A single cell RNA-seq analysis of cardiac fibroblasts from pressure-overloaded mouse hearts
Ontology highlight
ABSTRACT: To verify the hypothesis that there are distinct subsets of cardiac fibroblasts, we performed single-cell RNA sequencing analysis of cardiac fibroblasts from pressure-overloaded ventricles.
Project description:We have generated scRNA-seq data from embryonic day (E)10.5 and E12.5 atrioventricular canals (primitive heart valves) to assess cellular diversity during the distinct epithelial-to-mesenchymal transitions (EMTs) from endocardium and epicardium that guide the formation of valve mesenchyme. Alongside, wildtype atrioventricular canals, we generated scRNA-seq data from Sox9 conditional knockouts (Sox9fl/fl;Tie2-cre) to explore the role of SOX9 in EMT. Atrioventricular canals were microdissected from cardiac chambers and outflow tract, enriching for heart valve progenitor lineages.
Project description:Induced pluripotent stem cells (iPSC) derived from fibroblasts of two healthy individuals were differentiated into NPC. Cells were profiled by scRNA-seq.
Project description:Background. Ageing is one of the main risk factors of cardiovascular disease. Pericytes are capillary-associated mural cells involved in the maintenance and stability of the vascular network. In the heart, the consequences of ageing on cardiac pericytes are unknown. Methods. In this study, we have combined single nucleus RNA sequencing and histological analysis to determine the effects of ageing on cardiac pericytes. Furthermore, we have conducted in vivo and in vitro analysis of RGS5 loss of function and finally have perfomed pericytes-fibroblasts co-culture studies to understand the effect of RGS5 loss of function in pericytes on the neighbouring fibroblasts. Results. We showed that ageing reduces the pericyte area and coverage. Single nucleus RNA sequencing analysis further revealed that the expression of the Regulator of G protein signalling 5 (Rgs5) is reduced in old cardiac pericytes. In vivo and in vitro studies showed that the deletion of RGS5 induces morphological changes and a pro-fibrotic gene expression signature characterized by the expression of different extracellular matrix components and growth factors like TGFB2 and PDGFB in pericytes. Indeed, the culture of fibroblasts with the supernatant of RGS5 deficient pericytes induced their activation characterized by the increased expression of α smooth muscle actin in a TFGβ2 dependent mechanism. Conclusions. Our results identify RGS5 as a crucial regulator of pericyte function during cardiac ageing. The deletion of RGS5 causes cardiac dysfunction and induces myocardial fibrosis, one of the hallmarks of cardiac ageing.
Project description:Pathological cardiac hypertrophy is a leading cause of heart failure. The understanding of disease mechanisms is primarily based on experimental models, but knowledge of the full repertoire of cardiac cells and their gene expression profiles in the human heart is missing. Here, using large-scale single-nucleus transcriptomes, we highlight the transcriptional response of cardiomyocytes to pressure overload in humans with stenosis of the aortic valve and disclose major alterations in cellular cross-talk. Cardiomyocytes showed a reduction of incoming connections with endothelial cells and fibroblasts. Particularly Eph receptor tyrosine kinases, including the predominant EPHB1 gene, were significantly down-regulated in cardiomyocytes of the hypertrophied heart. We compared 5 AS patients to healthy patients from the human heart cell ATLAS (https://www.heartcellatlas.org). This repository contains the processed files from the single nuclei RNA-SEQ.
Project description:Study of the emergence of the rare 2C like cell population upon Retinoic Acid treatment. Transcriptionally characterise the different cell populations emerging at different timepoints upon Retinoic Acid treatment and identify genes driving cell fate decisions.
Project description:CIARA (Cluster Independent Algorithm for the identification of RAre cell types) is tested on mouse embryonic stem cell treated for 24h with Retinoic Acid.
Project description:Hematopoietic mutations in epigenetic regulators like DNA methyltransferase 3 alpha (DNMT3A) drive clonal hematopoiesis of indeterminate potential (CHIP) and are associated with adverse prognosis in patients with heart failure (HF). The interactions between CHIP-mutated cells and other cardiac cell types remain unknown. Here, we identify fibroblasts as potential interaction partners of CHIP-mutated monocytes using combined transcriptomic data from peripheral blood mononuclear cells of HF patients with and without CHIP and cardiac tissue. We demonstrate that DNMT3A inactivation augments macrophage-to-cardiac fibroblasts interactions and induces cardiac fibrosis in mice and humans. Mechanistically, DNMT3A inactivation increases the release of heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) to activate cardiac fibroblasts. These findings not only identify a novel pathway of DNMT3A CHIP-driver mutation-induced instigation and progression of HF, but may also provide a rationale for the development of new anti-fibrotic strategies.
Project description:Myocarditis is a heart condition that causes inflammation and results in the loss of heart muscle cells, often leading to fibrosis (scarring) of the heart tissue and heart failure. However, the molecular mechanisms underlying immune cell control and maintenance of tissue integrity in the inflamed cardiac microenvironment remain elusive. Based on our finding that bone morphogenic protein-4 (BMP4) serum concentration was reduced in myocarditis patients in combination with comprehensive single cell and single nucleus RNA sequencing analyses of inflamed murine and human myocardial tissue indicated that BMP4 gradients maintain cardiac tissue homeostasis. Indeed, restoration of BMP signaling through antibody-mediated neutralization of the BMP-inhibitors GREM1 and GREM2 reduced CD4+ T cell-mediated myocardial inflammation and blocked disease progression by reduction of adverse fibrotic remodelling. These results unveil a key function of the BMP4-GREM1/2 axis as promising approach for treating myocardial inflammation and the serious complications of cardiac fibrosis and heart failure.
Project description:Single cell RNA-seq was performed on healthy mouse skin fibroblasts. This data along with single cell transcriptomics datasets of fibroblasts from other healthy tissues was used to construct a steady state mouse fibroblast atlas.