Genome-wide association analyses identify new Brugada syndrome risk loci and highlight a new mechanism of sodium channel regulation in disease susceptibility.

Brugada syndrome (BrS) is a cardiac arrhythmia disorder associated with sudden death in young adults. With the exception of SCN5A, encoding the cardiac sodium channel Na1.5, susceptibility genes remain largely unknown. Here we performed a genome-wide association meta-analysis comprising 2,820 unrelated cases with BrS and 10,001 controls, and identified 21 association signals at 12 loci (10 new). Single nucleotide polymorphism (SNP)-heritability estimates indicate a strong polygenic influence. Polygenic risk score analyses based on the 21 susceptibility variants demonstrate varying cumulative contribution of common risk alleles among different patient subgroups, as well as genetic associations with cardiac electrical traits and disorders in the general population. The predominance of cardiac transcription factor loci indicates that transcriptional regulation is a key feature of BrS pathogenesis. Furthermore, functional studies conducted on MAPRE2, encoding the microtubule plus-end binding protein EB2, point to microtubule-related trafficking effects on Na1.5 expression as a new underlying molecular mechanism. Taken together, these findings broaden our understanding of the genetic architecture of BrS and provide new insights into its molecular underpinnings

Etude in vitro et in vivo de la régulation de la mort radioinduite des cellules endothéliales par la Sphingosine-1-Phosphate

Protéger la vascularisation de la mort radioinduite est d un intérêt majeur en radioprotection tissulaire. En développant un modèle d étude de la radiosensibilité des cellules endothéliales, nous avons prouvé que les cellules HMEC-1 subissent 2 voies de mort après une irradiation à 15 Gy: une apoptose prémitotique précoce induite par la génération du céramide, et une mort mitotique tardive induite par les dommages de l ADN. La Sphingosine-1-Phosphate (S1P), un métabolite du céramide, protège les HMEC-1 seulement de la mort précoce, mais pas de la mort mitotique. Nous avons confirmé in vivo que la S1P protège de l apoptose radioinduite due à la génération du céramide. Cette radioprotection est partiellement dépendante de l activation des récepteurs S1Ps. La distinction entre ces 2 types de mort pourrait permettre de définir de nouveaux radioprotecteurs spécifiques des tissus sains dont l endothélium est quiescent, mais pas des cellules endothéliales tumorales qui sont pro-angiogéniques.Protecting the vasculature from radiation-induced death is a major concern in tissue radioprotection. Developping a model of endothelial cells radiosensitivity, we proved that HMEC-1 undergo 2 waves of death after exposure to 15 Gy: an early premitotic apoptosis dependent of ceramide generation and a delayed DNA damage-induced mitotic death. Sphingosine-1-Phosphate (S1P), a ceramide antagonist, protects HMEC-1 only from early apoptosis, but not from mitotic death. We confirmed in vivo the S1P radioprotection from ceramide-mediated radio-induced apoptosis, and that S1P radioprotection is partially mediated by S1Ps receptors. Segregation between these 2 types of death may give the opportunity to define a new class of radioprotectors for normal tissue where quiescent endothelium represent the most sensitive target, while excluding malignant tumor containing pro-proliferating angiogenic endothelial cells, sensitive to mitotic death.NANTES-BU Médecine pharmacie (441092101) / SudocPARIS-BIUP (751062107) / SudocSudocFranceF

RhoA GTPase regulates radiation-induced alterations in endothelial cell adhesion and migration

International audienceEndothelial cells of the microvasculature are major target of ionizing radiation, responsible of the radiationinduced vascular early dysfunctions. Molecular signaling pathways involved in endothelial responses to ionizing radiation, despite being increasingly investigated, still need precise characterization. Small GTPase RhoA and its effector ROCK are crucial signaling molecules involved in many endothelial cellular functions. Recent studies identified implication of RhoA/ROCK in radiation-induced increase in endothelial permeability but other endothelial functions altered by radiation might also require RhoA proteins. Human microvascular endothelial cells HMEC-1, either treated with Y-27632 (inhibitor of ROCK) or invalidated for RhoA by RNA interference were exposed to 15 Gy. We showed a rapid radiation-induced activation of RhoA, leading to a deep reorganisation of actin cytoskeleton with rapid formation of stress fibers. Endothelial early apoptosis induced by ionizing radiation was not affected by Y-27632 pre-treatment or RhoA depletion. Endothelial adhesion to fibronectin and formation of focal adhesions increased in response to radiation in a RhoA/ROCK-dependent manner. Consistent with its pro-adhesive role, ionizing radiation also decreased endothelial cells migration and RhoA was required for this inhibition. These results highlight the role of RhoA GTPase in ionizing radiation-induced deregulation of essential endothelial functions linked to actin cytoskeleton

Plasma membrane reorganization links acid sphingomyelinase/ceramide to p38 MAPK pathways in endothelial cells apoptosis

International audienceThe p38 MAPK signaling pathway is essential in the cellular response to stress stimuli, in particular in the endothelial cells that are major target of external stress. The importance of the bioactive sphingolipid ceramide generated by acid sphingomyelinase is also firmly established in stress-induced endothelial apoptotic cell death. Despite a suggested link between the p38 MAPK and ceramide pathways, the exact molecular events of this connection remain elusive. In the present study, by using two different activators of p38 MAPK, namely anisomycin and ionizing radiation, we depicted how ceramide generated by acid sphingomyelinase was involved in p38 MAPK-dependent apoptosis of endothelial cells. We first proved that both anisomycin and ionizing radiation conducted to apoptosis through activation of p38 MAPK in human microvascular endothelial cells HMEC-1. We then found that both treatments induced activation of acid sphin-gomyelinase and the generation of ceramide. This step was required for p38 MAPK activation and apoptosis. We finally showed that irradiation, as well as treatment with exogenous C 16-ceramide or bacterial sphingomyelinase, induced in en-dothelial cells a deep reorganization of the plasma membrane with formation of large lipid platforms at the cell surface, leading to p38 MAPK activation and apoptosis in endothelial cells. Altogether, our results proved that the plasma membrane reorganization leading to ceramide production is essential for stress-induced activation of p38 MAPK and apoptosis in endothelial cells and established the link between the acid sphingomyelinase/ceramide and p38 MAPK pathways

Rare coding variants in CTSO , a potential new actor of arterial remodeling, are associated to familial intracranial aneurysm

Background Intracranial aneurysm (IA) is a common cerebrovascular abnormality characterized by localized dilation and wall thinning in intracranial arteries, that frequently leads to fatal vascular rupture. The mechanisms underlying IA formation, growth and rupture are mostly unknown, and while increasing evidence suggest a genetic component of IA, identification of specific genes or causal molecular pathways remains largely inconclusive and only a small fraction of the risk attributable to genetics for IA in the general population. Methods: Here, we combined whole exome sequencing and identity-by -descent analyses with functional investigations to identify rare IA predisposing variants in familial forms of IA and understand their contribution to the pathophysiology of IA. Results We identified two rare missense variants in the CTSO gene shared by all the affected relatives in two large pedigrees with multiple IA-affected relative. CTSO encodes for the cysteine-type papain-like cathepsin CTSO. Functional analyses revealed that CTSO acts as an extracellular protease controlling vascular smooth muscle cell migration and adhesion to the extracellular matrix. CTSO depletion, as well as expression of the two CTSO variants, which were poorly secreted, led to increase the amount of fibronectin. This effect is associated with a marked increase in VSMC stiffness which was rescued by exogenous CTSO. Conclusions This report identifies rare CTSO variants in familial IA patients and suggests that the increased susceptibility to IA induced by these variants is likely related to their primary effects on the vascular tissue, and more particularly on the media layer of the wall of cerebral arteries

0185 : Genetic screening identifies a high proportion of mutations in patients with idiopathic ventricular fibrillation and sudden cardiac death

International audienceIntroduction Several gene defects are associated with idiopathic ventricular fibrillation (IVF) and sudden cardiac death (SCD). The recent development of NGS-based mutation screening provides a unique opportunity to estimate extensively the spectrum and prevalence of rare variants in genes associated with cardiac diseases. Methods Cohort 1 was composed of 75 patients resuscitated from cardiac arrest due to IVF. All patients have undergone a complete clinical cardiac examination including 12 lead-ECG, cardiac echography, coronography and exercise test. Cohort 2 was composed of 99 victims of SCD related to ventricular fibrillation younger than 45 years old and without explanation for the SCD at the time of the reanimation. Genetic screening was based on the use of the HaloPlex(tm) Target Enrichment System (Agilent Technologies) prior to HiSeq sequencing (Illumina). The custom kit designed for this study covers 163 genes previously reported as involved in cardiac arrhythmias, conduction defect and cardiomyopathies. Results In cohort 1, the mean age was 36±10 years with a male predominance (52 males, 69%). In cohort 2, the mean age was 37±7 years with a male predominance (76 males, 79%). In cohort 1, we identified 50 putative mutations in 35 patients (47%). In cohort 2, we identified 30 putative mutations in 24 patients (24%). Conclusion Our study identified mutations in almost 50 % of IVF patients after a complete cardiac evaluation. These results suggest that molecular analysis must be part of the work up in this kind of patients. In young patients affected by unexplained sudden death, the molecular analyses are less contributive probably because of a more important percentage of patients affected by ischemic cardiomyopathies

Prospective, multicenter, controlled study of quality of life, psychological adjustment process and medical outcomes of patients receiving a preemptive kidney transplant compared to a similar population of recipients after a dialysis period of less than three years – The PreKit-QoL study protocol

International audienceBackground: Treatment of end stage renal disease has an impact on patients' physical and psychological health, including quality of life (QoL). Nowadays, it is known that reducing the dialysis period has many advantages regarding QoL and medical outcomes. Although preemptive transplantation is the preferred strategy to prevent patients undergoing dialysis, its psychological impact is unknown. Moreover, transplantation can be experienced in a completely different manner among patients who were on dialysis and those who still had a functioning kidney at the time of surgery. Longitudinal data are often collected to allow analyzing the evolution of patients' QoL over time using questionnaires. Such data are often difficult to interpret due to the patients' changing standards, values, or conceptualization of what the questionnaire is intended to measure (e.g. QoL). This phenomenon is referred to as response shift and is often linked to the way the patients might adapt or cope with their disease experience. Whether response shift is experienced in a different way among patients who were on dialysis and those who still had a functioning kidney at time of surgery is unknown and will be studied in the PreKit-QoL study (trial registration number: NCT02154815). Understanding the psychological impact of pre-emptive transplantation is an important issue since it can be associated with long-term patient and graft survival

Testing the burden of rare variation in arrhythmia-susceptibility genes provides new insights into molecular diagnosis for Brugada syndrome

International audienceThe Brugada syndrome (BrS) is a rare heritable cardiac arrhythmia disorder associated with ventricular fibrillation and sudden cardiac death. Mutations in the SCN5A gene have been causally related to BrS in 20-30% of cases. Twenty other genes have been described as involved in BrS, but their overall contribution to disease prevalence is still unclear. This study aims to estimate the burden of rare coding variation in arrhythmia-susceptibility genes among a large group of patients with BrS. We have developed a custom kit to capture and sequence the coding regions of 45 previously reported arrhythmia-susceptibility genes and applied this kit to 167 index cases presenting with a Brugada pattern on the electrocardiogram as well as 167 individuals aged over 65-year old and showing no history of cardiac arrhythmia. By applying burden tests, a significant enrichment in rare coding variation (with a minor allele frequency below 0.1%) was observed only for SCN5A, with rare coding variants carried by 20.4% of cases with BrS versus 2.4% of control individuals (P = 1.4 × 10(-7)). No significant enrichment was observed for any other arrhythmia-susceptibility gene, including SCN10A and CACNA1C. These results indicate that, except for SCN5A, rare coding variation in previously reported arrhythmia-susceptibility genes do not contribute significantly to the occurrence of BrS in a population with European ancestry. Extreme caution should thus be taken when interpreting genetic variation in molecular diagnostic setting, since rare coding variants were observed in a similar extent among cases versus controls, for most previously reported BrS-susceptibility gene

Burden of rare variants in arrhythmogenic cardiomyopathy with right dominant form associated genes provides new insights for molecular diagnosis and clinical management.

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