Cardiomyocyte-specific adenylyl cyclase type-8 overexpression induces cell-autonomous activation of RelA and non-cell-autonomous myocardial and systemic inflammation

Using a model of cardiac AC8 overexpression that lead to accelerated cardiac aging, fibrosis and immune system overactivation, this study used scRNA-seq to investigate the signaling pathways dysregulated in cardiac cells to have a better understanding of the biological processes invoved in cardiac aging. A single cell suspension was made from fresh cardiac tissue obtained from AC8 overexpressing mice and littermate controls, An equal number of FACS-sorted CD45+CD31- (leukocytes), CD45-CD31- (resident mesenchymal cells), and CD45-CD31+ (endothelial cells) cells were mixed and processed to generate single-cell cDNA libraries using the 10X Genomics platform Cells were sequenced at a target read depth of 75,000 reads/cell using a NovaSeq 6000 sequencer (Illumina). Single-cell counts were generated from unaligned reads by sequence alignment and de-multiplexing using Cell Ranger from 10x Genomics.