Combining germline, tissue and liquid biopsy analysis by comprehensive genomic profiling to improve the yield of actionable variants in a real-world cancer cohort.

BACKGROUND: Comprehensive next-generation sequencing is widely used for precision oncology and precision prevention approaches. We aimed to determine the yield of actionable gene variants, the capacity to uncover hereditary predisposition and liquid biopsy appropriateness instead of, or in addition to, tumor tissue analysis, in a real-world cohort of cancer patients, who may benefit the most from comprehensive genomic profiling. METHODS: Seventy-eight matched germline/tumor tissue/liquid biopsy DNA and RNA samples were profiled using the Hereditary Cancer Panel (germline) and the TruSight Oncology 500 panel (tumor tissue/cfDNA) from 23 patients consecutively enrolled at our center according to at least one of the following criteria: no available therapeutic options; long responding patients potentially fit for other therapies; rare tumor; suspected hereditary cancer; primary cancer with high metastatic potential; tumor of unknown primary origin. Variants were annotated for OncoKB and AMP/ASCO/CAP classification. RESULTS: The overall yield of actionable somatic and germline variants was 57% (13/23 patients), and 43.5%, excluding variants previously identified by somatic or germline routine testing. The accuracy of tumor/cfDNA germline-focused analysis was demonstrated by overlapping results of germline testing. Five germline variants in BRCA1, VHL, CHEK1, ATM genes would have been missed without extended genomic profiling. A previously undetected BRAF p.V600E mutation was emblematic of the clinical utility of this approach in a patient with a liver undifferentiated embryonal sarcoma responsive to BRAF/MEK inhibition. CONCLUSIONS: Our study confirms the clinical relevance of performing extended parallel tumor DNA and cfDNA testing to broaden therapeutic options, to longitudinally monitor cfDNA during patient treatment, and to uncover possible hereditary predisposition following tumor sequencing in patient care.

High BRAF variant allele frequencies are associated with distinct pathological features and responsiveness to target therapy in melanoma patients.

BACKGROUND: BRAF mutant melanoma patients are commonly treated with anti-BRAF therapeutic strategies. However, many factors, including the percentage of BRAF-mutated cells, may contribute to the great variability in patient outcomes. PATIENTS AND METHODS: The BRAF variant allele frequency (VAF; defined as the percentage of mutated alleles) of primary and secondary melanoma lesions, obtained from 327 patients with different disease stages, was assessed by pyrosequencing. The BRAF mutation rate and VAF were then correlated with melanoma pathological features and patients' clinical characteristics. Kaplan-Meier curves were used to study the correlations between BRAF VAF, overall survival (OS), and progression-free survival (PFS) in a subset of 62 patients treated by anti-BRAF/anti-MEK therapy after metastatic progression. RESULTS: A highly heterogeneous BRAF VAF was identified (3%-90%). Besides being correlated with age, a higher BRAF VAF level was related to moderate lymphocytic infiltration (P = 0.017), to melanoma thickness according to Clark levels, (level V versus III, P = 0.004; level V versus IV, P = 0.04), to lymph node metastases rather than cutaneous (P = 0.04) or visceral (P = 0.03) secondary lesions. In particular, a BRAF VAF >25% was significantly associated with a favorable outcome in patients treated with the combination of anti-BRAF/anti-MEK drug (OS P = 0.04; PFS P = 0.019), retaining a significant value as an independent factor for the OS and the PFS in the multivariate analysis (P = 0.014 and P = 0.003, respectively). CONCLUSION: These results definitively support the role of the BRAF VAF as a potential prognostic and predictive biomarker in melanoma patients in the context of BRAF inhibition.

Determination of selected trace elements in patients affected by dementia.

The relationship between dementia and trace elements is widely debated. Neurotoxicity of aluminium is well recognized. The purpose of the study was to evaluate serum levels of a few trace elements and a few serum proteins in demented subjects. The study was carried out on 452 women, age range 73-88 years. Thirty one of them were affected by dementia of Alzheimer type in early or middle stages. The diagnosis had been performed by history, physical and instrumental examinations, as well as by psychometric tests. The following parameters were determined: iron, zinc, copper, serum albumin, transferrin, ceruloplasmin. Iron, copper and zinc are somewhat lower in demented subjects than in controls, but the differences are statistically not significant. The slightly diminished levels of serum albumin and transferrin in the study group may be related to a mild malnutrition. The reduction of the proteins binding trace elements, particularly of transferrin, could cause a higher bioavailability of neurotoxic trace elements such as aluminium.

[Changes in the electrocardiogram in the elderly patient. The limits between normality and pathology]. / Modificazioni dell'elettrocardiogramma nell'anziano. Limiti tra normalità e patologia.

Normal electrocardiographic criteria in the elderly are not well defined. The prevalence of electrocardiographic abnormalities is three times higher in subjects over 85 years than in subjects 65-69 years old. With aging the heart rate does not vary during rest and sinus rhythm is prevalent, although sinus pauses, overall at night, are frequently seen. First degree atrio-ventricular block is present in 8.1%-19% of the elderly. 11% of subjects over 70 years suffer from left anterior hemiblock. 4.3% of subjects over 65 years and 7% of those over 85 present a right bundle branch block. 1.7% of subjects older than 65 years and 9% of those older than 85 are affected by a left bundle branch block. Atrial ectopic contractions are present in 8.8% of subjects over 60 years and in all older subjects. Atrial fibrillation is more common as age increases, being found in 2% of under 75s, in 5% of all subjects over this age, in 14% over 85 years and in 27% of patients hospitalized or institutionalized aged over 90 years. The prevalence of ventricular ectopic contractions varies from 76% in studies performed with baseline electrocardiogram to 96% in studies performed with portable monitoring electrocardiogram. Major ST-T wave alterations are present in 6.3%-13% of the elderly. In 340 patients over 80 years, hospitalized for diseases, other than cardiovascular, we found atrial fibrillation in 27.9% of subjects, ectopic beats in 26.2%, first degree atrioventricular block in 8.5%, right bundle branch block in 11.2%, left bundle branch block in 5.9%, left anterior hemiblock in 8.2%, ST-T wave alterations in 23.8% of the population.