Dataset Information

Arrythmogenic right ventricular acrdiomayopathy (PKP2-cko in mice and Human biopsies)

ABSTRACT: Summary: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inherited cardiac disorder. It is classified as the second most common cause of unexpected sudden death by cardiac arrest in the young. ARVC has devastating psychosocial consequences, especially as many patients are young adults, and current therapy is limited to extreme exercise restriction and defibrillator implantation. More than 40% of the reported genetic variants linked to ARVC reside in a gene called PKP2 , which encodes for the plakophilin-2 (PKP2) protein. The pathogenic mechanisms linking this protein to ARVC remain to be elucidated, which is the focus of this project. I aim to identify exactly which proteins in the heart are dysregulated and to identify why exercise is detrimental for carriers of PKP2-deficient hearts. Our approach is based on state-of-the-art mass spectrometry (MS) technologies that allow us to measure all proteins in the heart simultaneously. We used both human heart biopsy material as well as a murine disease model I anticipate to elucidate novel roles of PKP2 that are of utmost importance for understanding ARVC pathogenesis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Heart

DISEASE(S): Arrhythmogenic Right Ventricular Cardiomyopathy

SUBMITTER: Navratan Bagwan

LAB HEAD: Alicia Lundby

PROVIDER: PXD025523 | Pride | 2022-11-04

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	20200501_EXPL1_nLC14_LD_QC_MNT_ARCV_fr01.raw	Raw
	20200501_EXPL1_nLC14_LD_QC_MNT_ARCV_fr02.raw	Raw
	20200501_EXPL1_nLC14_LD_QC_MNT_ARCV_fr03.raw	Raw
	20200501_EXPL1_nLC14_LD_QC_MNT_ARCV_fr04.raw	Raw
	20200501_EXPL1_nLC14_LD_QC_MNT_ARCV_fr05.raw	Raw

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Publications

Loss of Nuclear Envelope Integrity and Increased Oxidant Production Cause DNA Damage in Adult Hearts Deficient in PKP2: A Molecular Substrate of ARVC.

Pérez-Hernández Marta M van Opbergen Chantal J M CJM Bagwan Navratan N Vissing Christoffer Rasmus CR Marrón-Liñares Grecia M GM Zhang Mingliang M Torres Vega Estefania E Sorrentino Andrea A Drici Lylia L Sulek Karolina K Zhai Ruxu R Hansen Finn B FB Christensen Alex H AH Boesgaard Søren S Gustafsson Finn F Rossing Kasper K Small Eric M EM Davies Michael J MJ Rothenberg Eli E Sato Priscila Y PY Cerrone Marina M Jensen Thomas Hartvig Lindkær THL Qvortrup Klaus K Bundgaard Henning H Delmar Mario M Lundby Alicia A

Circulation 20220812 11

<h4>Background</h4>Arrhythmogenic right ventricular cardiomyopathy (ARVC) is characterized by high propensity to life-threatening arrhythmias and progressive loss of heart muscle. More than 40% of reported genetic variants linked to ARVC reside in the <i>PKP2</i> gene, which encodes the PKP2 protein (plakophilin-2).<h4>Methods</h4>We describe a comprehensive characterization of the ARVC molecular landscape as determined by high-resolution mass spectrometry, RNA sequencing, and transmission elect ...[more]

PMID: 35959657

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Project description:Introduction: Arrhythmogenic Cardiomyopathy (AC) is an inherited disease that is caused by desmosome protein mutation with plakophilin 2 (PKP2) mutation being most common. AC is known to cause sporadic ventricular arrhythmias, myocyte damage, and subepicardial fibrosis. Given the origin of fibrosis from epicardium, we wanted to understand how epicardium-derived cells (EDCs) contribute to the AC pathogenesis. Hypothesis: EDCs propagate the pro-inflammatory signaling that contributes to AC pathogenesis. Methods: We developed transgenic mice that lack PKP2 in cardiomyocytes (PKP2-cKO) or in both cardiomyocyte and epicardial cells (PKP2-ceKO) via the tissue-specific expression of tamoxifen-inducible Cre recombinase. Tamoxifen injected mice expressing Cre in cardiomyocytes, but with no floxed PKP2 gene, was used as a control. EDCs were traced with Cre-dependent Green Fluorescence Protein (GFP). Non-myocyte populations were isolated 21 days post-tamoxifen injection for single cell RNA-sequencing (scRNA-seq) using 10X Genomics platform. Perfused hearts were separated into left and right ventricle for qRT-PCR. Immunohistochemistry was used for cardiac structure and cellular composition. Echocardiography was performed to measure cardiac physiology. Results: The scRNA-seq analysis identified an epicardium-derived fibroblast population that secretes pro-inflammatory cytokines, including CCL2, CCL7, Thbs1, and PTX3. While pro-inflammatory EDCs are found in both PKP2-cKO and PKP2-ceKO mice, they were most numerous in PKP2-ceKO mice. Importantly, macrophages and B cells accumulated in both PKP2-cKO and PKP2-ceKO mice compared to controls. qRT-PCR confirmed that the expression of genes encoding fibrosis markers (POSTN, Col1A1, Col1A2, and Col3A1), pro-inflammatory fibroblast markers (CCL2, CCL7, Thbs1, PTX3), macrophage markers (F4/80, Cx3cr1, Spp1, Trem2, and CCR2), and B cell markers (CD19 and CD79b) were significantly enriched in both PKP2-cKO and PKP2-ceKO mice compared to control. Our expression and histologic data also revealed an exaggerated pro-inflammatory response in PKP2-ceKO mice, that progresses from the right to bi-ventricular predominance. However, echocardiography showed no significant difference in cardiac function between PKP2-cKO and PKP2-ceKO, and B cell depletion did not alter disease progression. Conclusion: A subset of epicardium-derived fibroblasts in PKP2 KO mice secretes pro-inflammatory cytokines that contributes to cardiomyocyte damage and cardiac fibrosis in AC.

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