Genetic control of mRNA splicing as a potential mechanism for incomplete penetrance of rare coding variants.

Exonic variants present some of the strongest links between genotype and phenotype. However, these variants can have significant inter-individual pathogenicity differences, known as variable penetrance. In this study, we propose a model where genetically controlled mRNA splicing modulates the pathogenicity of exonic variants. By first cataloging exonic inclusion from RNA-sequencing data in GTEx V8, we find that pathogenic alleles are depleted on highly included exons. Using a large-scale phased whole genome sequencing data from the TOPMed consortium, we observe that this effect may be driven by common splice-regulatory genetic variants, and that natural selection acts on haplotype configurations that reduce the transcript inclusion of putatively pathogenic variants, especially when limiting to haploinsufficient genes. Finally, we test if this effect may be relevant for autism risk using families from the Simons Simplex Collection, but find that splicing of pathogenic alleles has a penetrance reducing effect here as well. Overall, our results indicate that common splice-regulatory variants may play a role in reducing the damaging effects of rare exonic variants.

Clinical predictors of acute hyponatremia following LARIAT ligation of the left atrial appendage.

INTRODUCTION: Hyponatremia commonly follows percutaneous exclusion of the left atrial appendage (LAA) with the LARIAT suture delivery device. The aim of this study was to evaluate for clinical predictors hyponatremia following ligation of the appendage with the LARIAT device. METHODS AND RESULTS: A retrospective analysis was conducted on 61 consecutive patients (average age 69.7 ± 9.8 years, 55.7% male) who underwent successful appendage ligation with the LARIAT device. Acute hyponatremia (AH) was defined as a drop in serum sodium (Na) by greater than or equal to 4 mmol/L within 48 hours of ligation while exaggerated acute hyponatremia (EAH) was defined as a drop greater than or equal to 10 mmol/L. Among all patients, there was a significant decrease in [Na] at 24 hours (3.26 ± 2.77 mmol/L) and 48 hours (4.98 ± 3.74 mmol/L). Thirty-two patients (52.4%) had AH while six patients (9.8%) experienced EAH. A body mass index (BMI) of less than 28.4 kg/m2 was associated with AH (P = .037) while a BMI < 25 kg/m2 was associated with EAH (P = .021). A linear regression analysis comparing the maximum sodium decrease to the indexed left atrial (LA) diameter found that for every 1 cm/m2 increase in indexed LA diameter, there was a 2.5 mEq/L decrease in serum sodium (P = .04). CONCLUSIONS: Hyponatremia frequently occurs following LAA ligation with the LARIAT device. A low BMI < 25 kg/m2 is associated with a drop in serum sodium of greater than 10 mmol/L while increasing indexed LA diameter predicts any AH. AH is also associated with a significantly lower systolic blood pressure 48 to 72 hours post LAA exclusion with the LARIAT device.

Premature battery depletion due to compromised low-voltage capacitor in a family of defibrillators.

BACKGROUND: Boston Scientific (Marlborough, MA, USA) implantable cardioverter-defibrillators (ICDs) and cardiac resynchronization therapy defibrillators (CRT-Ds) manufactured between 2008 and 2014 are potentially subject to premature battery depletion through a low-voltage capacitor malfunction occurring as a result of hydrogen buildup within the device. Although some of these devices are currently under advisory, other devices manufactured during this timeframe carry a lower risk of the same malfunction. These same devices are known to have superior longevity in general, and the overall mean lifespan of the devices remains long. METHODS: All patients implanted or followed at our two centers who experienced premature battery depletion and had a Boston Scientific ICD or CRT-D potentially at risk for low-voltage capacitor malfunction were studied retrospectively. RESULTS: Nineteen out of 838 patients (2.3%) with devices potentially at risk have had premature battery depletion: 5.7% of those under advisory and 1.1% of those not under advisory. None of our patients had compromised therapy, and all had >27 days of projected battery longevity remaining. CONCLUSIONS: Undetected premature battery depletion in this population of ICDs has the potential to expose a patient to an interval of time where the device is unable to provide therapy. However, with enrollment in remote monitoring, regular follow-up, and attention to audible alerts, the risk of therapy loss due to low-voltage state can be effectively mitigated. For these reasons, prophylactic generator replacement is not recommended.

Patient-independent human induced pluripotent stem cell model: A new tool for rapid determination of genetic variant pathogenicity in long QT syndrome.

BACKGROUND: Commercial genetic testing for long QT syndrome (LQTS) has rapidly expanded, but the inability to accurately predict whether a rare variant is pathogenic has limited its clinical benefit. Novel missense variants are routinely reported as variant of unknown significance (VUS) and cannot be used to screen family members at risk for sudden cardiac death. Better approaches to determine the pathogenicity of VUS are needed. OBJECTIVE: The purpose of this study was to rapidly determine the pathogenicity of a CACNA1C variant reported by commercial genetic testing as a VUS using a patient-independent human induced pluripotent stem cell (hiPSC) model. METHODS: Using CRISPR/Cas9 genome editing, CACNA1C-p.N639T was introduced into a previously established hiPSC from an unrelated healthy volunteer, thereby generating a patient-independent hiPSC model. Three independent heterozygous N639T hiPSC lines were generated and differentiated into cardiomyocytes (CM). Electrophysiological properties of N639T hiPSC-CM were compared to those of isogenic and population control hiPSC-CM by measuring the extracellular field potential (EFP) of 96-well hiPSC-CM monolayers and by patch clamp. RESULTS: Significant EFP prolongation was observed only in optically stimulated but not in spontaneously beating N639T hiPSC-CM. Patch-clamp studies revealed that N639T prolonged the ventricular action potential by slowing voltage-dependent inactivation of CaV1.2 currents. Heterologous expression studies confirmed the effect of N639T on CaV1.2 inactivation. CONCLUSION: The patient-independent hiPSC model enabled rapid generation of functional data to support reclassification of a CACNA1C VUS to likely pathogenic, thereby establishing a novel LQTS type 8 mutation. Furthermore, our results indicate the importance of controlling beating rates to evaluate the functional significance of LQTS VUS in high-throughput hiPSC-CM assays.

Genetic ACE I/D polymorphism and recurrence of atrial fibrillation after catheter ablation.

BACKGROUND: The angiotensin-converting enzyme (ACE) deletion allele, ACE D, is associated with increased cardiac ACE activity, cardiac fibrosis, and adverse outcomes in cardiovascular disease and has been linked with failure of antiatrial fibrillation (anti-AF) drug treatment. This study tested the hypothesis that the ACE gene insertion/deletion polymorphism associates with AF recurrence after catheter ablation. METHODS AND RESULTS: In 238 consecutive patients (69% male; mean age, 58±11 years) undergoing catheter ablation of paroxysmal (59%) or persistent (41%) AF, the ACE insertion/deletion polymorphism was genotyped using polymerase chain reaction. After a blanking period of 3 months, AF recurrence (defined as any atrial arrhythmia lasting ≥30 s) was detected using serial 7-day Holter ECG recordings after 3, 6, and 12 months. AF recurrence was observed in 39% and was associated with persistent AF, longer history of AF, previous antiarrhythmic drug use, previous use of diuretics, increased left atrial diameter, increased left ventricular end-diastolic diameter, additional linear ablation lesions, and ACE DD polymorphism. In multivariable analysis, left atrial diameter (odds ratio, 1.111; 95% confidence interval, 1.040-1.187; P=0.002) and ACE DD genotype (odds ratio, 2.251; 95% confidence interval, 1.056-4.798; P=0.036) remained predictors for AF recurrence. CONCLUSIONS: Left atrial enlargement and the ACE DD polymorphism are predictors for AF recurrence after catheter ablation. The association between the ACE DD polymorphism and AF recidivism supports the use of genetic data for predicting response to AF therapies and highlights the role of fibrosis in AF development.