Fig. 7: Single-cardiomyocyte RNA-seq and plasma proteome analysis of patients with heart failure.

a UMAP plot of single-cell transcriptomes of cardiomyocytes isolated from control subjects (CTL) or patients with heart failure (HF) (n = 85 cells for CTL, n = 678 for HF). b Trajectory analysis on the UMAP plot in a. Clusters classified by graph-based clustering are shown by colors. Trajectories identified by the Slingshot algorithm are also shown. c Bar graph showing the distribution of the clusters in b. d M2 expression on the UMAP plot (left) and its dynamics along pseudotime (upper right). Enriched GO terms are also shown (lower right). e Heatmap showing the expression levels of selected M2 genes during the trajectories. f Boxplot of the M2 expression. Data represent box plots and individual data points. Box plots show the median (center line), first and third quartiles (box edges), while the whiskers going from each quartile to the minimum or maximum. n = 300 for C1, n = 267 for C2, n = 196 for C3. g M1 expression on the UMAP plot (left) and its dynamics during trajectories (upper right). Enriched GO terms and enrichment P-values are also shown (lower right). h Heatmap showing the expression levels of selected M1 genes during the trajectories. i Boxplot of the M1 expression. Data represent box plots and individual data points. Box plots show the median (center line), first and third quartiles (box edges), while the whiskers going from each quartile to the minimum or maximum. n = 300 for C1, n = 267 for C2, n = 196 for C3. j Scatter plot showing the ME expression of M1 and M2 for each cell. k M44 expression on the UMAP plot (left) and its dynamics along pseudotime (upper right). Enriched GO terms are also shown (lower right). l Heatmap showing the expression levels of selected M44 genes during the trajectories. m Boxplot of the M44 expression. Data represent box plots and individual data points. Box plots show the median (center line), first and third quartiles (box edges), while the whiskers going from each quartile to the minimum or maximum. n = 300 for C1, n = 267 for C2, n = 196 for C3. n Boxplot of plasma protein levels of IGFBP7 (upper) and NT-proBNP (lower). RFU, relative fluorescence unit. HF, heart failure. HTx, heart transplantation. Data represent box plots and individual data points. Box plots show the median (center line), first and third quartiles (box edges), while the whiskers going from each quartile to the minimum or maximum. n = 768 for Control, n = 85 for non-advanced HF, n = 30 for advanced HF before HTx, n = 30 for advanced HF after HTx. o Mean decrease in accuracy (in order of decreasing accuracy from top to bottom) of proteins involved in classification between manifest and advanced heart failure as assigned by the random forest classifier. p Receiver-operating characteristic (ROC) curves based on plasma protein levels of NT-proBNP and IGFBP7 for predicting the incidence of heart failure. Area under the curve is also shown for each marker. NT-proBNP had significantly better ability to diagnose heart failure (P-value = 1.109e-07). Areas under the ROC curve were calculated using logistic regression. q ROC curves based on plasma protein levels of NT-proBNP and IGFBP7 for discriminating patients with advanced heart failure. Area under the curve is also shown for each marker. Although not statistically significant, IGFBP7 had a higher power to determine the severity of heart failure (P-value = 0.1063). Areas under the ROC curve were calculated using logistic regression.