BACKGROUND: Melanoma is the most aggressive skin cancer with ability to recur also after early-stage tumor surgery. The aim was to identify early-stage melanoma patients at high risk of recurrence using liquid biopsy, estimating of mutated BRAF ctDNA and the level of tumor marker S100B in plasma. METHODS: Eighty patients were enrolled in the study. BRAF V600E mutation was determined in FFPE tissue and plasma samples using ultrasensitive ddPCR with pre-amplification. The level of S100B was determined in plasma by immunoassay chemiluminescent method. RESULTS: The best prediction of melanoma recurrence after surgery was observed in patients with combined high level of S100B (S100Bhigh) and ctDNA BRAFV600E (BRAFmut) in preoperative (57.1% vs. 12.5%, p = 0.025) as well as postoperative blood samples (83.3% vs. 14.3%, resp., p = 0.001) in comparison with low S100B and BRAF wild-type. Similarly, patients with preoperative and postoperative S100Bhigh and BRAFmut experienced worse prognosis (DFI p = 0.05, OS p = 0.131 and DFI p = 0.001, OS = 0.001, resp.). CONCLUSION: We observed the benefit of the estimation of combination of S100B and ctDNA BRAFmut in peripheral blood for identification of patients at high risk of recurrence and unfavorable prognosis. SIGNIFICANCE: There is still no general consensus on molecular markers for deciding the appropriateness of adjuvant treatment of early-stage melanoma. We have shown for the first time that the combined determination of the ctDNA BRAFmut oncogene (liquid biopsy) and the high level of tumor marker S100B in pre- and postoperative plasma samples can identify patients with the worst prognosis and the highest risk of tumor recurrence. Therefore, modern adjuvant therapy would be appropriate for these patients with resectable melanoma, regardless of disease stage.

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

Department of Immunochemical Diagnostics University Hospital Pilsen Pilsen Czech Republic

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

Department of Pathology University Hospital Pilsen Pilsen Czech Republic

Department of Pharmacology Faculty of Medicine in Pilsen Charles University Pilsen Czech Republic

Department of Plastic Surgery University Hospital Pilsen Pilsen Czech Republic

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Davis L. E., Shalin S. C., and Tackett A. J., “Current State of Melanoma Diagnosis and Treatment,” Cancer Biology & Therapy 20, no. 11 (2019): 1366–1379. PubMed PMC

von Schuckmann L. A., Hughes M. C. B., Ghiasvand R., et al., “Risk of Melanoma Recurrence After Diagnosis of a High‐Risk Primary Tumor,” Journal of the American Medical Association Dermatology 155, no. 6 (2019): 688–693. PubMed PMC

Saad M., Castellano E., and Tarhini A. A., “Clinical Updates in Neoadjuvant Immunotherapy for Melanoma Before Surgery,” Expert Review of Clinical Immunology 1–17 (2023): 927–943. PubMed

Bagheri M. S., Polivka J., Treskova I., et al., “Preoperative Plasma miRNA Levels Predict Prognosis in Early‐Stage Malignant Melanoma,” Anticancer Research 43, no. 2 (2023): 695–706. PubMed

Simetić L., Blažičević K., Međugorac K., Golčić M., and Herceg D., “Relative Change in S100 as a Biomarker of Survival in Patients With Metastatic Melanoma Treated With Pembrolizumab,” Anticancer Research 40, no. 4 (2020): 2157–2163. PubMed

Ertekin S. S., Podlipnik S., Ribero S., et al., “Monthly Changes in Serum Levels of S100B Protein as a Predictor of Metastasis Development in High‐Risk Melanoma Patients,” Journal of the European Academy of Dermatology and Venereology 34, no. 7 (2020): 1482–1488. PubMed

Deckers E. A., Wevers K. P., Muller Kobold A. C., et al., “S‐100B as an Extra Selection Tool for FDG PET/CT Scanning in Follow‐Up of AJCC Stage III Melanoma Patients,” Journal of Surgical Oncology 120, no. 6 (2019): 1031–1037. PubMed PMC

Frauchiger A. L., Dummer R., and Mangana J., “Serum S100B Levels in Melanoma,” Methods in Molecular Biology 1929 (2019): 691–700. PubMed

Allgöwer C., Kretz A.‐L., von Karstedt S., Wittau M., Henne‐Bruns D., and Lemke J., “Friend or Foe: S100 Proteins in Cancer,” Cancers (Basel) 12, no. 8 (2020): 2037. PubMed PMC

Harpio R. and Einarsson R., “S100 Proteins as Cancer Biomarkers With Focus on S100B in Malignant Melanoma,” Clinical Biochemistry 37, no. 7 (2004): 512–518. PubMed

Xiong T.‐F., Pan F.‐Q., and Li D., “Expression and Clinical Significance of S100 Family Genes in Patients With Melanoma,” Melanoma Research 29, no. 1 (2019): 23–29. PubMed PMC

Bresnick A. R., Weber D. J., and Zimmer D. B., “S100 Proteins in Cancer,” Nature Reviews. Cancer 15, no. 2 (2015): 96–109. PubMed PMC

Ricciardi E., Giordani E., Ziccheddu G., et al., “Metastatic Melanoma: Liquid Biopsy as a New Precision Medicine Approach,” International Journal of Molecular Sciences 24, no. 4 (2023): 4014. PubMed PMC

Tivey A., Britton F., Scott J.‐A., Rothwell D., Lorigan P., and Lee R., “Circulating Tumour DNA in Melanoma‐Clinic Ready?,” Current Oncology Reports 24, no. 3 (2022): 363–373. PubMed PMC

Kamińska P., Buszka K., Zabel M., Nowicki M., Alix‐Panabières C., and Budna‐Tukan J., “Liquid Biopsy in Melanoma: Significance in Diagnostics, Prediction and Treatment Monitoring,” International Journal of Molecular Sciences 22, no. 18 (2021): 9714. PubMed PMC

Polivka J., Pesta M., and Janku F., “Testing for Oncogenic Molecular Aberrations in Cell‐Free DNA‐Based Liquid Biopsies in the Clinic: Are We There Yet?,” Expert Review of Molecular Diagnostics 15, no. 12 (2015): 1631–1644. PubMed PMC

Pantel K. and Speicher M. R., “The Biology of Circulating Tumor Cells,” Oncogene 35, no. 10 (2016): 1216–1224. PubMed

Gouda M. A., Polivka J., Huang H. J., et al., “Ultrasensitive Detection of BRAF Mutations in Circulating Tumor DNA of Non‐metastatic Melanoma,” ESMO Open 7, no. 1 (2022): 100357. PubMed PMC

Svedman F. C., Pillas D., Taylor A., Kaur M., Linder R., and Hansson J., “Stage‐Specific Survival and Recurrence in Patients With Cutaneous Malignant Melanoma in Europe – A Systematic Review of the Literature,” Clinical Epidemiology 8 (2016): 109–122. PubMed PMC

Pesta M., Shetti D., Kulda V., et al., “Applications of Liquid Biopsies in Non‐Small‐Cell Lung Cancer,” Diagnostics (Basel) 12, no. 8 (2022): 1799. PubMed PMC

Larkin J., Del Vecchio M., Mandalá M., et al., “Adjuvant Nivolumab Versus Ipilimumab in Resected Stage III/IV Melanoma: 5‐Year Efficacy and Biomarker Results From CheckMate 238,” Clinical Cancer Research 29, no. 17 (2023): 3352–3361. PubMed PMC

Eggermont A. M. M., Blank C. U., Mandala M., et al., “Longer Follow‐Up Confirms Recurrence‐Free Survival Benefit of Adjuvant Pembrolizumab in High‐Risk Stage III Melanoma: Updated Results From the EORTC 1325‐MG/KEYNOTE‐054 Trial,” Journal of Clinical Oncology 38, no. 33 (2020): 3925–3936. PubMed PMC

Dummer R., Hauschild A., Santinami M., et al., “Five‐Year Analysis of Adjuvant Dabrafenib Plus Trametinib in Stage III Melanoma,” New England Journal of Medicine 383, no. 12 (2020): 1139–1148. PubMed

Luke J. J., Rutkowski P., Queirolo P., et al., “Pembrolizumab Versus Placebo as Adjuvant Therapy in Completely Resected Stage IIB or IIC Melanoma (KEYNOTE‐716): A Randomised, Double‐Blind, Phase 3 Trial,” Lancet 399, no. 10336 (2022): 1718–1729. PubMed

Bettegowda C., Sausen M., Leary R. J., et al., “Detection of Circulating Tumor DNA in Early‐ and Late‐Stage Human Malignancies,” Science Translational Medicine 6, no. 224 (2014): 224ra24. PubMed PMC

Flaherty K. T., “A Twenty Year Perspective on Melanoma Therapy,” Pigment Cell & Melanoma Research 36, no. 6 (2023): 563–575. PubMed

Zheng Y., Sun H., Cong L., et al., “Prognostic Value of ctDNA Mutation in Melanoma: A Meta‐Analysis,” Journal of Oncology 2021 (2021): 6660571. PubMed PMC

Lee J. H., Saw R. P., Thompson J. F., et al., “Pre‐Operative ctDNA Predicts Survival in High‐Risk Stage III Cutaneous Melanoma Patients,” Annals of Oncology 30, no. 5 (2019): 815–822. PubMed PMC

Tan L., Sandhu S., Lee R. J., et al., “Prediction and Monitoring of Relapse in Stage III Melanoma Using Circulating Tumor DNA,” Annals of Oncology 30, no. 5 (2019): 804–814. PubMed PMC

Mocellin S., Zavagno G., and Nitti D., “The Prognostic Value of Serum S100B in Patients With Cutaneous Melanoma: A Meta‐Analysis,” International Journal of Cancer 123, no. 10 (2008): 2370–2376. PubMed

Janka E. A., Várvölgyi T., Sipos Z., et al., “Predictive Performance of Serum S100B Versus LDH in Melanoma Patients: A Systematic Review and Meta‐Analysis,” Frontiers in Oncology 11 (2021): 772165. PubMed PMC

Wagner N. B., Forschner A., Leiter U., Garbe C., and Eigentler T. K., “S100B and LDH as Early Prognostic Markers for Response and Overall Survival in Melanoma Patients Treated With Anti‐PD‐1 or Combined Anti‐PD‐1 Plus Anti‐CTLA‐4 Antibodies,” British Journal of Cancer 119, no. 3 (2018): 339–346. PubMed PMC

Karonidis A., Mantzourani M., Gogas H., and Tsoutsos D., “Serum S100B Levels Correlate With Stage, N Status, Mitotic Rate and Disease Outcome in Melanoma Patients Independent to LDH,” Journal of the Balkan Union of Oncology 22, no. 5 (2017): 1296–1302. PubMed

Dumitraşcu G., Constantin C., Manda G., et al., “Serum Markers in Skin Melanoma—Preliminary Study,” Roumanian Archives of Microbiology and Immunology 68, no. 3 (2009): 125–135. PubMed

Lugović L., Situm M., Buljan M., Poduje S., and Sebetić K., “Results of the Determination of Serum Markers in Patients With Malignant Melanoma,” Collegium Antropologicum 31, no. Suppl 1 (2007): 7–11. PubMed