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• W2023008559 abstract "circulating tumor cells melanoma-associated chondroitin sulfate proteoglycan TO THE EDITOR We thank Ulmer and Fierlbeck, 2011Ulmer A. Fierlbeck G. Detecting Circulating Melanoma Cells.J Invest Dermatol. 2011; 131: 1774-1775Abstract Full Text Full Text PDF PubMed Scopus (1) Google Scholar for their interest in our work. We fully agree that the detection and monitoring of circulating melanoma cells is a rapidly expanding field of potential clinical relevance for cutaneous melanoma patients. In the specific clinical context of cutaneous melanoma, PCR is by far the most commonly used technique for the detection of circulating tumor cells (CTC) and multimarker quantitative PCR is considered to be the state-of-the-art quantitative method. In this respect, the 2004 study by Ulmer et al., 2004Ulmer A. Schmidt-Kittler O. Fischer J. et al.Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells.Clin Cancer Res. 2004; 10: 531-537Crossref PubMed Scopus (100) Google Scholar is of interest, but not of fully appreciable clinical impact for the following reasons. First, the series lumped together, for unknown reasons, 164 patients with cutaneous or uveal melanoma with either localized or metastatic disease, but the number of cutaneous melanoma patients analyzed was not reported. As cutaneous and uveal melanoma are distinct tumor entities clinically, morphologically, genetically, or prognostically, the significance of the data is questionable. Second, CTC were enriched by immunomagnetic cell sorting using a single-murine monoclonal antibody against the melanoma-associated chondroitin sulfate proteoglycan (MCSP). MCSP is expressed in over 85% of primary and metastatic melanoma lesions, with similar levels of expression among lentigo maligna, nodular, and superficial spreading melanomas. However, a more variable and generally lower expression has been reported in acral lentiginous melanomas, spanning from 30% (Kageshita et al., 1993Kageshita T. Kuriya N. Ono T. et al.Association of high molecular weight melanoma-associated antigen expression in primary acral lentiginous melanoma lesions with poor prognosis.Cancer Res. 1993; 53: 2830-2833PubMed Google Scholar) to 53.6% (Nishi et al., 2010Nishi H. Inoue Y. Kageshita T. et al.The expression of human high molecular weight melanoma-associated antigen in acral lentiginous melanoma.Biosci Trends. 2010; 4: 86-89PubMed Google Scholar). Thus, MCSP-negative melanoma circulating cells may escape detection by the method described by Ulmer et al., 2004Ulmer A. Schmidt-Kittler O. Fischer J. et al.Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells.Clin Cancer Res. 2004; 10: 531-537Crossref PubMed Scopus (100) Google Scholar. In addition, MCSP is not detectable in cells of hematopoietic origin, including healthy peripheral blood cells (Campoli et al., 2004Campoli M.R. Chang C.C. Kageshita T. et al.Human high molecular weight-melanoma-associated antigen (HMW-MAA): a melanoma cell surface chondroitin sulfate proteoglycan (MSCP) with biological and clinical significance.Crit Rev Immunol. 2004; 24: 267-296Crossref PubMed Scopus (142) Google Scholar; Kitago et al., 2009Kitago M. Koyanagi K. Nakamura T. et al.mRNA expression and BRAF mutation in circulating melanoma cells isolated from peripheral blood with high molecular weight melanoma-associated antigen-specific monoclonal antibody beads.Clin Chem. 2009; 55: 757-764Crossref PubMed Scopus (55) Google Scholar). Thus, the observation by Ulmer et al., 2004Ulmer A. Schmidt-Kittler O. Fischer J. et al.Immunomagnetic enrichment, genomic characterization, and prognostic impact of circulating melanoma cells.Clin Cancer Res. 2004; 10: 531-537Crossref PubMed Scopus (100) Google Scholar that granulocytes showed positive staining for MCSP (as illustrated in Figure 1c) should have raised doubts concerning the specificity of the monoclonal antibody. Third, to prove the malignant nature of the MPCS-positive cells, genomic aberrations were demonstrated in isolated CTC from seven patients by single-cell comparative genomic hybridization. Although of interest, such high-throughput, high-cost analysis may be suitable for research settings, but its application in a routine clinical setting is questionable. Currently, there is no consensus on the best method to identify CTC in cutaneous melanoma, and only a case-by-case comparison of techniques may demonstrate which method is superior in terms of specificity and sensitivity. The choice of appropriate markers is a challenge, as antigens exclusively expressed by CTC and not shared by other circulating non-tumor or blood cells are scarce. In our study (De Giorgi et al., 2010De Giorgi V. Pinzani P. Salvianti F. et al.Application of a filtration- and isolation-by size technique for the detection of circulating tumor cells in cutaneous melanoma.J Invest Dermatol. 2010; 130: 2440-2447Abstract Full Text Full Text PDF PubMed Scopus (125) Google Scholar), we chose the CTC analysis associated with immunochemistry and molecular analysis because this represents the gold standard for routine diagnosis of cancer. It should be noted also that melanocytic differentiation biomarkers such as MART-1 and HMB-45 are commonly used to detect both primary and metastatic melanoma, including lymph nodal metastasis. Ulmer and Fierlbeck, 2011Ulmer A. Fierlbeck G. Detecting Circulating Melanoma Cells.J Invest Dermatol. 2011; 131: 1774-1775Abstract Full Text Full Text PDF PubMed Scopus (1) Google Scholar are not convinced that the cells stained by S-100 protein and HMB-45, reported in Figure 1, fulfill the morphological criteria for tumor cells. We disagree with their comment because the nucleocytoplasmic features of the cell reported in Figure 1 are similar to those of the melanoma cell line SK-Mel 28 and, more importantly, because the intensity of the staining and absence of background staining are of sufficient diagnostic value. Similarly, the “non-stained debris” around the HMB45-positive melanoma cells are simply fibrin depositions. Morphological criteria for the identification of CTC (e.g., cell size ≥16μm, nucleocytoplasmic ratio ≥50%, irregular nuclear shape, hyperchromatic nucleus, and basophilic cytoplasm) were originally defined for identification of epithelial cells using the method of isolation by size of epithelial tumor cells (Vona et al., 2000Vona G. Sabile A. Louha M. et al.Isolation by size of epithelial tumor cells: a new method for the immunomorphological and molecular characterization of circulatingtumor cells.Am J Pathol. 2000; 156: 57-63Abstract Full Text Full Text PDF PubMed Scopus (816) Google Scholar). Whereas epithelial cells in the isolation by size of epithelial tumour cells assay tend to form cell aggregates (Pinzani et al., 2006Pinzani P. Salvadori B. Simi L. et al.Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer: correlation with real-time reverse transcriptase-polymerase chain reaction results and feasibility of molecular analysis by laser microdissection.Hum Pathol. 2006; 37: 711-718Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar), in our experience CTC from cutaneous melanoma usually appear as isolated individual cells and are recognizable as either nonpigmented large epithelioid or smaller cells with a roundish to oval shape and indented nucleus. In the case of cells of uncertain morphological recognition, including small size cells, filters underwent further immunohistochemical identification with S-100 protein, HMB-45, MART-1, or CD45 to support an accurate identification of melanoma cells. More than one marker may be required to conclusively identify cells on the filters as CTC, as loss of antigen expression may occur." @default.
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• W2023008559 title "Circulating Tumor Cells in Cutaneous Melanoma" @default.
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