Knock Down of Plakophillin 2 Dysregulates Adhesion Pathway through Upregulation of miR200b and Alters the Mechanical Properties in Cardiac Cells

Puzzi, Luca; Borin, Daniele; Gurha, Priyatansh; Lombardi, Raffaella; Martinelli, Valentina; Weiss, Marek; Andolfi, Laura; Lazzarino, Marco; Mestroni, Luisa; Marian, Ali J.; Sbaizero, Orfeo

doi:10.3390/cells8121639

Cited by 20 publications

(16 citation statements)

References 43 publications

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“…The graphs show that the median value of the Young's modulus measured in HL-1 PKP2 cells was higher than that measured in HL-1 R735X cells ( Figure 1B). Puzzi et al using AFM in HL-1 cells showed that PKP2 knock-down was also associated with decreased cellular stiffness, as indicated by decreased Young's modulus, when comparing to control groups (Puzzi et al, 2019).These results support the observation that R735X cells lack most stress and F-actin fibers ( Figure 1C). Images from the top central area of cells revealed a scarcity of F-actin filaments in the area above the nucleus of cells expressing the R735X mutant (46.05%±0.73% in PKP2 cells vs. 26.05%±1.48% in R735X cells; p<0.0001, ****; n=11) ( Figure 1C).…”

Section: Mutant R735x Expression Alters Actin Cytoskeleton and Cellulsupporting

confidence: 78%

“…It is known that PKP2 loss impairs cortical actin remodeling and normal desmosome assembly (Godsel et al, 2010), which compromises cell mechanical properties like cellular stiffness (Puzzi et al, 2019). We measured cell stiffness in intact HL-1 cardiac cells by acquiring force-volume maps by atomic force microscopy (AFM).…”

Section: Mutant R735x Expression Alters Actin Cytoskeleton and Cellulmentioning

confidence: 99%

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SARS-CoV-2 protein Nsp1 alters actomyosin cytoskeleton and phenocopies arrhythmogenic cardiomyopathy-related PKP2 mutant

Márquez-López

Roche-Molina

Garcı́a-Quintáns

et al. 2020

Preprint

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Mutations in desmosomal Plakophilin-2 (PKP2) are the most prevalent drivers of arrhythmogenic cardiomyopathy (ACM) and a common cause of sudden cardiac death in young athletes. However, partner proteins that elucidate PKP2 cellular mechanism to understand cardiac dysfunction in ACM are mostly unknown. Here we identify the actin-based motor proteins Myh9 and Myh10 as key PKP2 interactors, and demonstrate that the expression of the ACM-related PKP2 mutant R735X alters actin fiber organization and cell mechanical stiffness. We also show that SARS-CoV-2 Nsp1 protein acts similarly to this known pathogenic R735X mutant, altering the actomyosin component distribution on cardiac cells. Our data reveal that the viral Nsp1 hijacks PKP2 into the cytoplasm and mimics the effect of delocalized R735X mutant. These results demonstrate that cytoplasmic PKP2, wildtype or mutant, induces the collapse of the actomyosin network, since shRNA-PKP2 knockdown maintains the cell structure, validating a critical role of PKP2 localization in the regulation of actomyosin architecture. The fact that Nsp1 and PKP2 mutant R735X share similar phenotypes also suggests that direct SARS-CoV-2 heart infection could induce a transient ACM-like disease in COVID-19 patients, which may contribute to right ventricle dysfunction, observed in patients with poor survival prognosis.HighlightsThe specific cardiac isoform Plakophilin-2a (PKP2) interacts with Myh9 and Myh10.PKP2 delocalization alters actomyosin cytoskeleton component organization. SARS-CoV-2 Nsp1 protein hijacks PKP2 from the desmosome into the soluble fraction where it is downregulated.Viral Nsp1 collapses the actomyosin cytoskeleton and phenocopies the arrhythmogenic cardiomyopathy-related mutant R735X.

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Section: Mutant R735x Expression Alters Actin Cytoskeleton and Cellulsupporting

confidence: 78%

Section: Mutant R735x Expression Alters Actin Cytoskeleton and Cellulmentioning

confidence: 99%

SARS-CoV-2 protein Nsp1 alters actomyosin cytoskeleton and phenocopies arrhythmogenic cardiomyopathy-related PKP2 mutant

Márquez-López

Roche-Molina

Garcı́a-Quintáns

et al. 2020

Preprint

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“…The molecular basis of ECM organization and remodeling in ACM is still under-investigated. Recently few papers identified a signature of ACM cardiac cell microRNAs, known to be involved in ECM turnover and mechanosensing (Rainer et al, 2018;Puzzi et al, 2019).…”

Section: Cardiac Extracellular Matrix Regulationmentioning

confidence: 99%

Fibrosis in Arrhythmogenic Cardiomyopathy: The Phantom Thread in the Fibro-Adipose Tissue

et al. 2020

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Arrhythmogenic cardiomyopathy (ACM) is an inherited heart disorder, predisposing to malignant ventricular arrhythmias leading to sudden cardiac death, particularly in young and athletic patients. Pathological features include a progressive loss of myocardium with fibrous or fibro-fatty substitution. During the last few decades, different clinical aspects of ACM have been well investigated but still little is known about the molecular mechanisms that underlie ACM pathogenesis, leading to these phenotypes. In about 50% of ACM patients, a genetic mutation, predominantly in genes that encode for desmosomal proteins, has been identified. However, the mutation-associated mechanisms, causing the observed cardiac phenotype are not always clear. Until now, the attention has been principally focused on the study of molecular mechanisms that lead to a prominent myocardium adipose substitution, an uncommon marker for a cardiac disease, thus often recognized as hallmark of ACM. Nonetheless, based on Task Force Criteria for the diagnosis of ACM, cardiomyocytes death associated with fibrous replacement of the ventricular free wall must be considered the main tissue feature in ACM patients. For this reason, it urges to investigate ACM cardiac fibrosis. In this review, we give an overview on the cellular effectors, possible triggers, and molecular mechanisms that could be responsible for the ventricular fibrotic remodeling in ACM patients.

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“…Results of in vitro and in vivo studies indicate that abnormalities in biomechanical properties of the desmosome, as well as of the cross-talk between the desmosomes and other cellular structures such as the cytoskeleton, nucleus, gap junctions, and ion channels, contribute to the pathogenesis of ACM [ 1 , 17 , 18 , 19 , 41 , 81 , 82 , 83 , 84 ].…”

Section: Molecular Pathogenic Pathwaysmentioning

confidence: 99%

Established and Emerging Mechanisms in the Pathogenesis of Arrhythmogenic Cardiomyopathy: A Multifaceted Disease

Gao

Puthenvedu

Lombardi

et al. 2020

IJMS

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Arrhythmogenic cardiomyopathy (ACM) is a heritable myocardial disease that manifests with cardiac arrhythmias, syncope, sudden cardiac death, and heart failure in the advanced stages. The pathological hallmark of ACM is a gradual replacement of the myocardium by fibroadiposis, which typically starts from the epicardium. Molecular genetic studies have identified causal mutations predominantly in genes encoding for desmosomal proteins; however, non-desmosomal causal mutations have also been described, including genes coding for nuclear proteins, cytoskeleton componentsand proteins involved in excitation-contraction coupling. Despite the poor prognosis, currently available treatments can only partially control symptoms and to date there is no effective therapy for ACM. Inhibition of the canonical Wnt/β-catenin pathway and activation of the Hippo and the TGF-β pathways have been implicated in the pathogenesis of ACM. Yet, our understanding of the molecular mechanisms involved in the development of the disease and the cell source of fibroadiposis remains incomplete. Elucidation of the pathogenesis of the disease could facilitate targeted approaches for treatment. In this manuscript we will provide a comprehensive review of the proposed molecular and cellular mechanisms of the pathogenesis of ACM, including the emerging evidence on abnormal calcium homeostasis and inflammatory/autoimmune response. Moreover, we will propose novel hypothesis about the role of epicardial cells and paracrine factors in the development of the phenotype. Finally, we will discuss potential innovative therapeutic approaches based on the growing knowledge in the field.

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Knock Down of Plakophillin 2 Dysregulates Adhesion Pathway through Upregulation of miR200b and Alters the Mechanical Properties in Cardiac Cells

Cited by 20 publications

References 43 publications

SARS-CoV-2 protein Nsp1 alters actomyosin cytoskeleton and phenocopies arrhythmogenic cardiomyopathy-related PKP2 mutant

SARS-CoV-2 protein Nsp1 alters actomyosin cytoskeleton and phenocopies arrhythmogenic cardiomyopathy-related PKP2 mutant

Fibrosis in Arrhythmogenic Cardiomyopathy: The Phantom Thread in the Fibro-Adipose Tissue

Established and Emerging Mechanisms in the Pathogenesis of Arrhythmogenic Cardiomyopathy: A Multifaceted Disease

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