Farinacci Maura, Krahn Thomas, Dinh Wilfried, Volk Hans-Dieter, Düngen Hans-Dirk, Wagner Josephine, Konen Timo, von Ahsen Oliver
Institute for Medical Immunology Charité University of Medicine Berlin Germany.
Berlin-Brandenburg Center for Regenerative Therapies Charité University of Medicine Berlin Germany.
Res Pract Thromb Haemost. 2018 Oct 26;3(1):49-58. doi: 10.1002/rth2.12158. eCollection 2019 Jan.
Endothelial dysfunction is involved in several cardiovascular diseases. Elevated levels of circulating endothelial cells (CECs) and low levels of endothelial progenitor cells (EPCs) have been described in different cardiovascular conditions, suggesting their potential use as diagnostic biomarkers for endothelial dysfunction. Compared to typical peripheral blood leukocyte subsets, CECs and EPCs occur at very low frequency. The reliable identification and characterization of CECs and EPCs is a prerequisite for their clinical use, however, a validated method to this purpose is still missing but a key for rare cell events.
To establish a validated flow cytometric procedure in order to quantify CECs and EPCs in human whole blood.
In the establishment phase, the assay sensitivity, robustness, and the sample storage conditions were optimized as prerequisite for clinical use. In a second phase, CECs and EPCs were analyzed in heart failure with preserved (HFpEF) and reduced (HFrEF) ejection fraction, in arterial hypertension (aHT), and in diabetic nephropathy (DN) in comparison to age-matched healthy controls.
The quantification procedure for CECs and EPCs showed high sensitivity and reproducibility. CEC values resulted significantly increased in patients with DN and HFpEF in comparison to healthy controls. CEC quantification showed a diagnostic sensitivity of 90% and a sensitivity of 68.0%, 70.4%, and 66.7% for DN, HFpEF, and aHT, respectively.
A robust and precise assay to quantify CECs and EPCs in pre-clinical and clinical studies has been established. CEC counts resulted to be a good diagnostic biomarker for DN and HFpEF.
内皮功能障碍与多种心血管疾病有关。在不同的心血管疾病状态下,循环内皮细胞(CECs)水平升高和内皮祖细胞(EPCs)水平降低已被描述,这表明它们有可能作为内皮功能障碍的诊断生物标志物。与典型的外周血白细胞亚群相比,CECs和EPCs的出现频率非常低。可靠地识别和表征CECs和EPCs是它们临床应用的前提条件,然而,目前仍缺少用于此目的的经过验证的方法,但这是罕见细胞事件研究的关键。
建立一种经过验证的流式细胞术方法,以定量检测人全血中的CECs和EPCs。
在方法建立阶段,作为临床应用的前提条件,对检测灵敏度、稳健性和样本储存条件进行了优化。在第二阶段,与年龄匹配的健康对照相比,对射血分数保留的心力衰竭(HFpEF)和射血分数降低的心力衰竭(HFrEF)、动脉高血压(aHT)和糖尿病肾病(DN)患者的CECs和EPCs进行了分析。
CECs和EPCs的定量检测方法显示出高灵敏度和可重复性。与健康对照相比,DN和HFpEF患者的CEC值显著升高。CEC定量检测对DN、HFpEF和aHT的诊断灵敏度分别为90%、68.0%、70.4%和66.7%。
已建立一种稳健且精确的方法,用于在临床前和临床研究中定量检测CECs和EPCs。CEC计数是DN和HFpEF的良好诊断生物标志物。