Antje Ebert1,2,3, Amit U Joshi4, Sandra Andorf5,6, Yuanyuan Dai1,2,3, Shrivatsan Sampathkumar1,2,3, Haodong Chen1,7, Yingxin Li1,7, Priyanka Garg1,7, Karl Toischer2,3, Gerd Hasenfuss2,3, Daria Mochly-Rosen4, Joseph C Wu1,7. 1. From the Stanford Cardiovascular Institute (A.E., Y.D., S.S., H.C., Y.L., P.G., J.C.W.), Stanford University School of Medicine, CA. 2. Department of Cardiology and Pneumology, Heart Center, University of Göttingen, Germany (A.E., Y.D., S.S., K.T., G.H.). 3. German Center for Cardiovascular Research, Partner Site, Göttingen, Germany (A.E., Y.D., S.S., K.T., G.H.). 4. Department of Chemical and Systems Biology (A.U.J., D.M.-R.), Stanford University School of Medicine, CA. 5. Division of Pulmonary and Critical Care Medicine, Department of Medicine (S.A.), Stanford University School of Medicine, CA. 6. Sean N. Parker Center for Allergy and Asthma Research (S.A.), Stanford University School of Medicine, CA. 7. Division of Cardiology, Department of Medicine (H.C., Y.L., P.G., J.C.W.), Stanford University School of Medicine, CA.
Abstract
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.
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.
Entities:
Keywords:
homeostasis; induced pluripotent stem cells; mitochondria; proteasome; regenerative medicine
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