Contractile Defect Caused by Mutation in MYBPC3 Revealed under Conditions Optimized for Human PSC-Cardiomyocyte Function.
Contractile Defect Caused by Mutation in MYBPC3 Revealed under Conditions Optimized for Human PSC-Cardiomyocyte Function.
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DOI：
10.1016/j.celrep.2015.09.025
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发表时间：
2015-10-27
期刊：
影响因子：
8.8
通讯作者：
Mummery CL
中科院分区：
文献类型：
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作者：
Birket MJ;Ribeiro MC;Kosmidis G;Ward D;Leitoguinho AR;van de Pol V;Dambrot C;Devalla HD;Davis RP;Mastroberardino PG;Atsma DE;Passier R;Mummery CL
最大限度地提高人多能干细胞来源的心肌细胞 (hPSC-CM) 的基线功能对于其在心脏毒性和疾病模型中的有效应用至关重要。在这里，我们的目的是确定能够促进足够的功能水平的因素，以便在肥厚型心肌病 (HCM) 的人类诱导多能干细胞 (hiPSC) 模型中进行稳健的单细胞收缩性测量。一个简单的筛选揭示了甲状腺激素、IGF-1 和糖皮质激素类似物地塞米松对 hPSC-CM 的电生理学、生物能学和收缩力产生的协同作用。在这种优化条件下，MYBPC3（一种编码肌球蛋白结合蛋白 C 的基因，突变时会导致 HCM）发生突变的 hiPSC-CM 表现出比对照组显着更低的收缩力产生。通过在对照 hPSC-CM 中直接敲低 MYBPC3 来重现这一点，支持单倍体不足的机制。使用人体细胞对这种疾病进行体外建模是确定 HCM 治疗干预措施的重要一步。 T3+IGF-1+ 地塞米松改善 hPSC 心肌细胞的电生理学这些因素协同增强生物能和收缩力的产生具有 HCM 引起突变的心肌细胞有收缩缺陷 Birket 等人。确定协同改善人类多能干细胞 (hiPSC) 来源的心肌细胞功能的因素组合。优化系统有助于识别肥厚型心肌病 (HCM) 模型中的收缩力缺陷，这种疾病影响约 1:500 的人口。
Maximizing baseline function of human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) is essential for their effective application in models of cardiac toxicity and disease. Here, we aimed to identify factors that would promote an adequate level of function to permit robust single-cell contractility measurements in a human induced pluripotent stem cell (hiPSC) model of hypertrophic cardiomyopathy (HCM). A simple screen revealed the collaborative effects of thyroid hormone, IGF-1 and the glucocorticoid analog dexamethasone on the electrophysiology, bioenergetics, and contractile force generation of hPSC-CMs. In this optimized condition, hiPSC-CMs with mutations in MYBPC3, a gene encoding myosin-binding protein C, which, when mutated, causes HCM, showed significantly lower contractile force generation than controls. This was recapitulated by direct knockdown of MYBPC3 in control hPSC-CMs, supporting a mechanism of haploinsufficiency. Modeling this disease in vitro using human cells is an important step toward identifying therapeutic interventions for HCM. T3+IGF-1+ dexamethasone improves the electrophysiology of hPSC cardiomyocytes These factors synergistically enhance bioenergetics and contractile force generation Cardiomyocytes with HCM-causing mutations have a contractile defect Birket et al. identify a combination of factors that cooperatively improve the function of human pluripotent stem cell (hiPSC)-derived cardiomyocytes. Optimizing the system facilitated the identification of a contraction force defect in a model of hypertrophic cardiomyopathy (HCM), a disease affecting ∼1:500 of the population.