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Background: The Baveno VI consensus proposed a dual liver stiffness (LS) by transient elastography threshold of 15 kPa for excluding and diagnosing compensated advanced chronic liver disease (cACLD) in the absence of other clinical signs. Herein, we aimed to validate these criteria in a real-world multicentre study. Methods: We included 5,648 patients (mean age 51 ± 13 years, 53% males) from 10 European liver centres who had a liver biopsy and LS measurement within 6 months. We included patients with chronic hepatitis C (n = 2,913, 52%), non-alcoholic fatty liver disease (NAFLD, n = 1,073, 19%), alcohol-related liver disease (ALD, n = 946, 17%) or chronic hepatitis B (n = 716, 13%). cACLD was defined as fibrosis stage ≥F3. Results: Overall, 3,606 (66%) and 987 (18%) patients had LS 15 kPa, respectively, while cACLD was histologically confirmed in 1,772 (31%) patients. The cut-offs of 15 kPa showed 75% sensitivity and 96% specificity to exclude and diagnose cACLD, respectively. Examining the ROC curve, a more optimal dual cut-off at 12 kPa, with 91% sensitivity and 92% specificity for excluding and diagnosing cACLD (AUC 0.87; 95% CI 0.86–0.88; p 15 kPa dual cut-offs, we found that the 12 kPa dual cut-offs have better diagnostic accuracy in cACLD. Lay summary: The term compensated advanced chronic liver disease (cACLD) was introduced in 2015 to describe the spectrum of advanced fibrosis and cirrhosis in asymptomatic patients. It was also suggested that cACLD could be diagnosed or ruled out based on specific liver stiffness values, which can be non-invasively measured by transient elastography. Herein, we assessed the suggested cut-off values and identified alternative values that offered better overall accuracy for diagnosing or ruling out cACLD. © 2020 European Association for the Study of the Liver

BACKGROUND: No prognostic tools are established for alcohol-related liver disease (ALD). Collagen proportionate area (CPA) measurement is a technique that quantifies fibrous tissue in liver biopsies using digital image analysis.AIM: To assess the predictive value of CPA on hepatic decompensation and liver-related mortality in ALD METHODS: In a multicentre cohort study, we included 386 patients with biopsy-verified ALD and with long-term follow-up. In the development cohort of 276 patients, we assessed the predictors of hepatic decompensation and liver-related death in standard and competing risk multivariable Cox regression analyses. The results were validated in an independent prospective cohort of 110 patients, where CPA was also correlated with liver stiffness measurement (LSM).RESULTS: In the development cohort, 231 (84%) patients had early/compensated ALD (non-cirrhotic or compensated cirrhosis) and 45 (16%) had decompensated cirrhosis. In the validation cohort, all patients had early/compensated ALD. Independent predictors of liver-related mortality were higher CPA values (HR = 1.04, 95% CI 1.02-1.04) and advanced fibrosis (HR = 2.80, 95% CI 1.29-6.05) with similar results in standard and competing risk multivariable Cox regression analysis. In early/compensated ALD, CPA was the only independent predictor of hepatic decompensation and liver-related death (HR = 1.08, 95% CI 1.06-1.11). In the prospective cohort, we validated that CPA independently predicts hepatic decompensation in early/compensated ALD. The predictive power of CPA and LSM was equally strong.CONCLUSIONS: CPA predicts liver-related mortality in ALD and hepatic decompensation and/or liver-related death in early/compensated ALD. Traditional histological assessment may benefit from the addition of CPA to the evaluation of ALD.

Background Patients with inflammatory bowel disease (IBD) have varying disease courses with phases of inactive disease and flares of active disease with impairment of the intestinal epithelial barrier leading to tissue damage. Due to the large surface area, basement membrane is abundant in the intestinal tissue and its constituents, such as laminins, play an important role in the intestinal epithelial homeostasis. The laminin gamma 1 chain is highly abundant along the entire crypt-villus axis in the basement membrane. We investigated if a serum biomarker of the basement membrane (LG1M) could serve as surrogate biomarkers for disease activity in a rat in vivo colitis model and in IBD patients. Methods Serum from male Sprague Dawley rats that received 5–6% dextran sulfate sodium (DSS) (n = 21) or regular drinking water (n = 9) for 5 days was included and scored for disease activity index (DAI). In addition, serum from 44 Crohn’s disease (CD) patients with inactive (n = 20) or active (n = 24) disease and healthy subjects (n = 20) was included in this study. A competitive ELISA for matrix metalloproteinase (MMP) 9 mediated degradation of the laminin gamma 1 chain (LG1M) was used to estimate the level of laminin degradation in serum. Sections of the distal colon from the rats were assessed histologically for structural changes. Results LG1M were significantly elevated in CD patients serum compared with healthy subjects (p < 0.001; p < 0.001), but with lower levels in active CD compared inactive CD (Figure 1A). The findings of LG1M were confirmed in the in vivo colitis model, in which the levels of LG1M in DSS rat serum with high DAI were significantly lower (p = 0.004) compared with controls (Figure 1B). LG1M correlated negatively to DAI (r2 = 0.2153; p < 0.01) (Figure 1C). The structure of the distal colonic tissue in DSS rats was disrupted compared with controls with loss of crypt architecture and total surface area (Figure 1D–E).

Background Stricturing disease remains one of the biggest complications leading to intestinal resection in Crohn’s disease (CD), and affecting 30–50% of patients with CD. Intestinal strictures are caused by fibrosis development, as a result of increased collagen deposition. Intestinal fibroblasts and myofibroblasts are the main effector cells for intestinal fibrosis development and intestinal subepithelial myofibroblasts have shown to produce significantly elevated levels of type XVI collagen in CD patients. Thus, we investigated a novel serum biomarker, quantifying type XVI collagen (C16-C), as a biomarker for intestinal fibrosis in CD patients. Methods Serum from CD patients (n = 44) and healthy subjects (n = 50) was included. The Montreal classification for CD disease behaviour (B1, non-stricturing/non-penetrating: n = 20; B2: stricturing, n = 11; B3: penetrating n = 13) and disease location (L1, ileum: n = 14; L2, colon: n = 5; L3, ileum + colon: n = 25) was applied. The patients were classified as having inactive disease (n = 20) or active disease (n = 24) based on the Crohn’s disease Activity Index (CDAI) score. Competitive ELISA was applied for the quantification of C16-C in serum from CD patients (total serum samples: n = 94). Results The biomarker C16-C was significantly elevated in patients with CD compared with healthy donors (p < 0.001, AUC: 0.81). CD patients with strictures (B2) demonstrated significantly elevated serum levels of C16-C compared with CD patient without strictures (B1 and B3), and healthy donors. Furthermore, the diagnostic accuracy to separate CD patients with strictures (B2) from CD patient without strictures (B1 and B3) was 76% (p < 0.01, AUC: 0.76) and healthy donors 96% (p < 0.001, AUC 0.82). In addition, C16-C was also elevated in CD patients with ileum or ileum+colon disease involvement compared with only colon involvement and healthy donors (p < 0.05). There was no significant difference between patients with active or inactive disease.

The TRAIL pathway can mediate apoptosis of hepatic stellate cells to promote the resolution of liver fibrosis. However, TRAIL has the capacity to bind to regulatory receptors in addition to death-inducing receptors; their differential roles in liver fibrosis have not been investigated. Here we have dissected the contribution of regulatory TRAIL receptors to apoptosis resistance in primary human hepatic stellate cells (hHSC). hHSC isolated from healthy margins of liver resections from different donors expressed variable levels of TRAIL-R2/3/4 (but negligible TRAIL-R1) ex vivo and after activation. The apoptotic potential of TRAIL-R2 on hHSC was confirmed by lentiviral-mediated knockdown. A functional inhibitory role for TRAIL-R3/4 was revealed by shRNA knockdown and mAb blockade, showing that these regulatory receptors limit apoptosis of hHSC in response to both oligomerised TRAIL and NK cells. A close inverse ex vivo correlation between hHSC TRAIL-R4 expression and susceptibility to apoptosis underscored its central regulatory role. Our data provide the first demonstration of non-redundant functional roles for the regulatory TRAIL receptors (TRAIL-R3/4) in a physiological setting. The potential for these inhibitory TRAIL receptors to protect hHSC from apoptosis opens new avenues for prognostic and therapeutic approaches to the management of liver fibrosis.

Hepatic tissue engineering using decellularized scaffolds is a potential therapeutic alternative to conventional transplantation. However, scaffolds are usually obtained using decellularization protocols that destroy the extracellular matrix (ECM) and hamper clinical translation. We aim to develop a decellularization technique that reliably maintains hepatic microarchitecture and ECM components. Isolated rat livers were decellularized by detergent-enzymatic technique with (EDTA-DET) or without EDTA (DET). Histology, DNA quantification and proteomics confirmed decellularization with further DNA reduction with the addition of EDTA. Quantification, histology, immunostaining, and proteomics demonstrated preservation of extracellular matrix components in both scaffolds with a higher amount of collagen and glycosaminoglycans in the EDTA-DET scaffold. Scanning electron microscopy and X-ray phase contrast imaging showed microarchitecture preservation, with EDTA-DET scaffolds more tightly packed. DET scaffold seeding with a hepatocellular cell line demonstrated complete repopulation in 14 days, with cells proliferating at that time. Decellularization using DET preserves microarchitecture and extracellular matrix components whilst allowing for cell growth for up to 14 days. Addition of EDTA creates a denser, more compact matrix. Transplantation of the scaffolds and scaling up of the methodology are the next steps for successful hepatic tissue engineering.