Expression of Melan-A/MART-1 in primary melanoma cell cultures has prognostic implication in metastatic melanoma patients

11034 Background: Skp2, a known oncogene, is overexpressed in several types of tumors and is associated with worse recurrence rate and overall survival in primary melanoma patients. Moreover, the anti-proliferative effects of Skp2 siRNA on various tumor cell lines have prompted the preclinical testing of Skp2 small molecule inhibitors. In this study, we assessed the clinical relevance and molecular mechanism(s) underlying Skp2 overexpression in metastatic melanoma patients. Methods: Skp2 protein levels were measured in 122 metastatic melanoma specimens using immunohistochemistry (IHC), and the association between Skp2 overexpression and post-recurrence survival was examined. Moreover, 22 cell lines (2 normal primary melanocytes, 2 primary immortal melanocytes, 4 primary melanoma cell lines, and 18 metastatic melanoma cell lines) were evaluated for Skp2 genomic amplification using Single Nucleotide Polymorphism (SNP) arrays (Affymetrix 6.0) and Skp2 gene expression using mRNA arrays (Affymetrix U133A 2.0) and quantitative RT-PCR. We also screened 18 cell lines for Skp2 mutation by sequencing. Results: Skp2 overexpression, defined as >25% tumor cells, was associated with shorter 3-yr post-recurrence survival (37%) compared to Skp2 expression ≤25% (55%) (HR=1.89, 95%, CI= 1.04, 3.42, p=0.04). Skp2 overexpression was significantly associated with the site of melanoma metastasis: visceral (n= 12; 89%), lymph node (n=49; 36%), brain (n=15; 14%), and soft-tissue (n=36; 6%) (p<0.001). SNP array revealed genomic amplification at the Skp2 locus in 6 (33%) metastatic cell lines and one primary melanoma cell line. Skp2 genomic amplification was associated with increased transcript expression. No Skp2 mutations were identified. Conclusions: Skp2 protein overexpression is associated with worse prognosis in metastasis in melanoma. Our results also support that gene amplification, rather than a Skp2 gene mutation, may be the major mechanism responsible for Skp2 aberrant expression in metastatic melanoma. No significant financial relationships to disclose.

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Can the plasma PD-1 levels predict the presence and efficiency of tumor-infiltrating lymphocytes in patients with metastatic melanoma?

Therapeutic Advances in Medical Oncology ◽

10.1177/1758835919848872 ◽

2019 ◽

Vol 11 ◽

pp. 175883591984887 ◽

Cited By ~ 5

Author(s):

Lorena Incorvaia ◽

Giuseppe Badalamenti ◽

Gaetana Rinaldi ◽

Juan Lucio Iovanna ◽

Daniel Olive ◽

...

Keyword(s):

Immune Response ◽

Overall Survival ◽

Survival Rate ◽

Metastatic Melanoma ◽

Braf Mutation ◽

Primary Melanoma ◽

Tumor Infiltrating Lymphocytes ◽

Enzyme Linked Immunosorbent Assay ◽

Melanoma Patients ◽

Infiltrating Lymphocytes

Background: The immune response in melanoma patients is locally affected by presence of tumor-infiltrating lymphocytes (TILs), generally divided into brisk, nonbrisk, and absent. Several studies have shown that a greater presence of TILs, especially brisk, in primary melanoma is associated with a better prognosis and higher survival rate. Patients and Methods: We investigated by enzyme-linked immunosorbent assay (ELISA) the correlation between PD-1 levels in plasma and the presence/absence of TILs in 28 patients with metastatic melanoma. Results: Low plasma PD-1 levels were correlated with brisk TILs in primary melanoma, whereas intermediate values correlated with the nonbrisk TILs, and high PD-1 levels with absent TILs. Although the low number of samples did not allow us to obtain a statistically significant correlation between the plasma PD-1 levels and the patients’ overall survival depending on the absence/presence of TILs, the median survival of patients having brisk type TILs was 5 months higher than that of patients with absent and nonbrisk TILs. Conclusions: This work highlights the ability of measuring the plasma PD-1 levels in order to predict the prognosis of patients with untreated metastatic melanoma without a BRAF mutation at the time of diagnosis.

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Can the plasma PD-1 levels predict the presence and efficiency of tumor-infiltrating lymphocytes in metastatic melanoma patients?

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e14035 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e14035-e14035

Author(s):

Daniele Fanale ◽

Lorena Incorvaia ◽

Gaetana Rinaldi ◽

Lidia Terruso ◽

Giuseppe Badalamenti ◽

...

Keyword(s):

Metastatic Melanoma ◽

Primary Tumor ◽

Primary Melanoma ◽

Tumor Infiltrating Lymphocytes ◽

Rank Test ◽

Expression Levels ◽

Kaplan Meier ◽

Melanoma Patients ◽

Infiltrating Lymphocytes ◽

First Time

e14035 Background: The immune response to melanoma has been shown to be locally affected by presence of tumor-infiltrating lymphocytes (TILs), generally divided into brisk (infiltrating the entire base of the invasive tumor), non-brisk (infiltrating only focally) and absent. Several studies showed that greater presence of TILs, especially brisk, in primary melanoma is associated with a better prognosis and a higher survival rate. Since recent studies revealed an association between PD-1/PD-L1 expression levels and tumor response, the aim of our study was to investigate the correlation between plasma PD-1 and presence/absence/class of TILs in metastatic melanoma patients. Methods: The plasma PD-1 levels were analyzed in 28 patients with metastatic melanoma using a specific ELISA assay. The characterization of TILs in tumor tissue was performed by immunohistochemistry. Statistical analysis was assessed using t-Student and ANOVA tests. Survival curves were estimated by using the Kaplan-Meier method and log-rank test to evaluate significant differences among them. Results: 16 out of 28 patients showed the presence of TILs in primary tumor, 10 of which brisk and 6 nonbrisk. The plasma PD-1 levels were analyzed in relation to the presence/absence of TILs (p = 0.022), brisk TILs versus nonbrisk/absent TILs (p = 0.014), and brisk vs nonbrisk vs absent TILs (p = 0.032). In particular, low plasma PD-1 levels have been shown to be associated with brisk TILs in primary melanoma, intermediate values with nonbrisk TILs, and high expression with absent TILs. Although the low number of samples did not allow us to obtain a statistically significant association between the plasma PD-1 expression levels and overall survival (OS) depending on the absence or presence of TILs (brisk/nonbrisk), however the median survival of patients having brisk TILs was five months higher than other 2 groups of patients with absent and nonbrisk TILs, respectively. Conclusions: This work highlights, for the first time, the potential ability of using the plasma PD-1 levels to predict prognosis also in patients with metastatic melanoma at diagnosis for which it is not possible to identify the primary tumor.

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Clinicohistopathological Characteristics of Malignant Melanoma in the Gall Bladder: A Case Report and Review of the Literature

Case Reports in Pathology ◽

10.1155/2018/6471923 ◽

2018 ◽

Vol 2018 ◽

pp. 1-4

Author(s):

Schmidt Adrian ◽

Caspar Clemens ◽

Schmidt-Weiss Elisabeth ◽

Stadlmann Sylvia

Keyword(s):

Case Report ◽

Malignant Melanoma ◽

Gall Bladder ◽

Metastatic Melanoma ◽

Primary Melanoma ◽

Review Of The Literature ◽

Melanoma Patients

Objective. Primary gall bladder melanoma is a rare and controversial entity. So far, only 36 cases are documented in the literature. Metastatic melanoma targeting the gall bladder, however, has been reported to occur in about 15–20% of melanoma patients and is much more common. Methods. Based on the case of a 58-year-old woman presenting with multiple melanoma nodules in the gall bladder, we searched in the available literature in PubMed for articles describing a “primary melanoma of the gallbladder” regardless of language used. Results. We detected 33 papers that described 36 cases of primary gall bladder melanoma between 1907 and 2017. From different criteria distinguishing primary and secondary gall bladder melanoma, generally, the following were accepted: (1) exclusion of previous primary melanoma, (2) absence of synchronous involvement of other sites, (3) unicity of the lesion, (4) polypoid or papillary shape of the lesion, and (5) presence of junctional melanocitary component. Review of the 36 published cases revealed that only about one-fourth of them fulfilled all the five criteria. Conclusion. Primary gall bladder melanoma is even rarer than described in the literature, and the question whether this entity really exists remains open.

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Comprehensive upstream and downstream regulatory analyses identify miR-675-3p as a potential prognostic biomarker in melanoma

Human Cell ◽

10.1007/s13577-020-00473-0 ◽

2021 ◽

Author(s):

Cai-Chou Zhao ◽

Hao Guo ◽

Ying Wang ◽

Jiu-Hong Li

Keyword(s):

Cell Cycle ◽

Cell Lines ◽

Melanoma Cell ◽

Peripheral Blood ◽

Melanoma Patient ◽

Target Genes ◽

Prognostic Biomarker ◽

Primary Melanoma ◽

Melanoma Patients ◽

Melanoma Cell Lines

AbstractThis study assessed miR-675-3p-related regulatory mechanisms in melanoma and the clinical relevance of such regulatory activities. We downloaded miRNA mature strand expression RNA-Seq, phenotypic, and DNA methylation data pertaining to the TCGA Melanoma cohort. Differentially expressed miRNAs (DEMs) between metastatic and primary melanoma patient tissues were then identified, and miR-675-3p expression in melanoma patient peripheral blood was confirmed using the GSE20994 GEO dataset, while its expression in melanoma cell lines was evaluated via qRT-RCR. The clinical and prognostic implications of miR-675-3p in melanoma were assessed, and miR-675-3p target genes were identified using bioinformatics tools. Functional roles of this miRNA were explored via Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. We identified 3 and 22 miRNAs that were up- and downregulated, respectively, in metastatic melanoma samples relative to primary melanoma samples. Upregulation of miR-675-3p was associated with poorer overall patient survival, tumor histologic grade, and Clark's level. Consistently, miR-675-3p was also overexpressed in the peripheral blood of melanoma patients relative to healthy controls, and in melanoma cell lines relative to control cells. Gene regulatory networks indicated that 32 transcription factors control miR-675-3p expression, and that it, in turn, regulates 10 target genes. KEGG analyses indicated that these genes were associated with cell cycle, transcriptional misregulation in cancer, TGF-beta signaling, and HIF-1 signaling pathways. Gain-of-function assays revealed that miR-675-3p could promote cell proliferation via accelerating cell cycle progression. Western blotting results indicated that miR-675-3p could active TGF-beta and HIF-1 signaling. Through upstream and downstream analyses of miR-675-3p-related regulatory activity, we confirmed that this miRNA participates in key melanoma-related processes and offers value as a prognostic biomarker in melanoma patients.

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Cytogenetic, immunohistochemical and molecular analysys of p53 gene by TGGE and DGGE gel electrophoresis in 14 primary melanoma cell cultures

Cancer Genetics and Cytogenetics ◽

10.1016/0165-4608(92)90414-4 ◽

1992 ◽

Vol 63 (2) ◽

pp. 116

Author(s):

P. Grammatico ◽

S. Costanzi ◽

E. Vigneti ◽

M. Plane ◽

M. Roccella ◽

...

Keyword(s):

Cell Cultures ◽

Melanoma Cell ◽

Gel Electrophoresis ◽

Primary Melanoma ◽

P53 Gene

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Second primary melanoma in advanced melanoma patients treated with anti-PD-1 monoclonal antibodies.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e22025 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e22025-e22025

Author(s):

Elisa Funck-Brentano ◽

Estelle Charvet ◽

Louise Chaplain ◽

Amelie Gantzer ◽

Oula Kassem ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Metastatic Melanoma ◽

Braf Inhibitor ◽

Primary Melanoma ◽

Advanced Melanoma ◽

Heterozygous Mutation ◽

Second Primary ◽

Exon 2 ◽

Mutational Status ◽

Melanoma Patients

e22025 Background: Development of a second primary melanoma (SCPM) has not been reported in melanoma patients treated with anti-PD-1 monoclonal antibodies (mAb), in contrast with those reported in BRAF-inhibitor-treated patients. Our aim was to report arising SCPM in patients with advanced melanoma treated with anti-PD-1 therapy. Methods: Retrospective study, conducted in 2 referral centres, including advanced melanoma patients who developed a SCPM after anti-PD-1 mAb initiation, between September 2010 and May 2019. BRAF or NRAS mutational status was assessed by targeted NGS panels, real-time PCR, and immunohistochemistry. Results: Among a total of 509 patients treated with anti-PD-1 mAb, 4 had a SCPM (incidence: 0.8%; 95%CI: 0.02-1.57%). All patients were treated with nivolumab, in first (N = 3) or second line after progression with BRAF + MEK inhibitors (N = 1). No immune-related adverse event greater than grade 2 according to Common Terminology Criteria for Adverse Events version 5.0. was observed in these 4 cases; a vitiligo-like depigmentation (grade 1) was observed in two patients. The median time from the first nivolumab infusion to the SCPM diagnosis was 17.5 months (range: 5-21). All patients developed the SCPM after achieving a complete response. Nivolumab administration had been discontinued (4 months prior) in one patient. Histology revealed 4 superficial spreading melanomas (SSM): one invasive (without BRAFV600 mutation) and 3 intraepidermal melanomas (2 with a BRAFV600E mutation and one with a NRASQ61H mutation). 3 patients had risk factors for developing multiple melanomas: a dysplastic nevus syndrome, a high number of nevi (≥100 nevi), and a family history of melanoma in first-degree relatives and constitutional heterozygous mutation of exon 2 of the CDKN2A gene. Occurrence of SPCM did not alter advanced melanoma treatment. With a median follow-up of 29 months [range: 18-41] from the first anti-PD-1 mAb infusion, all patients had prolonged CR, and treatment was discontinued in all patients, without relapse after a median 11.5 months [0-18] off therapy. The median duration of nivolumab treatment was 15.5 months [10-24]. Conclusions: Although anti-PD-1 mAb could theoretically decrease the risk of developing another melanoma in metastatic melanoma patients, we found 4 such cases, highlighting the importance of regular clinical screenings for new primary melanoma in patients with metastatic melanoma even when responsive to anti-PD-1 therapy. Immune checkpoint inhibitors do not totally prevent the risk of occurrence a SCPM.

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20. MELANOMA CELL INTRINSIC GABAA RECEPTOR ENHANCEMENT POTENTIATES RADIATION AND IMMUNE CHECKPOINT INHIBITOR RESPONSE BY PROMOTING DIRECT AND T CELL-MEDIATED ANTI-TUMOR ACTIVITY

Neuro-Oncology Advances ◽

10.1093/noajnl/vdaa073.010 ◽

2020 ◽

Vol 2 (Supplement_2) ◽

pp. ii3-ii3

Author(s):

Soma Sengupta ◽

Tahseen Nasti ◽

Milota Kaluzova ◽

Laura Kallay ◽

Johannes Melms ◽

...

Keyword(s):

Metastatic Melanoma ◽

Melanoma Cell ◽

Immune Checkpoint ◽

Immune Checkpoint Inhibitor ◽

Tumor Volume ◽

Checkpoint Inhibitor ◽

Checkpoint Inhibitors ◽

Expression Of Genes ◽

Melanoma Patients ◽

Anti Tumor Activity

Abstract Most metastatic melanoma patients exhibit poor and variable response to radiotherapy and targeted therapies, including immune checkpoint inhibitors. There is a need for therapeutics that can potentiate existing treatments to positively impact clinical outcomes of metastatic melanoma patients. We reanalyzed melanoma TCGA transcriptomes and identified, as linked to previously defined molecular subgroups, enhanced expression of genes coding for subunits of the Type A GABA receptor (GABAAR), a chloride ion channel and major inhibitory neurotransmitter receptor. Using whole-cell patch clamp electrophysiology, we find that melanoma cells possess GABAARs that control membrane permeability to anions. Select benzodiazepines, by enhancing GABAAR mediated anion transport, depolarize melanoma cell mitochondrial membrane potential and impair cell viability in vitro. Using a syngeneic melanoma mouse model, we find that a benzodiazepine promotes reduction in tumor volume when administered alone and potentiated radiation or immune checkpoint inhibitor α-PD-L1. When a benzodiazepine is combined with concurrent α-PD-L1 and a sub-lethal radiation dose, there is near complete loss of tumor, beyond what is observed for benzodiazepine with radiation or α-PD-L1. Mechanistically, benzodiazepine with radiation or α-PD-L1 results in ipsilateral and an abscopal tumor volume reduction commensurate with enhanced infiltration into the tumor milieu of polyfunctional CD8 T-cells. There is also an increased expression of genes with roles in the cytokine-cytokine receptor and p53 signaling pathways. This study provides evidence for melanoma cell GABAARs as a therapeutic vulnerability with benzodiazepines promoting both direct and immune-mediated anti-tumor activity.

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Long Noncoding RNA HOTAIR Is Associated with Motility, Invasion, and Metastatic Potential of Metastatic Melanoma

BioMed Research International ◽

10.1155/2013/251098 ◽

2013 ◽

Vol 2013 ◽

pp. 1-7 ◽

Cited By ~ 72

Author(s):

Lihua Tang ◽

Wei Zhang ◽

Bing Su ◽

Bo Yu

Keyword(s):

Lymph Node ◽

Cell Motility ◽

Metastatic Melanoma ◽

Melanoma Cell ◽

Noncoding Rna ◽

Primary Melanoma ◽

Melanoma Metastasis ◽

Gelatin Matrix ◽

Gelatinase Activity

Metastatic melanoma, the primary cause of skin cancer-related death, warrants new therapeutic approaches that target the regulatory machinery at molecular level. While long noncoding RNAs (lncRNAs) are dysregulated in a number of cancer types, limited data are available on the expression and function of lncRNAs in melanoma metastasis. The primary objective of this study was to investigate the role of 6 metastasis-related lncRNAs in pairs of primary melanoma and matched lymph node metastatic tissues. Among the tested lncRNAs, HOTAIR was the most highly expressed in lymph node metastasis. The role of HOTAIR in melanoma cell motility and invasion was further evaluated by knocking down HOTAIR with siRNAs. Knockdown of HOTAIR resulted in the reduction of motility and invasion of human melanoma cell line A375, as assessed by wound healing assay and Matrigel-based invasion assay. siHOTAIR also suppressed the degradation of gelatin matrix, suggesting that HOTAIR promotes gelatinase activity. Together, our study shows that HOTAIR is overexpressed in metastatic tissue, which is associated with the ability of HOTAIR to promote melanoma cell motility and invasion. These data indicate that lncRNAs may be involved in the metastasis of melanoma and provide support for further evaluation of lncRNAs in melanoma.

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