The specialized myocytes of the ventricular conduction system (VCS) coordinate ventricular contraction and are critical for efficient pumping by the heart. Impaired VCS conduction is characteristic of inherited forms of cardiac conduction disorders. Here we show that the Iroquois homeobox 3 (Irx3) transcription factor is preferentially expressed in the developing and mature VCS. Loss of Irx3 in mice results in slowed VCS conduction and prolonged QRS duration with right bundle branch block, caused by reduction (42%) in VCS-specific connexin 40 (Cx40) expression and VCS fiber hypoplasia, absent in littermate controls. Therefore, we show that the role of Irx3 in the heart is two-fold, whereby Irx3 (1) indirectly regulates Cx40 gene expression, by repressing a repressor of Cx40 transcript, and (2) controls VCS maturation, possibly in an Nkx2-5-dependent manner. To our knowledge, this is the first report of a role for Irx3 in regulating the development and function of the VCS.

The specialized myocytes of the ventricular conduction system (VCS) coordinate ventricular contraction and are critical for efficient pumping by the heart. Impaired VCS conduction is characteristic of inherited forms of cardiac conduction disorders. Here we show that the Iroquois homeobox 3 (Irx3) transcription factor is preferentially expressed in the developing and mature VCS. Loss of Irx3 in mice results in slowed VCS conduction and prolonged QRS duration with right bundle branch block, caused by reduction (42%) in VCS-specific connexin 40 (Cx40) expression and VCS fiber hypoplasia, absent in littermate controls. Therefore, we show that the role of Irx3 in the heart is two-fold, whereby Irx3 (1) indirectly regulates Cx40 gene expression, by repressing a repressor of Cx40 transcript, and (2) controls VCS maturation, possibly in an Nkx2-5-dependent manner. To our knowledge, this is the first report of a role for Irx3 in regulating the development and function of the VCS.

La LVNC est une cardiomyopathie rare, caractérisée par une hypertrabéculation et de profonds replis du ventricule gauche. A ce jour, nous ne savons toujours pas si la LVNC résulte d'un défaut se produisant durant le développement cardiaque et si sa gravité dépend du stade embryonnaire auquel l'arrêt de la compaction se produit. Notre objectif a été d'étudier l'évolution pathologique de la LVNC en caractérisant les défauts fonctionnels et en identifiant les mécanismes moléculaires dans des modèles de souris présentant un développement anormal des trabécules ventriculaires. Pour établir un modèle de LVNC, nous avons généré des souris KO conditionnel pour Nkx2.5 grâce au système Flox/loxP inductible par injection de tamoxifène qui active la recombinaison Cre. Nous avons ainsi supprimé l'allèle Nkx2.5 dans l'oreillette et les cardiomyocytes dérivant des trabécules. Nous avons choisi de sipprimé Nkx2.5 au stade embryonnaire E10 quand le trabécule s'accroît, au stade E14 quand il se compacte, ou juste après la naissance quand le cœur a terminé son processus de compaction. En résumé, nous avons réussi à générer différents modèles de LVNC, dans lesquelles nous avons pu étudier cette pathologie, en supprimant le facteur de transcription Nkx2.5 dans les oreillettes et les cardiomyocytes dérivés des trabécules. Nous avons également confirmé que la sévérité de la LVNC dépend du stade de développement du trabécule auquel le défaut se produit. Peu de publications décrivent à ce jour les mécanismes responsables de l'état inflammatoire observé dans la LVNC, nos résultats sont donc prometteurs pour de futures recherches dans cette voie. / LVNC is a rare cardiomyopathy, characterized by hypertrabeculation and deep trabecular recesses in the left ventricle. It is still unclear whether LVNC results from a defect occurring during cardiac development. One hypothesis to consider is that the severity of LVNC depends on which embryonic stage the arrest of myocardial compaction occurs. Our aim was to study the pathological evolution of LVNC by characterizing functional defects and identifying molecular mechanisms in mouse models with abnormal ventricular trabeculae development. To establish a LVNC mouse model, we generated specific Nkx2.5 conditional knockout mice to delete Nkx2.5 allele in atria and trabecular derived cardiomyocytes at embryonic stages when trabeculae arise (at around E10), or start to compact (at around E14), or at neonatal stages (after birth) when the heart is almost finish compaction step. After all, we were successful in generating several LVNC mouse models by the conditional deletion of Nkx2.5 transcription factor in atria and trabecular derived cardiomyocytes. These mouse models are suitable for studying LVNC pathology. We also confirmed the hypothesis that the severity of LVNC depends on stages when disturbances in the trabecular development occur. Hypertrabeculation, cardiac conduction defects, decreased ejection fraction, and existence of fibrosis are robustly observed following deletion at E10.5/11.5 meaning that the deletion at early stage of trabecular development causes the most severe pathological phenotype of LVNC. There had been just a few publications showing inflammation in LVNC heart, which could be a very good finding for future researches.