No description available.

No description available.

Aberrations in conduction or repolarization are established prerequisites for arrhythmogenesis. The following dissertation investigates how reducing either ephaptic (EpC) or gap junctional (GJ) coupling between cardiomyocytes can modulate cardiac conduction, repolarization, or the relationship between these two phenomena. Our lab has previously demonstrated that EpC can be modified in the ventricular epicardium using ionic and osmotic challenges to the Langendorff-perfused heart. In the first series of experiments, we show that reducing EpC via treatment with mannitol or hyponatremia can unmask conduction deficits that are otherwise below the resolution of detection in Scn5a+/- mice. Interestingly, we also observe that combination of the two treatments resolves severe conduction delay due to hyponatremia in the heterozygous animal. These data suggest it may be valuable to pursue the use of mannitol or hyponatremia as novel diagnostics for sodium channel loss of function diseases. The importance of extracellular perfusate is also highlighted by the second investigation, which evaluates how sodium and calcium concentration modulate repolarization in the context of hyperkalemia, a common comorbidity of hospitalized patients that increases the risk of arrhythmia. Calcium may potentially play a role in modulating APD adaptation to pacing rate in the context of this disease state, though more research is needed to clarify the exact mechanism of this effect. Finally, we investigate the relationship between conduction and repolarization in the epicardium, and conclude that this relationship does not appear to be dictated by the degree of cell-cell coupling in the myocardium, but instead is driven by endogenous gradients of action potential duration within the tissue. Taken together, these data demonstrate ways in which both conduction and repolarization are sensitive to modulations of EpC, though we also find that the relationship between these two phenomena is not influenced by such changes in electrical coupling. / Doctor of Philosophy / The ability of the heart to function as a pump is dependent on the successful coordination of electrical activity throughout the heart. Disruptions to this intricate electrical system result in cardiac arrhythmias, which in turn prevent the heart from effectively perfusing the body with oxygenated blood. The present dissertation investigates ways in which we can modulate cell-cell communication within the heart, and how this may in turn influence disease states with a high propensity for arrhythmia. We show that reducing electrical coupling between cells using simple interventions like reducing serum sodium or increasing osmolarity may be a viable technique for diagnosing "concealed" disease states (i.e. disease states that are asymptomatic for much of a patient's life). We then explore ways in which elevated serum potassium, known as hyperkalemia, may alter the heart's ability to recover from electrical activation (repolarization). Finally, we show that the relationship between cardiac activation and repolarization is not as dependent on cell-cell communication as was once thought. Taken together, this dissertation provides evidence that transiently disrupting cell-cell communication may hold promise for development of diagnostics for some congenital cardiac diseases, and yet does not appear to disrupt the relationship between electrical conduction and repolarization across the heart.

Thèse (M.Sc.)--Université Laval, 2004. / Bibliogr. Publié aussi en version électronique.

Abstract Brugada syndrome is an inherited arrhythmia disorder that predisposes to sudden cardiac death. It is characterized by its distinct ECG pattern. The purpose of this thesis was to study the phenotype and genotype characteristics of subjects with Brugada syndrome type ECG. The first study population consisted of 2479 young male Air Force applicants and 542 healthy middle-aged subjects. The 12-lead ECG was analyzed to assess the prevalence and prognosis of Brugada pattern in Finnish population. The second population consisted of 168 patients with AF. The ECGs of the patients with family history of lone AF were analysed in order to characterize the ECG features of familial AF. The third population consisted of 200 patients with Brugada syndrome and their ECGs were analyzed for detection of distinct ECG characteristics. In a substudy, the H558R variant was genotyped and the clinical presentation of this variant was evaluated. The clinical characteristics were collected of 47 patients with induced Brugada ECG during fever or medication. The prevalence of type 2 or 3 Brugada ECG was 0.61% in the young population and 0.55% in the middle-aged Finnish population. In a retrospective analysis, none of the Brugada ECG carriers had died. In the AF study, the prevalence of type 2 or 3 Brugada ECG was significantly higher among the subjects with lone AF compared to the healthy controls (p < 0.001). Many of the Brugada ECG carriers had a family history (> 30% of first-degree relatives) of AF. In patients with Brugada syndrome, the prolonged QRS duration was associated with previous symptoms. The R allele carriers in H558R variant had a trend towards less symptoms (p = 0.067) and had less conduction disturbances in 12-lead ECG than the HH genotype carriers (p < 0.05 in all ECG analysis). Among the subjects with induced Brugada ECG, 51% exhibited arrhythmic symptoms during the medical condition that had provoked the ECG pattern. In conclusion, type 2 and 3 Brugada ECGs were found to be benign in the Finnish population since no mortality occurred during an extensive follow-up period. On the other hand, these ECG abnormalities seem to be a marker of familial AF. Among patients with the Brugada syndrome, a prolongation of QRS is associated with prior symptoms. The variant H558R R allele seems to be a protecting genetic modulator. Induced Brugada ECG is a medical emergency since the patients are at high risk of sudden cardiac death.

Le syndrome du QT long (LQTs) et le syndrome de Brugada (BS) sont deux maladies cardiaques héréditaires, pouvant causer la mort subite en relation avec des torsades de pointe dues à une fibrillation ventriculaire. SCN5A est le gène codant pour la sous-unité a du canal Na4 dépendant du voltage, exprimé dans le cœur humain. Objectif : Dans la présente étude, deux mutations (R1193Q et F1344S) dans le gène SCN5A ont été identifiées. La mutation R1193Q a été trouvée chez deux patients après le test de provocation à la procainamide par voie intraveineuse. Un des patients est atteint de BS et l'autre est atteint à la fois de BS et de LQTs. La mutation F1344S a été trouvée chez un autre patient atteint uniquement de BS. Méthode : Les canaux mutants ont été exprimés dans un système d'expression de mammifère (lignée cellulaire tsA 201) et les propriétés biophysiques ont été étudiées avec la technique de patch clamp en configuration cellule entière. Résultats : L'analyse des séquences montre un changement de G à A à la position 3587 sur l'exon 20. La mutation RI 193Q montre un déplacement négatif de 5mV de l'inactivation et un courant Na+ persistant. Le déplacement vers des potentiels négatifs de l'inactivation est responsable d'une perte de fonction causant l'élévation du segment ST chez le patient atteint de BS. La présence du courant persistant est responsable du gain de fonction causant l'augmentation de l'intervalle QT chez le patient avec LQTs. La mutation F1344S montre une manifestation typique de BS sur l'électrocardiogramme pendant une période de fièvre. Le séquençage a révélé un changement de T à C à la position 4031 sur l'exon 23. L'analyse biophysique montre une perte de fonction due au déplacement de la courbe d'activation vers des potentiels positifs à 23°C et ce déplacement est exacerbé à 40.5°C avec une pente plus lente. Conclusion : Les différentes manifestations cliniques de ces deux mutations sont la conséquence des anormalités électrophysiologiques distinctes du canal Na+ cardiaque mutant décrites dans cette étude. Pour les patients porteurs de R1193Q, la prescription de quelques drogues pouvant provoquer le LQTs et le BS doit être limitée. Pour éviter les épisodes de fibrillation ventriculaire, l'hyperthermie doit être contrôlée chez les patients porteurs de la mutation F1344S. / Long QT syndrome (LQTs) and Brugada syndrome (BS) are two distinct hereditary cardiac diseases, causing sudden cardiac death related to torsade de pointes and ventricular tachycardia (VT) / ventricular fibrillation (VF). SCN5A is the gene encoding the principal voltage-gated Na+ channel a-subunit, which is only expressed in the human heart. Objective: In present study, two mutations (R1193Q and F1344S) in SCN5A have been identified. RI 193Q mutation was found in two patients, one with BS and another with an overlap of BS and LQTs after the intravenous procainamide test. F1344S mutation was found in another patient with BS. Method: The mutant channels were expressed in a mammalian expression System (tsA 201 cell Une) and the biophysical properties were studied by the patch clamp technique with whole cell configuration. Results: The sequence analysis showed a G to A base change at position 3587 in exon 20. The R1193Q mutation produced a negative shift for 5 mV of inactivation and a persistent Na+ current. The negative shift of inactivation is responsible for loss of the function, leading to ST segment elevation in BS. The persistent Na+ current is responsible for gain of function, causing QT interval prolongation in LQTs. The F1344S mutation presented a typical BS manifestation on ECG during fever period. The sequencing analysis revealed a T to C base change at position 4031 in exon 23. The biophysical analysis showed loss of function due to significantly positive shift of activation at both 23°C and 40.5°C, but the shift was more important at 40.5°C with a slower slope factor. Conclusion: The different clinical manifestations of these two mutations probably derive from the distinct electrophysiological abnormalities of the mutant cardiac Na+ channels reported in this study. For patients with RI 193Q mutation, some drugs that likely precipitate LQTs and BS should be limited to prescribe. To prevent from VT/VF events, hyperthermia should be effectively controlled in the patients with F1344S mutation.

El Síndrome de Brugada (SB) se caracteriza por una elevación del segmento ST en las derivaciones precordiales derechas del ECG. Tiene una base genética estando ligado a mutaciones en el canal de Na+ del corazón (SCN5A) y otros genes recientemente relacionados. El estudio de secuenciación directa del gen SCN5A en nuestra población llevó a la identificación de mutaciones causales o probablemente causales en un 10% de los casos, inferior al publicado en la literatura. En los casos índice en los que no se detectó una variante en su secuencia se realizó el estudio de grandes reordenamientos génicos sin identificarse ninguno mediante la técnica de Multiplex Ligation Probe Amplification. Los pacientes de nuestra cohorte portadores de mutación presentan con mayor frecuencia ECG tipo I espontáneo y más casos de muerte súbita familiar que los pacientes de SB con genotipo negativo. El estudio genético de las familias de los portadores de mutación ha permitido identificar a un grupo significativo de portadores sin expresión fenotípica.

Reproduction de : Thèse de doctorat : Médecine. Génétique moléculaire : Nantes : 2008. / Bibliogr.

Le syndrome de Brugada (BrS) est une cardiopathie héréditaire à transmission autosomique dominante, se manifestant par une anomalie de l'ECG et un risque accru de mort subite. Les mutations retrouvées dans la sous-unité α du canal sodique cardiaque Nav1.5 chez certains patients entraînent un défaut d'adressage membranaire de ces canaux. Ceux-ci restent alors séquestrés dans des compartiments intracellulaires. L'étude de ces mutants se réduisant souvent à l'utilisation de traitements correcteurs, les mécanismes de rétention impliqués restent encore méconnus. L'objectif de ce travail est d'étudier des mutants Nav1.5 présentant un défaut d'adressage en tenant compte non seulement de l'hétérozygotie des patients BrS mais également de la présence de la sous unité régulatrice β1 prédominante dans le cœur. Des études fonctionnelles et biochimiques mettent en évidence un effet dominant négatif exercé par les mutants R1432G, L325R et S910L sur la densité de courant INa sauvage (WT). Cet effet nécessite la présence de la sous-unité β1 et passe par l'altération de l'adressage membranaire des formes WT. Ceci est la conséquence d'une interaction physique entre des sous-unités α mutantes et WT. D'autre part, les mutants étudiés présentent un profil de maturation lié aux N-glycosylations qui différent de celui des canaux WT. Nos données suggèrent que ces canaux peuvent emprunter (i) la voie classique d'adressage dans leur forme mature (ii) la voie dite non conventionnelle lorsqu'ils sont partiellement glycosylés. En conclusion, ces travaux mettent en évidence le rôle de la sous-unité β1 ainsi que l'implication des N-glycosylations dans la modulation de l'adressage des canaux Nav1.5 / Brugada syndrome (BrS) is an inherited autosomal dominant cardiac channelopathy characterized by abnormal ECG pattern and an increased risk of sudden cardiac death. Several mutations on the cardiac sodium channel Nav1.5 which are responsible for BrS lead to misfolded proteins that do not traffic properly to the plasma membrane and are instead retained in intracellular compartments. Although pharmacological rescue is commonly used to characterize misfolded mutants, underlying cellular retention mechanisms remain unclear. The aim of this work is to investigate trafficking defective Nav1.5 mutants considering BrS patient heterozygosity and the presence of the regulatory β1-subunit which is largely expressed in cardiac tissue. By combining electrophysiology and biochemical approaches, we show that three distinct mutants, R1432G, L325R and S910L, exert a strong dominant negative effect upon wild-type (WT) sodium current density. Our data indicate that this effect requires the presence of the β1-subunit and is mediated by disruption of membrane trafficking of WT channels. Co-immunoprecipitation experiments demonstrate a physical interaction between mutant and WT α-subunits occurring only when the β1-subunit was present. Furthermore, we investigate the maturation pattern of Na channels. Our data show distinct N-glycosylated states between WT and mutant channels, suggesting that Nav1.5 α-subunits traffic (i) via unconventional secretion pathway as a partially glycosylated product, (ii) through the classical secretory pathway for mature fully-glycosylated form. This work highlights that β1-subunit and N-linked glycosylation process play key roles in modulating Nav1.5 trafficki

En utilisant la méthode de génétique inverse, la mutation causale d'un bloc de conduction cardiaque familial a été localisée sur le bras long du chromosome 19 en 13.3 dans une grande famille libanaise. Après avoir testé 12 gènes candidats, nous avons trouvé 3 mutations différentes dans trois familles indépendantes de bloc de conduction cardiaque isolé. Les conséquences des 3 mutations ont été explorées par des études électrophysiologiques. Il s'agit dans les 3 cas d'un gain de fonction. Puis, une cohorte de 248 patients atteints de syndrome de Brugada a été étudiée. Onze mutations du gène TRPM4 ont été trouvées chez 20 patients. Les conséquences électrophysiologiques des mutations étaient diverses. Ensuite, des cohortes de patients atteints de syndrome du QT long, de fibrillation auriculaire ou de cardiomyopathie dilaté ont été étudiées. Nous avons également trouvé des mutations ou des variants prédisposants du gène TRPM4 chez ces patients. Une étude électrophysiologique doit être réalisée pour comprendre le rôle de TRPM4 dans ces autres pathologies. Enfin, notre étude immunohistochimique a démontré que TRPM4 est fortement exprimé dans le système de conduction cardiaque mais aussi plus faiblement dans les cardiomyocytes auriculaires et ventriculaires communs. Ce travail a permis d'impliquer pour la première fois le gène TRPM4 dans des maladies humaines. Ce travail donne les bases pour comprendre le rôle du canal TRPM4 dans le fonctionnement cardiaque. C'est le préalable nécessaire avant de pouvoir développer de nouvelles thérapeutiques dans le futur / By using reverse genetics, the causal mutation of a familial cardiac conduction block was localized to the long arm of chromosome 19 in 13.3. After screening 12 candidate genes, we found 3 different mutations in three independent families with isolated cardiac conduction block. The consequences of these 3 mutations were explored by electrophysiological studies. In all 3 mutations it was a gain of function. Then, a cohort of 248 patients with a Brugada syndrome was studied. Eleven mutations were found in the TRPM4 gene in 20 patients. The electrophysiological consequences of these mutations were diverse. Then, cohorts of patients with long QT syndrome, atrial fibrillation, and dilated cardiomyopathy were studied. We found also mutations or predisposing variants in these patients. An electrophysiology study should be conducted to understand the role of TRPM4 in these other pathologies. Finally, our immunohistochemical study showed that TRPM4 is highly expressed in the cardiac conduction system but also although with less intensity in common auricular and ventricular cardiomyocytes. This work implied for the first time the TRPM4 gene in human diseases. This work provides the basis to understand the role of the TRPM4 channel in cardiac function. This is a prerequisite to be able to develop novel therapies in the future