The cytoprotective impact of yes-associated protein 1 after ischemia-reperfusion injury in AC16 human cardiomyocytes

Abstract

The Hippo-signaling pathway is a mechanism implicated in cardiomyocyte cytoprotection and regeneration after a myocardial infarction. Yes-associated protein 1, the main effector protein of this pathway, acts as a co-transcriptional activator to promote cardiomyocyte proliferation and survival. However, the biological mechanisms by which yes-associated protein 1 protects the heart post-MI are currently unknown. Here, we propose that yes-associated protein 1 plays a critical role in cardiomyocyte cytoprotection after simulated ischemia-reperfusion injury. AC16 human cardiomyocytes were infected with lentiviral plasmids containing normal human yes-associated protein 1 and a constitutively active form of YAP, YAP1S127A. Cells were exposed to ischemia-reperfusion injury using a hypoxic chamber. Hippo-signaling characterization after ischemia-reperfusion injury was performed via Western blotting and reverse transcriptase polymerase chain reaction. Cell viability, apoptosis, and cellular hypertrophy were assessed as a measure of cytoprotection. The GSK3β inhibitor CHIR99021 was used to investigate cross-talk between Hippo and Wnt-signaling and their role in cytoprotection after ischemia-reperfusion-injury. Ischemia-reperfusion injury resulted in significant decreased expression of the non-phosphorylated Hippo signaling kinases MST1 and LATS1, along with decreased expression of YAP/TAZ. Overexpression of yes-associated protein 1 improved cellular viability, while reducing hypertrophy and apoptosis via the ATM/ATR DNA damage response pathway. Activation of β-catenin in YAP-infected cardiomyocytes synergistically reduced cellular hypertrophy after ischemia-reperfusion-injury. Our findings indicate that yes-associated protein 1 is cytoprotective in AC16 human cardiomyocytes after ischemia-reperfusion injury, which may be mediated by co-activation of the canonical Wnt/β-catenin pathway. Thus, activation of yes-associated protein 1 may be a novel therapeutic to repair the infarcted myocardium. Impact statement Genetically engineering the cells of the heart after myocardial infarction to display a more regenerative phenotype is a promising therapy for heart failure patients. Here, we support a regenerative role for yes-associated protein 1, the main effector protein of the Hippo signaling pathway, in AC16 human cardiomyocytes as a potential therapeutic gene target for cardiac repair after myocardial infarction.

Molecular Mechanism of Hippo–YAP1/TAZ Pathway in Heart Development, Disease, and Regeneration

“…Activated YAP1 has a protective effect on IR-damaged heart. Lentivirusgenerated YAP1 overexpression in AC16 human CMs decreases CM apoptosis, cell hypertrophy, and generation of reactive oxygen species (ROS), which protects CMs from IR injury (Khan et al, 2019). Interestingly, downregulation of the hypertrophy effect of YAP1 is not consistent with the results of previous studies that showed that YAP1 promotes CM hypertrophy after IR stress (Yang et al, 2015;Zhou et al, 2015).…”

Cardioprotective Effect of Echinatin Against Ischemia/Reperfusion Injury: Involvement of Hippo/Yes-Associated Protein Signaling

“…Decreased expression of MST1, LATS1, and YAP/YAZ was also reported in AC16 cells subjected to H/R. Moreover, upregulation of YAP1 attenuated MI/R damage in AC16 human cardiomyocytes by co-activation of the Wnt/β-catenin pathway ( Khan et al, 2019 ). Additionally, blockage of the Hippo/YAP signaling pathway diminished the protective effect of melatonin on MI/R-induced cardiomyocyte death and mitochondrial damage, indicating the involvement of Hippo/YAP signaling in MI/R injury ( Ma and Dong, 2019 ).…”

Human Cell Modeling for Cardiovascular Diseases

“…Emerging technologies will allow in the future an increasingly precise understanding of the pathophysiology of CVDs and consequent therapeutic advancements. Arrhythmogenic Cardiomyopathy Pathogenic mechanism [8] BMCs Arrhythmogenic Cardiomyopathy Pathogenic mechanism [10] Heart failure - [11] CMs Heart failure Pathogenic mechanism [14] Hypertrophic Cardiomyopathy Drug discovery [16,17] C-MSCs Myocardial ischemia Pathogenic mechanism [24] Arrhythmogenic Cardiomyopathy Pathogenic mechanism/Involvement in disease pathogenesis [25] c-kit + C-MSCs Atrial Fibrillation Pathogenic mechanism [31] FAPs Arrhythmogenic Cardiomyopathy Involvement in disease pathogenesis [35] ECs Hypertension Pathogenic mechanism [39,43] Atherosclerosis Pathogenic mechanism/Involvement in disease pathogenesis [41] Pathogenic mechanism [42] VSMCs Hypertension Involvement in disease pathogenesis [48] Pathogenic mechanism [49] Atherosclerosis Differentiation phenotypes [46] Pathogenic mechanism/Drug discovery [47] AC 16 Hypertrophic Cardiomyopathy Drug discovery [51] Myocardial ischemia Pathogenic mechanism [52] hESC-CMs Hypertrophic Cardiomyopathy Pathogenic mechanism [64,65] hiPSC-CMs Atrial Fibrillation Pathogenic mechanism [76] Hypertrophic Cardiomyopathy Drug discovery [78] Long QT Syndrome Pathogenic mechanism/Drug discovery [79] hiPSC-ECs Hypertension Drug discovery [81] Moyamoya disease Pathogenic mechanism [82] hiPSC-SMCs Supravalvular aortic stenosis syndrome Pathogenic mechanism [83] Bicuspid Aortic Valve-related Thoracic Aortic Aneurysm Pathogenic mechanism/Drug discovery [84] VSMCs and CD14 + cells co-culture Atherosclerosis Pathogenic mechanism [88] [89] ECs and VSMCs co-culture Atherosclerosis Pathogenic mechanism [90]...…”

“…Another work proposed AC 16 CMs as an ischemia/reperfusion (IR) injury model to evaluate the cytoprotective role of yes-associated protein 1 (YAP1), the main effector of Hippo-signaling pathway. After simulated IR injury using a hypoxic chamber, AC 16 cells overexpressing YAP1 showed a reduction in apoptosis, hypertrophy, and generation of ROS, hinting to the potentiality of YAP1 as a therapeutic target after myocardial infarction [ 52 ].…”