Additional file 1 of A universal predictive and mechanistic urinary peptide signature in acute kidney injury

Additional file 1: Supplementary Figure S1: S100A9 expression after epithelial injury. A. Urinary calprotectin (S100A8/A9) abundance 4 hours after cardiac bypass-surgery. B–F. Blood urea nitrogen (B), mRNA S100a9 (C), S100A9 immunostaining (D-E) and mRNA Kim1 (F) in sham mice and after bilateral renal ischemia/reperfusion (hours 6, 24 and 48). G–J. mRNA expression of S100a9 and Kim1 in MCT cells submitted to interleukin-1b (IL1β, 10 ng/mL) or tumor necrosis factor-1a (TNFa, 10 ng/mL) (G-H) or hypoxia (I-J). AKI, acute kidney injury; BUN, blood urea nitrogen; Norm, normoxia; Hyp, hypoxia. Supplementary Figure S2: Performances of the peptide-based signature to identify AKI that developed within the first 2 days following cardiac surgery. ROC curves with corresponding AUROC and 95% confidence intervals of the local clinical score (blue, pointed), the full 204 peptides-based score (red), the urinary NGAL level (yellow, pointed) and the nephrocheck ([IGFBP7].[TIMP2] product) in the validation cohort. Supplementary Figure S3: Reduction and combination of the peptide-based signature. A. ROC curves with corresponding AUROC and 95% confidence intervals of the local clinical score (blue, pointed), the full 204 peptides-based score (red), the reduced 17 peptides-based score (black, dashed) and the combination of local clinical and full peptide-based score in the validation cohort. B. List of peptides included in the reduced 17-peptides signature according to their parental protein. LMAN2, Lectin mannose binding 2 ; MGP, Matrix gla protein. Supplementary Figure S4: Performances of the 204 peptides-based signature and the reference urinary biomarker NGAL for AKI prediction in the external ICU validation cohort. A. ROC curves with corresponding AUROC and 95% confidence intervals of the 204 peptides-based score and the reference urinary biomarker NGAL to predict AKI after ICU admission. B. ROC curves with corresponding AUROC and 95% confidence intervals of the 204 peptides-based score and reference urinary biomarker NGAL to predict the development of AKI within seven days after admission. Supplementary Figure S5: Performances of the 204 peptides-based score for in-hospital mortality prediction. Odds-ratio (OR) of in-hospital mortality were calculated with unadjusted, Euroscore-II-adjusted or propensity score-adjusted logistic regression. Supplementary Table S1: Correlations between clinical characteristics and the 204-peptides-based score. BMI, body mass index; PAOD, peripheral artery obliterans disease; COPD, chronic obstructive pulmonary disease; LVEF, left ventricular ejection fraction; eGFR, estimated glomerular filtration rate; CBP, cardiac bypass; RBC, red blood cells. Supplementary Table S2: Performance of the peptide-based score to predict acute kidney injury in the external ICU validation cohort, according to the cause of admission to the intensive care unit. ICU, intensive care unit; AUROC, area under the receiver operating characteristics curve. Supplementary file S1: Methodology. Urinary peptidomics and statistical analyses.