Transient outward current (I to) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome

John R. Giudicessi, Dan Ye, David J. Tester, Lia Crotti, Alessandra Mugione, Vladislav V. Nesterenko, Richard M. Albertson, Charles Antzelevitch, Peter J. Schwartz, Michael John Ackerman

Research output: Contribution to journalArticle

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Abstract

Background: Brugada syndrome (BrS) is a sudden death-predisposing genetic condition characterized electrocardiographically by ST segment elevation in the leads V 1-V 3. Given the prominent role of the transient outward current (I to) in BrS pathogenesis, we hypothesized that rare gain-of-function mutations in KCND3 may serve as a pathogenic substrate for BrS. Methods: Comprehensive mutational analysis of KCND3-encoded Kv4.3 (I to) was conducted using polymerase chain reaction, denaturing high performance liquid chromatography, and direct sequencing of DNA derived from 86 unrelated BrS1-8 genotype-negative BrS patients. DNA from 780 healthy individuals was examined to assess allelic frequency for nonsynonymous variants. Putative BrS-associated Kv4.3 mutations were engineered and coexpressed with wild-type KChIP2 in HEK293 cells. Wild-type and mutant I to ion currents were recorded using whole-cell patch clamp. Results: Two BrS1-8 genotype-negative cases possessed novel Kv4.3 missense mutations. Both Kv4.3-L450F and Kv4.3-G600R were absent in 1,560 reference alleles and involved residues highly conserved across species. Both Kv4.3-L450F and Kv4.3-G600R demonstrated a gain-of-function phenotype, increasing peak I to current density by 146.2% (n = 15, P <.05) and 50.4% (n = 15, P <.05), respectively. Simulations using a Luo-Rudy II action potential (AP) model demonstrated the stable loss of the AP dome as a result of the increased I to maximal conductance associated with the heterozygous expression of either L450F or G600R. Conclusions: These findings provide the first molecular and functional evidence implicating novel KCND3 gain-of-function mutations in the pathogenesis and phenotypic expression of BrS, with the potential for a lethal arrhythmia being precipitated by a genetically enhanced I to current gradient within the right ventricle where KCND3 expression is the highest.

Original languageEnglish (US)
Pages (from-to)1024-1032
Number of pages9
JournalHeart Rhythm
Volume8
Issue number7
DOIs
StatePublished - Jul 2011

Fingerprint

Shal Potassium Channels
Brugada Syndrome
Mutation
Action Potentials
Genotype
HEK293 Cells
Missense Mutation
Sudden Death
DNA Sequence Analysis
Heart Ventricles
Cardiac Arrhythmias
Alleles
High Pressure Liquid Chromatography
Ions
Phenotype
Polymerase Chain Reaction
DNA

Keywords

  • Brugada syndrome
  • Genetic diseases
  • Ion channels
  • Ito current
  • J-wave syndromes
  • Kv4.3 channels
  • Sudden cardiac death

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

Transient outward current (I to) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome. / Giudicessi, John R.; Ye, Dan; Tester, David J.; Crotti, Lia; Mugione, Alessandra; Nesterenko, Vladislav V.; Albertson, Richard M.; Antzelevitch, Charles; Schwartz, Peter J.; Ackerman, Michael John.

In: Heart Rhythm, Vol. 8, No. 7, 07.2011, p. 1024-1032.

Research output: Contribution to journalArticle

Giudicessi, JR, Ye, D, Tester, DJ, Crotti, L, Mugione, A, Nesterenko, VV, Albertson, RM, Antzelevitch, C, Schwartz, PJ & Ackerman, MJ 2011, 'Transient outward current (I to) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome', Heart Rhythm, vol. 8, no. 7, pp. 1024-1032. https://doi.org/10.1016/j.hrthm.2011.02.021
Giudicessi, John R. ; Ye, Dan ; Tester, David J. ; Crotti, Lia ; Mugione, Alessandra ; Nesterenko, Vladislav V. ; Albertson, Richard M. ; Antzelevitch, Charles ; Schwartz, Peter J. ; Ackerman, Michael John. / Transient outward current (I to) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome. In: Heart Rhythm. 2011 ; Vol. 8, No. 7. pp. 1024-1032.
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abstract = "Background: Brugada syndrome (BrS) is a sudden death-predisposing genetic condition characterized electrocardiographically by ST segment elevation in the leads V 1-V 3. Given the prominent role of the transient outward current (I to) in BrS pathogenesis, we hypothesized that rare gain-of-function mutations in KCND3 may serve as a pathogenic substrate for BrS. Methods: Comprehensive mutational analysis of KCND3-encoded Kv4.3 (I to) was conducted using polymerase chain reaction, denaturing high performance liquid chromatography, and direct sequencing of DNA derived from 86 unrelated BrS1-8 genotype-negative BrS patients. DNA from 780 healthy individuals was examined to assess allelic frequency for nonsynonymous variants. Putative BrS-associated Kv4.3 mutations were engineered and coexpressed with wild-type KChIP2 in HEK293 cells. Wild-type and mutant I to ion currents were recorded using whole-cell patch clamp. Results: Two BrS1-8 genotype-negative cases possessed novel Kv4.3 missense mutations. Both Kv4.3-L450F and Kv4.3-G600R were absent in 1,560 reference alleles and involved residues highly conserved across species. Both Kv4.3-L450F and Kv4.3-G600R demonstrated a gain-of-function phenotype, increasing peak I to current density by 146.2{\%} (n = 15, P <.05) and 50.4{\%} (n = 15, P <.05), respectively. Simulations using a Luo-Rudy II action potential (AP) model demonstrated the stable loss of the AP dome as a result of the increased I to maximal conductance associated with the heterozygous expression of either L450F or G600R. Conclusions: These findings provide the first molecular and functional evidence implicating novel KCND3 gain-of-function mutations in the pathogenesis and phenotypic expression of BrS, with the potential for a lethal arrhythmia being precipitated by a genetically enhanced I to current gradient within the right ventricle where KCND3 expression is the highest.",
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AU - Ye, Dan

AU - Tester, David J.

AU - Crotti, Lia

AU - Mugione, Alessandra

AU - Nesterenko, Vladislav V.

AU - Albertson, Richard M.

AU - Antzelevitch, Charles

AU - Schwartz, Peter J.

AU - Ackerman, Michael John

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N2 - Background: Brugada syndrome (BrS) is a sudden death-predisposing genetic condition characterized electrocardiographically by ST segment elevation in the leads V 1-V 3. Given the prominent role of the transient outward current (I to) in BrS pathogenesis, we hypothesized that rare gain-of-function mutations in KCND3 may serve as a pathogenic substrate for BrS. Methods: Comprehensive mutational analysis of KCND3-encoded Kv4.3 (I to) was conducted using polymerase chain reaction, denaturing high performance liquid chromatography, and direct sequencing of DNA derived from 86 unrelated BrS1-8 genotype-negative BrS patients. DNA from 780 healthy individuals was examined to assess allelic frequency for nonsynonymous variants. Putative BrS-associated Kv4.3 mutations were engineered and coexpressed with wild-type KChIP2 in HEK293 cells. Wild-type and mutant I to ion currents were recorded using whole-cell patch clamp. Results: Two BrS1-8 genotype-negative cases possessed novel Kv4.3 missense mutations. Both Kv4.3-L450F and Kv4.3-G600R were absent in 1,560 reference alleles and involved residues highly conserved across species. Both Kv4.3-L450F and Kv4.3-G600R demonstrated a gain-of-function phenotype, increasing peak I to current density by 146.2% (n = 15, P <.05) and 50.4% (n = 15, P <.05), respectively. Simulations using a Luo-Rudy II action potential (AP) model demonstrated the stable loss of the AP dome as a result of the increased I to maximal conductance associated with the heterozygous expression of either L450F or G600R. Conclusions: These findings provide the first molecular and functional evidence implicating novel KCND3 gain-of-function mutations in the pathogenesis and phenotypic expression of BrS, with the potential for a lethal arrhythmia being precipitated by a genetically enhanced I to current gradient within the right ventricle where KCND3 expression is the highest.

AB - Background: Brugada syndrome (BrS) is a sudden death-predisposing genetic condition characterized electrocardiographically by ST segment elevation in the leads V 1-V 3. Given the prominent role of the transient outward current (I to) in BrS pathogenesis, we hypothesized that rare gain-of-function mutations in KCND3 may serve as a pathogenic substrate for BrS. Methods: Comprehensive mutational analysis of KCND3-encoded Kv4.3 (I to) was conducted using polymerase chain reaction, denaturing high performance liquid chromatography, and direct sequencing of DNA derived from 86 unrelated BrS1-8 genotype-negative BrS patients. DNA from 780 healthy individuals was examined to assess allelic frequency for nonsynonymous variants. Putative BrS-associated Kv4.3 mutations were engineered and coexpressed with wild-type KChIP2 in HEK293 cells. Wild-type and mutant I to ion currents were recorded using whole-cell patch clamp. Results: Two BrS1-8 genotype-negative cases possessed novel Kv4.3 missense mutations. Both Kv4.3-L450F and Kv4.3-G600R were absent in 1,560 reference alleles and involved residues highly conserved across species. Both Kv4.3-L450F and Kv4.3-G600R demonstrated a gain-of-function phenotype, increasing peak I to current density by 146.2% (n = 15, P <.05) and 50.4% (n = 15, P <.05), respectively. Simulations using a Luo-Rudy II action potential (AP) model demonstrated the stable loss of the AP dome as a result of the increased I to maximal conductance associated with the heterozygous expression of either L450F or G600R. Conclusions: These findings provide the first molecular and functional evidence implicating novel KCND3 gain-of-function mutations in the pathogenesis and phenotypic expression of BrS, with the potential for a lethal arrhythmia being precipitated by a genetically enhanced I to current gradient within the right ventricle where KCND3 expression is the highest.

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