Faculty of Medical Sciences, Translational and Clinical Research Institute, Newcastle University, Newcastle upon Tyne, United Kingdom.
Department of Haematology and Oncology, University of Regensburg, Regensburg, Germany.
Front Immunol. 2021 Mar 15;12:639171. doi: 10.3389/fimmu.2021.639171. eCollection 2021.
Acute graft vs. host disease (aGvHD) is a frequent complication following allogeneic haematopoeitic transplantation (HSCT). Despite recent advances, there are no universally accepted biomarkers to determine development of aGvHD. MicroRNAs miR-146a and miR-155 have been previously associated with aGvHD and show promise as clinically translatable biomarkers. In this study, we performed comprehensive expression profiling of miR-146a, miR-155, and miR-155 expression in aGvHD target tissue and biofluids and relate expression to post-HSCT outcomes. MicroRNA expression was assessed by qRT-PCR in gastrointestinal ( = 31) and skin ( = 31) biopsies as well as serum (exploratory cohort = 34, verification cohort = 81, diagnostic cohort = 65) and urine (exploratory cohort = 30, verification cohort = 56, diagnostic cohort = 20) biofluids, including extracellular vesicle (EV) cohorts (serum EV = 15, urine EV = 30). Expression was related to aGvHD incidence, severity and overall survival. In GI samples, expression of miR-155 ( = 0.03) and miR-146a ( = 0.03) was higher at aGvHD onset compared to patients with no GvHD. In skin biopsies, expression of miR-155 ( = 0.004) was upregulated in aGvHD patients compared to normal control skin. Expression of miR-146a was higher in aGvHD compared to no aGvHD biopsies ( = 0.002). In serum, miR-155 ( = 0.03) and miR-146a ( = 0.02) expression was higher at day 14 (D14), while in urine expression was elevated at D7 post-HSCT in patients who developed aGvHD compared to those disease-free. This was verified in an independent serum (miR-155 = 0.005, miR-146a = 0.003) and urine (miR-155 = 0.02, miR-146a = 0.04) cohort, where both microRNAs were also associated with aGvHD by ROC analysis. In serum and urine samples taken at the time of aGvHD symptoms, expression of miR-155 and miR-146a was also elevated (serum miR-155 = 0.03, miR-146a < 0.001; urine miR-155 = 0.02, miR-146a = 0.02). In contrast, miR-146a and miR-155 were downregulated at D14 in serum EVs and at D7 in urine EVs in patients who developed aGvHD compared to those that remained disease-free, in both an exploratory (serum miR-155 = 0.02, miR-146a = 0.06; urine miR-155 = 0.02, miR-146a = 0.07) and an independent cohort (serum miR-155 = 0.01, miR-146a = 0.02). These results further support a role for miR-155 and miR-146a as non-invasive, clinically relevant biomarkers for aGvHD. However, the link between their involvement in generalized inflammation and in specific pathophysiology requires further investigation at a systemic level.
急性移植物抗宿主病（aGvHD）是异基因造血干细胞移植（HSCT）后常见的并发症。尽管最近取得了进展，但尚无普遍接受的生物标志物来确定 aGvHD 的发展。miR-146a 和 miR-155 以前与 aGvHD 相关，并有望成为具有临床转化价值的生物标志物。在这项研究中，我们对 aGvHD 靶组织和生物流体中的 miR-146a、miR-155 表达进行了全面的表达谱分析，并将表达与 HSCT 后结果相关联。通过 qRT-PCR 评估了胃肠（= 31）和皮肤（= 31）活检以及血清（探索性队列= 34、验证性队列= 81、诊断性队列= 65）和尿液（探索性队列= 30、验证性队列= 56、诊断性队列= 20）生物流体中的 microRNA 表达，包括细胞外囊泡（EV）队列（血清 EV= 15、尿液 EV= 30）。表达与 aGvHD 的发生率、严重程度和总生存相关。在 GI 样本中，与无 GvHD 患者相比，aGvHD 发病时 miR-155（= 0.03）和 miR-146a（= 0.03）的表达更高。在皮肤活检中，与正常对照皮肤相比，aGvHD 患者的 miR-155（= 0.004）表达上调。与无 aGvHD 活检相比，miR-146a 在 aGvHD 中表达更高（= 0.002）。在血清中，miR-155（= 0.03）和 miR-146a（= 0.02）在第 14 天（D14）表达更高，而在尿液中，在 HSCT 后第 7 天（D7），与无疾病患者相比，发生 aGvHD 的患者的表达升高。在一个独立的血清（miR-155= 0.005、miR-146a= 0.003）和尿液（miR-155= 0.02、miR-146a= 0.04）队列中得到了验证，通过 ROC 分析，这两种 microRNA 也与 aGvHD 相关。在出现 aGvHD 症状时采集的血清和尿液样本中，miR-155 和 miR-146a 的表达也升高（血清 miR-155= 0.03，miR-146a<0.001；尿液 miR-155= 0.02，miR-146a= 0.02）。相比之下，与无疾病患者相比，在发生 aGvHD 的患者中，血清 EV 中 miR-155 和 miR-146a 在 D14 下调，尿液 EV 中 miR-155 和 miR-146a 在 D7 下调，在探索性（血清 miR-155= 0.02，miR-146a= 0.06；尿液 miR-155= 0.02，miR-146a= 0.07）和独立队列（血清 miR-155= 0.01，miR-146a= 0.02）中均如此。这些结果进一步支持 miR-155 和 miR-146a 作为非侵入性、临床相关的 aGvHD 生物标志物的作用。然而，它们在全身性炎症和特定病理生理学中的参与之间的联系需要在系统水平上进一步研究。
