Fig. 2: Identification of epithelial JUN activity as a key switch in immune-mediated kidney disease.
From: Pathology-oriented multiplexing enables integrative disease mapping
a, Schematic overview for the proof-of-concept experiment in a mouse model of immune-mediated kidney disease before (acute injury) and after (CGN) pathological lesion formation (n = 10 mice; ROIs = 40). NTS, nephrotoxic serum; details of the antibody panel are provided in Supplementary Table 1. b, Spatiotemporal distribution of colour-coded clusters. c, Examples of interpretable clusters (C28, C21, C4 and C7) of biological significance. Each dot represents an ROI, which was used as an independent observation (n = 11 ROIs for controls, n = 11 ROIs for acute injury and n = 18 ROIs for CGN) and red bars represent medians and inter-quartile ranges. Mes, mesangial. d, Identification of C21 (with pJUN as a top contributor) as a key regulated pathomechanism before and after lesion formation. e, Images of the spatiotemporal distribution of C21 (left) and cell-specific frequency (right) among tubular epithelial cells and PECs. f, Treatment with a JNK inhibitor (JNKi) reduces the PDGF-mediated collective migration of murine PECs in vitro. In ‘collective migration’, error bars represent upper and lower limits. Data are from four biological replicates. Veh, vehicle. g, Confirmation of pJUN expression in PECs during different lesion stages among human kidney biopsy samples (n = 12 patients and n = 3 healthy individuals), which was also associated with CD44 co-expression. h, Schematic overview of the use of a JNKi as a preventive strategy (before NTS) and a therapeutic strategy (7 days after NTS) during the progression of immune-mediated kidney disease in a rat model of CGN. i,j, Proteinuria (n = 4 rats for all groups) and glomerular damage (n = 4 rats for day 0, n = 6 rats for all other groups; red bars represent medians and interquartile ranges) show a direct preventive (i) and therapeutic (j) effect of the JNKi. k, Using expression of CD44 as a readout of PEC activation, we confirmed the effect of the JNKi on PEC activation (using all rats available from i and j). Differential cluster abundance analysis used a two-sided t-test. Cluster composition analysis relied on a two-sided t-test with Holm–Šidák correction. For other comparisons, two-sided Mann–Whitney, Kruskal–Wallis with Dunn, analysis of variance (ANOVA) with Dunnett T3 or ANOVA with Holm–Šidák tests were used depending on the number of comparisons. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 or not significant (NS). Scale bars, 50 µm (c,e,g,k). Diagrams in a, f and h were created using BioRender (https://biorender.com).
