A rare HCN4 variant with combined sinus bradycardia, left atrial dilatation, and hypertrabeculation/left ventricular noncompaction phenotype

Alonso-Fernández-Gatta, Marta; Gallego‐Delgado, María; Caballero, Ricardo; Villacorta, Eduardo; Díaz-Peláez, Elena; García‐Berrocal, Belén; Crespo-García, Teresa; Izquierdo, Beatriz Plata; Marcos-Vadillo, Elena; García-Cuenllas, Luisa; Barreiro-Pérez, Manuel; Isidoro‐García, María; Tamargo-Menéndez, Juan; Delpón, Eva; Sánchez, Pedro L.

doi:10.1016/j.rec.2020.06.019

Cited by 9 publications

(17 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The first one was the p.R524Q HCN4 variant, that increases the channel affinity for cAMP and was described in an Italian family with symptomatic sinus tachyarrhythmia (Baruscotti et al, 2017). Conversely, many loss-of-function heterozygous mutations in the HCN4 gene have been associated with sinus bradycardia that can be accompanied, or not, with atrial fibrillation, atrioventricular block, structural diseases such as noncompaction cardiomyopathy, and even QT prolongation (Alonso-Fernández-Gatta et al, 2021; Cambon-Viala et al, 2021; Macri et al, 2014; Milanesi et al, 2006; Nof et al, 2007; Rivolta et al, 2020; Schweizer et al, 2010; Ueda et al, 2004; Verkerk and Wilders, 2015). As mentioned, there has been important controversy regarding the role of HCN4 isoforms in generating human I f and in pacemaking of sinoatrial node.…”

Section: Discussionmentioning

confidence: 99%

“…CHO cells were grown in Ham-F12 medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin (Caballero et al, 2010; Caballero et al, 2017; Pérez-Hernández et al, 2018; Alonso-Fernández-Gatta et al, 2021; Crespo-García et al, 2023). HEK-293 cells were cultured in Dulbecco’s modified Eagle’s (DMEM) medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin as previously described (Caballero et al, 2010; Crespo-García et al, 2023).…”

Section: Methodsmentioning

confidence: 99%

“…After settling to the bottom of the chamber, cells were perfused at ≈1 mL/min with external solution (see the composition below). Macroscopic currents were recorded at room temperature (21-23ºC) by means of the whole-cell patch-clamp technique using Axopatch-200B patch clamp amplifiers and pCLAMP software (Molecular Devices, San José, CA, USA) (Caballero et al, 2010; Caballero et al, 2017; Pérez-Hernández et al, 2018; Alonso-Fernández-Gatta et al, 2021; Nieto-Marín et al, 2022; Crespo-García et al, 2023) Recording pipettes were pulled from 1.0 mm o.d. borosilicate capillary tubes (GD1, Narishige Co., Ltd, Tokio, Japan) using a programmable patch micropipette puller (Model P-2000 Brown-Flaming, Sutter Instruments Co., Novato, CA, USA) and were heat-polished with a microforge (Model MF-83, Narishige).…”

Section: Methodsmentioning

confidence: 99%

“…The cultures were passed every 2-5 days using a brief trypsin treatment. For macroscopic current recordings, subconfluent cultures were transiently transfected with cDNA encoding WT or mutated HCN4 channels (1.6 µg) together with cDNA encoding the CD8 antigen (0.5 µg) by using X-tremeGENE™ HP (Roche Diagnostics, Rotkreuz, Switzerland) in CHO cells or Lipofectamine 2000 (Thermofisher Scientific, Waltham, MA, USA) in HEK-293 cells, with manufacturer instructions followed (Caballero et al, 2010;Caballero et al, 2017;Pérez-Hernández et al, 2018;Alonso-Fernández-Gatta et al, 2021;Crespo-García et al, 2023). In some experiments, WT and p.V240M HCN4 were cotransfected (WT+p.V240M) at a 0.5:0.5 ratio (0.8 µg each).…”

Section: Next-generation and Sanger Sequencingmentioning

confidence: 99%

See 3 more Smart Citations

A rare gain of function HCN4 gene mutation is responsible for inappropriate sinus tachycardia in a Spanish family

Cámara-Checa

Perín

Rubio-Alarcón

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Background. In a family with inappropriate sinus tachycardia (IST) we identified a novel mutation (p.V240M) of the hyperpolarization-activated cyclic nucleotide-gated type 4 (HCN4) channel, which contributes to the pacemaker current (If) in human sinoatrial node cells. Here we clinically study the family and functionally analyze the p.V240M variant. Methods. Macroscopic (IHCN4) and single-channel currents were recorded using patch-clamp in cells expressing human native (WT) and/or p.V240M HCN4 channels. Results. All p.V240M mutation carriers exhibited IST (mean heart rate 113[7] bpm, n=9), that in adults, was accompanied by cardiomyopathy. IHCN4 generated by p.V240M channels either alone or in combination with WT was significantly greater than that generated by WT channels. The variant, which lies in the N-terminal HCN domain, increased single-channel conductance and opening frequency and probability of HCN4 channels. Conversely, it did not modify channel sensitivity for cAMP and ivabradine or the level of expression at the membrane. Treatment with ivabradine based on functional data reversed the IST and the cardiomyopathy of the carriers. Conclusions. The p.V240M gain-of-function variant increases If during diastole, which explains the IST of the carriers. The results demonstrate the importance of the unique HCN domain in HCN4 which stabilizes the channels in the closed state. Funding. Ministerio de Ciencia e Innovación (PID2020-118694RB-I00); Comunidad Autónoma de Madrid (P2022/BMD-7229), European Structural and Investment Funds); and Instituto de Salud Carlos III (CIBERCV; CB16/11/00303).

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Next-generation and Sanger Sequencingmentioning

confidence: 99%

See 2 more Smart Citations

A rare gain of function HCN4 gene mutation is responsible for inappropriate sinus tachycardia in a Spanish family

Cámara-Checa

Perín

Rubio-Alarcón

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…In families with affected subjects, the main channels to be mutated in SSS are Nav1.5 and HCN, whose genes SCN5A and HCN are responsible for the generation of I Na and I f currents, as observed by genetic screening [144][145][146][147][148][149][150]. It is noteworthy to mention that I Na alterations are not only manifest in SSS, but also in other cardiac arrhythmias and might often generate a mixed clinical picture (as an example, [151,152]).…”

Section: Sss Due To Genetic Mutationsmentioning

confidence: 99%

Inherited and Acquired Rhythm Disturbances in Sick Sinus Syndrome, Brugada Syndrome, and Atrial Fibrillation: Lessons from Preclinical Modeling

Iop

Iliceto

Civieri

et al. 2021

Cells

View full text Add to dashboard Cite

Rhythm disturbances are life-threatening cardiovascular diseases, accounting for many deaths annually worldwide. Abnormal electrical activity might arise in a structurally normal heart in response to specific triggers or as a consequence of cardiac tissue alterations, in both cases with catastrophic consequences on heart global functioning. Preclinical modeling by recapitulating human pathophysiology of rhythm disturbances is fundamental to increase the comprehension of these diseases and propose effective strategies for their prevention, diagnosis, and clinical management. In silico, in vivo, and in vitro models found variable application to dissect many congenital and acquired rhythm disturbances. In the copious list of rhythm disturbances, diseases of the conduction system, as sick sinus syndrome, Brugada syndrome, and atrial fibrillation, have found extensive preclinical modeling. In addition, the electrical remodeling as a result of other cardiovascular diseases has also been investigated in models of hypertrophic cardiomyopathy, cardiac fibrosis, as well as arrhythmias induced by other non-cardiac pathologies, stress, and drug cardiotoxicity. This review aims to offer a critical overview on the effective ability of in silico bioinformatic tools, in vivo animal studies, in vitro models to provide insights on human heart rhythm pathophysiology in case of sick sinus syndrome, Brugada syndrome, and atrial fibrillation and advance their safe and successful translation into the cardiology arena.

show abstract

Establishment and identification of cardiomyocyte arhGEF18 gene conditional knockout mice

Wan

Yuan

2023

Pediatric Discovery

View full text Add to dashboard Cite

Left ventricular noncompaction (LVNC) is a heterogeneous disorder with unclear genetic causes. The arhGEF18 gene is a guanine nucleotide exchange factor related to the Rho pathway and a possible predisposing gene for LVNC. In this study, a mouse model of arhGEF18 gene conditional knockout (cKO) in cardiomyocytes was established using the Cre‐LoxP system to provide an animal model for the genetic study of LVNC. ArhGEF18 cKO mice were obtained by crossing arhGEF18f/+/myh6‐Cre+ and arhGEF18f/f mice. The mRNA and protein expression levels of arhGEF18 in different tissues were verified. The myocardial cytoskeleton and polar protein expression, the survival rate, cardiac function, and cardiac histology of model mice were determined. ArhGEF18 cKO mice were successfully established. ArhGEF18 was confirmed to be specifically knocked out in the myocardial tissue of arhGEF18f/f/myh6‐Cre+ mice at the mRNA and protein levels, and the knockout rate was approximately 75%. The relative expression levels of cytoskeleton and cell polarity proteins such as α/β‐tubulin, Scribble, Crb2, and PAR3 in the cKO group were significantly lower than those in the control group (p < 0.05). During monitoring, the survival rate, cardiac systolic function, and myocardial structure of arhGEF18f/f/myh6‐Cre+ mice were not significantly different from those of control mice. A mouse model of cardiomyocyte arhGEF18 gene cKO was successfully established, and it was confirmed that knocking out arhGEF18 changed cytoskeleton and cell polar protein expression levels in the myocardium. However, there was no obvious LVNC phenotype in the constructed arhGEF18 cKO mice.

show abstract

A rare HCN4 variant with combined sinus bradycardia, left atrial dilatation, and hypertrabeculation/left ventricular noncompaction phenotype

Cited by 9 publications

References 25 publications

A rare gain of function HCN4 gene mutation is responsible for inappropriate sinus tachycardia in a Spanish family

A rare gain of function HCN4 gene mutation is responsible for inappropriate sinus tachycardia in a Spanish family

Inherited and Acquired Rhythm Disturbances in Sick Sinus Syndrome, Brugada Syndrome, and Atrial Fibrillation: Lessons from Preclinical Modeling

Establishment and identification of cardiomyocyte arhGEF18 gene conditional knockout mice

Contact Info

Product

Resources

About