Project description:Single-cell RNA sequencing was performed on embryonic Drosophila heart cells. Analysis of single-cell RNA sequence (scRNA-seq) data at timepoints prior to migration of cardiac progenitor cells through to heart tube closure (embryonic stages 13, 14-early, 14-late, 15 and 16) revealed several interesting findings. We found specification of cardiac cell types takes place early, before stage 13, with biggest changes in transcriptomic profiles detected once cells had settled at the midline for further cardioblast maturation. Throughout development, our data identified multiple cell types, covering cardioblasts and five types of pericardial cells including a neural cardiac cell type. The scRNA-seq data further revealed a combination of first and second heart fields during heart development in fly. Further, we uncovered new cell type-specific markers discerning the different cardiac cell types. And we identified signaling pathways key to heart cell maturation, which are conserved from fly to human.
Project description:Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) primarily infects the respiratory tract, but pulmonary and cardiac complications occur in severe coronavirus disease 2019 (COVID-19). To elucidate molecular mechanisms in the lung and heart, we conductedpaired experiments in human stem cell-derived lung alveolar type II (AT2) epithelial cell and cardiac cultures infected with SARS-CoV-2.With CRISPR-Cas9-mediated knockout of ACE2, we demonstrated that angiotensin-converting enzyme 2 (ACE2) was essential for SARSCoV-2 infection of both cell types but that further processing in lung cells required TMPRSS2, while cardiac cells required the endosomalpathway. Host responses were significantly different; transcriptome profiling and phosphoproteomics responses depended strongly onthe cell type. We identified several antiviral compounds with distinct antiviral and toxicity profiles in lung AT2 and cardiac cells, highlighting the importance of using several relevant cell types for evaluation of antiviral drugs. Our data provide new insights into rationaldrug combinations for effective treatment of a virus that affects multiple organ systems.
Project description:Development of specialized cell types and structures in the vertebrate heart is regulated by spatially-restricted molecular pathways. Disruptions in these pathways can cause severe congenital cardiac malformations or functional defects. To better understand these pathways and how they regulate cardiac development and function we used tomo-seq, combining high-throughput RNA sequencing with tissue sectioning, to establish a genome-wide expression dataset with high spatial resolution for the developing zebrafish heart. Analysis of the dataset revealed over 1100 genes differentially expressed in sub-compartments. Pacemaker cells in the sinoatrial region induce heart contractions, but little is known about the mechanisms underlying their development and function. Using our transcriptome map, we identified spatially restricted Wnt/β-catenin signaling activity in pacemaker cells, which was controlled by Islet-1 activity. Moreover, Wnt/β-catenin signaling at a specific developmental stage in the myocardium controls heart rate by regulating pacemaker cellular response to parasympathetic stimuli. Thus, this high-resolution transcriptome map incorporating all cell types in the embryonic heart can expose spatially-restricted molecular pathways critical for specific cardiac functions.
Project description:We report bulk RNA-sequencing, ChIP-seq, and ATAC-seq of endothelial cells harvested from heart and lung of multiple mouse strains investigating the role of KLF2 and KLF4 in endothelial transcription
Project description:Heart biopsies of explanted organ after heart failure. Same patient, 3 different sample types. 1,2,6 - from paraffin tissue, Thermo fisher kit 3,4,5 - from paraffin tissue, Qiagen kit 7,8,9 - fresh tissue with Qiagen kit