scholarly journals Isolation of extracellular vesicles improves the detection of mutant DNA from plasma of metastatic melanoma patients

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Davide Zocco ◽  
Simona Bernardi ◽  
Mauro Novelli ◽  
Chiara Astrua ◽  
Paolo Fava ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Braf Inhibitor ◽  
Extracellular Vesicle ◽  
Therapeutic Monitoring ◽  
Clinical Samples ◽  
Outer Side ◽  
Alternative Source ◽  
Gene Copies ◽  
Melanoma Patients ◽  
Promising Source

Abstract Detection of BRAFV600E within cell free tumor DNA (ctDNA) is emerging as a promising means to improve patients’ stratification or enable BRAF inhibitor (BRAFi) therapeutic monitoring in a minimally invasive manner. Here, we investigated whether extracellular vesicle-(EV)-associated-DNA (EV-DNA) has value as an alternative source of circulating BRAFV600E. To do so, we identified a clinical practice-compatible protocol for the isolation of EV-DNA and assessed BRAF gene status on plasma samples from metastatic melanoma patients at the beginning and during BRAFi therapy. This protocol uses a peptide with high affinity for EVs and it has been found to recover more mutant DNA from plasma than standard ultracentrifugation. Molecular analyses revealed that mutant DNA is largely unprotected from nuclease digestion, interacting with the outer side of the EV membrane or directly with the peptide. When used on clinical samples, we found that the protocol improves the detection of BRAFV600E gene copies in comparison to the reference protocol for ctDNA isolation. Taken together, these findings indicate that EVs are a promising source of mutant DNA and should be considered for the development of next-generation liquid biopsy approaches.

Relevance of MIA and S100 serum tumor markers to monitor BRAF inhibitor therapy in metastatic melanoma patients

2014 ◽  
Vol 429 ◽  
pp. 168-174 ◽  
Author(s):  
Miguel F. Sanmamed ◽  
Sara Fernández-Landázuri ◽  
Carmen Rodríguez ◽  
María D. Lozano ◽  
José I. Echeveste ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Tumor Markers ◽  
Braf Inhibitor ◽  
Inhibitor Therapy ◽  
Serum Tumor Markers ◽  
Melanoma Patients

Relevance of MIA and S-100 to monitor BRAF inhibitor (iBRAF) therapy in metastatic melanoma patients.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. 9074-9074
Author(s):  
Miguel F. Sanmamed ◽  
Sara Fernandez-Landazuri ◽  
Eduardo Castanon ◽  
Jose Echeveste ◽  
Maria D. Lozano ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Tumor Markers ◽  
Direct Sequencing ◽  
Braf Inhibitor ◽  
Braf V600e Mutation ◽  
Braf V600e ◽  
Best Response ◽  
Response Table ◽  
S 100 ◽  
Melanoma Patients

9074 Background: MIA and S-100 have been proposed as tumor markers for patients with melanoma, but they are not widely accepted. BRAF V600E mutation has been reported in more than 50% of melanomas. Recently, selective BRAF inhibitors have proved to be more active than DTIC in first line treatment of BRAF V600E melanoma patients. The aim of the present work is to evaluate the utility of MIA and S-100 during iBRAF treatment. Methods: BRAF V600E mutation was analyzed in 77 patients with metastatic melanoma by automated direct sequencing in tumor DNA. Tumor markers (MIA, S-100 and LDH) were studied in serum from all patients. Sixteen of these patients received iBRAF therapy (11 Vemurafenib, 5 Dabrafenib) and tumor markers were analyzed sequentially: baseline, best response and progression. MIA and S-100 were determined by immunometric methods and LDH by a spectrophotometric assay. The cut-off points were MIA=9 ug/L, S-100=0.1 ug/L, and LDH=290 U/L. Non-parametric statistical analysis was performed. Results: Forty-three patients had BRAF V600E mutation and 34 were wild type (WT). The percentage of cases with MIA above the cut-off in patients with V600E mutation was significantly higher than in the WT group (76.3% vs. 52.9%; p<0.05), while the frequency of elevated S-100 and LDH was similar. Among patients treated with iBRAF, the response rate was 87.5% (5 CR, 9PR). In responding patients, MIA and S100 levels decreased dramatically, but not LDH (Table). At the time of this report, thirteen patients have progressed. Upon progression, MIA and S-100 increased significantly above levels achieved at best response (Table). Conclusions: Serum MIA and S-100 are potentially useful markers in the clinical and follow-up management of patients receiving iBRAF therapy. Validation in a larger series is needed. [Table: see text]

scholarly journals Plasma lncRNA expression profile as a prognostic tool in BRAF-mutant metastatic melanoma patients treated with BRAF inhibitor

Oncotarget ◽  
2019 ◽  
Vol 10 (39) ◽  
pp. 3879-3893 ◽  
Author(s):  
Tomasz Kolenda ◽  
Piotr Rutkowski ◽  
Michał Michalak ◽  
Katarzyna Kozak ◽  
Kacper Guglas ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Expression Profile ◽  
Braf Inhibitor ◽  
Prognostic Tool ◽  
Lncrna Expression ◽  
Melanoma Patients ◽  
Braf Mutant

BKM120 combined with vemurafenib in vemurafenib-refractory BRAF mutant metastatic melanoma: Two cases.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. e20010-e20010 ◽  
Author(s):  
Alain Patrick Algazi ◽  
Christian Posch ◽  
Susana Ortiz-Urda ◽  
Alyson Cockerill ◽  
Pamela N. Munster ◽  
...  
Keyword(s):  
Metastatic Melanoma ◽  
Disease Progression ◽  
Braf Inhibitor ◽  
Study Drug ◽  
Braf V600e ◽  
Response To Treatment ◽  
Mixed Response ◽  
Pi3k Activation ◽  
Melanoma Patients ◽  
Braf Mutant

e20010 Background: PTEN loss and PI3K activation are common in BRAF mutant metastatic melanoma, and PI3K activation has been implicated as a cause of acquired resistance to BRAF inhibitors. Two vemurafenib refractory were treated with the potent BRAF inhibitor, vemurafenib, and the pan-class I PI3K inhibitor BKM120. Methods: The study enrolled BRAF-V600E/K mutant metastatic melanoma patients with an ECOG PS ≥ 2 and adequate organ function. Vemurafenib refractory BRAF-V600E/K mutant melanoma patients started both vemurafenib twice daily and BKM120 daily on cycle 1, day 1 after a vemurafenib washout of at least 14 days. Serial biopsies prior to treatment, on cycle 1 day 15, and at progression were obtained for pharmacodynamics analysis in patients with visible or palpable tumors. Results: 3 BRAF inhibitor refractory patients were treated on study with vemurafenib 720 mg PO bid and BKM120 60 mg PO daily. One patient was inevaluable due to non-compliance and had minimal exposure to study drug. Pre-treatment biopsy specimens were available in the remaining 2 vemurafenib-refractory patients. Both patient expressed PTEN at baseline and had demonstrable pAKT and pS6 staining. One patient had a mixed response to treatment with a 35.9% reduction in target lesions and two new small subcutaneous lesions. This patient also developed dose limiting febrile neutropenia on trial. The second patient tolerated treatment well but had widespread disease progression at 8 weeks. Conclusions: Combination therapy with vemurafenib and BKM120 in BRAF-V600E/K mutant melanoma led to substantial regression of several tumors in a PTEN+ patient with prior disease progression on vemurafenib alone. A phase I dose escalation trial in ongoing. Clinical trial information: NCT01512251.

Pretreatment features and risk of death from BRAF-positive metastatic melanoma patients treated with vemurafenib.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e21028-e21028
Author(s):  
Dorota Malgorzata Kwapisz
Keyword(s):  
Metastatic Melanoma ◽  
Braf Mutation ◽  
Performance Status ◽  
Elevated Serum ◽  
Braf Inhibitor ◽  
Poor Performance ◽  
Poor Performance Status ◽  
Disease Symptoms ◽  
Risk Of Death ◽  
Melanoma Patients

e21028 Background: Clinicopathologic features of BRAF-positive metastatic melanoma patients were identified. No study to date has examined influence on risk of death features at baseline before starting treatment with vemurafenib. The purpose of the study is to identify demographic, clinical, biochemical, and tumor-related features at baseline before treatment with vemurafenib that are related with increase risk of death in BRAF-positive metastatic melanoma patients. Methods: 323 consecutive patients with metastatic melanoma tested for BRAF mutation were analysed. The presence of BRAF gene V600 mutation was evaluated by the cobas 4800 BRAF V600 Mutation Test. The pretreatment features were collected prospectively after inclusion patients for BRAF inhibitor treatment but before treatment was started. Results: 204 patients had a BRAF mutation. Of these, 115 were treated with vemurafenib. The median time of treatment with vemurafenib was 198 days. Death occurred more frequently among patients with: pretreatment disease symptoms (p = 0.003), ECOG PS ≥1 (p = 0.03), CNS metastases at baseline (p = 0.049) [χ2 test]. The pretreatment elevated serum LDH level (p < 0.0005), leukocytosis (p = 0.042), and hipoalbuminemia (p = 0.002) had an impact on higher incidence of death. AJCC stage 3 was reported more frequently in patients who have died (AJCC 1-2 vs. AJCC 3; p = 0.025, χ2 test). Higher incidence of death in patients with ≥4 localisations of metastases (different organs) was noted (1-3 vs 4-8; p = 0.001, χ2test). The maximum metastasis dimension was greater in the group of patients who have died (p = 0.009; Mann-Whitney test). In the analysed population, all patients whose pretreatment metastasis dimension exceeded 56 mm have died. Age, BMI, and sex were not related with higher risk of death. Conclusions: Pretreatment features may identify patients at higher risk of death. Those with features at baseline such as: disease symptoms, poor performance status, elevated LDH, leukocytosis, hipoalbuminemia, increased metastatic tumor burden, especially when brain is one of the metastatic localisation are in higher risk of death among all patients treated with vemurafenib.

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

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.

scholarly journals Baseline Genomic Features in BRAFV600-Mutated Metastatic Melanoma Patients Treated with BRAF Inhibitor + MEK Inhibitor in Routine Care

Cancers ◽  
2019 ◽  
Vol 11 (8) ◽  
pp. 1203 ◽  
Author(s):  
Baptiste Louveau ◽  
Fanelie Jouenne ◽  
Coralie Reger de Moura ◽  
Aurelie Sadoux ◽  
Barouyr Baroudjian ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Metastatic Melanoma ◽  
Clinical Benefit ◽  
Real Life ◽  
Braf Inhibitor ◽  
Phase Iii ◽  
Line Treatment ◽  
Genomic Features ◽  
Dna Alterations ◽  
Melanoma Patients

In BRAFV600mut metastatic melanoma, the combination of BRAF and MEK inhibitors (BRAFi, MEKi) has undergone multiple resistance mechanisms, limiting its clinical benefit and resulting in the need for response predicting biomarkers. Based on phase III clinical trial data, several studies have previously explored baseline genomic features associated with response to BRAFi + MEKi. Using a targeted approach that combines the examination of mRNA expression and DNA alterations in a subset of genes, we performed an analysis of baseline genomic alterations involved in MAPK inhibitors’ resistance in a real-life cohort of BRAFV600mut metastatic melanoma patients. Twenty-seven patients were included in this retrospective study, and tumor samples were analyzed when the BRAFi + MEKi therapy was initiated. The clinical characteristics of our cohort were consistent with previously published studies. The BRAFi + MEKi treatment was initiated in seven patients as a following-line treatment, and had a specific transcriptomic profile exhibiting 14 genes with lower mRNA expression. However, DNA alterations in CCND1, RB1, and MET were only observed in patients who received BRAFi + MEKi as the first-line treatment. Furthermore, KIT mRNA expression was significantly higher in patients showing clinical benefit from the combined therapy, emphasizing the tumor-suppressor role of KIT already described within the context of BRAF-mutant melanoma.

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