CD31+ Extracellular Vesicles From Patients With Type 2 Diabetes Shuttle a miRNA Signature Associated With Cardiovascular Complications
CD31+ Extracellular Vesicles From Patients With Type 2 Diabetes Shuttle a miRNA Signature Associated With Cardiovascular Complications
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DOI：
10.2337/db20-0199
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发表时间：
2021-01-01
期刊：
影响因子：
7.7
通讯作者：
Ceriello, Antonio
中科院分区：
文献类型：
--
作者：
Prattichizzo, Francesco;De Nigris, Valeria;Ceriello, Antonio
需要创新的生物标志物来改善2型糖尿病（T2 DM）患者的管理。血液循环miRNAs已被提出作为检测T2 DM并发症的潜在工具，但缺乏组织特异性等原因阻碍了它们向临床环境的转化。已经提出细胞外囊泡（EV）穿梭的miRNA作为替代方法。在这里，我们采用了一种基于免疫磁珠的方法来分离血浆CD 31（+）EV，以收获来自与T2 DM并发症相关的组织的囊泡。表面标志物表征显示，CD 31（+）EV对血小板和活化内皮细胞的一系列典型标志物也呈阳性。在表征之后，我们定量了11种与血管性能相关的候选miRNAs，并在一个大型（n = 218）横断面队列中被CD 31（+）EV穿梭，这些患者被分类为无并发症的T2 DM患者、有并发症的T2 DM患者和对照受试者。我们发现，10种检测的miRNA受T2 DM影响，而由miR-146 a、-320a、-422a和-451a组成的签名有效地鉴定了具有并发症的T2 DM患者。此外，另一个CD 31（+）EV穿梭的miRNA标签，即，miR-155、-320a、-342-3p、-376和-422a检测了既往有重大不良心血管事件的T2 DM患者。这些miRNAs中的许多与临床变量显著相关，这些临床变量在并发症的发展中发挥关键作用。此外，我们发现来自T2 DM患者的CD 31（+）EV能够促进选定的炎性mRNA的表达，即，CCL 2、IL-1 α和TNF α，当体外给予内皮细胞时。总体而言，这些数据表明，血浆CD 31（+）EV的miRNA货物在很大程度上受到T2 DM和相关并发症的影响，鼓励进一步研究探索这些改变的诊断潜力和功能作用。
Innovative biomarkers are needed to improve the management of patients with type 2 diabetes mellitus (T2DM). Blood circulating miRNAs have been proposed as a potential tool to detect T2DM complications, but the lack of tissue specificity, among other reasons, has hampered their translation to clinical settings. Extracellular vesicle (EV)-shuttled miRNAs have been proposed as an alternative approach. Here, we adapted an immunomagnetic bead-based method to isolate plasma CD31(+) EVs to harvest vesicles deriving from tissues relevant for T2DM complications. Surface marker characterization showed that CD31(+) EVs were also positive for a range of markers typical of both platelets and activated endothelial cells. After characterization, we quantified 11 candidate miRNAs associated with vascular performance and shuttled by CD31(+) EVs in a large (n = 218) cross-sectional cohort of patients categorized as having T2DM without complications, having T2DM with complications, and control subjects. We found that 10 of the tested miRNAs are affected by T2DM, while the signature composed by miR-146a, -320a, -422a, and -451a efficiently identified T2DM patients with complications. Furthermore, another CD31(+) EV-shuttled miRNA signature, i.e., miR-155, -320a, -342-3p, -376, and -422a, detected T2DM patients with a previous major adverse cardiovascular event. Many of these miRNAs significantly correlate with clinical variables held to play a key role in the development of complications. In addition, we show that CD31(+) EVs from patients with T2DM are able to promote the expression of selected inflammatory mRNAs, i.e., CCL2, IL-1 alpha, and TNF alpha, when administered to endothelial cells in vitro. Overall, these data suggest that the miRNA cargo of plasma CD31(+) EVs is largely affected by T2DM and related complications, encouraging further research to explore the diagnostic potential and the functional role of these alterations.