title: Transient outward current (I(to)) gain-of-function mutations in the KCND3-encoded Kv4.3 potassium channel and Brugada syndrome.
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.
authors.affiliation: Department of Medicine (Division of Cardiovascular Diseases), Department of Pediatrics (Division of Pediatric Cardiology), and Department of Molecular Pharmacology and Experimental Therapeutics, Windland Smith Rice Sudden Death Genomics Laboratory, Mayo Clinic, Rochester, Minnesota, USA. ackerman.michael@mayo.edu
authors.forename: John R
authors.initials: JR
authors.lastname: Giudicessi
authors.forename: Dan
authors.initials: D
authors.lastname: Ye
authors.forename: David J
authors.initials: DJ
authors.lastname: Tester
authors.forename: Lia
authors.initials: L
authors.lastname: Crotti
authors.forename: Alessandra
authors.initials: A
authors.lastname: Mugione
authors.forename: Vladislav V
authors.initials: VV
authors.lastname: Nesterenko
authors.forename: Richard M
authors.initials: RM
authors.lastname: Albertson
authors.forename: Charles
authors.initials: C
authors.lastname: Antzelevitch
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Schwartz
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Ackerman
mesh_terms: D000200:Action Potentials / Q000235:genetics; D000328:Adult; D000483:Alleles; D053840:Brugada Syndrome / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics*; D004562:Electrocardiography; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D018408:Patch-Clamp Techniques; D016133:Polymerase Chain Reaction; D051672:Shal Potassium Channels / Q000235:genetics* / Q000378:metabolism; D055815:Young Adult
chemical_list: C497021:KCND3 protein, human; D051672:Shal Potassium Channels; D004247:DNA