TY - JOUR
T1 - BRAF mutation analysis in circulating free tumor DNA of melanoma patients treated with BRAF inhibitors
AU - Gonzalez-Cao, Maria
AU - Mayo-De-Las-Casas, Clara
AU - Molina-Vila, Miguel A.
AU - De Mattos-Arruda, Leticia
AU - Muñoz-Couselo, Eva
AU - Manzano, Jose L.
AU - Cortes, Javier
AU - Berros, Jose P.
AU - Drozdowskyj, Ana
AU - Sanmamed, Miguel
AU - Gonzalez, Alvaro
AU - Alvarez, Carlos
AU - Viteri, Santiago
AU - Karachaliou, Niki
AU - Algarra, Salvador Martin
AU - Bertran-Alamillo, Jordi
AU - Jordana-Ariza, Nuria
AU - Rosell, Rafael
N1 - Publisher Copyright:
© 2015 Wolters Kluwer Health, Inc. All rights reserved.
PY - 2015
Y1 - 2015
N2 - BRAFV600E is a unique molecular marker for metastatic melanoma, being the most frequent somatic point mutation in this malignancy. Detection of BRAFV600E in blood could have prognostic and predictive value and could be useful for monitoring response to BRAF-targeted therapy. We developed a rapid, sensitive method for the detection and quantification of BRAFV600E in circulating free DNA (cfDNA) isolated from plasma and serum on the basis of a quantitative 5 Œ-nuclease PCR (Taqman) in the presence of a peptide.nucleic acid. We validated the assay in 92 lung, colon, and melanoma archival serum and plasma samples with paired tumor tissue (40 wild-type and 52 BRAFV600E). The correlation of cfDNA BRAFV600E with clinical parameters was further explored in 22 metastatic melanoma patients treated with BRAF inhibitors. Our assay could detect and quantify BRAFV600E in mixed samples with as little as 0.005% mutant DNA (copy number ratio 1 : 20 000), with a specificity of 100% and a sensitivity of 57.7% in archival serum and plasma samples. In 22 melanoma patients treated with BRAF inhibitors, the median progression-free survival was 3.6 months for those showing BRAFV600E in pretreatment cfDNA compared with 13.4 months for those in whom the mutation was not detected (P=0.021). Moreover, the median overall survival for positive versus negative BRAFV600E tests in pretreatment cfDNA differed significantly (7 vs. 21.8 months, P=0.017). This finding indicates that the sensitive detection and accurate quantification of low-abundance BRAFV600E alleles in cfDNA using our assay can be useful for predicting treatment outcome.
AB - BRAFV600E is a unique molecular marker for metastatic melanoma, being the most frequent somatic point mutation in this malignancy. Detection of BRAFV600E in blood could have prognostic and predictive value and could be useful for monitoring response to BRAF-targeted therapy. We developed a rapid, sensitive method for the detection and quantification of BRAFV600E in circulating free DNA (cfDNA) isolated from plasma and serum on the basis of a quantitative 5 Œ-nuclease PCR (Taqman) in the presence of a peptide.nucleic acid. We validated the assay in 92 lung, colon, and melanoma archival serum and plasma samples with paired tumor tissue (40 wild-type and 52 BRAFV600E). The correlation of cfDNA BRAFV600E with clinical parameters was further explored in 22 metastatic melanoma patients treated with BRAF inhibitors. Our assay could detect and quantify BRAFV600E in mixed samples with as little as 0.005% mutant DNA (copy number ratio 1 : 20 000), with a specificity of 100% and a sensitivity of 57.7% in archival serum and plasma samples. In 22 melanoma patients treated with BRAF inhibitors, the median progression-free survival was 3.6 months for those showing BRAFV600E in pretreatment cfDNA compared with 13.4 months for those in whom the mutation was not detected (P=0.021). Moreover, the median overall survival for positive versus negative BRAFV600E tests in pretreatment cfDNA differed significantly (7 vs. 21.8 months, P=0.017). This finding indicates that the sensitive detection and accurate quantification of low-abundance BRAFV600E alleles in cfDNA using our assay can be useful for predicting treatment outcome.
KW - BRAF
KW - Melanoma
KW - Plasma
KW - Serum
KW - cfDNA
UR - http://www.scopus.com/inward/record.url?scp=84946499416&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84946499416&partnerID=8YFLogxK
U2 - 10.1097/CMR.0000000000000187
DO - 10.1097/CMR.0000000000000187
M3 - Article
C2 - 26366702
AN - SCOPUS:84946499416
SN - 0960-8931
VL - 25
SP - 486
EP - 495
JO - Melanoma research
JF - Melanoma research
IS - 6
ER -