Original articleCardiac potassium channel dysfunction in sudden infant death syndrome
Introduction
Life-threatening arrhythmias including the congenital long QT syndrome (LQTS) have been proposed to contribute to some cases of the sudden infant death syndrome (SIDS), the leading cause of death among infants 1 month to 1 year of age [1], [2], [3]. Supporting this theory are recent observations that mutations in genes responsible for LQTS are found in approximately 5–10% of SIDS cases [4], [5], [6]. Congenital LQTS is an inherited disorder associated with mutations in genes predominantly encoding subunits of cardiac ion channels [7]. The disorder is caused by impaired ventricular repolarization leading to prolongation of the QT interval and an increased risk for life-threatening ventricular arrhythmias. Other mutations in some of the same genes (KCNH2, KCNQ1) can enhance repolarization and cause shortening of the QT interval [8], [9], [10]. In the short QT syndrome (SQTS), there is also increased risk for cardiac arrhythmias and sudden death [11], [12].
Data linking cardiac potassium channel mutations with SIDS include anecdotal reports and population surveys. In 2001, Schwartz and colleagues reported discovery of a de novo KCNQ1 mutation (P117L) in a SIDS victim [13]. The same mutation was also observed to segregate with autosomal dominant LQTS at reduced penetrance in an unrelated Italian family [13]. Another missense KCNQ1 variant (H105L) without functional consequences was discovered among 41 German SIDS cases [14]. Two novel KCNH2 mutations (K101E, P1157L) have also been reported in single SIDS probands [5], [15], and in one case this finding led to discovery of undiagnosed LQTS in family members [15]. But, there have been no studies demonstrating that any of these mutant channels are dysfunctional.
Ackerman et al. reported a systematic survey of autopsied SIDS cases for mutations in the LQTS genes, an effort that originally identified two SCN5A mutations in 93 subjects [4]. The study later reported novel mutations in cardiac potassium channel genes (KCNH2-G294V, KCNQ1-T600M) occurring in two SIDS victims [5]. Three other potassium channel variants were identified, but these alleles were also observed in ethnically matched population controls, albeit rarely. Most recently, the largest series of cases screened for mutations in arrhythmia-susceptibility genes revealed novel missense KCNH2 and KCNQ1 mutations in Norwegian SIDS victims [6]. Importantly, the electrophysiological consequences of these novel SIDS-associated potassium channel mutations were previously unknown.
We report here the functional characterization of novel KCNH2 and KCNQ1 mutations identified in the Norwegian SIDS study. Our findings demonstrated a diversity of functional phenotypes that revealed plausible explanations for sudden death in this clinical setting.
Section snippets
Mutagenesis and heterologous expression
Four single KCNH2 variants (V279M, R273Q, R885C, S1040G), one compound genotype (K897T/R954C) and a KCNQ1 mutation (I274V) were constructed in HERG or KCNQ1 cDNA vectors, respectively, using recombinant PCR mutagenesis (primer sequences available upon request). The final constructs were assembled in bicistronic mammalian expression plasmids (pIRES2-EGFP or pIRES2-DsRed, BD Biosciences-Clontech, Palo Alto, CA) to enable monitoring of transfection success by co-expression of fluorescent proteins.
Results
We examined the biophysical properties of four single missense KCNH2 variants, one compound KCNH2 genotype, and a missense KCNQ1 mutation using whole-cell patch clamp recording of recombinant potassium channels heterologously expressed in cultured mammalian cells. All mutations were originally identified in a large, retrospective series of 201 Norwegian SIDS cases that were screened for genetic variants in the major LQTS genes [6]. Three KCNH2 alleles (V279M, R885C, and S1040G) exhibited
Discussion
Sudden unexplained death of an infant is a tragic event. In the absence of an obvious cause of death including intentional injury as demonstrated by a negative autopsy and a thorough death scene investigation, a diagnosis of sudden infant death syndrome (SIDS) can be considered [23]. Although the diagnosis of SIDS is one of exclusion, ongoing efforts to define preventable causes have great importance for understanding the pathogenesis of this syndrome and ultimately for reducing infant
Acknowledgments
We thank Drs. Jonathan Silva and Yoram Rudy for helpful discussions regarding action potential modeling. This work was supported in part by NIH grant HL083374 (A.L.G.) and T.E.R. was supported by an institutional postdoctoral training grant (T32-GM007569).
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