Single Construct Suppression and Replacement Gene Therapy for the Treatment of All CALM1 -, CALM2 -, and CALM3 -Mediated Arrhythmia Disorders.
Autor: | Hamrick SK; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Kim CSJ; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Tester DJ; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Gencarelli M; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Tobert KE; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Gluscevic M; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN., Ackerman MJ; Department of Molecular Pharmacology and Experimental Therapeutics (Windland Smith Rice Sudden Death Genomics Laboratory) (S.K.H., C.S.J.K., D.J.T., M. Gencarelli, K.E.T., M. Gluscevic, M.J.A.), Mayo Clinic, Rochester, MN.; Department of Cardiovascular Medicine (Division of Heart Rhythm Services, Windland Smith Rice Genetic Heart Rhythm Clinic) (M.J.A.), Mayo Clinic, Rochester, MN.; Department of Pediatric and Adolescent Medicine (Division of Pediatric Cardiology) (M.J.A.), Mayo Clinic, Rochester, MN. |
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Jazyk: | angličtina |
Zdroj: | Circulation. Arrhythmia and electrophysiology [Circ Arrhythm Electrophysiol] 2024 Aug; Vol. 17 (8), pp. e012036. Date of Electronic Publication: 2024 Jul 29. |
DOI: | 10.1161/CIRCEP.123.012036 |
Abstrakt: | Background: CaM (calmodulin)-mediated long-QT syndrome is a genetic arrhythmia disorder (calmodulinopathies) characterized by a high prevalence of life-threatening ventricular arrhythmias occurring early in life. Three distinct genes ( CALM1 , CALM2 , and CALM3 ) encode for the identical CaM protein. Conventional pharmacotherapies fail to adequately protect against potentially lethal cardiac events in patients with calmodulinopathy. Methods: Five custom-designed CALM1 -, CALM2 -, and CALM3 -targeting short hairpin RNAs (shRNAs) were tested for knockdown (KD) efficiency using TSA201 cells and reverse transcription-quantitative polymerase chain reaction. A dual-component suppression and replacement (SupRep) CALM gene therapy (CALM-SupRep) was created by cloning into a single construct CALM1 -, CALM2 -, and CALM3-specific shRNAs that produce KD (suppression) of each respective gene and a shRNA-immune CALM1 cDNA (replacement). CALM1-F142L, CALM2-D130G, and CALM3-D130G induced pluripotent stem cell-derived CMs were generated from patients with CaM-mediated long-QT syndrome. A voltage-sensing dye was used to measure action potential duration at 90% repolarization (APD90). Results: Following shRNA KD efficiency testing, a candidate shRNA was identified for CALM1 (86% KD), CALM2 (71% KD), and CALM3 (94% KD). The APD90 was significantly prolonged in CALM2-D130G (647±9 ms) compared with CALM2-WT (359±12 ms; P <0.0001). Transfection with CALM -SupRep shortened the average APD90 of CALM2-D130G to 457±19 ms (66% attenuation; P <0.0001). Additionally, transfection with CALM -SupRep shortened the APD90 of CALM1-F142L (665±9 to 410±15 ms; P <0.0001) and CALM3-D130G (978±81 to 446±6 ms; P <0.001). Conclusions: We provide the first proof-of-principle suppression-replacement gene therapy for CaM-mediated long-QT syndrome. The CALM-SupRep gene therapy shortened the pathologically prolonged APD90 in CALM1 -, CALM2 -, and CALM3-variant CaM-mediated long-QT syndrome induced pluripotent stem cell-derived CM lines. The single CALM-SupRep construct may be able to treat all calmodulinopathies, regardless of which of the 3 CaM-encoding genes are affected. Competing Interests: Dr Ackerman is a consultant for Abbott, Boston Scientific, Bristol Myers Squibb, Daiichi Sankyo, Invitae, Medtronic, Tenaya Therapeutics, and UpToDate. Dr Ackerman and Mayo Clinic are involved in an equity/royalty relationship with AliveCor, Anumana, ARMGO Pharma, Pfizer, and Thryv Therapeutics. However, none of these entities have contributed to this study in any manner. The other authors report no conflicts. |
Databáze: | MEDLINE |
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