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Single cell RNA transcriptome analysis of Epicardial Derived Cells in the Desmoplakin haplo-insufficient Arrhythmogenic Cardiomyopathy mouse

ABSTRACT: Single cell RNA transcriptome analysis of Epicardial Derived Cells in the Desmoplakin haplo-insufficient Arrhythmogenic Cardiomyopathy mouse

PROVIDER: PRJNA634559 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR11832973_1.fastq.gz	Fastqsanger.gz
	SRR11832973_2.fastq.gz	Fastqsanger.gz
	SRR11832974_1.fastq.gz	Fastqsanger.gz
	SRR11832974_2.fastq.gz	Fastqsanger.gz
	SRR11832975_1.fastq.gz	Fastqsanger.gz

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Exercise Reverses Dysregulated Gene Expression in a Desmoplakin Haplo-insufficient Mouse Model of Arrhythmogenic Cardiomyopathy

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Single cell RNA transcriptome analysis of Epicardial Derived Cells in the Desmoplakin haplo-insufficient Arrhythmogenic Cardiomyopathy mouse

Project description:To delineate the role of the epicardium, the initial site of presentation of the ACM phenotype, the Dsp gene, encoding desmosome protein desmoplakin (DSP) was conditionally deleted in the mouse epicardial cells under the transcriptional regulation of Wilms tumor 1 (Wt1) locus. Single cell RNA sequencing (scRNA-Seq) of ~ 40,000 tagged epicardial-derived cells (EDCs), isolated using the dual reporter R26mT/mG mice, showed an increased number of EDCs expressing unique molecular identifiers of fibroblasts as well as a unique subset of epicardial-derived fibroblasts expressing pro-fibrotic genes in the Wt1-Cre: R26mT/mG: Dsp mice. Analysis of pseudo-bulk RNA-Seq data identified ~ 800 differentially expressed genes (DEGs), which were predominantly involved in epithelial-mesenchymal transition (EMT). The DEGs were the targets of TGFB1 among other trophic/mitotic factors, whose activation was verified by immunohistochemistry and quantification of transcript levels of selected TGBF1 pathway genes. Retro-mapping of the DEGs to the cell clusters, identified by scRNA-Seq, denoted epicardial-derived fibroblasts and to a lesser extent the epithelial cells as the main sources of paracrine factors in the myocardium of Wt1-Cre: R26mT/mG: Dsp mice.

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Epicardial Differentiation Drives Fibro-fatty Remodeling in Arrhythmogenic Cardiomyopathy [scRNA-seq]

Project description:We performed single cell RNA sequencing (scRNAseq) on epicardial cells generated from human induced pluripotent stem cells (hiPSCs) of an arrhythmogenic cardiomyopathy patient and their PKP2 reverted isogenic control. The goal of this experiment was to analyze the process of epicardial to fibro-fatty celluar differentiation in diseased cells and compare the cellular transcirptomes of diseased and healthy cells.

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Exercise Reverses Dysregulated Gene Expression in a Desmoplakin Haplo-insufficient Mouse Model of Arrhythmogenic Cardiomyopathy

Project description:Mutations in desmosome genes cause arrhyythmogenic cardiomyopathy (ACM). Using desmoplakin haploinsufficient mouse model of ACM, we evaluated the effects of treadmill exercise on cardiac myocyte transcritional profile. Strand specific, ribosome depleted paired end RNA sequencing of cardiac myocytes from wild type and Myh6-Cre:DspW/F were generate using Illumina HiSeq 4000.

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Epicardial Differentiation Drives Fibro-fatty Remodeling in Arrhythmogenic Cardiomyopathy [bulk RNA-seq]

Project description:We performed bulk cell RNA sequencing (RNAseq) on epicardial cells generated from human induced pluripotent stem cells (hiPSCs) of an arrhythmogenic cardiomyopathy patient and their PKP2 reverted isogenic control. The goal of this experiment was to compare the transcriptomes of diseased and healthy cells.

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Modeling arrhythmogenic cardiomyopathy fattyfibro pathology through the use of PKP2-deficient human iPSC-derived epicardial cells

Project description:Arrhythmogenic cardiomyopathy (ACM) is an inherited cardiac disease characterized by progressive fatty-fibro replacement of the ventricular myocardium leading to arrhythmias and an increased risk of sudden cardiac death. To date, the cell types and signaling mechanisms involved in fatty-fibro infiltration of the myocardium have yet to be fully resolved. However, given that fatty-fibro replacement is initiated within the subepicardial layer, epicardial cells are predicted to contribute to the development of this pathology. To investigate the epicardial contributions to the development of ACM, we generated human induced pluripotent stem cell (hiPSC) lines from ACM patients harboring either a PKP2 1849C>T or a PKP2 2013delC mutation alongside CRISPR/Cas9 gene-edited isogenic control lines and a PKP2 knockout line. Epicardial cells derived from patient and knockout conditions displayed enhanced epithelial-to-mesenchymal transition characteristics, dysregulated Wnt and interferon signaling, increased lipid accumulation, and an enhanced fibrotic phenotype when converted to epicardial-derived fibroblasts. Transcriptomic analysis revealed upregulation of insulin growth factor 2 (IGF2) in ACM hiPSC-EPCs as well as in human ACM heart tissue. Subsequent treatment with exogenous IGF2 enhanced expression of the adipogenic transcription factors CEBPA and PPARG in hiPSC-EPCs, suggesting insulin growth factor signaling contributes to ACM fatty-fibro remodeling.

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