BACKGROUND: Implementation of adjuvant therapies in non-metastatic melanoma improved treatment outcomes in some patients; however, adjuvant therapy can be associated with significant cost and risk of toxicity. Therefore, there is an unmet need to better identify patients at high risk of recurrence. PATIENTS AND METHODS: We carried out an ultrasensitive droplet digital PCR (ddPCR)-based detection of BRAFV600E-mutated circulating tumor DNA (ctDNA) from blood samples prospectively collected before surgery, 1 hour after surgery, and then serially during follow-up. RESULTS: In 80 patients (stages ≤III), BRAFV600E mutations were detected in 47.2% of tissue, in 37.7% of ctDNA samples collected before surgery, and in 25.9% of ctDNA samples collected 1 hour after surgery. Patients with detected ctDNA in blood collected 1 hour after surgery compared to patients without detected ctDNA had higher likelihood of melanoma recurrence (P < 0.001) and shorter median disease-free survival (P = 0.001) and overall survival (P = 0.003). CONCLUSIONS: Ultrasensitive ddPCR can detect ctDNA in pre- and post-surgical blood samples from patients with resectable melanoma. Detection of ctDNA in post-surgical samples is associated with inferior treatment outcomes.

Biomedical Center Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic; Department of Biology Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic

Department of Dermatovenerology University Hospital Pilsen Pilsen Czech Republic

Department of Histology and Embryology Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic; Biomedical Center Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic; Department of Neurology University Hospital Pilsen Pilsen Czech Republic

Department of Investigational Cancer Therapeutics The University of Texas MD Anderson Cancer Center Houston USA

Department of Pathology University Hospital Pilsen Pilsen Czech Republic

Department of Plastic Surgery University Hospital Pilsen Pilsen Czech Republic

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Siegel R.L., Miller K.D., Fuchs H.E., Jemal A. Cancer statistics, 2021. CA Cancer J Clin. 2021;71(1):7–33. PubMed

Von Schuckmann L.A., Hughes M.C.B., Ghiasvand R., et al. Risk of melanoma recurrence after diagnosis of a high-risk primary tumor. JAMA Dermatol. 2019;155(6):688–693. PubMed PMC

Eggermont A.M.M., Blank C.U., Mandala M., et al. Adjuvant pembrolizumab versus placebo in resected stage III melanoma. N Engl J Med. 2018;378(19):1789–1801. PubMed

Weber J., Mandala M., Del Vecchio M., et al. Adjuvant nivolumab versus ipilimumab in resected stage III or IV melanoma. N Engl J Med. 2017;377(19):1824–1835. PubMed

Long G.V., Hauschild A., Santinami M., et al. Adjuvant dabrafenib plus trametinib in stage III BRAF-mutated melanoma. N Engl J Med. 2017;377(19):1813–1823. PubMed

Hodis E., Watson I.R., Kryukov G.V., et al. A landscape of driver mutations in melanoma. Cell. 2012;150(2):251–263. PubMed PMC

Bettegowda C., Sausen M., Leary R.J., et al. Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci Transl Med. 2014;6(224):224ra24. PubMed PMC

Amin M.B., Edge S.B., Greene F.L., et al., editors. AJCC Cancer Staging Manual. 8th ed. Springer International Publishing; New York City, NY: 2016.

Che G., Huang B., Xie Z., et al. Trends in incidence and survival in patients with melanoma, 1974-2013. Am J Cancer Res. 2019;9(7):1396–1414. PubMed PMC

Ossio R., Roldán-Marín R., Martínez-Said H., Adams D.J., Robles-Espinoza C.D. Melanoma: a global perspective. Nat Rev Cancer. 2017;17(7):393–394. PubMed

Garbe C., Keim U., Eigentler T.K., et al. Time trends in incidence and mortality of cutaneous melanoma in Germany. J Eur Acad Dermatol Venereol. 2019;33(7):1272–1280. PubMed

Dawson S.-J., Tsui D.W.Y., Murtaza M., et al. Analysis of circulating tumor DNA to monitor metastatic breast cancer. N Engl J Med. 2013;368(13):1199–1209. PubMed

Hodi F.S., O'Day S.J., McDermott D.F., et al. Improved survival with ipilimumab in patients with metastatic melanoma. N Engl J Med. 2010;363(8):711–723. PubMed PMC

Tan L., Sandhu S., Lee R.J., et al. Prediction and monitoring of relapse in stage III melanoma using circulating tumor DNA. Ann Oncol. 2019;30(5):804–814. PubMed PMC

Huang R.S.P., Xiao J., Pavlick D.C., et al. Circulating cell-free DNA yield and circulating-tumor DNA quantity from liquid biopsies of 12 139 cancer patients. Clin Chem. 2021;67:1554–1566. PubMed

Chen K., Zhao H., Shi Y., et al. Perioperative dynamic changes in circulating tumor DNA in patients with lung cancer (DYNAMIC) Clin Cancer Res. 2019;25(23):7058–7067. PubMed

Xi L., Pham T.H.T., Payabyab E.C., Sherry R.M., Rosenberg S.A., Raffeld M. Circulating tumor DNA as an early indicator of response to T-cell transfer immunotherapy in metastatic melanoma. Clin Cancer Res. 2016;22(22):5480–5486. PubMed PMC

Forthun R.B., Hovland R., Schuster C., et al. ctDNA detected by ddPCR reveals changes in tumour load in metastatic malignant melanoma treated with bevacizumab. Sci Rep. 2019;9(1):17471. PubMed PMC

Lee J.H., Long G.V., Menzies A.M., et al. Association between circulating tumor DNA and pseudoprogression in patients with metastatic melanoma treated with anti–programmed cell death 1 antibodies. JAMA Oncol. 2018;4(5):717–721. PubMed PMC

Gray E.S., Rizos H., Reid A.L., et al. Circulating tumor DNA to monitor treatment response and detect acquired resistance in patients with metastatic melanoma. Oncotarget. 2015;6(39):42008–42018. PubMed PMC

McEvoy A.C., Warburton L., Al-Ogaili Z., et al. Correlation between circulating tumour DNA and metabolic tumour burden in metastatic melanoma patients. BMC Cancer. 2018;18(1):726. PubMed PMC

Seremet T., Jansen Y., Planken S., et al. Undetectable circulating tumor DNA (ctDNA) levels correlate with favorable outcome in metastatic melanoma patients treated with anti-PD1 therapy. J Transl Med. 2019;17(1):303. PubMed PMC

Santiago-Walker A., Gagnon R., Mazumdar J., et al. Correlation of BRAF mutation status in circulating-free DNA and tumor and association with clinical outcome across four BRAFi and MEKi clinical trials. Clin Cancer Res. 2016;22(3):567–574. PubMed

Lee J.H., Long G.V., Boyd S., et al. Circulating tumour DNA predicts response to anti-PD1 antibodies in metastatic melanoma. Ann Oncol. 2017;28(5):1130–1136. PubMed

Michielin O., van Akkooi A.C.J., Ascierto P.A., Dummer R., Keilholz U., ESMO Guidelines Committee Cutaneous melanoma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2019;30(12):1884–1901. PubMed

Parikh A.R., Van Seventer E.E., Siravegna G., et al. Minimal residual disease detection using a plasma-only circulating tumor DNA assay in patients with colorectal cancer. Clin Cancer Res. 2021;27(20):5586–5594. PubMed PMC

Kasi P.M., Le A.D., Barrett A. Comparative landscape of actionable somatic alterations in advanced cholangiocarcinoma from circulating tumor and tissue-based DNA profiling. J Clin Oncol. 2021;39(3_suppl):342.

Mack P., Minichielle K., Redman M., et al. LUNGMAP Master Protocol (LUNGMAP): concordance between plasma ctDNA and tissue molecular analysis. J Thorac Oncol. 2021;16(3):S163–S164.

Gerlinger M., Rowan A.J., Horswell S., et al. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. N Engl J Med. 2012;366(10):883–892. PubMed PMC

Predictive Significance of Combined Plasmatic Detection of BRAF Mutations and S100B Tumor Marker in Early-Stage Malignant Melanoma