Following surgery requiring the use of a double-lumen endobronchial tube, a patient immediately complained of persistent severe hoarseness. On the third day after the operation, fibreoptic laryngoscopy revealed posterolateral dislocation of the left arytenoid cartilage. By the sixth day of the operation, a slight improvement was observed in the hoarseness without treatment and a spontaneous recovery of arytenoid cartilage dislocation was expected. The patient did not consent to surgical treatment, and therefore a conservative therapy was selected. Ten weeks after the operation, it was found that the dislocated left arytenoid cartilage had spontaneously repositioned and the patient regained his normal voice. The causes and treatment options are discussed.
Background
The A341V mutation in the pore-forming KCNQ1 subunit of the slowly activating delayed-rectifier potassium current (IKs) underlies a common form of the long QT syndrome, and is associated with an unusually severe phenotype. However, there is controversy regarding the underlying mechanism responsible for the clinically observed phenotype. We investigated the biophysical characteristics of A341V in a cardiac environment by utilizing a cardiac cell line, and in particular the impact of the KCNE1 β -subunit.
Methods
Whole-cell current were recorded from transiently transfected HL-1 cells, a cardiac cell line. Mutant KCNQ1 and KCNE1 were constructed by site-directed mutagenesis.
Results
The A341V mutant resulted in a non-functional channel when expressed alone. When co-expressed with wild type KCNE1, A341V produced a slowly activating current, with a smaller current density, slower rates of activation, and a depolarized shift in its activation curve compared to the wild type KCNQ1+KCNE1. Confocal microscopy confirmed the surface expression of GFP-tagged A341V, suggesting a functionally defective protein. A T58A mutation in KCNE1 abolished functional restoration of A341V. Under heterozygous conditions, the expression of A341V+KCNQ1+KCNE1 reduced but did not abolish the electrophysiological changes observed in A341V+KCNE1. A dominant negative effect of A341V was also observed. Action potential simulations revealed that the A341V mutation is arrhythmogenic.
Conclusions
The KCNE1 β-subunit partially rescued the non-functional A341V mutant, with electrophysiological properties distinct from the wild type IKs.
General Significance
The severity of the A341V phenotype may be due to a combination of a significant suppression of the IKs with altered biophysical characteristics.
Background
The impact of volatile anesthetics on patients with inherited long QT syndrome
(LQTS) is not well understood. This is further complicated by the different genotypes
underlying LQTS. No studies have reported on the direct effects of volatile anesthetics
on specific LQTS-associated mutations. We investigated the effects of isoflurane on a
common LQTS-Type 1 mutation, A341V, with an unusually severe phenotype.
Methods
Whole-cell potassium currents (IKs) were recorded from HEK293 and HL-1 cells
transiently expressing/co-expressing wild-type KCNQ1 (α-subunit), mutant KCNQ1,
wild-type KCNE1 (β-subunit), and fusion KCNQ1+KCNE1. Current was
monitored in the absence and presence of clinically relevant concentration of isoflurane
(0.54 ± 0.05 mM, 1.14 vol%). Computer simulations determined the
resulting impact on the cardiac action potential.
Results
Isoflurane had significantly greater inhibitory effect on A341V+KCNE1
(62.2 ± 3.4%, n=8), than on wild-type KCNQ1+KCNE1 (40.7
± 4.5%; n=9) in transfected HEK293 cells. Under heterozygous
conditions, isoflurane inhibited A341V+KCNQ1+KCNE1 by 65.2 ±
3.0% (n=13), and wild-type KCNQ1+KCNE1 (2:1 ratio) by 32.0
± 4.5% (n=11). A341V exerted a dominant negative effect on IKs.
Similar differential effects of isoflurane were also observed in experiments using the
cardiac HL-1 cells. Mutations of the neighboring F340 residue significantly attenuated
the effects of isoflurane, and fusion proteins revealed the modulatory effect of KCNE1.
Action potential simulations revealed a stimulation-frequency dependent effect of
A341V.
Conclusions
The LQTS-associated A341V mutation rendered the IKs channel more sensitive to
the inhibitory effects of isoflurane compared to wild-type IKs in transfected cell
lines; F340 is a key residue for anesthetic action.
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