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Detection of cell-free circulating BRAFV 600E by droplet digital polymerase chain reaction in patients with and without melanoma under dermatological surveillance.

在皮肤科监测下,通过液滴数字聚合酶链反应检测黑色素瘤和非黑色素瘤患者的无细胞循环 BRAFV 600E。

  • 影响因子:2.45
  • DOI:10.1111/bjd.18147
  • 作者列表:"Calbet-Llopart N","Potrony M","Tell-Martí G","Carrera C","Barreiro A","Aguilera P","Podlipnik S","Puig S","Malvehy J","Puig-Butillé JA
  • 发表时间:2020-02-01
Abstract

BACKGROUND:The p.V600E mutation in the BRAF protein is the most frequent mutation in cutaneous melanoma and is a recurrent alteration found in common benign naevi. Analysis of the cell-free BRAF c.1799T>A, p.V600E mutation (cfBRAFV600E ) in plasma has emerged as a biomarker for monitoring prognosis and treatment response in patients with melanoma. OBJECTIVES:To quantify cfBRAFV600E levels in plasma from patients with melanoma and from patients without melanoma undergoing regular follow-up of their melanocytic lesions, in order to assess the clinical significance of the test. METHODS:We quantified cfBRAFV600E by droplet digital polymerase chain reaction in plasma from 146 patients without melanoma undergoing continuous dermatological screening, from 26 stage III and seven stage IV patients with BRAF-mutant melanoma, and from 32 patients with melanoma who were free of disease for 3 or more years. RESULTS:Among disease-free patients and individuals without melanoma, 52% presented a high naevus count (> 50) and 49% had clinically atypical naevi. cfBRAFV600E was detected in 71% of patients with stage IV melanoma and 15% with stage III, and in 1·4% of individuals without melanoma. No cfBRAFV600E mutation was detected in disease-free patients with melanoma. Individuals without melanoma had lower cfBRAFV600E levels than patients with melanoma. We established a variant allelic frequency of 0·26% or 5 copies mL-1 of cfBRAFV600E as the optimal cutoff value for identifying patients with melanoma with > 99% specificity. CONCLUSIONS:This study suggests that naevus-related factors do not influence the detection of cfBRAFV600E in individuals without melanoma, and supports the clinical diagnostic value of plasma cfBRAFV600E quantification in patients with melanoma. What's already known about this topic? The analysis of the BRAF c.1799T>A (p.V600E) mutation in cell-free (cf)DNA has emerged as a potential biomarker for monitoring prognosis and treatment response in patients with metastatic BRAFV600E melanoma. The BRAFV600E alteration is a common genetic alteration found in benign proliferations such as melanocytic naevi. No information exists about the impact of the number of common acquired naevi or the presence of clinically atypical naevi in cfBRAFV600E detection in an individual. What does this study add? The cfBRAFV600E mutation is detected in plasma from a reduced number of individuals without melanoma undergoing continuous dermatological follow-up. A high number of naevi or the presence of clinically atypical naevi are factors that do not influence cfBRAFV600E detection in an individual. Both total cfBRAF concentration and cfBRAFV600E frequency are effective biomarkers in patients with advanced melanoma but not in patients at early stages or with micrometastases. What is the translational message? Detection of cfBRAFV600E in an individual is not influenced by naevus-related factors. cfBRAFV600E is a robust and reliable biomarker that can be used in dermatological surveillance programmes.

摘要

背景: BRAF蛋白中的p.V600E突变是皮肤黑色素瘤中最常见的突变,是在普通良性naevi中发现的复发性改变。血浆中无细胞BRAF c.1799T>A分析,p.V600E突变 (cfBRAFV600E) 已成为监测黑色素瘤患者预后和治疗反应的生物标志物。 目的: 定量黑素瘤患者和黑素细胞病变定期随访的非黑素瘤患者血浆中cfBRAFV600E水平,以评估试验的临床意义。 方法: 我们通过液滴数字聚合酶链反应对 146 例接受连续皮肤科筛查的无黑色素瘤患者,26 例 ⅲ 期和 7 例 ⅳ 期BRAF突变黑色素瘤患者的血浆中cfBRAFV600E进行定量。并且来自 32 例 3 年或更长时间无病的黑色素瘤患者。 结果: 在无病患者和非黑色素瘤个体中,52% 出现高痣计数 (> 50),49% 出现临床不典型痣。cfBRAFV600E在 71% 的IV期黑色素瘤患者和 15% 的III期患者中检测到,在 1 · 4% 的无黑色素瘤个体中检测到。在无病黑色素瘤患者中未检测到cfBRAFV600E突变。无黑色素瘤的个体cfBRAFV600E水平低于黑色素瘤患者。我们建立了cfBRAFV600E mL-1 0 · 26% 或 5 拷贝的变异等位基因频率作为识别特异性> 99% 的黑色素瘤患者的最佳临界值。 结论: 本研究提示naevus相关因素不影响非黑色素瘤个体cfBRAFV600E的检测,支持血浆cfBRAFV600E定量在黑色素瘤患者中的临床诊断价值。关于这个话题已经知道了什么?BRAF c.1799T>A的分析 (p。v600E) 无细胞 (cf)DNA突变已成为监测转移性BRAFV600E黑色素瘤患者预后和治疗反应的潜在生物标志物。BRAFV600E改变是在黑素细胞痣等良性增生中发现的常见遗传改变。没有关于个体cfBRAFV600E检测中常见获得性naevi数量或临床非典型naevi存在的影响的信息。这项研究增加了什么?CfBRAFV600E突变是在接受连续皮肤科随访的无黑色素瘤个体数量减少的血浆中检测到的。大量naevi或临床上不典型naevi的存在是不影响个体cfBRAFV600E检测的因素。CfBRAF总浓度和cfBRAFV600E频率在晚期黑色素瘤患者中都是有效的生物标志物,但在早期或微转移患者中不是。什么是翻译信息?在个体中检测cfBRAFV600E不受naevus相关因素的影响。cfBRAFV600E是一种稳健可靠的生物标志物,可用于皮肤科监测方案。

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