TY - JOUR
T1 - Impairment of the ER/mitochondria compartment in human cardiomyocytes with PLN p.Arg14del mutation
AU - Cuello, Friederike
AU - Knaust, Anika E.
AU - Saleem, Umber
AU - Loos, Malte
AU - Raabe, Janice
AU - Mosqueira, Diogo
AU - Laufer, Sandra
AU - Schweizer, Michaela
AU - van der Kraak, Petra
AU - Flenner, Frederik
AU - Ulmer, B. rbel M.
AU - Braren, Ingke
AU - Yin, Xiaoke
AU - Theofilatos, Konstantinos
AU - Ruiz-Orera, Jorge
AU - Patone, Giannino
AU - Klampe, Birgit
AU - Schulze, Thomas
AU - Piasecki, Angelika
AU - Pinto, Yigal
AU - Vink, Aryan
AU - Hübner, Norbert
AU - Harding, Sian
AU - Mayr, Manuel
AU - Denning, Chris
AU - Eschenhagen, Thomas
AU - Hansen, Arne
N1 - Funding Information: We would like to acknowledge Prof. Hendrik Milting for contributing RNA samples of DCM patients carrying a PLN p. Arg14del mutation. We greatly appreciate the assistance of Kristin Hartman (UKE mouse pathology core facility) and UKE FACS Core unit and the team approach of hiPSC and CRISPR/Cas9 group at IEPT/UKE. We are grateful to expert technical assistance provided by Emanuela Szpotovicz. This study was supported by the British Heart Foundation RM/13/30157, European Research Council (ERC‐AG IndivuHeart), Deutsche Forschungsgemeinschaft (DFG Es 88/12‐1, DFG HA 3423/5‐1, DFG CU 53/5‐1), the Werner‐Otto‐Stiftung (7/92 to FC), the Deutsche Stiftung für Herzforschung (F/19/19 to FC), the British National Centre for the Replacement Refinement & Reduction of Animals in Research (NC3Rs CRACK‐IT grant 35911‐259146), the German Ministry of Education and Research (BMBF) and the Centre for Cardiovascular Research (DZHK), and the Freie und Hansestadt Hamburg. M. Mayr is a British Heart Foundation (BHF) Chair Holder (CH/16/3/32406) with BHF program grant support (RG/16/14/32397) and is part of the Marie Skłodowska‐Curie Innovative Training Network TRAIN‐HEART ( http://train‐heart.eu ), and JR was supported by a fellowship of the Studienstiftung des deutschen Volkes. Open Access funding enabled and organized by Projekt DEAL. Funding Information: We would like to acknowledge Prof. Hendrik Milting for contributing RNA samples of DCM patients carrying a PLN p. Arg14del mutation. We greatly appreciate the assistance of Kristin Hartman (UKE mouse pathology core facility) and UKE FACS Core unit and the team approach of hiPSC and CRISPR/Cas9 group at IEPT/UKE. We are grateful to expert technical assistance provided by Emanuela Szpotovicz. This study was supported by the British Heart Foundation RM/13/30157, European Research Council (ERC-AG IndivuHeart), Deutsche Forschungsgemeinschaft (DFG Es 88/12-1, DFG HA 3423/5-1, DFG CU 53/5-1), the Werner-Otto-Stiftung (7/92 to FC), the Deutsche Stiftung f?r Herzforschung (F/19/19 to FC), the British National Centre for the Replacement Refinement & Reduction of Animals in Research (NC3Rs CRACK-IT grant 35911-259146), the German Ministry of Education and Research (BMBF) and the Centre for Cardiovascular Research (DZHK), and the Freie und Hansestadt Hamburg. M. Mayr is a British Heart Foundation (BHF) Chair Holder (CH/16/3/32406) with BHF program grant support (RG/16/14/32397) and is part of the Marie Sk?odowska-Curie Innovative Training Network TRAIN-HEART (http://train-heart.eu), and JR was supported by a fellowship of the Studienstiftung des deutschen Volkes. Open Access funding enabled and organized by Projekt DEAL. Publisher Copyright: © 2021 The Authors. Published under the terms of the CC BY 4.0 license Copyright: Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/6/7
Y1 - 2021/6/7
N2 - The phospholamban (PLN) p.Arg14del mutation causes dilated cardiomyopathy, with the molecular disease mechanisms incompletely understood. Patient dermal fibroblasts were reprogrammed to hiPSC, isogenic controls were established by CRISPR/Cas9, and cardiomyocytes were differentiated. Mutant cardiomyocytes revealed significantly prolonged Ca2+ transient decay time, Ca2+-load dependent irregular beating pattern, and lower force. Proteomic analysis revealed less endoplasmic reticulum (ER) and ribosomal and mitochondrial proteins. Electron microscopy showed dilation of the ER and large lipid droplets in close association with mitochondria. Follow-up experiments confirmed impairment of the ER/mitochondria compartment. PLN p.Arg14del end-stage heart failure samples revealed perinuclear aggregates positive for ER marker proteins and oxidative stress in comparison with ischemic heart failure and non-failing donor heart samples. Transduction of PLN p.Arg14del EHTs with the Ca2+-binding proteins GCaMP6f or parvalbumin improved the disease phenotype. This study identified impairment of the ER/mitochondria compartment without SR dysfunction as a novel disease mechanism underlying PLN p.Arg14del cardiomyopathy. The pathology was improved by Ca2+-scavenging, suggesting impaired local Ca2+ cycling as an important disease culprit.
AB - The phospholamban (PLN) p.Arg14del mutation causes dilated cardiomyopathy, with the molecular disease mechanisms incompletely understood. Patient dermal fibroblasts were reprogrammed to hiPSC, isogenic controls were established by CRISPR/Cas9, and cardiomyocytes were differentiated. Mutant cardiomyocytes revealed significantly prolonged Ca2+ transient decay time, Ca2+-load dependent irregular beating pattern, and lower force. Proteomic analysis revealed less endoplasmic reticulum (ER) and ribosomal and mitochondrial proteins. Electron microscopy showed dilation of the ER and large lipid droplets in close association with mitochondria. Follow-up experiments confirmed impairment of the ER/mitochondria compartment. PLN p.Arg14del end-stage heart failure samples revealed perinuclear aggregates positive for ER marker proteins and oxidative stress in comparison with ischemic heart failure and non-failing donor heart samples. Transduction of PLN p.Arg14del EHTs with the Ca2+-binding proteins GCaMP6f or parvalbumin improved the disease phenotype. This study identified impairment of the ER/mitochondria compartment without SR dysfunction as a novel disease mechanism underlying PLN p.Arg14del cardiomyopathy. The pathology was improved by Ca2+-scavenging, suggesting impaired local Ca2+ cycling as an important disease culprit.
KW - endoplasmic reticulum
KW - engineered heart tissue
KW - human-induced pluripotent stem cells
KW - mitochondria
KW - phospholamban p.Arg14del
UR - http://www.scopus.com/inward/record.url?scp=85105933409&partnerID=8YFLogxK
U2 - https://doi.org/10.15252/emmm.202013074
DO - https://doi.org/10.15252/emmm.202013074
M3 - Article
C2 - 33998164
SN - 1757-4676
VL - 13
JO - EMBO molecular medicine
JF - EMBO molecular medicine
IS - 6
M1 - e13074
ER -