diabetes mellitus with DR (T2DM-DR), T2DM without DR (T2DM-no-DR), and healthy controls. We validated differential circulating miRNAs in two phases using qRT-PCR assays. RNA-seq analysis identified 7 differential circulating miRNAs between T2DM-DR and T2DM-no-DR and 47 differential miRNAs between T2DM-DR and healthy subjects. Two-stage analysis verified that a profile of five serum miRNAs (hsa-let-7a-5p, hsa-miR-novel-chr5_15976, hsa-miR-28-3p, has-miR-151a-5p, has-miR-148a-3p) was significantly associated with T2DM-DR. Receiver-operator-characteristic analyses showed that a panel of three miRNAs (hsa-let-7a-5p, hsa-miR-novel-chr5_15976, and hsa-miR-28-3p) presented 0.92 sensitivity and 0.94 specificity for distinguishing T2DM-DR from T2DM-no-DR, and 0.93 sensitivity and 0.86 specificity for differentiating early-stage T2DM-DR (NPDR) from late-stage DR (PDR). Lentivirus-mediated overexpression of hsa-let-7a-5p in human retinal microvascular endothelial cells (HRMECs) significantly promoted proliferation rates of HRMECs. In conclusion, the three-miRNA signature from serum may serve as a noninvasive diagnostic biomarker for DR. Furthermore, we showed that DR-associated miRNAs may be involved in the pathogenesis of DR, at least in part, through modifying proliferation of HRMECs.