ACTN2 Mutant Causes Proteopathy in Human iPSC-Derived Cardiomyocytes
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ACTN2 Mutant Causes Proteopathy in Human iPSC-Derived Cardiomyocytes. / Zech, Antonia T L; Prondzynski, Maksymilian; Singh, Sonia R; Pietsch, Niels; Orthey, Ellen; Alizoti, Erda; Busch, Josefine; Madsen, Alexandra; Behrens, Charlotta S; Meyer-Jens, Moritz; Mearini, Giulia; Lemoine, Marc D; Krämer, Elisabeth; Mosqueira, Diogo; Virdi, Sanamjeet; Indenbirken, Daniela; Depke, Maren; Salazar, Manuela Gesell; Völker, Uwe; Braren, Ingke; Pu, William T; Eschenhagen, Thomas; Hammer, Elke; Schlossarek, Saskia; Carrier, Lucie.
in: CELLS-BASEL, Jahrgang 11, Nr. 17, 2745, 02.09.2022.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - ACTN2 Mutant Causes Proteopathy in Human iPSC-Derived Cardiomyocytes
AU - Zech, Antonia T L
AU - Prondzynski, Maksymilian
AU - Singh, Sonia R
AU - Pietsch, Niels
AU - Orthey, Ellen
AU - Alizoti, Erda
AU - Busch, Josefine
AU - Madsen, Alexandra
AU - Behrens, Charlotta S
AU - Meyer-Jens, Moritz
AU - Mearini, Giulia
AU - Lemoine, Marc D
AU - Krämer, Elisabeth
AU - Mosqueira, Diogo
AU - Virdi, Sanamjeet
AU - Indenbirken, Daniela
AU - Depke, Maren
AU - Salazar, Manuela Gesell
AU - Völker, Uwe
AU - Braren, Ingke
AU - Pu, William T
AU - Eschenhagen, Thomas
AU - Hammer, Elke
AU - Schlossarek, Saskia
AU - Carrier, Lucie
PY - 2022/9/2
Y1 - 2022/9/2
N2 - Genetic variants in α-actinin-2 (ACTN2) are associated with several forms of (cardio)myopathy. We previously reported a heterozygous missense (c.740C>T) ACTN2 gene variant, associated with hypertrophic cardiomyopathy, and characterized by an electro-mechanical phenotype in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Here, we created with CRISPR/Cas9 genetic tools two heterozygous functional knock-out hiPSC lines with a second wild-type (ACTN2wt) and missense ACTN2 (ACTN2mut) allele, respectively. We evaluated their impact on cardiomyocyte structure and function, using a combination of different technologies, including immunofluorescence and live cell imaging, RNA-seq, and mass spectrometry. This study showed that ACTN2mut presents a higher percentage of multinucleation, protein aggregation, hypertrophy, myofibrillar disarray, and activation of both the ubiquitin-proteasome system and the autophagy-lysosomal pathway as compared to ACTN2wt in 2D-cultured hiPSC-CMs. Furthermore, the expression of ACTN2mut was associated with a marked reduction of sarcomere-associated protein levels in 2D-cultured hiPSC-CMs and force impairment in engineered heart tissues. In conclusion, our study highlights the activation of proteolytic systems in ACTN2mut hiPSC-CMs likely to cope with ACTN2 aggregation and therefore directs towards proteopathy as an additional cellular pathology caused by this ACTN2 variant, which may contribute to human ACTN2-associated cardiomyopathies.
AB - Genetic variants in α-actinin-2 (ACTN2) are associated with several forms of (cardio)myopathy. We previously reported a heterozygous missense (c.740C>T) ACTN2 gene variant, associated with hypertrophic cardiomyopathy, and characterized by an electro-mechanical phenotype in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Here, we created with CRISPR/Cas9 genetic tools two heterozygous functional knock-out hiPSC lines with a second wild-type (ACTN2wt) and missense ACTN2 (ACTN2mut) allele, respectively. We evaluated their impact on cardiomyocyte structure and function, using a combination of different technologies, including immunofluorescence and live cell imaging, RNA-seq, and mass spectrometry. This study showed that ACTN2mut presents a higher percentage of multinucleation, protein aggregation, hypertrophy, myofibrillar disarray, and activation of both the ubiquitin-proteasome system and the autophagy-lysosomal pathway as compared to ACTN2wt in 2D-cultured hiPSC-CMs. Furthermore, the expression of ACTN2mut was associated with a marked reduction of sarcomere-associated protein levels in 2D-cultured hiPSC-CMs and force impairment in engineered heart tissues. In conclusion, our study highlights the activation of proteolytic systems in ACTN2mut hiPSC-CMs likely to cope with ACTN2 aggregation and therefore directs towards proteopathy as an additional cellular pathology caused by this ACTN2 variant, which may contribute to human ACTN2-associated cardiomyopathies.
KW - Actinin/genetics
KW - Cardiomyopathy, Hypertrophic/metabolism
KW - Humans
KW - Induced Pluripotent Stem Cells/metabolism
KW - Myocytes, Cardiac/metabolism
KW - Sarcomeres/metabolism
U2 - 10.3390/cells11172745
DO - 10.3390/cells11172745
M3 - SCORING: Journal article
C2 - 36078153
VL - 11
JO - CELLS-BASEL
JF - CELLS-BASEL
SN - 2073-4409
IS - 17
M1 - 2745
ER -