Molecular and Functional Characterization of Rare CACNA1C Variants in Sudden Unexplained Death in the Young.

Abstract

INTRODUCTION:

Perturbations in the CACNA1C-encoded L-type calcium channel α-subunit have been linked recently to heritable arrhythmia syndromes, including Timothy syndrome, Brugada syndrome, early repolarization syndrome, and long QT syndrome. These heritable arrhythmia syndromes may serve as a pathogenic basis for autopsy-negative sudden unexplained death in the young (SUDY). However, the contribution of CACNA1C mutations to SUDY is unknown.

OBJECTIVE:

We set out to determine the spectrum, prevalence, and pathophysiology of rare CACNA1C variants in SUDY.

METHODS:

Mutational analysis of CACNA1C was conducted in 82 SUDY cases using polymerase chain reaction, denaturing high-performance liquid chromatography, and direct sequencing. Identified variants were engineered using site-directed mutagenesis, and heterologously expressed in TSA-201 or HEK293 cells.

RESULTS:

Two SUDY cases (2.4%) harbored functional variants in CACNA1C. The E850del and N2091S variants involve highly conserved residues and localize to the II-III linker and C-terminus, respectively. Although observed in publically available exome databases, both variants confer abnormal CaV 1.2 electrophysiological characteristics. Examination of the electrophysiological properties revealed the E850del mutation in CACNA1C led to a 95% loss-of-function in ICa , and the N2091S variant led to a 105% gain-of-function in ICa. Additionally, N2091S led to minor kinetic alterations including a -3.4 mV shift in V1/2 of activation.

CONCLUSION:

This study provides molecular and functional evidence that rare CACNA1C genetic variants may contribute to the underlying pathogenic basis for some cases of SUDY in either a gain or loss-of-function mechanism.

• Authors: Brittan S Sutphin, Nicole J Boczek, Héctor Barajas-Martínez, Dan Hu, Dan Ye, David J Tester, Charles Antzelevitch, Michael J Ackerman
• Journal: Congenital heart disease (Congenit Heart Dis) Vol. 11 Issue 6 Pg. 683-692 (Dec 2016) ISSN: 1747-0803 [Electronic] United States
• PMID: 27218670 (Publication Type: Journal Article)
• Copyright: © 2016 Wiley Periodicals, Inc.

Chemical References

• CACNA1C protein, human
• Calcium Channels, L-Type

Topics

• Adolescent
• Adult
• Age Factors
• Arrhythmias, Cardiac (diagnosis, genetics, mortality, physiopathology)
• Calcium Channels, L-Type (genetics, metabolism)
• Child
• Child, Preschool
• DNA Mutational Analysis
• Death, Sudden, Cardiac (etiology)
• Female
• Genetic Predisposition to Disease
• HEK293 Cells
• Humans
• Infant
• Kinetics
• Male
• Membrane Potentials
• Mutagenesis, Site-Directed
• Mutation
• Patch-Clamp Techniques
• Phenotype
• Risk Factors
• Transfection
• Young Adult