Detection rate and prognostic value of circulating tumor cells and circulating tumor DNA in metastatic uveal melanoma

Abstract

Circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) have been recently investigated in several cancer types, but their respective clinical significance remains to be determined. In our prospective study, we compared the detection rate and the prognostic value of these two circulating biomarkers in patients with metastatic uveal melanoma. GNAQ=GNA11 mutations were characterized in archived tumor tissue. Using a highly sensitive and mutation-specific bidirectional pyrophosphorolysisactivated polymerization (bi-PAP) technique, GNAQ c.626A>T, GNAQ c.626A>C and GNA11 c.626A>T copy numbers were quantified in plasma from 12 mL of blood. CTCs were detected at the same time in 7.5 mL of blood by the CellSearchV R technique. Patient characteristics and outcome were prospectively collected. CTCs (1) were detected in 12 of the 40 included patients (30%, range 1-20). Among the 26 patients with known detectable mutations, ctDNA was detected and quantified in 22 (84%, range 4-11,421 copies=mL). CTC count and ctDNA levels were associated with the presence of miliary hepatic metastasis (p 5 0.004 and 0.03, respectively), with metastasis volume (p 5 0.005 and 0.004) and with each other (p < 0.0001). CTC count and ctDNA levels were both strongly associated with progression-free survival (p 5 0.003 and 0.001) and overall survival (p 5 0.0009 and <0.0001). In multivariate analyses, ctDNA appeared to be a better prognostic marker than CTC. In conclusion, ctDNA and CTC are correlated and both have poor prognostic significance. CTC detection can be performed in every patient but, in patients with detectable mutations, ctDNA was more frequently detected than CTC and has possibly more prognostic value.Uveal melanoma is a rare cancer, with a reported incidence of two to eight new cases per million per year in Europe 1 and the United States.2 Specific mutations are found in this particular type of melanoma: >80% of uveal melanoma express mutually exclusive somatic mutations in two paralog proto-oncogenes, GNAQ 3 and GNA11, 4 which encode a-subunits of heterotrimeric G-proteins involved in the MEK-ERK signaling pathway. 5 In both genes, most mutations occur at nucleotide 626 encoding a glutamine at codon 209 (Q209). The presence of these mutations does not influence the risk of metastasis in patients. 4 Uveal melanoma metastases develop mostly in the liver through hematogenous spread of cancer cells. Despite improvement of diagnosis and treatment of the primary eye tumor, there is no effective treatment of metastatic disease; the prognosis of patients with metastatic uveal melanoma is limited, and although a few patients experience extended survival, the median overall survival (OS) following metastases detection is less than 1 year. 6 Circulating tumor cells (CTCs), which are cancer cells detected in patient blood, may correspond to cancer "seeds" that initiate metastatic relapse.7 Over the past two decades, both molecular and cytological detection techniques have

Circulating DNA as a Strong Multimarker Prognostic Tool for Metastatic Colorectal Cancer Patient Management Care

“…Several reports have already described the potential impact of ccfDNA on the prognosis of patient outcome in terms of either OS or progression-free survival (6,9,28,29). Most of these previous publications were oriented to analyze the correlation between an increase in the total level of ccfDNA with a decrease in PFS and OS.…”

Neueste technologische Entwicklungen für die Analyse von zirkulierender Tumor-DNA

“…In einem direkten Vergleich zwischen dem Therascreen ARMS assay(Qiagen)-einvonderFDA zugelassener diagnostischer Test für die Detektion von EGFR-und KRAS-Mutationen aus FFPE-Material -und der castPCR (Life Technologies) für KRAS, erreichte die castPCR eine vergleichbar gute Sensitivität und Spezifität [32]. Eine ganze Reihe von ähnlichen, etwas exotischeren Anwendungen wurde bereits publiziert, allerdings hat bisher keine davon eine breite Anwendung gefunden [33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48].…”

Circulating Tumor Cells, DNA, and mRNA: Potential for Clinical Utility in Patients With Melanoma

“…In melanoma management, the utility of genetic and molecular characterization is equally important, if not more so, because of the increased availability of targeted agents against BRAF and mitogen-activated protein kinase kinase (MEK), as well as other immunotherapies [10][11][12][13][14]. Genetic characterization has been widely achieved in melanoma CTP studies ( RBC depletion), cytospin, and IHC Near statistically significant association between increased CTC counts and decreased OS (p 5 .12) [85] Marker independent: Oncoquick plus telomerase probe Trend toward statistically significant association between increased CTC counts and disease progression (p 5 .21) [27] ctDNA Bidirectional PAP Low levels of ctDNA associated with longer PFS and OS in univariate and multivariate analysis [22] Methylation-specific PCR Hypermethylation of RASSF1A associated with shorter OS in univariate and multivariate analysis [77] PCR Loss of heterozygosity at DNA microsatellites associated with disease progression and OS [78,82,84] mRNA MART-1, MAGEA3, PAX3 .0 biomarker detected at pretreatment significantly associated with decreased DFS and OS in multivariate analysis [56] Serial presence of CTCs (.0 biomarker detected) significantly associated with decreased DFS and OS MART-1, MAGEA3, GalNAc-T Baseline CTC level ($2 biomarkers detected) significantly associated with decreased DFS, recurrence-free survival, and melanoma-specific survival in multivariate analysis [62] MART-1, GalNAc-T, MAGEA3, PAX3 Increased number of biomarkers significantly associated with decreased relapse-free survival and OS [54] Tyrosinase, p97, MUC-18, MAGEA3 Number of markers correlated with disease stage. Increased number of positive markers significantly associated with disease recurrence.…”

“…Another important application of CTPs as prognostic biomarkers lies in the correlation between CTPs and traditional prognosticators such as stage and disease volume [22,[53][54][55]. Of particular use to clinical melanoma management isthe ability to further risk stratify patients with stage II (sentinel lymph node negative) disease into groups of low and high risk for recurrence.…”

“…Marker-dependent strategies use melanomaspecific surface antigens and immunomagnetic beads to positively select for melanoma CTCs in blood. Surface markers such as high molecular weight melanoma-associated antigen (HMW-MAA), also known as melanoma-associated chondroitin sulfate proteoglycan (MCSP), CD146 or melanoma cell adhesion molecule (MCAM), and ATP-binding cassette subfamily B member 5 (ABCB5) are used either in isolation or combination to facilitate CTC capture [16][17][18][19][20][21][22]. In contrast, marker-independent strategies capitalize on CTCs' physical properties of size and density.…”