Whole-genome sequencing reveals complex mechanisms of intrinsic resistance to BRAF inhibition.

BACKGROUND: BRAF is mutated in ∼42% of human melanomas (COSMIC. http://www.sanger.ac.uk/genetics/CGP/cosmic/) and pharmacological BRAF inhibitors such as vemurafenib and dabrafenib achieve dramatic responses in patients whose tumours harbour BRAF(V600) mutations. Objective responses occur in ∼50% of patients and disease stabilisation in a further ∼30%, but ∼20% of patients present primary or innate resistance and do not respond. Here, we investigated the underlying cause of treatment failure in a patient with BRAF mutant melanoma who presented primary resistance. METHODS: We carried out whole-genome sequencing and single nucleotide polymorphism (SNP) array analysis of five metastatic tumours from the patient. We validated mechanisms of resistance in a cell line derived from the patient's tumour. RESULTS: We observed that the majority of the single-nucleotide variants identified were shared across all tumour sites, but also saw site-specific copy-number alterations in discrete cell populations at different sites. We found that two ubiquitous mutations mediated resistance to BRAF inhibition in these tumours. A mutation in GNAQ sustained mitogen-activated protein kinase (MAPK) signalling, whereas a mutation in PTEN activated the PI3 K/AKT pathway. Inhibition of both pathways synergised to block the growth of the cells. CONCLUSIONS: Our analyses show that the five metastases arose from a common progenitor and acquired additional alterations after disease dissemination. We demonstrate that a distinct combination of mutations mediated primary resistance to BRAF inhibition in this patient. These mutations were present in all five tumours and in a tumour sample taken before BRAF inhibitor treatment was administered. Inhibition of both pathways was required to block tumour cell growth, suggesting that combined targeting of these pathways could have been a valid therapeutic approach for this patient

Overview of the radiographers’ practice in 65 healthcare centers using digital mammography systems in Portugal

Purpose: To assess current practices in digital mammography (DM) in Portuguese healthcare providers using digital systems. To investigate compliance with European standards regarding mean glandular dose and quality control practice and to identify optimisation needs. Methods: Two questionnaires, targeted at breast radiographers and chief radiographers, were designed and applied in 65 imaging departments offering DM. Questions fielded were focused on the staff profile and technical/ clinical practice. Results: Prior to starting their activity in DM, 70% (82 out of 118) of the respondents received training in DM. The practice in 29 out of 59 providers was established by the manufacturers’ recommendations for image acquisition. Variations were observed between radiographers who belong to the same provider namely the selection of exposure parameters such as the target-filter combination and automatic mode. The use of the manual exposure mode was reported for imaging breast implants (44%) and surgical specimens (22%). The main causes of repeat examinations were skin folding (21%) and absence of pectoral muscle (PM) (20%). Conclusions: The study revealed opportunities to optimise radiographers’ practice in DM regarding the selection of exposure parameters. A robust and consistent training programme in DM and established local protocols can help to reduce the variations observed and improve clinical practice