Impact of functional studies on exome sequence variant interpretation in early-onset cardiac conduction system diseases

Aims The genetic cause of cardiac conduction system disease (CCSD) has not been fully elucidated. Whole-exome sequencing (WES) can detect various genetic variants; however, the identification of pathogenic variants remains a challenge. We aimed to identify pathogenic or likely pathogenic variants in CCSD patients by using WES and 2015 American College of Medical Genetics and Genomics (ACMG) standards and guidelines as well as evaluating the usefulness of functional studies for determining them. Methods and Results We performed WES of 23 probands diagnosed with early-onset (<65 years) CCSD and analyzed 117 genes linked to arrhythmogenic diseases or cardiomyopathies. We focused on rare variants (minor allele frequency < 0.1%) that were absent from population databases. Five probands had protein truncating variants in EMD and LMNA which were classified as “pathogenic” by 2015 ACMG standards and guidelines. To evaluate the functional changes brought about by these variants, we generated a knock-out zebrafish with CRISPR-mediated insertions or deletions of the EMD or LMNA homologs in zebrafish. The mean heart rate and conduction velocities in the CRISPR/Cas9-injected embryos and F2 generation embryos with homozygous deletions were significantly decreased. Twenty-one variants of uncertain significance were identified in 11 probands. Cellular electrophysiological study and in vivo zebrafish cardiac assay showed that 2 variants in KCNH2 and SCN5A, 4 variants in SCN10A, and 1 variant in MYH6 damaged each gene, which resulted in the change of the clinical significance of them from “Uncertain significance” to “Likely pathogenic” in 6 probands. Conclusions Of 23 CCSD probands, we successfully identified pathogenic or likely pathogenic variants in 11 probands (48%). Functional analyses of a cellular electrophysiological study and in vivo zebrafish cardiac assay might be useful for determining the pathogenicity of rare variants in patients with CCSD. SCN10A may be one of the major genes responsible for CCSD. Translational Perspective Whole-exome sequencing (WES) may be helpful in determining the causes of cardiac conduction system disease (CCSD), however, the identification of pathogenic variants remains a challenge. We performed WES of 23 probands diagnosed with early-onset CCSD, and identified 12 pathogenic or likely pathogenic variants in 11 of these probands (48%) according to the 2015 ACMG standards and guidelines. In this context, functional analyses of a cellular electrophysiological study and in vivo zebrafish cardiac assay might be useful for determining the pathogenicity of rare variants, and SCN10A may be one of the major development factors in CCSD

Structural Stability of Lexical Semantic Spaces: Nouns in Chinese and French

Many studies in the neurosciences have dealt with the semantic processing of words or categories, but few have looked into the semantic organization of the lexicon thought as a system. The present study was designed to try to move towards this goal, using both electrophysiological and corpus-based data, and to compare two languages from different families: French and Mandarin Chinese. We conducted an EEG-based semantic-decision experiment using 240 words from eight categories (clothing, parts of a house, tools, vehicles, fruits/vegetables, animals, body parts, and people) as the material. A data-analysis method (correspondence analysis) commonly used in computational linguistics was applied to the electrophysiological signals. The present cross-language comparison indicated stability for the following aspects of the languages' lexical semantic organizations: (1) the living/nonliving distinction, which showed up as a main factor for both languages; (2) greater dispersion of the living categories as compared to the nonliving ones; (3) prototypicality of the \emph{animals} category within the living categories, and with respect to the living/nonliving distinction; and (4) the existence of a person-centered reference gradient. Our electrophysiological analysis indicated stability of the networks at play in each of these processes. Stability was also observed in the data taken from word usage in the languages (synonyms and associated words obtained from textual corpora).Comment: 17 pages, 4 figure

Response inhibition is linked to emotional devaluation: behavioural and electrophysiological evidence

To study links between the inhibition of motor responses and emotional evaluation, we combined electrophysiological measures of prefrontal response inhibition with behavioural measures of affective evaluation. Participants first performed a Go-Nogo task in response to Asian and Caucasian faces (with race determining their Go or Nogo status), followed by a trustworthiness rating for each face. Faces previously seen as Nogo stimuli were rated as less trustworthy than previous Go stimuli. To study links between the efficiency of response inhibition in the Go-Nogo task and subsequent emotional evaluations, the Nogo N2 component was quantified separately for faces that were later judged to be high versus low in trustworthiness. Nogo N2 amplitudes were larger in response to low-rated as compared to high-rated faces, demonstrating that trial-by-trial variations in the efficiency of response inhibition triggered by Nogo faces, as measured by the Nogo N2 component, co-vary with their subsequent affective evaluation. These results suggest close links between inhibitory processes in top-down motor control and emotional responses

Preserving neural function under extreme scaling

Important brain functions need to be conserved throughout organisms of extremely varying sizes. Here we study the scaling properties of an essential component of computation in the brain: the single neuron. We compare morphology and signal propagation of a uniquely identifiable interneuron, the HS cell, in the blowfly (Calliphora) with its exact counterpart in the fruit fly (Drosophila) which is about four times smaller in each dimension. Anatomical features of the HS cell scale isometrically and minimise wiring costs but, by themselves, do not scale to preserve the electrotonic behaviour. However, the membrane properties are set to conserve dendritic as well as axonal delays and attenuation as well as dendritic integration of visual information. In conclusion, the electrotonic structure of a neuron, the HS cell in this case, is surprisingly stable over a wide range of morphological scales

International Guillain-Barré Syndrome Outcome Study (IGOS): protocol of a prospective observational cohort study on clinical and biological predictors of disease course and outcome in Guillain-Barré syndrome

Guillain-Barré syndrome (GBS) is an acute polyradiculoneuropathy with a highly variable clinical presentation, course, and outcome. The factors that determine the clinical variation of GBS are poorly understood which complicates the care and treatment of individual patients. The protocol of the ongoing International GBS Outcome Study (IGOS), a prospective, observational, multi-centre cohort study that aims to identify the clinical and biological determinants and predictors of disease onset, subtype, course and outcome of GBS is presented here. Patients fulfilling the diagnostic criteria for GBS, regardless of age, disease severity, variant forms, or treatment, can participate if included within two weeks after onset of weakness. Information about demography, preceding infections, clinical features, diagnostic findings, treatment, course and outcome is collected. In addition, cerebrospinal fluid and serial blood samples for serum and DNA is collected at standard time points. The original aim was to include at least 1000 patients with a follow-up of 1-3 years. Data are collected via a web-based data entry system and stored anonymously. IGOS started in May 2012 and by January 2017 included more than 1400 participants from 143 active centres in 19 countries across 5 continents. The IGOS data/biobank is available for research projects conducted by expertise groups focusing on specific topics including epidemiology, diagnostic criteria, clinimetrics, electrophysiology, antecedent events, antibodies, genetics, prognostic modelling, treatment effects and long-term outcome of GBS. The IGOS will help to standardize the international collection of data and biosamples for future research of GBS. ClinicalTrials.gov Identifier: NCT01582763

Name agreement in picture naming: An ERP study

Name agreement is the extent to which different people agree on a name for a particular picture. Previous studies have found that it takes longer to name low name agreement pictures than high name agreement pictures. To examine the effect of name agreement in the online process of picture naming, we compared event-related potentials (ERPs) recorded whilst 19 healthy, native English speakers silently named pictures which had either high or low name agreement. A series of ERP components was examined: P1 approximately 120ms from picture onset, N1 around 170ms, P2 around 220ms, N2 around 290ms, and P3 around 400ms. Additionally, a late time window from 800 to 900ms was considered. Name agreement had an early effect, starting at P1 and possibly resulting from uncertainty of picture identity, and continuing into N2, possibly resulting from alternative names for pictures. These results support the idea that name agreement affects two consecutive processes: first, object recognition, and second, lexical selection and/or phonological encoding

The European Network for Translational Research in Atrial Fibrillation (EUTRAF): objectives and initial results.

Atrial fibrillation (AF) is the most common sustained arrhythmia in the general population. As an age-related arrhythmia AF is becoming a huge socio-economic burden for European healthcare systems. Despite significant progress in our understanding of the pathophysiology of AF, therapeutic strategies for AF have not changed substantially and the major challenges in the management of AF are still unmet. This lack of progress may be related to the multifactorial pathogenesis of atrial remodelling and AF that hampers the identification of causative pathophysiological alterations in individual patients. Also, again new mechanisms have been identified and the relative contribution of these mechanisms still has to be established. In November 2010, the European Union launched the large collaborative project EUTRAF (European Network of Translational Research in Atrial Fibrillation) to address these challenges. The main aims of EUTRAF are to study the main mechanisms of initiation and perpetuation of AF, to identify the molecular alterations underlying atrial remodelling, to develop markers allowing to monitor this processes, and suggest strategies to treat AF based on insights in newly defined disease mechanisms. This article reports on the objectives, the structure, and initial results of this network

Virtual cardiac monolayers for electrical wave propagation

The complex structure of cardiac tissue is considered to be one of the main determinants of an arrhythmogenic substrate. This study is aimed at developing the first mathematical model to describe the formation of cardiac tissue, using a joint in silico-in vitro approach. First, we performed experiments under various conditions to carefully characterise the morphology of cardiac tissue in a culture of neonatal rat ventricular cells. We considered two cell types, namely, cardiomyocytes and fibroblasts. Next, we proposed a mathematical model, based on the Glazier-Graner-Hogeweg model, which is widely used in tissue growth studies. The resultant tissue morphology was coupled to the detailed electrophysiological Korhonen-Majumder model for neonatal rat ventricular cardiomyocytes, in order to study wave propagation. The simulated waves had the same anisotropy ratio and wavefront complexity as those in the experiment. Thus, we conclude that our approach allows us to reproduce the morphological and physiological properties of cardiac tissue