Proteasome-Dependent Regulation of Distinct Metabolic States During Long-Term Culture of Human iPSC-Derived Cardiomyocytes
Proteasome-Dependent Regulation of Distinct Metabolic States During Long-Term Culture of Human iPSC-Derived Cardiomyocytes
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DOI：
10.1161/circresaha.118.313973
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发表时间：
2019-06-21
影响因子：
20.1
通讯作者：
Wu, Joseph C.
中科院分区：
文献类型：
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作者：
Ebert, Antje;Joshi, Amit U.;Wu, Joseph C.
理由：目前，人类诱导多能干细胞衍生心肌细胞（iPSC-CMs）的不成熟表现对其在疾病建模、药物筛选和再生医学中的应用构成了挑战。长期培养可以实现iPSC-CMs的部分成熟。然而，在iPSC-CMs的长期培养过程中，对控制功能变化、代谢输出和细胞稳态的分子信号通路知之甚少。目的：通过iPSC-CMs的长期培养，我们旨在识别和表征在发育过程中控制心脏细胞功能和代谢转变的关键信号事件。方法和结果：我们将转录组测序与途径网络定位结合在iPSC-CMs中，这些iPSC-CMs培养到晚时间点（200天），与中时间点（90天）和早时间点（30天）进行比较。长期培养的iPSC-CMs的转录组学景观允许绘制成熟iPSC-CMs发育过程中不同的代谢阶段。线粒体代谢的时间差异控制被发现由cAMP/PKA（蛋白激酶A）和蛋白酶体依赖的信号事件调节。PKA/蛋白酶体依赖的信号级联由Hsp90（热休克蛋白90）介导下游，Hsp90反过来调节线粒体呼吸链蛋白及其代谢输出。在长期培养过程中，该回路被发现启动iPSC-CM代谢的上调，导致细胞收缩性增加，在第200天的时间点达到最大值。结论：我们的研究结果揭示了PKA/蛋白酶体和hsp90依赖性信号通路调节线粒体呼吸链蛋白并决定心肌细胞能量产生和功能输出。这些发现为iPSC-CMs在发育过程中控制代谢稳态的信号通路提供了更深入的见解。
Rationale: The immature presentation of human induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) is currently a challenge for their application in disease modeling, drug screening, and regenerative medicine. Long-term culture is known to achieve partial maturation of iPSC-CMs. However, little is known about the molecular signaling circuitries that govern functional changes, metabolic output, and cellular homeostasis during long-term culture of iPSC-CMs. Objective: We aimed to identify and characterize critical signaling events that control functional and metabolic transitions of cardiac cells during developmental progression, as recapitulated by long-term culture of iPSC-CMs. Methods and Results: We combined transcriptomic sequencing with pathway network mapping in iPSC-CMs that were cultured until a late time point, day 200, in comparison to a medium time point, day 90, and an early time point, day 30. Transcriptomic landscapes of long-term cultured iPSC-CMs allowed mapping of distinct metabolic stages during development of maturing iPSC-CMs. Temporally divergent control of mitochondrial metabolism was found to be regulated by cAMP/PKA (protein kinase A)- and proteasome-dependent signaling events. The PKA/proteasome-dependent signaling cascade was mediated downstream by Hsp90 (heat shock protein 90), which in turn modulated mitochondrial respiratory chain proteins and their metabolic output. During long-term culture, this circuitry was found to initiate upregulation of iPSC-CM metabolism, resulting in increased cell contractility that reached a maximum at the day 200 time point. Conclusions: Our results reveal a PKA/proteasome- and Hsp90-dependent signaling pathway that regulates mitochondrial respiratory chain proteins and determines cardiomyocyte energy production and functional output. These findings provide deeper insight into signaling circuitries governing metabolic homeostasis in iPSC-CMs during developmental progression.