Journal of Clinical Medicine

Research

11 pages, 15277 KiB

Open AccessArticle

Quantitative Detection of Disseminated Melanoma Cells by Trp-1 Transcript Analysis Reveals Stochastic Distribution of Pulmonary Metastases

by Lenka Kyjacova, Rafael Saup, Melanie Rothley, Anja Schmaus, Tabea Wagner, Anja Boßerhoff, Boyan K. Garvalov, Wilko Thiele and Jonathan P. Sleeman

J. Clin. Med. 2021, 10(22), 5459; https://doi.org/10.3390/jcm10225459 - 22 Nov 2021

Cited by 2 | Viewed by 1848

Abstract

A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification [...] Read more.

A better understanding of the process of melanoma metastasis is required to underpin the development of novel therapies that will improve patient outcomes. The use of appropriate animal models is indispensable for investigating the mechanisms of melanoma metastasis. However, reliable and practicable quantification of metastases in experimental mice remains a challenge, particularly if the metastatic burden is low. Here, we describe a qRT-PCR-based protocol that employs the melanocytic marker Trp-1 for the sensitive quantification of melanoma metastases in the murine lung. Using this protocol, we were able to detect the presence of as few as 100 disseminated melanoma cells in lung tissue. This allowed us to quantify metastatic burden in a spontaneous syngeneic B16-F10 metastasis model, even in the absence of visible metastases, as well as in the autochthonous Tg(Grm1)/Cyld^−/− melanoma model. Importantly, we also observed an uneven distribution of disseminated melanoma cells amongst the five lobes of the murine lung, which varied considerably from animal to animal. Together, our findings demonstrate that the qRT-PCR-based detection of Trp-1 allows the quantification of low pulmonary metastatic burden in both transplantable and autochthonous murine melanoma models, and show that the analysis of lung metastasis in such models needs to take into account the stochastic distribution of metastatic lesions amongst the lung lobes. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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20 pages, 3782 KiB

Open AccessArticle

Learning from Embryogenesis—A Comparative Expression Analysis in Melanoblast Differentiation and Tumorigenesis Reveals miRNAs Driving Melanoma Development

by Lisa Linck-Paulus, Lisa Lämmerhirt, Daniel Völler, Katharina Meyer, Julia C. Engelmann, Rainer Spang, Norbert Eichner, Gunter Meister, Silke Kuphal and Anja Katrin Bosserhoff

J. Clin. Med. 2021, 10(11), 2259; https://doi.org/10.3390/jcm10112259 - 24 May 2021

Cited by 5 | Viewed by 2245

Abstract

Malignant melanoma is one of the most dangerous tumor types due to its high metastasis rates and a steadily increasing incidence. During tumorigenesis, the molecular processes of embryonic development, exemplified by epithelial–mesenchymal transition (EMT), are often reactivated. For melanoma development, the exact molecular [...] Read more.

Malignant melanoma is one of the most dangerous tumor types due to its high metastasis rates and a steadily increasing incidence. During tumorigenesis, the molecular processes of embryonic development, exemplified by epithelial–mesenchymal transition (EMT), are often reactivated. For melanoma development, the exact molecular differences between melanoblasts, melanocytes, and melanoma cells are not completely understood. In this study, we aimed to identify microRNAs (miRNAs) that promote melanoma tumorigenesis and progression, based on an in vitro model of normal human epidermal melanocyte (NHEM) de-differentiation into melanoblast-like cells (MBrCs). Using miRNA-sequencing and differential expression analysis, we demonstrated in this study that a majority of miRNAs have an almost equal expression level in NHEMs and MBrCs but are significantly differentially regulated in primary tumor- and metastasis-derived melanoma cell lines. Further, a target gene analysis of strongly regulated but functionally unknown miRNAs yielded the implication of those miRNAs in many important cellular pathways driving malignancy. We hypothesize that many of the miRNAs discovered in our study are key drivers of melanoma development as they account for the tumorigenic potential that differentiates melanoma cells from proliferating or migrating embryonic cells. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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11 pages, 619 KiB

Open AccessArticle

Incidence of Melanoma in Catalonia, Spain, Is Rapidly Increasing in the Elderly Population. A Multicentric Cohort Study

by Sebastian Podlipnik, Cristina Carrera, Aram Boada, Nina Richarz, Joaquim Marcoval, Josep Ramón Ferreres, Domingo Bodet, Rosa María Martí, Sonia Segura, Mireia Sabat, Joan Dalmau, Mònica Quintana, Antoni Azon, Neus Curcó, Manel Formigon, María Rosa Olivella-Garcés, Pedro Zaballos, Joaquim Sola, Loida Galvany, Carola Baliu-Piqué, Marta Alegre, Paola Pasquali, Josep Malvehy, Susana Puig and on behalf of the Network of Melanoma Centres of Catalonia Show full author list Hide full author list

J. Clin. Med. 2020, 9(11), 3396; https://doi.org/10.3390/jcm9113396 - 23 Oct 2020

Cited by 17 | Viewed by 3580

Abstract

The incidence of melanoma has been increasing worldwide during recent decades. The objective of the study was to analyse the trends in incidence for in situ and invasive melanoma in the Spanish region of Catalonia during the period of 2008–2017. We designed a [...] Read more.

The incidence of melanoma has been increasing worldwide during recent decades. The objective of the study was to analyse the trends in incidence for in situ and invasive melanoma in the Spanish region of Catalonia during the period of 2008–2017. We designed a cross-sectional study with an age-period-cohort analysis of melanoma patient data from the Network of Melanoma Centres in Catalonia. Our database covered a population of over seven million and included a total of 8626 patients with incident melanoma. The main outcome measures were crude and age-standardised incidence rates to the European 2013 standard population. Joinpoint regression models were used to evaluate the population trends. We observed an increase in the age-standardised incidence rate (per 100,000 population) of all melanoma subtypes from 11.56 in 2008 to 13.78 in 2017 with an average annual percent change (AAPC) of 3.5%. This incidence increase was seen exclusively in the older population. Moreover, the stratified analysis showed a statistically significant increase in the age-standardised incidence rate for invasive (AAPC 2.1%) and in situ melanoma (AAPC 6.5%). In conclusion, the incidence of melanoma has continued to increase in the elderly population over recent decades, with a rapidly increasing trend of in situ melanomas and the lentigo maligna subtype. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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26 pages, 4430 KiB

Open AccessArticle

Identification of microRNAs Targeting the Transporter Associated with Antigen Processing TAP1 in Melanoma

by Maria-Filothei Lazaridou, Chiara Massa, Diana Handke, Anja Mueller, Michael Friedrich, Karthikeyan Subbarayan, Sandy Tretbar, Reinhard Dummer, Peter Koelblinger and Barbara Seliger

J. Clin. Med. 2020, 9(9), 2690; https://doi.org/10.3390/jcm9092690 - 20 Aug 2020

Cited by 18 | Viewed by 2667

Abstract

The underlying molecular mechanisms of the aberrant expression of components of the HLA class I antigen processing and presentation machinery (APM) in tumors leading to evasion from T cell-mediated immune surveillance could be due to posttranscriptional regulation mediated by microRNAs (miRs). So far, [...] Read more.

The underlying molecular mechanisms of the aberrant expression of components of the HLA class I antigen processing and presentation machinery (APM) in tumors leading to evasion from T cell-mediated immune surveillance could be due to posttranscriptional regulation mediated by microRNAs (miRs). So far, some miRs controlling the expression of different APM components have been identified. Using in silico analysis and an miR enrichment protocol in combination with small RNA sequencing, miR-26b-5p and miR-21-3p were postulated to target the 3′ untranslated region (UTR) of the peptide transporter TAP1, which was confirmed by high free binding energy and dual luciferase reporter assays. Overexpression of miR-26b-5p and miR-21-3p in melanoma cells downregulated the TAP1 protein and reduced expression of HLA class I cell surface antigens, which could be reverted by miR inhibitors. Moreover, miR-26b-5p overexpression induced a decreased T cell recognition. Furthermore, an inverse expression of miR-26b-5p and miR-21-3p with TAP1 was found in primary melanoma lesions, which was linked with the frequency of CD8⁺ T cell infiltration. Thus, miR-26-5p and miR-21-3p are involved in the HLA class I-mediated immune escape and might be used as biomarkers or therapeutic targets for HLA class I^low melanoma cells. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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14 pages, 585 KiB

Open AccessArticle

Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients

by Maria Colombino, Carla Rozzo, Panagiotis Paliogiannis, Milena Casula, Antonella Manca, Valentina Doneddu, Maria Antonietta Fedeli, Maria Cristina Sini, Grazia Palomba, Marina Pisano, Paolo A. Ascierto, Corrado Caracò, Amelia Lissia, Antonio Cossu and Giuseppe Palmieri

J. Clin. Med. 2020, 9(8), 2430; https://doi.org/10.3390/jcm9082430 - 30 Jul 2020

Cited by 9 | Viewed by 2791

Abstract

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time [...] Read more.

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain reaction (rtPCR) or next-generation sequencing (NGS) assays in a real-life, hospital-based series of advanced MM patients. Consecutive patients with AJCC (American Joint Committee on Cancer) stage IIIC and IV MM from Sardinia, Italy, who were referred for molecular testing, were enrolled into the study. Initial screening was performed to assess the mutational status of the BRAF and NRAS genes, using the conventional methodologies recognized by the nationwide guidelines, at the time of the molecular classification, required by clinicians: at the beginning, Sanger-based sequencing (SS) and, after, pyrosequencing. The present study was then focused on BRAF mutation detecting approaches only. BRAF wild-type cases with available tissue and adequate DNA were further tested with rtPCR (Idylla™) and NGS assays. Globally, 319 patients were included in the study; pathogenic BRAF mutations were found in 144 (45.1%) cases examined with initial screening. The rtPCR detected 11 (16.2%) and 3 (4.8%) additional BRAF mutations after SS and pyrosequencing, respectively. NGS detected one additional BRAF-mutated case (2.1%) among 48 wild-type cases previously tested with pyrosequencing and rtPCR. Our study evidenced that rtPCR and NGS were able to detect additional BRAF mutant cases in comparison with conventional sequencing methods; therefore, we argue for the preferential utilization of the aforementioned assays (NGS and rtPCR) in clinical practice, to eradicate false-negative cases and improve the accuracy of BRAF detection. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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Review

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20 pages, 1743 KiB

Open AccessReview

An Update on the Role of Ubiquitination in Melanoma Development and Therapies

by Frédéric Soysouvanh, Serena Giuliano, Nadia Habel, Najla El-Hachem, Céline Pisibon, Corine Bertolotto and Robert Ballotti

J. Clin. Med. 2021, 10(5), 1133; https://doi.org/10.3390/jcm10051133 - 08 Mar 2021

Cited by 6 | Viewed by 2630

Abstract

The ubiquitination system plays a critical role in regulation of large array of biological processes and its alteration has been involved in the pathogenesis of cancers, among them cutaneous melanoma, which is responsible for the most deaths from skin cancers. Over the last [...] Read more.

The ubiquitination system plays a critical role in regulation of large array of biological processes and its alteration has been involved in the pathogenesis of cancers, among them cutaneous melanoma, which is responsible for the most deaths from skin cancers. Over the last decades, targeted therapies and immunotherapies became the standard therapeutic strategies for advanced melanomas. However, despite these breakthroughs, the prognosis of metastatic melanoma patients remains unoptimistic, mainly due to intrinsic or acquired resistances. Many avenues of research have been investigated to find new therapeutic targets for improving patient outcomes. Because of the pleiotropic functions of ubiquitination, and because each step of ubiquitination is amenable to pharmacological targeting, much attention has been paid to the role of this process in melanoma development and resistance to therapies. In this review, we summarize the latest data on ubiquitination and discuss the possible impacts on melanoma treatments. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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21 pages, 1002 KiB

Open AccessReview

Melanoma Single-Cell Biology in Experimental and Clinical Settings

by Hans Binder, Maria Schmidt, Henry Loeffler-Wirth, Lena Suenke Mortensen and Manfred Kunz

J. Clin. Med. 2021, 10(3), 506; https://doi.org/10.3390/jcm10030506 - 01 Feb 2021

Cited by 7 | Viewed by 3916

Abstract

Cellular heterogeneity is regarded as a major factor for treatment response and resistance in a variety of malignant tumors, including malignant melanoma. More recent developments of single-cell sequencing technology provided deeper insights into this phenomenon. Single-cell data were used to identify prognostic subtypes [...] Read more.

Cellular heterogeneity is regarded as a major factor for treatment response and resistance in a variety of malignant tumors, including malignant melanoma. More recent developments of single-cell sequencing technology provided deeper insights into this phenomenon. Single-cell data were used to identify prognostic subtypes of melanoma tumors, with a special emphasis on immune cells and fibroblasts in the tumor microenvironment. Moreover, treatment resistance to checkpoint inhibitor therapy has been shown to be associated with a set of differentially expressed immune cell signatures unraveling new targetable intracellular signaling pathways. Characterization of T cell states under checkpoint inhibitor treatment showed that exhausted CD8⁺ T cell types in melanoma lesions still have a high proliferative index. Other studies identified treatment resistance mechanisms to targeted treatment against the mutated BRAF serine/threonine protein kinase including repression of the melanoma differentiation gene microphthalmia-associated transcription factor (MITF) and induction of AXL receptor tyrosine kinase. Interestingly, treatment resistance mechanisms not only included selection processes of pre-existing subclones but also transition between different states of gene expression. Taken together, single-cell technology has provided deeper insights into melanoma biology and has put forward our understanding of the role of tumor heterogeneity and transcriptional plasticity, which may impact on innovative clinical trial designs and experimental approaches. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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Jump to: Research, Review

10 pages, 850 KiB

Open AccessBrief Report

NF1-Dependent Transcriptome Regulation in the Melanocyte Lineage and in Melanoma

by Lionel Larribère and Jochen Utikal

J. Clin. Med. 2021, 10(15), 3350; https://doi.org/10.3390/jcm10153350 - 29 Jul 2021

Cited by 2 | Viewed by 1710

Abstract

The precise role played by the tumor suppressor gene NF1 in melanocyte biology and during the transformation into melanoma is not completely understood. In particular, understanding the interaction during melanocyte development between NF1 and key signaling pathways, which are known to be reactivated [...] Read more.

The precise role played by the tumor suppressor gene NF1 in melanocyte biology and during the transformation into melanoma is not completely understood. In particular, understanding the interaction during melanocyte development between NF1 and key signaling pathways, which are known to be reactivated in advanced melanoma, is still under investigation. Here, we used RNAseq datasets from either situation to better understand the transcriptomic regulation mediated by an NF1 partial loss of function. We found that NF1 mutations had a differential impact on pluripotency and on melanoblast differentiation. In addition, major signaling pathways such as VEGF, senescence/secretome, endothelin, and cAMP/PKA are likely to be upregulated upon NF1 loss of function in both melanoblasts and metastatic melanoma. In sum, these data bring new light on the transcriptome regulation of the NF1-mutated melanoma subgroup and will help improve the possibilities for specific treatment. Full article

(This article belongs to the Special Issue New Advances in Melanoma)

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