Christiaan C Veerman1, Svitlana Podliesna1, Rafik Tadros1, Elisabeth M Lodder1, Isabella Mengarelli1, Berend de Jonge1, Leander Beekman1, Julien Barc1, Ronald Wilders1, Arthur A M Wilde1, Bastiaan J Boukens1, Ruben Coronel1, Arie O Verkerk1, Carol Ann Remme1, Connie R Bezzina2. 1. From the Department of Clinical and Experimental Cardiology, Heart Center, Academic Medical Center, Amsterdam, The Netherlands (C.C.V., S.P., R.T., E.M.L., I.M., L.B., A.A.M.W., R.C., A.O.V., C.A.R., C.R.B.); Department of Medicine, Cardiovascular Genetics Center, Montreal Heart Institute, Canada (R.T.); Université de Montréal, Canada (R.T.); Department of Medical Biology, Academic Medical Center, Amsterdam, The Netherlands (B.d.J., R.W., B.J.B., A.O.V.); INSERM, CNRS, Université de Nantes, L'institut du Thorax, Nantes, France (J.B.); Princess Al-Jawhara Al-Brahim Centre of Excellence in Research of Hereditary Disorders, Jeddah, Saudi Arabia (A.A.M.W.); and Electrophysiology and Heart Modeling Institute LIRYC, Université de Bordeaux, France (R.C.). 2. From the Department of Clinical and Experimental Cardiology, Heart Center, Academic Medical Center, Amsterdam, The Netherlands (C.C.V., S.P., R.T., E.M.L., I.M., L.B., A.A.M.W., R.C., A.O.V., C.A.R., C.R.B.); Department of Medicine, Cardiovascular Genetics Center, Montreal Heart Institute, Canada (R.T.); Université de Montréal, Canada (R.T.); Department of Medical Biology, Academic Medical Center, Amsterdam, The Netherlands (B.d.J., R.W., B.J.B., A.O.V.); INSERM, CNRS, Université de Nantes, L'institut du Thorax, Nantes, France (J.B.); Princess Al-Jawhara Al-Brahim Centre of Excellence in Research of Hereditary Disorders, Jeddah, Saudi Arabia (A.A.M.W.); and Electrophysiology and Heart Modeling Institute LIRYC, Université de Bordeaux, France (R.C.). c.r.bezzina@amc.uva.nl.
Abstract
RATIONALE: Genome-wide association studies previously identified an association of rs9388451 at chromosome 6q22.3 (near HEY2) with Brugada syndrome. The causal gene and underlying mechanism remain unresolved. OBJECTIVE: We used an integrative approach entailing transcriptomic studies in human hearts and electrophysiological studies in Hey2+/- (Hey2 heterozygous knockout) mice to dissect the underpinnings of the 6q22.31 association with Brugada syndrome. METHODS AND RESULTS: We queried expression quantitative trait locus data acquired in 190 human left ventricular samples from the genotype-tissue expression consortium for cis-expression quantitative trait locus effects of rs9388451, which revealed an association between Brugada syndrome risk allele dosage and HEY2 expression (β=+0.159; P=0.0036). In the same transcriptomic data, we conducted genome-wide coexpression analysis for HEY2, which uncovered KCNIP2, encoding the β-subunit of the channel underlying the transient outward current (Ito), as the transcript most robustly correlating with HEY2 expression (β=+1.47; P=2×10-34). Transcript abundance of Hey2 and the Ito subunits Kcnip2 and Kcnd2, assessed by quantitative reverse transcription-polymerase chain reaction, was higher in subepicardium versus subendocardium in both left and right ventricles, with lower levels in Hey2+/- mice compared with wild type. Surface ECG measurements showed less prominent J waves in Hey2+/- mice compared with wild-type. In wild-type mice, patch-clamp electrophysiological studies on cardiomyocytes from right ventricle demonstrated a shorter action potential duration and a lower Vmax in subepicardium compared with subendocardium cardiomyocytes, which was paralleled by a higher Ito and a lower sodium current (INa) density in subepicardium versus subendocardium. These transmural differences were diminished in Hey2+/- mice because of changes in subepicardial cardiomyocytes. CONCLUSIONS: This study uncovers a role of HEY2 in the normal transmural electrophysiological gradient in the ventricle and provides compelling evidence that genetic variation at 6q22.31 (rs9388451) is associated with Brugada syndrome through a HEY2-dependent alteration of ion channel expression across the cardiac ventricular wall.
RATIONALE: Genome-wide association studies previously identified an association of rs9388451 at chromosome 6q22.3 (near HEY2) with Brugada syndrome. The causal gene and underlying mechanism remain unresolved. OBJECTIVE: We used an integrative approach entailing transcriptomic studies in human hearts and electrophysiological studies in Hey2+/- (Hey2 heterozygous knockout) mice to dissect the underpinnings of the 6q22.31 association with Brugada syndrome. METHODS AND RESULTS: We queried expression quantitative trait locus data acquired in 190 human left ventricular samples from the genotype-tissue expression consortium for cis-expression quantitative trait locus effects of rs9388451, which revealed an association between Brugada syndrome risk allele dosage and HEY2 expression (β=+0.159; P=0.0036). In the same transcriptomic data, we conducted genome-wide coexpression analysis for HEY2, which uncovered KCNIP2, encoding the β-subunit of the channel underlying the transient outward current (Ito), as the transcript most robustly correlating with HEY2 expression (β=+1.47; P=2×10-34). Transcript abundance of Hey2 and the Ito subunits Kcnip2 and Kcnd2, assessed by quantitative reverse transcription-polymerase chain reaction, was higher in subepicardium versus subendocardium in both left and right ventricles, with lower levels in Hey2+/- mice compared with wild type. Surface ECG measurements showed less prominent J waves in Hey2+/- mice compared with wild-type. In wild-type mice, patch-clamp electrophysiological studies on cardiomyocytes from right ventricle demonstrated a shorter action potential duration and a lower Vmax in subepicardium compared with subendocardium cardiomyocytes, which was paralleled by a higher Ito and a lower sodium current (INa) density in subepicardium versus subendocardium. These transmural differences were diminished in Hey2+/- mice because of changes in subepicardial cardiomyocytes. CONCLUSIONS: This study uncovers a role of HEY2 in the normal transmural electrophysiological gradient in the ventricle and provides compelling evidence that genetic variation at 6q22.31 (rs9388451) is associated with Brugada syndrome through a HEY2-dependent alteration of ion channel expression across the cardiac ventricular wall.
Authors: Can Hasdemir; Jimmy Jyh-Ming Juang; Sedat Kose; Umut Kocabas; Mehmet N Orman; Serdar Payzin; Hatice Sahin; Candan Celen; Emin E Ozcan; Ching-Yu Julius Chen; Ramazan Gunduz; Oguzhan E Turan; Oktay Senol; Elena Burashnikov; Charles Antzelevitch Journal: Pacing Clin Electrophysiol Date: 2018-07-16 Impact factor: 1.976
Authors: Julien Barc; Rafik Tadros; Charlotte Glinge; David Y Chiang; Mariam Jouni; Floriane Simonet; Vincent Probst; Arthur A Wilde; Jean-Jacques Schott; Richard Redon; Connie R Bezzina; Sean J Jurgens; Manon Baudic; Michele Nicastro; Franck Potet; Joost A Offerhaus; Roddy Walsh; Seung Hoan Choi; Arie O Verkerk; Yuka Mizusawa; Soraya Anys; Damien Minois; Marine Arnaud; Josselin Duchateau; Yanushi D Wijeyeratne; Alison Muir; Michael Papadakis; Silvia Castelletti; Margherita Torchio; Cristina Gil Ortuño; Javier Lacunza; Daniela F Giachino; Natascia Cerrato; Raphaël P Martins; Oscar Campuzano; Sonia Van Dooren; Aurélie Thollet; Florence Kyndt; Andrea Mazzanti; Nicolas Clémenty; Arnaud Bisson; Anniek Corveleyn; Birgit Stallmeyer; Sven Dittmann; Johan Saenen; Antoine Noël; Shohreh Honarbakhsh; Boris Rudic; Halim Marzak; Matthew K Rowe; Claire Federspiel; Sophie Le Page; Leslie Placide; Antoine Milhem; Hector Barajas-Martinez; Britt-Maria Beckmann; Ingrid P Krapels; Johannes Steinfurt; Bo Gregers Winkel; Reza Jabbari; Moore B Shoemaker; Bas J Boukens; Doris Škorić-Milosavljević; Hennie Bikker; Federico Manevy; Peter Lichtner; Marta Ribasés; Thomas Meitinger; Martina Müller-Nurasyid; Jan H Veldink; Leonard H van den Berg; Philip Van Damme; Daniele Cusi; Chiara Lanzani; Sidwell Rigade; Eric Charpentier; Estelle Baron; Stéphanie Bonnaud; Simon Lecointe; Audrey Donnart; Hervé Le Marec; Stéphanie Chatel; Matilde Karakachoff; Stéphane Bézieau; Barry London; Jacob Tfelt-Hansen; Dan Roden; Katja E Odening; Marina Cerrone; Larry A Chinitz; Paul G Volders; Maarten P van de Berg; Gabriel Laurent; Laurence Faivre; Charles Antzelevitch; Stefan Kääb; Alain Al Arnaout; Jean-Marc Dupuis; Jean-Luc Pasquie; Olivier Billon; Jason D Roberts; Laurence Jesel; Martin Borggrefe; Pier D Lambiase; Jacques Mansourati; Bart Loeys; Antoine Leenhardt; Pascale Guicheney; Philippe Maury; Eric Schulze-Bahr; Tomas Robyns; Jeroen Breckpot; Dominique Babuty; Silvia G Priori; Carlo Napolitano; Carlo de Asmundis; Pedro Brugada; Ramon Brugada; Elena Arbelo; Josep Brugada; Philippe Mabo; Nathalie Behar; Carla Giustetto; Maria Sabater Molina; Juan R Gimeno; Can Hasdemir; Peter J Schwartz; Lia Crotti; Pascal P McKeown; Sanjay Sharma; Elijah R Behr; Michel Haissaguerre; Frédéric Sacher; Caroline Rooryck; Hanno L Tan; Carol A Remme; Pieter G Postema; Mario Delmar; Patrick T Ellinor; Steven A Lubitz; Jean-Baptiste Gourraud; Michael W Tanck; Alfred L George; Calum A MacRae; Paul W Burridge; Christian Dina Journal: Nat Genet Date: 2022-02-24 Impact factor: 41.307
Authors: Vincent Portero; Ronald Wilders; Simona Casini; Flavien Charpentier; Arie O Verkerk; Carol Ann Remme Journal: Front Physiol Date: 2018-03-12 Impact factor: 4.566