PURPOSE: Despite successful local tumor control, uveal melanoma (UM) patients may develop lethal metastases. To reliably identify circulating melanoma cells (CMC) in UM patients, we set out to test a new immunomagnetic enrichment assay and screened UM patients for the presence of CMC. We also determined whether we could find CMC in culture; for example, for future drug testing. METHODS: A dual-immunomagnetic enrichment assay using antibodies against two melanoma markers (NKI/C3 and NKI/beteb) was used to determine the presence of UM cells in blood. The sensitivity of the assay was determined by spiking normal blood with 92.1 cells (concentration range, 1-10(4) cells/mL). Isolated cells were characterized by immunocytochemistry directly after immunoenrichment and after a 2-week culture. The presence of CMC was determined in the peripheral blood of 31 patients with UM, and results were compared to clinical prognostic factors at the time of presentation. RESULTS: The CMC were detected in 93.5% (n = 29 of 31) of the patients with primary nonmetastatic UM at a median density of 3.5 cells/10 mL blood (range, 0-10.2 cells), as well as in blood cultures. No significant association was observed between the presence or number of CMC and any clinical prognostic factors. CONCLUSIONS: The improved dual-immunoenrichment assay enabled the detection of intact and viable CMC in the majority of UM patients. We also were able to identify CMC after short-term culturing. Molecular characterization of the CMC rather than the prevalence of these cells is expected to provide relevant information on the individual risk of metastasis. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: Despite successful local tumor control, uveal melanoma (UM) patients may develop lethal metastases. To reliably identify circulating melanoma cells (CMC) in UM patients, we set out to test a new immunomagnetic enrichment assay and screened UM patients for the presence of CMC. We also determined whether we could find CMC in culture; for example, for future drug testing. METHODS: A dual-immunomagnetic enrichment assay using antibodies against two melanoma markers (NKI/C3 and NKI/beteb) was used to determine the presence of UM cells in blood. The sensitivity of the assay was determined by spiking normal blood with 92.1 cells (concentration range, 1-10(4) cells/mL). Isolated cells were characterized by immunocytochemistry directly after immunoenrichment and after a 2-week culture. The presence of CMC was determined in the peripheral blood of 31 patients with UM, and results were compared to clinical prognostic factors at the time of presentation. RESULTS: The CMC were detected in 93.5% (n = 29 of 31) of the patients with primary nonmetastatic UM at a median density of 3.5 cells/10 mL blood (range, 0-10.2 cells), as well as in blood cultures. No significant association was observed between the presence or number of CMC and any clinical prognostic factors. CONCLUSIONS: The improved dual-immunoenrichment assay enabled the detection of intact and viable CMC in the majority of UM patients. We also were able to identify CMC after short-term culturing. Molecular characterization of the CMC rather than the prevalence of these cells is expected to provide relevant information on the individual risk of metastasis. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
Authors: Aaron Beasley; Timothy Isaacs; Muhammad A Khattak; James B Freeman; Richard Allcock; Fred K Chen; Michelle R Pereira; Kyle Yau; Jaqueline Bentel; Tersia Vermeulen; Leslie Calapre; Michael Millward; Melanie R Ziman; Elin S Gray Journal: JCO Precis Oncol Date: 2018-05-17
Authors: Charlotte Ness; Øystein Garred; Nils A Eide; Theresa Kumar; Ole K Olstad; Thomas P Bærland; Goran Petrovski; Morten C Moe; Agate Noer Journal: Mol Vis Date: 2017-10-03 Impact factor: 2.367
Authors: Manuel F Bande; Maria Santiago; Laura Muinelo-Romay; Maria Jose Blanco; Purificacion Mera; Carmela Capeans; Maria Pardo; Antonio Piñeiro Journal: BMC Res Notes Date: 2015-09-17