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Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes.

Matsa E, Dixon JE, Medway C, Georgiou O, Patel MJ, Morgan K, Kemp PJ, Staniforth A, Mellor I, Denning C - Eur. Heart J. (2013)

Bottom Line: Allele-specific RNA interference (RNAi) directed towards the mutated KCNH2 mRNA caused knockdown, while leaving the wild-type mRNA unaffected.These findings provide in vitro evidence that allele-specific RNAi can rescue diseased phenotype in LQTS cardiomyocytes.This is a potentially novel route for the treatment of many autosomal-dominant-negative disorders, including those of the heart.

View Article: PubMed Central - PubMed

Affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), University of Nottingham, Nottingham NG7 2RD, UK.

ABSTRACT

Aims: Long-QT syndromes (LQTS) are mostly autosomal-dominant congenital disorders associated with a 1:1000 mutation frequency, cardiac arrest, and sudden death. We sought to use cardiomyocytes derived from human-induced pluripotency stem cells (hiPSCs) as an in vitro model to develop and evaluate gene-based therapeutics for the treatment of LQTS.

Methods and results: We produced LQTS-type 2 (LQT2) hiPSC cardiomyocytes carrying a KCNH2 c.G1681A mutation in a IKr ion-channel pore, which caused impaired glycosylation and channel transport to cell surface. Allele-specific RNA interference (RNAi) directed towards the mutated KCNH2 mRNA caused knockdown, while leaving the wild-type mRNA unaffected. Electrophysiological analysis of patient-derived LQT2 hiPSC cardiomyocytes treated with mutation-specific siRNAs showed normalized action potential durations (APDs) and K(+) currents with the concurrent rescue of spontaneous and drug-induced arrhythmias (presented as early-afterdepolarizations).

Conclusions: These findings provide in vitro evidence that allele-specific RNAi can rescue diseased phenotype in LQTS cardiomyocytes. This is a potentially novel route for the treatment of many autosomal-dominant-negative disorders, including those of the heart.

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Related in: MedlinePlus

Isoprenaline and E4031 drug treatment in siRNA-treated long-QT syndrome-type 2 cardiomyocytes. (A) (i) Averaged and (ii) raw data of isoprenaline-induced arrhythmias in the form of early-afterdepolarizations observed only in control long-QT syndrome-type 2 cardiomyocytes (n = 10). (B). (i) Isoprenaline (100 nM, n = 10) or (ii) E4031 (1 µM, n = 5) treatment in A13-treated long-QT syndrome-type 2 cardiomyocytes did not cause early-afterdepolarizations, depicting rescue of long-QT syndrome-type 2-phenotype by RNAi-based therapy.
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EHT067F6: Isoprenaline and E4031 drug treatment in siRNA-treated long-QT syndrome-type 2 cardiomyocytes. (A) (i) Averaged and (ii) raw data of isoprenaline-induced arrhythmias in the form of early-afterdepolarizations observed only in control long-QT syndrome-type 2 cardiomyocytes (n = 10). (B). (i) Isoprenaline (100 nM, n = 10) or (ii) E4031 (1 µM, n = 5) treatment in A13-treated long-QT syndrome-type 2 cardiomyocytes did not cause early-afterdepolarizations, depicting rescue of long-QT syndrome-type 2-phenotype by RNAi-based therapy.

Mentions: A13-treated LQT2-myocytes did not develop isoprenaline-induced arrhythmias (n = 10; Figure 6B), suggesting patients under siRNA therapy could have increased tolerance to exercise or stress. This was in contrast to control LQT2 cardiomyocytes (n = 10, Figure 6A), and cardiomyocytes under combined Nicorandil and PD-118057 treatment which had enabled comparable APD decrease to siRNA treatment (Supplementary material online, Figure S5B). In addition, siRNA-treated myocytes showed no arrhythmias with E4031 treatment (n = 5, Figure 6B), which indicates that patients potentially undertaking this therapy could be less susceptible to drug-induced cardiac episodes when administered drugs with cardio-toxic QT-elongation side-effects.30 In summary, although focusing on one specific LQT2 mutation, the results presented here are evidence that allele-specific RNAi can rescue LQTS phenotype, in a humanized functionally relevant in vitro disease model.Figure 6


Allele-specific RNA interference rescues the long-QT syndrome phenotype in human-induced pluripotency stem cell cardiomyocytes.

Matsa E, Dixon JE, Medway C, Georgiou O, Patel MJ, Morgan K, Kemp PJ, Staniforth A, Mellor I, Denning C - Eur. Heart J. (2013)

Isoprenaline and E4031 drug treatment in siRNA-treated long-QT syndrome-type 2 cardiomyocytes. (A) (i) Averaged and (ii) raw data of isoprenaline-induced arrhythmias in the form of early-afterdepolarizations observed only in control long-QT syndrome-type 2 cardiomyocytes (n = 10). (B). (i) Isoprenaline (100 nM, n = 10) or (ii) E4031 (1 µM, n = 5) treatment in A13-treated long-QT syndrome-type 2 cardiomyocytes did not cause early-afterdepolarizations, depicting rescue of long-QT syndrome-type 2-phenotype by RNAi-based therapy.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3992427&req=5

EHT067F6: Isoprenaline and E4031 drug treatment in siRNA-treated long-QT syndrome-type 2 cardiomyocytes. (A) (i) Averaged and (ii) raw data of isoprenaline-induced arrhythmias in the form of early-afterdepolarizations observed only in control long-QT syndrome-type 2 cardiomyocytes (n = 10). (B). (i) Isoprenaline (100 nM, n = 10) or (ii) E4031 (1 µM, n = 5) treatment in A13-treated long-QT syndrome-type 2 cardiomyocytes did not cause early-afterdepolarizations, depicting rescue of long-QT syndrome-type 2-phenotype by RNAi-based therapy.
Mentions: A13-treated LQT2-myocytes did not develop isoprenaline-induced arrhythmias (n = 10; Figure 6B), suggesting patients under siRNA therapy could have increased tolerance to exercise or stress. This was in contrast to control LQT2 cardiomyocytes (n = 10, Figure 6A), and cardiomyocytes under combined Nicorandil and PD-118057 treatment which had enabled comparable APD decrease to siRNA treatment (Supplementary material online, Figure S5B). In addition, siRNA-treated myocytes showed no arrhythmias with E4031 treatment (n = 5, Figure 6B), which indicates that patients potentially undertaking this therapy could be less susceptible to drug-induced cardiac episodes when administered drugs with cardio-toxic QT-elongation side-effects.30 In summary, although focusing on one specific LQT2 mutation, the results presented here are evidence that allele-specific RNAi can rescue LQTS phenotype, in a humanized functionally relevant in vitro disease model.Figure 6

Bottom Line: Allele-specific RNA interference (RNAi) directed towards the mutated KCNH2 mRNA caused knockdown, while leaving the wild-type mRNA unaffected.These findings provide in vitro evidence that allele-specific RNAi can rescue diseased phenotype in LQTS cardiomyocytes.This is a potentially novel route for the treatment of many autosomal-dominant-negative disorders, including those of the heart.

View Article: PubMed Central - PubMed

Affiliation: Wolfson Centre for Stem Cells, Tissue Engineering and Modelling (STEM), University of Nottingham, Nottingham NG7 2RD, UK.

ABSTRACT

Aims: Long-QT syndromes (LQTS) are mostly autosomal-dominant congenital disorders associated with a 1:1000 mutation frequency, cardiac arrest, and sudden death. We sought to use cardiomyocytes derived from human-induced pluripotency stem cells (hiPSCs) as an in vitro model to develop and evaluate gene-based therapeutics for the treatment of LQTS.

Methods and results: We produced LQTS-type 2 (LQT2) hiPSC cardiomyocytes carrying a KCNH2 c.G1681A mutation in a IKr ion-channel pore, which caused impaired glycosylation and channel transport to cell surface. Allele-specific RNA interference (RNAi) directed towards the mutated KCNH2 mRNA caused knockdown, while leaving the wild-type mRNA unaffected. Electrophysiological analysis of patient-derived LQT2 hiPSC cardiomyocytes treated with mutation-specific siRNAs showed normalized action potential durations (APDs) and K(+) currents with the concurrent rescue of spontaneous and drug-induced arrhythmias (presented as early-afterdepolarizations).

Conclusions: These findings provide in vitro evidence that allele-specific RNAi can rescue diseased phenotype in LQTS cardiomyocytes. This is a potentially novel route for the treatment of many autosomal-dominant-negative disorders, including those of the heart.

Show MeSH
Related in: MedlinePlus