基于微阵列的黑色素瘤患者循环肿瘤 DNA 中 BRAF V600 突变分析。

Microarray-based analysis of the BRAF V600 mutations in circulating tumor DNA in melanoma patients.

机构信息

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, 32 Vavilova St., Russian Federation.

Russian Scientific Center of Roentgen Radiology, Ministry of Healthcare of the Russian Federation, 117997 Moscow, Profsoyuznaya St. 86, Russian Federation.

出版信息

Cancer Genet. 2021 Jan;250-251:25-35. doi: 10.1016/j.cancergen.2020.11.005. Epub 2020 Nov 20.

DOI:10.1016/j.cancergen.2020.11.005

PMID:33249369

Abstract

BACKGROUND

Circulating tumor DNA (ctDNA) holds great potential for cancer therapy and can provide diagnostic and prognostic information before and during treatment.

METHODS

Plasma DNA samples from 97 melanoma patients, 20 healthy donors and 3 patients with benign skin tumors were analyzed by microarray analysis and droplet digital PCR (ddPCR).

RESULTS

A microarray for simultaneous detection of six BRAF V600 mutations in ctDNA has been developed. The method allows the detection of 0.05% mutated DNA from WT DNA background. For paired samples (pre-surgery plasma and tumor tissue) isolated from 74 patients, the concordance of genotypes between tumor DNA and ctDNA was 65% (48/74). BRAF mutations in ctDNA were detected in 27/50 patients with BRAF-positive tumors and in 3/24 patients with BRAF wild-type tumors. The presence of ctDNA BRAF mutations in 23 plasma samples from melanoma patients undergoing therapy correlated significantly with tumor progression (P=0.005). The increase in cell-free DNA levels measured by ddPCR also correlated with disease progression (P=0.02). The concordance of results obtained by microarray identification of BRAF mutations and those obtained by ddPCR was 91%.

CONCLUSION

The novel microarray-based approach can be a useful non-invasive tool for accurate identification of ctDNA BRAF mutations to monitor disease progression.

摘要

背景

循环肿瘤 DNA（ctDNA）在癌症治疗中有很大的潜力，可以在治疗前后提供诊断和预后信息。

方法

通过微阵列分析和液滴数字 PCR（ddPCR）分析了 97 名黑色素瘤患者、20 名健康供体和 3 名良性皮肤肿瘤患者的血浆 DNA 样本。

结果

开发了一种用于同时检测 ctDNA 中六个 BRAF V600 突变的微阵列。该方法允许从 WT DNA 背景中检测到 0.05%的突变 DNA。对于从 74 名患者分离的 74 对术前血浆和肿瘤组织样本，肿瘤 DNA 和 ctDNA 之间基因型的一致性为 65%（48/74）。在 50 名 BRAF 阳性肿瘤患者中有 27 名和 24 名 BRAF 野生型肿瘤患者中有 3 名检测到 ctDNA BRAF 突变。23 名接受治疗的黑色素瘤患者的血浆样本中存在 ctDNA BRAF 突变与肿瘤进展显著相关（P=0.005）。通过 ddPCR 测量的游离细胞 DNA 水平的增加也与疾病进展相关（P=0.02）。微阵列鉴定 BRAF 突变和 ddPCR 获得的结果的一致性为 91%。