Fig. 5: OPN/SPP1, one of the main factors induced by Lecanemab treatment, promotes Aβ clearance.
a, Heatmap displaying the significance of enrichment of marker genes of previously reported microglial cell states16,26,27 in the pink WGCNA module, as assessed by a one-sided hypergeometric overlap test. Color intensity reflects enrichment significance, represented as −log10(FDR), with darker red indicating stronger enrichment. Respective BH Padj values for the enrichments are specified. The list of shared genes with significantly enriched states is outlined in Supplementary Table 3 (***Padj < 0.001). b, NES of WGCNA modules in Lecanemab TDs (40-μm hexbin pseudospots) with respect to distance to pathology, as identified by GSEA, performed on the DE analysis in Fig. 1e, with Padj values indicated. Two-sided P values were adjusted using the BH correction (**Padj< 0.01, ***Padj < 0.001). c, Cortical TDs in Lecanemab-treated mice colored based on their relative expression of genes belonging to the pink module (purple, low expression; yellow, high expression) and overlayed with plaque ROIs (white). ESs were obtained using Scanpy’s ‘score_genes()’ function. Please note the significant enrichment of the module in TDs close to Aβ plaques, as quantified in b. d, Representative confocal images of cortical X-34+ and 82E1+ plaques surrounded by OPN+ microglia in IgG1-treated, Lecanemab-treated and Lecanemab LALA-PG-treated mice. Scale bar = 25 µm. e,f, Quantification of the area of OPN+ area within IBA1+ cells around X-34+ (e) and 82E1+ plaques (f) in IgG1-treated and Lecanemab-treated mice; (e) one-way ANOVA (P = 0.0008) and (j) one-way ANOVA (P = 0.0007; IgG1, n = 10 mice; Lec, n = 12 mice; Lec LALA-PG, n = 9 mice). g, Quantification of the area covered by IBA1+ cells around X-34+ plaques; one-way ANOVA (NS) (IgG1, n = 10 mice; Lec, n = 12 mice; Lec LALA-PG, n = 9 mice). h, Schematic representation of the in vitro plaque clearance assay paradigm used to study Aβ clearance in response to OPN stimulation. Human-derived microglial cells were plated onto sagittal cryosections from 6-month-old AppNL-G-F mice, followed by the treatment with increasing concentrations of human OPN (17, 50, 150, 450 and 1,350 ng ml−1). After 3 days, Aβ plaque coverage was quantified using the pan-Aβ antibody 82E1. Panel h was created with BioRender.com. i, Representative confocal images of 82E1 (Aβ, cyan) and CD9 (human microglia, magenta) immunoreactivity in AppNL-G-F brain cryosections after OPN stimulation. Scale bar = 1 mm; inset = 200 µm. j, Quantification of 82E1 covered area (relative to no OPN) in sections plated with or without human microglia; modified chi-squared method (n = 3 independent experiments; P = 0.03). Graphs show mean ± s.e.m. and points represent individual animals (e–g) or independent experiments, with each being the average of one to four cryosections (j). Square symbols, males; triangles, females.
