No abstract

Somatic copy number variations (CNV; i.e., amplifications and deletions) have been implicated in the origin and development of multiple cancers and some of these aberrations are designated targets for therapies. Although FISH is still considered the gold standard for CNV detection, the increasing number of potentially druggable amplifications to be assessed makes a gene-by-gene approach time- and tissue-consuming. Here we investigated the potential of next generation sequencing (NGS) custom panels to simultaneously determine CNVs across FFPE solid tumor samples. DNA was purified from cell lines and FFPE samples and analyzed by NGS sequencing using a 20-gene custom panel in the GeneReader Platform®. CNVs were identified using an in-house algorithm based on the UMI read coverage. Retrospective validation of in-house algorithm to identify CNVs showed 97.1% concordance rate with the NGS custom panel. The prospective analysis was performed in a cohort of 243 FFPE samples from patients arriving at our hospital, which included 74 NSCLC tumors, 148 CRC tumors, and 21 other tumors. Of them, 33% presented CNVs by NGS and in 14 cases (5.9%) the CNV was the only alteration detected. We have identified CNV alterations in about one-third of our cohort, including FGFR1, CDK6, CDK4, EGFR, MET, ERBB2, BRAF, or KRAS. Our work highlights the need to include CNV testing as a part of routine NGS analysis in order to uncover clinically relevant gene amplifications that can guide the selection of therapies.

(1) Background: Extracellular vesicles (EVs) have emerged as crucial players in the communication between cells in both physiological and pathological scenarios. The functions of EVs are strongly determined by their molecular content, which includes all bioactive molecules, such as proteins, lipids, RNA, and, as more recently described, double-stranded DNA. It has been shown that in oncological settings DNA associated with EVs (EV-DNA) is representative of the genome of parental cells and that it reflects the mutational status of the tumor, gaining much attention as a promising source of biomarker mutant DNA. However, one of the challenges in studies of EV-DNA is the lack of standardization of protocols for the DNA extraction from EVs, as well as ways to assess quality control, which hinders its future implementation in clinics. (2) Methods: We performed a comprehensive comparison of commonly used approaches for EV-DNA extraction by assessing DNA quantity, quality, and suitability for downstream analyses. (3) Results: We here established strategic points to consider for EV-DNA preparation for mutational analyses, including qPCR and NGS. (4) Conclusions: We put in place a workflow that can be applied for the detection of clinically relevant mutations in the EV-DNA of cancer patients.

Migraine is a common neurological disorder considered the second most disabling condition worldwide. Its prevalence ranges from 2.6% to 21.7% in population studies. This study aimed to know the prevalence of diagnosed and undiagnosed migraine in the general Spanish adult population, their health care use, and factors related. A descriptive cross-sectional study was undertaken with 23,089 individuals >15 years from the 2017 Spanish National Health Survey. Three groups were defined: people diagnosed with migraine (DM), people reporting undiagnosed migraine (UM) and people without migraine. Sociodemographic, clinical and use of health resources data were collected. The scales Duke Social Support Index (DSSI) and General Health Questionnaire (GHQ-12) were used. Prevalence of DM and UM were determined with 95% confidence intervals. To determine the factors associated with DM and UM, a multinomial logistic regression model was used. The prevalence of DM was 8.6% (95%CI: 8.2–9), and UM, 0.9% (95%CI: 0.8–1). People with DM more frequently visited healthcare professionals (47.8%), required more supplementary tests (86.8), had a higher percentage of hospitalization (11.3%), and used emergency services (45.1%). Women had nearly three times the risk of DM and UM. Worse mental health was a risk factor for UM (OR = 1.20) and DM (OR = 1.18). The greater the work stress, the greater the risk of DM (OR = 1.12). An adequate monitoring and management of migraine in people with these characteristics could contribute to improving their quality of life and reducing costs in the system.