Fig. 2: Progressive metabolic evolution from AAH to IAC.

a Partial least squares discriminant analysis (PLS-DA) of the metabolomics data from AHH, AIS, MIA, and IAC patients. b Volcano plots of the significantly differential metabolites in pre-invasive group (AAH/AIS) versus IAC or in MIA versus IAC were shown. Two-sided Wilcoxon rank-sum tests followed by Benjamini-Hochberg (BH) multiple comparison test with false discovery rate (FDR) < 0.05 and fold change > 1.25 or <0.8. Metabolites significantly increased or decreased were colored in purple and green, respectively. c Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways enriched by significantly differential metabolites in pre-invasive group (AAH/AIS) versus IAC or in MIA versus IAC group. One-sided Fisher’s exact test followed by BH multiple comparison test with FDR < 0.05. d Clustering of metabolic trajectories using differential metabolites among AAH, AIS, MIA, and IAC. Two-sided Kruskal-Wallis tests followed by BH multiple comparison test with FDR < 0.1. e, f Dynamic alterations of metabolites in cluster 1 and cluster 2. The dots represent the mean log₂ relative abundance. Two-sided Kruskal-Wallis tests followed by BH multiple comparison test with FDR < 0.1. g Violin plots of metabolites in cluster 3 and cluster 4. The difference of metabolites among AHH, AIS, MIA, and IAC were evaluated using a two-sided Kruskal-Wallis test. Black dots represent population medians.