Suppression and Replacement Gene Therapy for KCNH2-Mediated Arrhythmias
BACKGROUND: KCNH2-mediated arrhythmia syndromes are caused by loss-of-function (type 2 long QT syndrome [LQT2]) or gain-of-function (type 1 short QT syndrome [SQT1]) pathogenic variants in the KCNH2-encoded Kv11.1 potassium channel, which is essential for the cardiac action potential.
METHODS: A dual-component "suppression-and-replacement" (SupRep) KCNH2 gene therapy was created by cloning into a single construct a custom-designed KCNH2 short hairpin RNA with ~80% knockdown (suppression) and a "short hairpin RNA-immune" KCNH2 cDNA (replacement). Induced pluripotent stem cell-derived cardiomyocytes and their CRISPR-Cas9 variant-corrected isogenic control (IC) induced pluripotent stem cell-derived cardiomyocytes were made for 2 LQT2- (G604S, N633S) and 1 SQT1- (N588K) causative variants. All variant lines were treated with KCNH2-SupRep or non-targeting control short hairpin RNA (shCT). The action potential duration (APD) at 90% repolarization (APD90) was measured using FluoVolt voltage dye.
RESULTS: KCNH2-SupRep achieved variant-independent rescue of both pathologic phenotypes. For LQT2-causative variants, treatment with KCNH2-SupRep resulted in shortening of the pathologically prolonged APD90 to near curative (IC-like) APD90 levels (G604S IC, 471±25 ms; N633S IC, 405±55 ms) compared with treatment with shCT (G604S: SupRep-treated, 452±76 ms versus shCT-treated, 550±41 ms; P<0.0001; N633S: SupRep-treated, 399±105 ms versus shCT-treated, 577±39 ms, P<0.0001). Conversely, for the SQT1-causative variant, N588K, treatment with KCNH2-SupRep resulted in therapeutic prolongation of the pathologically shortened APD90 (IC: 429±16 ms; SupRep-treated: 396±61 ms; shCT-treated: 274±12 ms).
CONCLUSIONS: We provide the first proof-of-principle gene therapy for correction of both LQT2 and SQT1. KCNH2-SupRep gene therapy successfully normalized the pathologic APD90, thereby eliminating the pathognomonic feature of both LQT2 and SQT1.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:15 |
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| Enthalten in: |
Circulation. Genomic and precision medicine - 15(2022), 6 vom: 17. Dez., Seite e003719 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Bains, Sahej [VerfasserIn] |
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| Links: |
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| Themen: |
ERG1 Potassium Channel |
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| Anmerkungen: |
Date Completed 22.12.2022 Date Revised 29.01.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1161/CIRCGEN.122.003719 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM347670105 |
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| 245 | 1 | 0 | |a Suppression and Replacement Gene Therapy for KCNH2-Mediated Arrhythmias |
| 264 | 1 | |c 2022 | |
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| 500 | |a Date Completed 22.12.2022 | ||
| 500 | |a Date Revised 29.01.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a BACKGROUND: KCNH2-mediated arrhythmia syndromes are caused by loss-of-function (type 2 long QT syndrome [LQT2]) or gain-of-function (type 1 short QT syndrome [SQT1]) pathogenic variants in the KCNH2-encoded Kv11.1 potassium channel, which is essential for the cardiac action potential | ||
| 520 | |a METHODS: A dual-component "suppression-and-replacement" (SupRep) KCNH2 gene therapy was created by cloning into a single construct a custom-designed KCNH2 short hairpin RNA with ~80% knockdown (suppression) and a "short hairpin RNA-immune" KCNH2 cDNA (replacement). Induced pluripotent stem cell-derived cardiomyocytes and their CRISPR-Cas9 variant-corrected isogenic control (IC) induced pluripotent stem cell-derived cardiomyocytes were made for 2 LQT2- (G604S, N633S) and 1 SQT1- (N588K) causative variants. All variant lines were treated with KCNH2-SupRep or non-targeting control short hairpin RNA (shCT). The action potential duration (APD) at 90% repolarization (APD90) was measured using FluoVolt voltage dye | ||
| 520 | |a RESULTS: KCNH2-SupRep achieved variant-independent rescue of both pathologic phenotypes. For LQT2-causative variants, treatment with KCNH2-SupRep resulted in shortening of the pathologically prolonged APD90 to near curative (IC-like) APD90 levels (G604S IC, 471±25 ms; N633S IC, 405±55 ms) compared with treatment with shCT (G604S: SupRep-treated, 452±76 ms versus shCT-treated, 550±41 ms; P<0.0001; N633S: SupRep-treated, 399±105 ms versus shCT-treated, 577±39 ms, P<0.0001). Conversely, for the SQT1-causative variant, N588K, treatment with KCNH2-SupRep resulted in therapeutic prolongation of the pathologically shortened APD90 (IC: 429±16 ms; SupRep-treated: 396±61 ms; shCT-treated: 274±12 ms) | ||
| 520 | |a CONCLUSIONS: We provide the first proof-of-principle gene therapy for correction of both LQT2 and SQT1. KCNH2-SupRep gene therapy successfully normalized the pathologic APD90, thereby eliminating the pathognomonic feature of both LQT2 and SQT1 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a genetic therapy | |
| 650 | 4 | |a induced pluripotent stem cells | |
| 650 | 4 | |a ion channel | |
| 650 | 4 | |a long QT syndrome | |
| 650 | 7 | |a ERG1 Potassium Channel |2 NLM | |
| 650 | 7 | |a KCNH2 protein, human |2 NLM | |
| 700 | 1 | |a Zhou, Wei |e verfasserin |4 aut | |
| 700 | 1 | |a Dotzler, Steven M |e verfasserin |4 aut | |
| 700 | 1 | |a Martinez, Katherine |e verfasserin |4 aut | |
| 700 | 1 | |a Kim, Cs John |e verfasserin |4 aut | |
| 700 | 1 | |a Tester, David J |e verfasserin |4 aut | |
| 700 | 1 | |a Ye, Dan |e verfasserin |4 aut | |
| 700 | 1 | |a Ackerman, Michael J |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:15 |g year:2022 |g number:6 |g day:17 |g month:12 |g pages:e003719 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1161/CIRCGEN.122.003719 |3 Volltext |
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