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MicroRNA regulation in blood cells of renal transplanted patients with interstitial fibrosis/tubular atrophy and antibody-mediated rejection

机译:肾移植患者间质纤维化/肾小管萎缩和抗体介导的排斥反应中血细胞中的microRNA调节

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摘要

Interstitial fibrosis/tubular atrophy (IFTA) is associated with reduced allograft survival, whereas antibody-mediated rejection (ABMR) is the major cause for renal allograft failure. To identify specific microRNAs and their regulation involved in these processes, total RNA from blood cells of 16 kidney transplanted (KTx) patients with ABMR, stable graft function (SGF) and with T-cell mediated rejection (TCMR) was isolated. MicroRNA expression was determined by high-throughput sequencing. Differentially expressed candidate microRNAs were analyzed with RT-PCR in patients with SGF (n = 53), urinary tract infection (UTI) (n = 17), borderline rejection (BL) (n = 19), TCMR (n = 40), ABMR (n = 22) and IFTA (n = 30). From the 301 detected microRNAs, 64 were significantly regulated between the three cohorts. Selected candidate microRNAs miR-223-3p, miR-424-3p and miR-145-5p distinguished TCMR and ABMR from SGF, but not from other pathologies. Most importantly, miR-145-5p expression in IFTA patients was significantly downregulated and displayed a high diagnostic accuracy compared to SGF alone (AUC = 0.891) and compared to SGF, UTI, BL, TCMR and ABMR patients combined (AUC = 0.835), which was verified by cross-validation. The identification of miR-145-5p as IFTA specific marker in blood constitutes the basis for evaluating this potentially diagnostic microRNA as biomarker in studies including high numbers of patients and different pathologies and also the further analysis of fibrosis causing etiologies after kidney transplantation.
机译:间质纤维化/肾小管萎缩(IFTA)与同种异体移植物的存活减少有关,而抗体介导的排斥反应(ABMR)是同种异体肾移植失败的主要原因。为了鉴定特定的microRNA及其参与这些过程的调控,从16例具有ABMR,稳定移植功能(SGF)和T细胞介导排斥(TCMR)的肾移植(KTx)患者的血细胞中分离了总RNA。通过高通量测序确定MicroRNA表达。应用RT-PCR对SGF(n = 53),尿路感染(UTI)(n = 17),边界排斥(BL)(n = 19),TCMR(n = 40), ABMR(n = 22)和IFTA(n = 30)。在这301个检测到的microRNA中,三个队列之间有64个被显着调节。选定的候选microRNA miR-223-3p,miR-424-3p和miR-145-5p将TCMR和ABMR与SGF区别开来,但与其他病理却没有区别。最重要的是,与单独使用SGF(AUC = 0.891)以及与SGF,UTI,BL,TCMR和ABMR合并患者(AUC = 0.835)相比,IFTA患者中的miR-145-5p表达显着下调并显示出较高的诊断准确性,通过交叉验证进行了验证。将miR-145-5p识别为血液中的IFTA特异性标志物,是在包括大量患者和不同病理学以及肾脏移植后进一步分析引起纤维化的病因的研究中评估这种具有潜在诊断意义的microRNA作为生物标志物的基础。

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