Generation of left ventricle-like cardiomyocytes with improved structural, functional, and metabolic maturity from human pluripotent stem cells.
Generation of left ventricle-like cardiomyocytes with improved structural, functional, and metabolic maturity from human pluripotent stem cells.
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DOI：
10.1016/j.crmeth.2023.100456
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发表时间：
2023-04-24
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由基因突变或损伤引起的左心室（LV）功能下降通常会导致衰弱和致命的心血管疾病。因此，LV心肌细胞是潜在的有价值的治疗靶点。人多能干细胞衍生的心肌细胞（hPSC-CM）既不是同质的，也不是功能成熟的，这降低了它们的效用。在这里，我们利用心脏发育知识来指导hPSC特异性地向LV心肌细胞分化。正确的中胚层模式化和视黄酸途径阻断对于产生接近同质的LV特异性hPSC-CM（hPSC-LV-CM）是必不可少的。这些细胞通过第一心野祖细胞转运并显示典型的心室动作电位。重要的是，与使用标准WNT-ON/WNT-OFF方案产生的年龄匹配的心肌细胞相比，hPSC-LV-CM表现出代谢增加，增殖减少，细胞结构和功能成熟度改善。类似地，由hPSC-LV-CM制成的工程心脏组织更好地组织，产生更高的力，并且跳动更慢，但可以起搏到生理水平。总之，我们表明，可以快速获得功能成熟的hPSC-LV-CM，而无需暴露于当前的成熟机制。左心室（LV）中胚层前体保留心房和右心室电位阻断视黄酸信号传导可防止心房逃逸LV心肌细胞迅速达到功能、结构和代谢成熟机械负荷诱导的成熟是累加性的而不是变革性的心肌梗死（MI）、室性心律失常、或药物诱导的心脏毒性是最常见的和使人衰弱的心血管疾病，经常导致器官功能障碍和衰竭。因此，这对于LV心血管疾病的建模和开发治疗它们的新疗法是非常理想的。在这里，我们开发了一种方法，以区分快速和有效的近同质群体的LV心肌细胞从人类多能干细胞（hPSC）。这些心肌细胞也比用标准WNT-ON/WNT-OFF方案产生的心肌细胞更成熟，因此使它们成为有吸引力的模型。Dark等人报道了可以从人多能干细胞快速衍生出接近同质的人左心室心肌细胞。与其他年龄匹配的心肌细胞培养物相比，这些细胞表现出增强的成熟度。这些细胞的同质性和成熟性使它们成为左心室发育和疾病的合适模型，并且应该能够进行更忠实的心脏毒性筛选。
Decreased left ventricle (LV) function caused by genetic mutations or injury often leads to debilitating and fatal cardiovascular disease. LV cardiomyocytes are, therefore, a potentially valuable therapeutical target. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are neither homogeneous nor functionally mature, which reduces their utility. Here, we exploit cardiac development knowledge to instruct differentiation of hPSCs specifically toward LV cardiomyocytes. Correct mesoderm patterning and retinoic acid pathway blocking are essential to generate near-homogenous LV-specific hPSC-CMs (hPSC-LV-CMs). These cells transit via first heart field progenitors and display typical ventricular action potentials. Importantly, hPSC-LV-CMs exhibit increased metabolism, reduced proliferation, and improved cytoarchitecture and functional maturity compared with age-matched cardiomyocytes generated using the standard WNT-ON/WNT-OFF protocol. Similarly, engineered heart tissues made from hPSC-LV-CMs are better organized, produce higher force, and beat more slowly but can be paced to physiological levels. Together, we show that functionally matured hPSC-LV-CMs can be obtained rapidly without exposure to current maturation regimes. Left ventricle (LV) mesoderm precursors retain atrial and right ventricle potential Blocking retinoic acid signaling prevents atrial escapee emergence LV cardiomyocytes quickly attain functional, structural, and metabolic maturity Mechanical load-induced maturity is additive rather than transformative Impaired LV-cardiomyocyte function caused by myocardial infarction (MI), ventricular arrhythmias, or drug-induced cardiotoxicity are among the most common and debilitating cardiovascular diseases, often leading to organ dysfunction and failure.In vitro models of LV cardiomyocytes are, therefore, highly desirable for modeling LV cardiovascular diseases and for developing novel therapies to treat them. Here, we developed a method to differentiate rapidly and efficiently near-homogeneous populations of LV cardiomyocytes from human pluripotent stem cells (hPSCs). These cardiomyocytes are also more mature than those generated with the standard WNT-ON/WNT-OFF protocol, thus making them an attractive model. Dark et al. report that near-homogeneous human left ventricle cardiomyocytes can be rapidly derived from human pluripotent stem cells. These cells exhibit enhanced maturity compared with other age-matched cardiomyocyte cultures. The homogeneity and maturity of these cells renders them a suitable model of left ventricle development and disease and should enable more faithful cardiotoxicity screens.