Search

Background Lung cancer is one of the most incident neoplastic diseases, and a leading cause of death for cancer worldwide. Knowledge of the incidence of druggable genetic alterations, their correlation with clinical and pathological features of the disease, and their interplay in cases of co-occurrence is crucial for selecting the best therapeutic strategies of patients with non-small cell lung cancer. In this real-life study, we describe the molecular epidemiology of genetic alterations in five driver genes and their correlations with the demographic and clinical characteristics of Sardinian patients with lung adenocarcinoma. Methods Data from 1440 consecutive Sardinian patients with a histologically proven diagnosis of lung adenocarcinoma from January 2011 through July 2016 were prospectively investigated. EGFR mutation analysis was performed for all of them, while KRAS and BRAF mutations were searched in 1047 cases; ALK alterations were determined with fluorescence in situ hybridization in 899 cases, and cMET amplifications in 788 cases. Results KRAS mutations were the most common genetic alterations involving 22.1% of the cases and being mutually exclusive with the EGFR mutations, which were found in 12.6% of them. BRAF mutations, ALK rearrangements, and cMET amplifications were detected in 3.2, 5.3, and 2.1% of the cases, respectively. Concomitant mutations were detected only in a few cases. Conclusions Almost all the genetic alterations studied showed a similar incidence in comparison with other Caucasian populations. Concomitant mutations were rare, and they probably have a scarce impact on the clinical management of Sardinians with lung adenocarcinoma. The low incidence of concomitant cMET amplifications at diagnosis suggests that these alterations are acquired in subsequent phases of the disease, often during treatment with TKIs.

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation testing plays a predominant role in the management of MM patients, because modern targeted therapies essentially consist of inhibitors of BRAF. BRAF V600 mutation must be detected using a FDA approved (USA) or CE IVD certified (Europe) test. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real time PCR rtPCR ) or next generation sequencing (NGS) assays in a real life, hospital based series of advanced MM patients.

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation testing plays a predominant role in the management of MM patients, because modern targeted therapies essentially consist of inhibitors of BRAF. BRAF V600 mutation must be detected using a FDA-approved (USA) or CE IVD certified (Europe) test. The aim of this study was to compare BRAF mutational testing performed by conventiona lnucleotide sequencing approaches with either real-time PCR (rtPCR )or next-generation sequencing (NGS) assays in a real-life,hospital-based series of advanced MM patients. Consecutive patients fromSardinia, Italy, with histological diagnosis of advanced MM (AJCC stages III Cand IV) and referred a the laboratory of Biomolecular Chemistry (ICB) of the National Research Council (CNR), Sassari, Italy for molecular testing from 2012 through 2019, were enrolled into the study.

Background Identification of somatic mutations in key oncogenes in melanoma is important to lead the effective and efficient use of personalized anticancer treatment. Conventional methods focus on few genes per run and, therefore, are unable to screen for multiple genes simultaneously. The use of Next-Generation Sequencing (NGS) technologies enables sequencing of multiple cancer-driving genes in a single assay, with reduced costs and DNA quantity needed and increased mutation detection sensitivity. Methods We designed a customized IMI somatic gene panel for targeted sequencing of actionable melanoma mutations; this panel was tested on three different NGS platforms using 11 metastatic melanoma tissue samples in blinded manner between two EMQN quality certificated laboratory. Results The detection limit of our assay was set-up to a Variant Allele Frequency (VAF) of 10% with a coverage of at least 200x. All somatic variants detected by all NGS platforms with a VAF ≥ 10%, were also validated by an independent method. The IMI panel achieved a very good concordance among the three NGS platforms. Conclusion This study demonstrated that, using the main sequencing platforms currently available in the diagnostic setting, the IMI panel can be adopted among different centers providing comparable results.

Background Cutaneous malignant melanoma (CMM) is one of the most common skin cancers worldwide. Limited information is available in the current scientific literature on the concordance of genetic alterations between primary and metastatic CMM. In the present study, we performed next-generation sequencing (NGS) analysis of the main genes participating in melanoma pathogenesis and progression, among paired primary and metastatic lesions of CMM patients, with the aim to evaluate levels of discrepancies in mutational patterns. Methods Paraffin-embedded tumor tissues of the paired lesions were retrieved from the archives of the institutions participating in the study. NGS was performed using a specific multiple-gene panel constructed by the Italian Melanoma Intergroup (IMI) to explore the mutational status of selected regions (343 amplicons; amplicon range: 125–175 bp; coverage 100%) within the main 25 genes involved in CMM pathogenesis; sequencing was performed with the Ion Torrent PGM System. Results A discovery cohort encompassing 30 cases, and a validation cohort including eleven Sardinian patients with tissue availability from both the primary and metachronous metastatic lesions were identified; the global number of analyzed tissue specimens was 90. A total of 829 genetic non-synonymous variants were detected: 101 (12.2%) were pathogenic/likely pathogenic, 131 (15.8%) were benign/likely benign, and the remaining 597 (72%) were uncertain/unknown significance variants. Considering the global cohort, the consistency in pathogenic/pathogenic like mutations was 76%. Consistency for BRAF and NRAS mutations was 95.2% and 85.7% respectively, without statistically significant differences between the discovery and validation cohort. Conclusions Our study showed a high level of concordance in mutational patterns between primary and metastatic CMM, especially when pathogenic mutations in driver genes were considered.

Cutaneous malignant melanoma is a heterogeneous disease, being the consequence of specific genetic alterations along several molecular pathways. Despite the increased knowledge about the biology and pathogenesis of melanoma, the incidence has grown markedly worldwide, making it extremely important to develop preventive measures. The beneficial role of correct nutrition and of some natural dietary compounds in preventing malignant melanoma has been widely demonstrated. This led to numerous studies investigating the role of several dietary attitudes, patterns, and supplements in the prevention of melanoma, and ongoing research investigates their impact in the clinical management and outcomes of patients diagnosed with the disease. This article is an overview of recent scientific advances regarding specific dietary compounds and their impact on melanoma development and treatment.

In our previous experience, a high consistency of BRAF and NRAS muta- tion patterns was observed between primary tumors and lymph node me- tastases in patients with advanced melanoma. Conversely, increasing rates of discrep- ancies in BRAF/NRAS mutation pat- terns were found between primary melanomas and metastases in other sites (brain or, mostly, skin). When the distribution of BRAF/NRAS mutations was evaluated in a larger cohort, the high rate of consistency in sequence variations of these two genes was further confirmed between primary melanomas and lymph node metastases.

The occurrence of multiple primary melanomas(MPM) is quite rare with an estimated incidence ranging from 0.2% to 8.6%(Buljan M et al.2010). However, the risk of developing a primary melanoma is increased in patients with a history of a previous melanoma compared with the general population (De Giorgi et al.,2012). Between 6 and 14% of MPM cases occur in individuals with a family history of disease (Ang CG et al.1998), suggesting a genetic component in the pathogenesis of MPM. Germline mutations in the CDKN2A gene have been widely reported as the most common cause of inherited susceptibility to melanoma (Palmieri G et al., 2009). In the literature,CDKN2A germline mutations have been identified in less than 2% of single primary melanoma cases, 8 to 12% of sporadic MPM cases and 47% of MPM patients with familial melanoma (De Giorgi V et al.,2014;Puig S et al.,2005). This article is protected by copyright

Background: Our group previously demonstrated a high consistency of BRAF and NRAS mutations between primary tumors and lymph node metastases in melanoma patients (JCO 2012; 30: 2522-9). To further verify the level of homogeneity during clonal expansion to lymph node sites, we here performed a more detailed analysis on a subset of those paired samples using a NGS technology. Methods: Genomic DNA was isolated from tumor tissues of 12 melanoma patients, using standard methods. Specimens were analyzed for mutations in 50 most common cancer genes with the Ion Torrent AmpliSeq Cancer Panel HotSpot V2 (CHPv2) on the Ion Torrent platform. All variants detected by NGS were confirmed through PCR-based Sanger sequencing. Results: Genomic DNA was isolated from paired metastatic lymph nodes either asynchronously (n = 8) or synchronously (n = 4) excised from melanoma patients with clinically-evident nodal involvement. Data from the runs on the Ion Ampliseq Cancer Hotspot panel indicated an average of about 1500 reads (range, 500-2000) per amplicon. Overall, one (8%) out of the 12 analyzed patients presented discrepancies in mutation patterns during disease progression to lymph nodes and between the distinct lymph nodal metastases. In particular, the differences were represented by BRAF and CDKN2A mutations, suggesting that changes - when occur (though in a very limited fraction of cases) - may affect the main genes controlling cell proliferation and survival in melanoma. Mutations in BRAF, NRAS, CDKN2A, and TP53 genes were confirmed to play a predominant role in melanoma pathogenesis. Conclusions: Even with a more comprehensive mutation analysis, our findings indicated a very low genetic heterogeneity in metastatic lymph nodes during disease progression in melanoma patients, confirming previously published data on BRAF and NRAS mutational status only.

Background: Mutation in KRAS gene has been extensively demonstrated to act as a predictor of response to EGFR-targeted agents in patients with colorectal cancer. Less is known about the significance of KRAS mutation as a prognostic factor of disease progression and survival, independently of anti-EGFR therapy. The aim of the present study is to evaluate the prognostic role of the KRAS mutational status in a cohort of Sardinian patients with colorectal cancer. Materials and methods: Five hundred and fifty-one consecutive Sardinian patients with histologically proven diagnosis of invasive colorectal carcinoma were included into the study, regardless of age at diagnosis and disease characteristics. Clinical and pathological disease features were confirmed by medical records, pathology reports, and cancer registry data. For mutational analysis, paraffin embedded tissue samples with at least 70% neoplastic cells were processed; genomic DNA was isolated from tissue sections and DNA quality assessed for each specimen. The coding sequences and splice junctions of exons 2, 3, and 4 in KRAS gene were screened for mutations by direct automated sequencing. Results: KRAS mutations were detected in 183/551 (33%) patients; three of them presented the coexistence of two KRAS mutations in the same primary tumor tissue. Among the 186 KRAS mutations identified, two thirds (125; 67%) were located in codon 12, about one fifth (36; 19.4%) in codon 13, and about one tenth (18; 9.7%) in codon 61. The remaining mutations (7; 3.8%) were detected in uncommonly-affected codons of the KRAS gene. No significant correlations between KRAS mutations and sex, age at diagnosis, anatomical location, and disease stage at the time of diagnosis were found. No prognostic values of KRAS mutations were found for either the time to progression as metastatic disease or the overall survival. When patients were stratified by both KRAS mutational status and sex, a significantly better metastasis-free survival was observed for KRAS-mutated male cases. Considering the gene positions of the identified KRAS mutations, no correlation with both the time to progression as metastatic disease and overall survival was observed. Conclusions: Our data suggest that KRAS mutations are correlated to a slower progression of the disease in males with colorectal cancer from Sardinia, irrespectively of the diagnosis age and the mutation position. Such findings were not confirmed considering the overall survival in both sexes.

The paper reports the results of an experimental investigation aimed at transferring aerobic granulation to a demonstrative SBBGR system (i.e., a submerged biofilter that operates in a "fill and draw" mode) for the treatment of municipal wastewater by financial support of the EU Life programme. The results show that following the generation of granular biomass during the start-up period, the SBBGR was able to remove 80-90% of COD, total suspended solids and TKN in primary effluent from a municipal wastewater treatment plant, independently of the hydraulic residence time investigated (i.e., from 12 down to 3 h). The pocess was characterised by a very high sludge age value (?c?150 d) that led to a biomass concentration as high as 40 gTSS/Lbed and a sludge production almost one magnitude order lower than commonly reported for conventional treatment plants.