Full-length dystrophin deficiency leads to contractile and calcium transient defects in human engineered heart tissues.
Full-length dystrophin deficiency leads to contractile and calcium transient defects in human engineered heart tissues.
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DOI：
10.1177/20417314221119628
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发表时间：
2022-01
影响因子：
8.2
通讯作者：
Mack, David L.
中科院分区：
文献类型：
--
作者：
Bremner, Samantha B.;Mandrycky, Christian J.;Leonard, Andrea;Padgett, Ruby M.;Levinson, Alan R.;Rehn, Ethan S.;Pioner, J. Manuel;Sniadecki, Nathan J.;Mack, David L.
关键词：
心肌病目前是杜氏肌营养不良症（DMD）患者死亡的主要原因，DMD是一种影响年轻男孩的严重神经肌肉疾病。动物模型已经提供了肌营养不良蛋白缺乏导致心肌病的机制的见解，但仍然需要开发DMD的人类模型来验证致病机制并确定治疗靶点。在这里，我们已经从CRISPR编辑的人类诱导多能干细胞衍生的心肌细胞（hiPSC-CM）开发了人类工程化心脏组织（EHT），hiPSC-CM表达缺乏部分肌动蛋白结合结构域的截短肌营养不良蛋白。3D EHT平台可以直接测量收缩力，同时监测Ca 2+瞬变，并评估肌原纤维结构。与同基因对照组相比，抗肌萎缩蛋白突变EHTs产生较少的收缩力以及延迟的力产生和松弛动力学。收缩功能障碍伴随着肌节长度减少，静息细胞质Ca 2+水平增加，Ca 2+释放和再摄取延迟，心率不规则性增加。转录组学分析显示肌营养不良蛋白缺陷和对照EHT之间存在明显差异，包括与Ca 2+稳态和细胞外基质组织相关的基因下调，与膜电位调节、心肌发育和心脏收缩相关的基因上调。这些发现表明，EHT平台提供了必要的线索，以揭示力产生的临床相关功能表型，以及对营养不良心脏功能中Ca 2+处理和转录组失调作用的机制见解，最终为疾病建模和药物发现的进一步研究提供了强大的平台。
Cardiomyopathy is currently the leading cause of death for patients with Duchenne muscular dystrophy (DMD), a severe neuromuscular disorder affecting young boys. Animal models have provided insight into the mechanisms by which dystrophin protein deficiency causes cardiomyopathy, but there remains a need to develop human models of DMD to validate pathogenic mechanisms and identify therapeutic targets. Here, we have developed human engineered heart tissues (EHTs) from CRISPR-edited, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) expressing a truncated dystrophin protein lacking part of the actin-binding domain. The 3D EHT platform enables direct measurement of contractile force, simultaneous monitoring of Ca2+ transients, and assessment of myofibril structure. Dystrophin-mutant EHTs produced less contractile force as well as delayed kinetics of force generation and relaxation, as compared to isogenic controls. Contractile dysfunction was accompanied by reduced sarcomere length, increased resting cytosolic Ca2+ levels, delayed Ca2+ release and reuptake, and increased beat rate irregularity. Transcriptomic analysis revealed clear differences between dystrophin-deficient and control EHTs, including downregulation of genes related to Ca2+ homeostasis and extracellular matrix organization, and upregulation of genes related to regulation of membrane potential, cardiac muscle development, and heart contraction. These findings indicate that the EHT platform provides the cues necessary to expose the clinically-relevant, functional phenotype of force production as well as mechanistic insights into the role of Ca2+ handling and transcriptomic dysregulation in dystrophic cardiac function, ultimately providing a powerful platform for further studies in disease modeling and drug discovery.
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DOI：
10.1073/pnas.93.17.9142
发表时间：
1996-08-20
影响因子：
11.1
作者：
Chang, WJ;Iannaccone, ST;Stull, JT
通讯作者：
Stull, JT
影响因子：
2.8
作者：
Eagle, M;Baudouin, SV;Bushby, K
通讯作者：
Bushby, K
DOI：
10.1073/pnas.0908540107
发表时间：
2010-01-26
影响因子：
11.1
作者：
Fauconnier, Jeremy;Thireau, Jerome;Lacampagne, Alain
通讯作者：
Lacampagne, Alain
影响因子：
5
作者：
Leonard A;Bertero A;Powers JD;Beussman KM;Bhandari S;Regnier M;Murry CE;Sniadecki NJ
通讯作者：
Sniadecki NJ
影响因子：
4.6
作者：
Grancharova T;Gerbin KA;Rosenberg AB;Roco CM;Arakaki JE;DeLizo CM;Dinh SQ;Donovan-Maiye RM;Hirano M;Nelson AM;Tang J;Theriot JA;Yan C;Menon V;Palecek SP;Seelig G;Gunawardane RN
通讯作者：
Gunawardane RN