Impairment of the ER/mitochondria compartment in human cardiomyocytes with PLN p.Arg14del mutation.
Impairment of the ER/mitochondria compartment in human cardiomyocytes with PLN p.Arg14del mutation.
复制标题
PLN p.Arg14del突变对人心肌细胞内质网/线粒体室的损伤。
DOI：
10.15252/emmm.202013074
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发表时间：
2021-06-07
影响因子：
11.1
通讯作者：
Hansen A
中科院分区：
文献类型：
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作者：
Cuello F;Knaust AE;Saleem U;Loos M;Raabe J;Mosqueira D;Laufer S;Schweizer M;van der Kraak P;Flenner F;Ulmer BM;Braren I;Yin X;Theofilatos K;Ruiz-Orera J;Patone G;Klampe B;Schulze T;Piasecki A;Pinto Y;Vink A;Hübner N;Harding S;Mayr M;Denning C;Eschenhagen T;Hansen A
磷蛋白（PLN） p.a g14del突变引起扩张性心肌病，其分子疾病机制尚不完全清楚。将患者真皮成纤维细胞重编程为hiPSC，通过CRISPR/Cas9建立等基因对照，并分化心肌细胞。突变心肌细胞表现出Ca2+瞬态衰减时间明显延长，Ca2+负荷依赖性不规则跳动模式和较低的力。蛋白质组学分析显示内质网（ER）、核糖体和线粒体蛋白减少。电镜显示内质网扩张，与线粒体密切相关的大脂滴。后续实验证实内质网/线粒体室受损。与缺血性心力衰竭和非衰竭供体心脏样本相比，PLN p.a g14del终末期心力衰竭样本显示，ER标记蛋白和氧化应激在核周聚集物中呈阳性。Ca2+结合蛋白GCaMP6f或小白蛋白转导PLN p.a g14del ets可改善疾病表型。本研究发现无SR功能障碍的ER/线粒体室损伤是PLN p.a g14del心肌病的一种新的疾病机制。Ca2+清除改善了病理，提示局部Ca2+循环受损是一个重要的疾病罪魁祸首。将磷蛋白（PLN） p.a g14del突变与扩张性心肌病联系起来的疾病机制尚不完全清楚。在这项研究中，使用患者来源的人诱导多能干细胞-心肌细胞来阐明这一分子机制。
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. The disease mechanism linking the phospholamban (PLN) p.Arg14del mutation to dilated cardiomyopathy is incompletely understood. In this study, patient‐derived human induced pluripotent stem cell‐cardiomyocytes were used to elucidate this molecular mechanism.