Abstract:
Heart failure contributes to Duchenne muscular dystrophy (DMD), which arises from mutations that ablate dystrophin, rendering the plasma membrane prone to disruption. Cardiomyocyte membrane breakdown in patients with DMD yields a serum injury profile similar to other types of myocardial injury with the release of creatine kinase and troponin isoforms. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are highly useful but can be improved. We generated hiPSC-CMs from a patient with DMD and subjected these cells to equibiaxial mechanical strain to mimic in vivo stress. Compared to healthy cells, DMD hiPSC-CMs demonstrated greater susceptibility to equibiaxial strain after 2 h at 10% strain. We generated an aptamer-based profile of proteins released from hiPSC-CMs both at rest and subjected to strain and identified a strong correlation in the mechanical stress-induced proteome from hiPSC-CMs and serum from patients with DMD. We exposed hiPSC-CMs to recombinant annexin A6, a protein resealing agent, and found reduced biomarker release in DMD and control hiPSC-CMs subjected to strain. Thus, the application of mechanical strain to hiPSC-CMs produces a model that reflects an in vivo injury profile, providing a platform to assess pharmacologic intervention.
摘要:
心力衰竭导致Duchenne型肌营养不良(DMD),这是由消融肌营养不良蛋白的突变引起的,使质膜容易破裂。DMD患者的心肌细胞膜破坏产生的血清损伤谱与其他类型的心肌损伤相似,并释放肌酐激酶和肌钙蛋白亚型。人诱导多能干细胞来源的心肌细胞(hiPSC-CM)是非常有用的,但可以被改进。我们产生了DMDhiPSC-CM,并使这些细胞经受等双轴机械应变以模拟体内应力。与健康细胞相比,DMDhiPSC-CM在10%应变下2小时后表现出对等双轴应变的更大敏感性。我们产生了基于适体的从hiPSC-CM在静止和经受应变时释放的蛋白质的谱,并且鉴定了来自hiPSC-CM和DMD患者血清的机械应力诱导的蛋白质组中的强相关性。我们将hiPSC-CM暴露于重组膜联蛋白A6,一种蛋白质再密封剂,并发现DMD和对照hiPSC-CM中生物标志物的释放减少。因此,机械应变对hiPSC-CM的应用产生了一个反映体内损伤特征的模型,提供评估药物干预的平台。