Medical Clinics I-Cardiology and Angiology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Department of Cardiology, Kerckhoff Clinic GmbH, 61231 Bad Nauheim, Germany.
Biomolecules. 2019 Oct 29;9(11):666. doi: 10.3390/biom9110666.
Extracellular vesicles are released by numerous cell types of the human body under physiological but also under pathophysiological conditions. They are important for cell-cell communication and carry specific signatures of peptides and RNAs. In this study, we aimed to determine whether extracellular vesicles isolated from patients with pulmonary hypertension show a disease specific signature of small non-coding RNAs and thus have the potential to serve as diagnostic and prognostic biomarkers. Extracellular vesicles were isolated from the serum of 23 patients with chronic thromboembolic pulmonary hypertension (CTEPH) and 23 controls using two individual methods: a column-based method or by precipitation. Extracellular vesicle- associated RNAs were analyzed by next-generation sequencing applying molecular barcoding, and differentially expressed small non-coding RNAs were validated by quantitative real-time polymerase chain reaction (qRT-PCR). We identified 18 microRNAs and 21 P-element induced wimpy testis (PIWI)-interacting RNAs (piRNAs) or piRNA clusters that were differentially expressed in CTEPH patients compared with controls. Bioinformatic analysis predicted a contribution of these piRNAs to the progression of cardiac and vascular remodeling. Expression levels of DQ593039 correlated with clinically meaningful parameters such as mean pulmonary arterial pressure, pulmonary vascular resistance, right ventricular systolic pressure, and levels of N-terminal pro-brain natriuretic peptide. Thus, we identified the extracellular vesicle- derived piRNA, DQ593039, as a potential biomarker for pulmonary hypertension and right heart disease.
细胞外囊泡在人体的许多细胞类型在生理和病理条件下都会释放。它们在细胞间通讯中起着重要作用，并携带肽和 RNA 的特定特征。在这项研究中，我们旨在确定从肺动脉高压患者中分离出的细胞外囊泡是否具有小非编码 RNA 的疾病特异性特征，因此具有作为诊断和预后生物标志物的潜力。使用两种不同的方法：基于柱的方法或沉淀法，从 23 例慢性血栓栓塞性肺动脉高压（CTEPH）患者和 23 例对照者的血清中分离细胞外囊泡。通过应用分子条形码的下一代测序分析细胞外囊泡相关 RNA，并通过定量实时聚合酶链反应（qRT-PCR）验证差异表达的小非编码 RNA。我们鉴定了 18 个 microRNAs 和 21 个 P 元素诱导的软弱睾丸（PIWI）相互作用 RNA（piRNA）或 piRNA 簇，与对照组相比，这些 piRNA 在 CTEPH 患者中差异表达。生物信息学分析预测这些 piRNA 对心脏和血管重构的进展有贡献。DQ593039 的表达水平与有意义的临床参数相关，如平均肺动脉压、肺血管阻力、右心室收缩压和 N 端脑钠肽前体水平。因此，我们鉴定了细胞外囊泡衍生的 piRNA，DQ593039，作为肺动脉高压和右心疾病的潜在生物标志物。
