TY - JOUR
T1 - Unfolded Protein Response as a Compensatory Mechanism and Potential Therapeutic Target in PLN R14del Cardiomyopathy
AU - Feyen, Dries A M
AU - Perea-Gil, Isaac
AU - Maas, Renee G C
AU - Harakalova, Magdalena
AU - Gavidia, Alexandra A
AU - Arthur Ataam, Jennifer
AU - Wu, Ting-Hsuan
AU - Vink, Aryan
AU - Pei, Jiayi
AU - Vadgama, Nirmal
AU - Suurmeijer, Albert J
AU - Te Rijdt, Wouter P
AU - Vu, Michelle
AU - Amatya, Prashila L
AU - Prado, Maricela
AU - Zhang, Yuan
AU - Dunkenberger, Logan
AU - Sluijter, Joost P G
AU - Sallam, Karim
AU - Asselbergs, Folkert W
AU - Mercola, Mark
AU - Karakikes, Ioannis
PY - 2021/8/3
Y1 - 2021/8/3
N2 - BACKGROUND: Phospholamban (PLN) is a critical regulator of calcium cycling and contractility in the heart. The loss of arginine at position 14 in PLN (R14del) is associated with dilated cardiomyopathy with a high prevalence of ventricular arrhythmias. How the R14 deletion causes dilated cardiomyopathy is poorly understood, and there are no disease-specific therapies.METHODS: We used single-cell RNA sequencing to uncover PLN R14del disease mechanisms in human induced pluripotent stem cells (hiPSC-CMs). We used both 2-dimensional and 3-dimensional functional contractility assays to evaluate the impact of modulating disease-relevant pathways in PLN R14del hiPSC-CMs.RESULTS: Modeling of the PLN R14del cardiomyopathy with isogenic pairs of hiPSC-CMs recapitulated the contractile deficit associated with the disease in vitro. Single-cell RNA sequencing revealed the induction of the unfolded protein response (UPR) pathway in PLN R14del compared with isogenic control hiPSC-CMs. The activation of UPR was also evident in the hearts from PLN R14del patients. Silencing of each of the 3 main UPR signaling branches (IRE1, ATF6, or PERK) by siRNA exacerbated the contractile dysfunction of PLN R14del hiPSC-CMs. We explored the therapeutic potential of activating the UPR with a small molecule activator, BiP (binding immunoglobulin protein) inducer X. PLN R14del hiPSC-CMs treated with BiP protein inducer X showed a dose-dependent amelioration of the contractility deficit in both 2-dimensional cultures and 3-dimensional engineered heart tissues without affecting calcium homeostasis.CONCLUSIONS: Together, these findings suggest that the UPR exerts a protective effect in the setting of PLN R14del cardiomyopathy and that modulation of the UPR might be exploited therapeutically.
AB - BACKGROUND: Phospholamban (PLN) is a critical regulator of calcium cycling and contractility in the heart. The loss of arginine at position 14 in PLN (R14del) is associated with dilated cardiomyopathy with a high prevalence of ventricular arrhythmias. How the R14 deletion causes dilated cardiomyopathy is poorly understood, and there are no disease-specific therapies.METHODS: We used single-cell RNA sequencing to uncover PLN R14del disease mechanisms in human induced pluripotent stem cells (hiPSC-CMs). We used both 2-dimensional and 3-dimensional functional contractility assays to evaluate the impact of modulating disease-relevant pathways in PLN R14del hiPSC-CMs.RESULTS: Modeling of the PLN R14del cardiomyopathy with isogenic pairs of hiPSC-CMs recapitulated the contractile deficit associated with the disease in vitro. Single-cell RNA sequencing revealed the induction of the unfolded protein response (UPR) pathway in PLN R14del compared with isogenic control hiPSC-CMs. The activation of UPR was also evident in the hearts from PLN R14del patients. Silencing of each of the 3 main UPR signaling branches (IRE1, ATF6, or PERK) by siRNA exacerbated the contractile dysfunction of PLN R14del hiPSC-CMs. We explored the therapeutic potential of activating the UPR with a small molecule activator, BiP (binding immunoglobulin protein) inducer X. PLN R14del hiPSC-CMs treated with BiP protein inducer X showed a dose-dependent amelioration of the contractility deficit in both 2-dimensional cultures and 3-dimensional engineered heart tissues without affecting calcium homeostasis.CONCLUSIONS: Together, these findings suggest that the UPR exerts a protective effect in the setting of PLN R14del cardiomyopathy and that modulation of the UPR might be exploited therapeutically.
KW - Adaptation, Physiological
KW - Biomarkers
KW - Calcium-Binding Proteins/genetics
KW - Cardiomyopathies/diagnosis
KW - Cardiomyopathy, Dilated/genetics
KW - Culture Media, Conditioned/metabolism
KW - Disease Management
KW - Disease Susceptibility
KW - Gene Expression Profiling
KW - Genetic Predisposition to Disease
KW - Humans
KW - Induced Pluripotent Stem Cells/metabolism
KW - Molecular Targeted Therapy
KW - Myocardial Contraction/drug effects
KW - Sequence Deletion
KW - Single-Cell Analysis
KW - Transcriptome
KW - Unfolded Protein Response
U2 - https://doi.org/10.1161/CIRCULATIONAHA.120.049844
DO - https://doi.org/10.1161/CIRCULATIONAHA.120.049844
M3 - Article
C2 - 33928785
SN - 0009-7322
VL - 144
SP - 382
EP - 392
JO - Circulation
JF - Circulation
IS - 5
ER -