Plasma Extracellular Vesicles Contain Protein Biomarkers for Capturing Stages of Metabolic Dysfunction-Associated Steatotic Liver Disease: A Preliminary Exploratory Study

Figures

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  Figure 1. Characterization of plasma extracellular vesicles. Nanoparticle tracking analysis (NTA) showing the size distribution and concentration of isolated plasma EVs, with the majority of particles ranging between 100–200 nm. Western blot (File S1) analysis confirms the expression of EV-enriched markers CD63, CD9, CD81, and TSG101, supporting the identity of the isolated EV population.

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  Figure 2. Volcano plot showing differentially abundant plasma EV proteins between participants with S1 vs. S3 (A) and boxplots of differentially abundant plasma EV proteins per steatosis grade; H4C1 (B), OIT3 (C), ANPEP (D), CCDC25 (E) and KLHL41 (F). EV, extracellular vesicles; S1, mild steatosis; S2, moderate steatosis; S3, advanced steatosis; H4C1, histone H4; OIT3, oncoprotein-induced transcript 3; ANPEP, alanyl aminopeptidase; CCDC25, coiled-coil domain-containing protein 25; KLHL41, Kelch-like protein 41.

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  Figure 3. Volcano plot showing differentially abundant plasma EV proteins between participants with MASH and those without steatohepatitis (A) and Receiver operating characteristic (ROC)-curve of GP1BA for distinguishing between MASH from MASLD without steatohepatitis (B). MASH, metabolic dysfunction-associated steatohepatitis; TPR, true positive rate; AUC, area under the curve; CI, confidence interval; FPR, false positive rate; GP1BA, glycoprotein Ib alpha chain.

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  Figure 4. Volcano plot showing differentially abundant plasma EV proteins between participants with F0-1 vs. ≥F3 (A) and boxplots of differentially abundant plasma EV proteins per fibrosis grade; IGHD (B), GNB1 (C) and C7 (D). F0, no fibrosis; F1, mild fibrosis; F2, significant fibrosis; F3, advanced fibrosis; F4, cirrhosis; IGHD, immunoglobulin heavy constant delta; GNB1, guanine nucleotide-binding protein subunit beta-1; C7, complement component 7.