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Journal: Molecular & Cellular Proteomics : MCP

Article Title: Urinary Proteomics Identifies ADAMDEC1 as a Non-invasive Biomarker of Unstable Carotid Atherosclerotic Plaques

doi: 10.1016/j.mcpro.2026.101624

Figure Lengend Snippet: Schematic overview of the study design. The study comprised four phases: cohort establishment, urinary proteomics and tissue-data integration, ADAMDEC1 prioritization and clinical validation, and cellular/experimental support. A discovery cohort (n = 179) and an independent validation cohort (n = 225) were established. Urinary proteomic profiling by data-independent acquisition mass spectrometry was integrated with published plaque tissue proteomic and transcriptomic datasets. ADAMDEC1 was prioritized through cross-referencing of urinary and tissue signatures and was subsequently evaluated by ELISA, receiver operating characteristic analysis, multivariable logistic regression, prognostic analysis, and assessment of associations with plaque vulnerability features. single-cell RNA sequencing, human plaque validation (immunohistochemistry / western blotting /IF), and THP-1 macrophage assays were used to define the cellular context and provide experimental support.IF, immunofluorescence.

Article Snippet: For functional stimulation assays, cells were treated with recombinant ADAMDEC1 (200 ng/ml; MedChemExpress) or lipopolysaccharide (LPS) (1000 ng/ml; Sigma-Aldrich) in complete medium.

Techniques: Biomarker Discovery, Data-independent acquisition, Mass Spectrometry, Enzyme-linked Immunosorbent Assay, Single Cell, RNA Sequencing, Immunohistochemistry, Western Blot, Immunofluorescence

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Urinary Proteomics Identifies ADAMDEC1 as a Non-invasive Biomarker of Unstable Carotid Atherosclerotic Plaques

doi: 10.1016/j.mcpro.2026.101624

Figure Lengend Snippet: Cross-omics prioritization and independent validation identify ADAMDEC1 as a urinary biomarker of unstable carotid plaques. A, schematic workflow outlining the strategy for integrating tissue-based proteomic and transcriptomic datasets with urinary proteomics to identify overlapping targets. B, UpSet plot revealing the intersection of DEPs across six independent published proteomic studies. The red box highlights ADAMDEC1 and PRKAR1A as recurrent candidate proteins identified in multiple plaque datasets. C and D, transcriptomic validation of ADAMDEC1 and PRKAR1A expression in unstable versus stable plaques using two external Gene Expression Omnibus datasets ( GSE28829 and GSE43292 ). ADAMDEC1 was upregulated, whereas PRKAR1A was downregulated in unstable plaques. E and F, ELISA-based quantification of urinary ADAMDEC1 and PRKAR1A in the independent external validation cohort (n = 225; 85 HCs, 57 stable CAPs, and 83 unstable CAPs). Urinary ADAMDEC1 levels showed a stepwise increase from HCs to stable and unstable CAPs, whereas PRKAR1A was higher in unstable CAPs than in stable CAPs but did not differ significantly between HCs and stable CAPs. G, receiver operating characteristic curve showing the diagnostic performance of urinary ADAMDEC1 for discriminating unstable from stable carotid plaques. H , Kaplan-Meier analysis of cerebrovascular event-free survival based on intraplaque ADAMDEC1 expression in the GSE21545 dataset (n = 126), showing that high ADAMDEC1 expression was associated with poorer prognosis. Data in (C and F) are presented as mean ± SEM. Statistical significance was determined using Student’s t test for (C and D) and the Kruskal–Wallis test followed by Dunn’s post hoc test for (E and F) . ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. AUC, area under the curve; CAP, carotid atherosclerotic plaque; DEP, differentially expressed protein; HC, healthy control.

Article Snippet: For functional stimulation assays, cells were treated with recombinant ADAMDEC1 (200 ng/ml; MedChemExpress) or lipopolysaccharide (LPS) (1000 ng/ml; Sigma-Aldrich) in complete medium.

Techniques: Biomarker Discovery, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Diagnostic Assay, Control

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Urinary Proteomics Identifies ADAMDEC1 as a Non-invasive Biomarker of Unstable Carotid Atherosclerotic Plaques

doi: 10.1016/j.mcpro.2026.101624

Figure Lengend Snippet: Association of urinary ADAMDEC1 level with plaque characteristics in the validation cohort

Article Snippet: For functional stimulation assays, cells were treated with recombinant ADAMDEC1 (200 ng/ml; MedChemExpress) or lipopolysaccharide (LPS) (1000 ng/ml; Sigma-Aldrich) in complete medium.

Techniques: Biomarker Discovery

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Urinary Proteomics Identifies ADAMDEC1 as a Non-invasive Biomarker of Unstable Carotid Atherosclerotic Plaques

doi: 10.1016/j.mcpro.2026.101624

Figure Lengend Snippet: ADAMDEC1 is enriched in inflammatory macrophages within advanced carotid plaques. A, UMAP visualization of the single-cell transcriptomic landscape in human carotid atherosclerotic plaques, identifying six major cell lineages. B, Dot plot showing the expression intensity and proportion of canonical marker genes used to annotate major cell populations. C, Feature plot showing that ADAMDEC1 expression is predominantly restricted to the macrophage cluster. D, Violin plots comparing ADAMDEC1 expression across major cell populations between AC and PA tissues, showing significantly higher expression in macrophages from AC tissues. E, Uniform Manifold Approximation and Projection projection of the macrophage subpopulation, stratified into AC inflammatory, AC noninflammatory, PA inflammatory, and PA noninflammatory subsets. F, distribution of inflammatory signature scores across macrophage subsets projected onto the Uniform Manifold Approximation and Projection embedding. G, feature plots showing preferential enrichment of ADAMDEC1 expression in inflammatory macrophage subsets, particularly in AC inflammatory macrophages. H , quantification of the proportion of ADAMDEC1 -positive cells across macrophage subsets, showing the highest proportion in AC inflammatory macrophages. I, Scatter plots showing positive correlations between ADAMDEC1 expression and the pro-inflammatory genes CCL3 , CD86 , HLA-DRA , and CCL4 in plaque macrophages. Pearson’s correlation coefficients (r) and p values are indicated. Data in ( D ) are presented as mean ± SEM. Statistical significance was determined using the Wilcoxon rank-sum test for ( D ). ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. AC, atherosclerotic core; PA, proximal adjacent tissue.

Article Snippet: For functional stimulation assays, cells were treated with recombinant ADAMDEC1 (200 ng/ml; MedChemExpress) or lipopolysaccharide (LPS) (1000 ng/ml; Sigma-Aldrich) in complete medium.

Techniques: Single Cell, Expressing, Marker

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Urinary Proteomics Identifies ADAMDEC1 as a Non-invasive Biomarker of Unstable Carotid Atherosclerotic Plaques

doi: 10.1016/j.mcpro.2026.101624

Figure Lengend Snippet: Human plaque validation and in vitro assays support an association between ADAMDEC1 and pro-inflammatory macrophage polarization. A, representative immunohistochemical staining of ADAMDEC1 in human carotid plaque specimens, showing stronger staining in AC than in PA regions. The scale bar represents 500 μm in the overview image and 50 μm in the enlarged panels. B, quantification of ADAMDEC1 immunoreactivity by integrated optical density /area, confirming increased staining intensity in AC compared with PA tissues. C, representative Western blot images showing higher ADAMDEC1 protein abundance in AC than in PA tissues. GAPDH was used as the loading control. D, quantification of western blotting results, showing increased relative ADAMDEC1 protein levels in AC tissues. E, IF staining showing colocalization of ADAMDEC1 with CD68-and CD86-positive macrophages in human carotid plaques. The scale bar represents 20 μm. F, representative flow cytometry plots showing macrophage polarization in THP-1 derived macrophages following rADAMDEC1 treatment or ADAMDEC1 knockdown under lipopolysaccharide stimulation. G, quantification of the proportion of M1-like macrophages, showing increased polarization following rADAMDEC1 treatment and reduced polarization following ADAMDEC1 silencing. H, representative IF images of CD86 expression in THP-1-derived macrophages after rADAMDEC1 treatment or ADAMDEC1 knockdown. The cale bar represents 50 μm. I, quantification of CD86 fluorescence intensity, showing enhanced CD86 expression after rADAMDEC1 treatment and reduced expression after ADAMDEC1 knockdown. Data in ( B , D , G , I ) are presented as mean ± SEM. Statistical significance was determined using Student’s t test for two-group comparisons ( B , D , G top , I top ) and one-way ANOVA followed by Tukey’s post hoc test for multi-group comparisons ( G bottom, I bottom ). ns p > 0.05, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. AC, atherosclerotic core; IF, immunofluorescence; PA, proximal adjacent tissue; rADAMDEC1, recombinant ADAMDEC1.

Article Snippet: For functional stimulation assays, cells were treated with recombinant ADAMDEC1 (200 ng/ml; MedChemExpress) or lipopolysaccharide (LPS) (1000 ng/ml; Sigma-Aldrich) in complete medium.

Techniques: Biomarker Discovery, In Vitro, Immunohistochemical staining, Staining, Western Blot, Quantitative Proteomics, Control, Flow Cytometry, Derivative Assay, Knockdown, Expressing, Fluorescence, Immunofluorescence, Recombinant