Fig. 7: A spatial transcriptomics-inspired drug repurposing approach.

a Flow chart showing gene numbers present in each step of our drug repurposing analysis. Eleven potential targets of 24 drugs were predicted in the tegument and ten targets of 37 drugs in the gut. b SMART analysis and alignment of catalytic domains of human and liver fluke PKCβ. The domain composition (C2 regulatory domain in front of the kinase domain) classifies both as conventional PKCs⁸⁹. The catalytic domain of human PKCβ, which is bound by ruboxistaurin, shows 73.36% identity to the Fasciola ortholog. The conserved ATP binding site (bold) and the activation loop residues (underlined) are marked. The main residues involved in ruboxistaurin binding obtained for the human sequence⁸⁹ are Lys349, Gly350 and Lys467 (gray). c Left: Heatmap showing the average expression of different protein kinase C (PKC) genes per cluster (mean of UMI counts, normalized and scaled). Right: Heatmap showing the average spot expression of those genes in the whole dataset (log1p normalized counts). Red rectangle marks PKCβ with tegumental expression. Please note: While the spatial plot is shown for only one representative section, the heatmap includes expression data from all four tissue sections in the dataset. Source data are provided as a Source Data file. d Spatial projection only (left) and overlay with H&E-stained tissue section (right) showing expression of PKCβ (D915_006901). Several positive spots can be seen along the tegument of the parasite. Mehlis’ gland is not contained in this tissue section. For spatial projections of other PKCs, see Supplementary Fig. 9d. Expression level encoded by color (gray = low, red = high). e–g Adult flukes were treated for 72 h with different concentrations of the PCKβ isoform-specific inhibitor ruboxistaurin (20–100 µM) or triclabendazole as positive control. Motility was assessed every 24 h. Control worms were treated with the inhibitor solvent DMSO. Representative images are shown in (e) and motility scores for all time points and concentrations in (f and g) (score 3 = normal, 2 = reduced, 1 = severely reduced, 0 = no motility). See also Supplementary Movies 1 and 2. Data represent the mean ± SEM of n = 4 (triclabendazole at 20 µM) or n = 6 flukes (others) from 2 (triclabendazole at 20 µM) or 3 independent experiments (others) with 2 worms per condition and experiment. Significant differences to controls are indicated with *p = 0.0333 and **p = 0.0022 (two-sided Mann-Whitney U test). Scale bars correspond to 5 mm.