Wewer Albrechtsen, N. | Search bioRxiv

bioRxiv2026

Early Identification of At-Risk Patients: ProteomicSignature Predicts Progression to DecompensatedCirrhosis

Background & AimsEarly identification of decompensation in patients with cirrhosis is important to enable timely detection, management of complications and for effective treatment. This study investigates the biology of decompensation and aim to identify protein biomarkers for identification of high-risk patients. MethodsThe primary analysis included plasma samples from 46 patients with metabolic dysfunction associated steatotic liver disease (MASLD) related cirrhosis. Plasma samples were depleted for the top 14 most abundant proteins and the proteome was measured by liquid chromatography tandem mass spectrometry. The dataset was divided into a training (14 compensated, 10 decompensated) and a test cohort of compensated patients (11 progressing to decompensation, 11 remaining compensated). Changes in protein levels were determined by ANCOVA and a prognostic model was developed using logistic regression. External validation was performed in an independent cohort of 120 patients with alcohol-related cirrhosis. Time-to-event analyses were conducted in this cohort using Cox regression. Results52 proteins involved in impaired hepatic function, fibrogenesis, immune activation, and metabolic changes were significantly different between compensated and decompensated patients. A prognostic model with four proteins (NBL1, LTBP4, APOC4, GHR), demonstrated predictive ability for future decompensation (AUC=0.93, 73% sensitivity, 100% specificity). In the external validation cohort, the model demonstrated generalizability (AUC=0.78, 72% sensitivity, 82% specificity). Validation cohort time-to-event analyses showed that higher baseline scores were associated with shorter time to liver-related events (HR 1.32; log-rank p = 0.027), underscoring the panels prognostic value. ConclusionOur study indicates that patients with decompensated cirrhosis are characterized by proteomic signatures of fibrogenesis and metabolic dysfunction. Capturing these signatures could help identify patients at risk of complications and potentially those eligible for aetiology directed treatment. Impact and ImplicationsAddressing a critical unmet need for early detection of cirrhosis decompensation, our proteomic study identifies a four-protein panel with predictive ability for decompensation. These findings hold significant implications for hepatologists, clinical researchers, and healthcare systems, offering a novel tool to enhance prognostication and refine treatment strategies, potentially facilitating targeted patient monitoring. However, considering the small discovery sample size and the distinct aetiology of the external validation cohort, further validation is essential before broad clinical integration. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=183 SRC="FIGDIR/small/709475v1_ufig1.gif" ALT="Figure 1"> View larger version (55K): org.highwire.dtl.DTLVardef@6620e2org.highwire.dtl.DTLVardef@f8dfe4org.highwire.dtl.DTLVardef@1331101org.highwire.dtl.DTLVardef@1a195ca_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

bioRxiv2023

Inhibition of dipeptidyl peptidase 4 allows accurate measurement of GLP-1 secretion in mice

Dipeptidyl peptidase (DPP)-4 and neprilysin (NEP) within few minutes degrade glucagon-like peptide 1 (GLP-1) in mice, generating small, inactive fragments of the peptide. Commercially available sandwich ELISA kits may not accurately detect intact GLP-1 (that is GLP-1[7-36]NH2) and these moieties, leading to underestimation of secretion and potentially misleading results. Single-site antibody approaches may pick up some fragments, yet require large plasma volumes and the accuracy is uncertain. We aimed to find a way to stabilize GLP-1 in mice allowing reliable measurement with sensitive commercially available ELISA kits. Non-anesthetized male C57Bl/6JRj mice were subjected to an oral glucose tolerance test (OGTT; 2 g/kg glucose via oral gavage). Blood was drawn from the retrobulbar plexus before and repeatedly during the OGTT and total and intact GLP-1 were measured by commercially available sandwich ELISA kits (Mercodia and Alpco, respectively). In blood samples taken 15 minutes after the glucose load, there were no increase in plasma GLP-1 concentration. There was a small insignificant increase in total GLP-1 (1-2 pmol/L) in samples taken at t=5 and t=10 minutes following the OGTT, but no rise in intact GLP-1. We then administered control (saline), or a DPP-4 inhibitor (valine pyrrolidide, 0.1 {micro}mol/g or sitagliptin, 10 mg/kg) with or without a NEP-inhibitor (sacubitril, 0.3 mg/kg) 30 minutes before the OGTT. In the four inhibitor groups, intact GLP-1 increased during the OGTT (levels ranging between 4.04 [SD 2.87] and 15.53 [SD 6.76] pmol/L). The combination of sitagliptin with sacubitril gave the largest increase in intact GLP-1 levels. Finally, after injecting male C57Bl/6JRj mice with a known dose of GLP-1(7-36)NH2, the peak GLP-1 levels were significantly higher during sitagliptin, but not with the combination of sitagliptin/sacubitril. Both inhibitor groups, however, showed prolonged half-life of the GLP-1 plasma disappearance. We conclude that for measurements of GLP-1 secretion in mice with commercially available sandwich ELISA kits, it is necessary to consider both timing of blood sampling and in vivo inhibition of DPP-4. The described approach allows improved estimates of GLP-1 secretion for future studies. It is a limitation that DPP-4 and NEP inhibition may have metabolic effects by stabilizing the intact GLP-1 peptide (e.g. influencing levels of insulin and glucagon).

physiology↗