Fig. 5: Human clinical validation of the plasma-to-tumour tissue integrative proteomic analysis pipeline.
a Schematic representation outlining the human clinical cohort study design. Plasma samples obtained from pre-operative high-grade glioma patients were matched with healthy controls, both then formed ex vivo coronas with liposome NPs. Snap-frozen tumour tissue from the same GB patients during tumour surgical resection were cryo-sectioned and annotated by a clinical pathologist before laser capture microdissection (Supplementary Fig. S4b). Both corona and tumour tissue samples were subjected to LC-MS/MS analysis. Created in BioRender. Hadjidemetriou, M. (2025) https://BioRender.com/k40i192. b Volcano plot illustrating the relationship between fold change and significance for DAPs identified by comparing GB patient-corona with healthy control-corona (n = 10 biological replicates). A total of n = 321 proteins (with an FDR-adjusted one-way ANOVA p-value < 0.05) were found to be differentially abundant, of which n = 272 proteins exhibited a fold change > 2. Filled dots represent 140 proteins that were also identified by LC:MS/MS analysis in human tumour tissue. Protein names with log2 fold change > 10 or -log(p-value) 6 are displayed. Full lists of proteins are shown in Supplementary Data 14–16. c Dot plot presents common KEGG pathways identified through independent enrichment analyses of human plasma DAPs and tumour tissue proteins, using a significance threshold of adjusted p-value < 0.001 by one-sided Fisher’s exact test. Pathways are classified into three groups according to the functional category in the KEGG pathway database. The colour of the dots represents the adjusted p-value, and the dot size indicates the gene ratio (genes involved/total number of genes). *Indicates the common enriched pathways between humans and mice. d Chord diagram connects the seven enriched pathways shared between humans and mice, with the n = 48 DAPs identified to be shared between the plasma and tumour tissue analyses. Human DAPs found to be shared with mouse plasma DAPs are represented in medium-grey colour. Human DAPs identified as common with mouse plasma and tumour tissue DAPs are depicted in dark-grey colour. The full list of the n = 48 DAPs is shown in Supplementary Fig. 6.
