Human-Induced Pluripotent Stem Cell–Derived Cardiomyocyte Model for TNNT2 Δ160E-Induced Cardiomyopathy

Author:

Kondo Takumi1ORCID,Higo Shuichiro2ORCID,Shiba Mikio1ORCID,Kohama Yasuaki1ORCID,Kameda Satoshi1,Tabata Tomoka1,Inoue Hiroyuki1,Okuno Shota1ORCID,Ogawa Shou1,Nakamura Satoki3,Takeda Maki4ORCID,Ito Emiko4,Li Junjun4ORCID,Liu Li4,Kuramoto Yuki15ORCID,Lee Jong-Kook16ORCID,Takashima Seiji7,Miyagawa Shigeru4ORCID,Sawa Yoshiki4ORCID,Hikoso Shungo1ORCID,Sakata Yasushi1ORCID

Affiliation:

1. Department of Cardiovascular Medicine (T.K., M.S., S.K., T.T., H.I., S. Okuno, S.Ogawa, Y.K., J.-K.L., S. Hikoso, Y.S.)

2. Department of Medical Therapeutics for Heart Failure (S. Higo)

3. Osaka Police Hospital, Osaka (S.N.).

4. Department of Cardiovascular Surgery (M.T., E.I., J.L., L.L., S.M., Y.S.)

5. National Hospital Organization, Osaka-Minami Medical Center, Kawachinagano, Osaka (Y.K.).

6. Department of Cardiovascular Regenerative Medicine (J.-K.L.)

7. Department of Medical Biochemistry (S.T.), Osaka University Graduate School of Medicine, Suita, Osaka.

Abstract

Background: The Δ160E mutation in TNNT2 , which encodes troponin T, is a rare pathogenic variant identified in patients with hypertrophic cardiomyopathy and is associated with poor prognosis. Thus, a convenient human model recapitulating the pathological phenotype caused by TNNT2 Δ160E is required for therapeutic development. Methods: We identified a heterozygous in-frame deletion mutation (c.478_480del, p.Δ160E) in TNNT2 in a patient with familial hypertrophic cardiomyopathy showing progressive left ventricular systolic dysfunction, leading to advanced heart failure. To investigate the pathological phenotype caused by Δ160E, we generated a set of isogenic induced pluripotent stem cells carrying the heterozygous Δ160E, homozygously corrected or homozygously introduced Δ160E using genome editing and differentiated them into cardiomyocytes (Hetero-Δ160E-, wild type-, and Homo-Δ160E-induced pluripotent stem cells [iPSC]-derived cardiomyocytes [iPSC-CMs]). Results: Hetero-Δ160E-iPSC-CMs exhibited prolonged calcium decay, relaxation impairment, and hypertrophy compared to wild type-iPSC-CMs. Notably, these phenotypes were further exacerbated in Homo-Δ160E-iPSC-CMs. Overexpression of R-GECO-fused Δ160E mutant troponin T prolonged decay time and time to peak of the myofilament-localized calcium transient in iPSC-CMs, indicating that sarcomeric calcium retention with Δ160E may affect intracellular calcium concentration. High-content imaging analysis detected remarkable nuclear translocation of NFATc1, especially in Homo-Δ160E-iPSC-CMs, indicating that the Δ160E mutation promotes hypertrophic signaling pathway in a dose-dependent manner. Increased phosphorylation of CaMKIIδ (calcium/calmodulin-dependent protein kinase IIδ) and phospholamban at Thr17 was observed in Homo- and Hetero-Δ160E-iPSC-CMs. Epigallocatechin-3-gallate, a calcium desensitizing compound, shortened prolonged calcium decay and relaxation duration in Δ160E-iPSC-CMs. Conclusions: Isogenic iPSC-CMs recapitulate the prolonged calcium decay, relaxation impairment, and subsequent calcium-regulated signaling pathways caused by the TNNT2 Δ160E mutation and can serve as a human model for therapeutic development to prevent hypertrophic cardiomyopathy pathology.

Publisher

Ovid Technologies (Wolters Kluwer Health)

Subject

General Medicine

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