Fibrosis Stage Research Articles

Design and evaluation of collagenase-loaded nanoparticles for mechanical intervention of orthotopic hepatocellular carcinoma in rat model

Altered tissue mechanics is vital for cancer development and malignancy, whether targeting the mechanical microenvironment could retard the initiation and progression of tumor is less explored. In this study, an orthotopic hepatocellular carcinoma (HCC) rat model was constructed to reproduce the mechanical and pathophysiological microenvironment of HCC development. LpMSN@CLG, a liver-targeted (L) and pH-sensitive (p) mesoporous silica nanoparticle (MSN) encapsulated with collagenase type-I (CLG), and DOX-LpMSN@CLG, on the basis of LpMSN@CLG, encapsulated with CLG and doxorubicin (DOX), were prepared to reduce matrix stiffness by degrading collagen in liver and HCC tumor. LpMSN@CLG, and DOX-LpMSN@CLG were respectively injected (i.v.) into rats at the stage of fibrosis and HCC, resulting in decreased collagen content in liver and HCC tissue, as well as reduced matrix stiffness. In addition, LpMSN@CLG treatment at the fibrosis stage retarded HCC initiation, and DOX-LpMSN@CLG treatment inhibited the growth of HCC tumor when compared with that of the rats treated by DOX alone, suggesting that reducing matrix stiffness in HCC tumor can improve the therapeutic efficacy of DOX. Taken together, our study demonstrated that mechanical intervention of tissue stiffness by CLG-loaded nanoparticles could retard the initiation and progression of HCC, suggesting the promising of mechanical intervention of hepatocellular carcinoma.

Exertional Desaturation and Ambulatory Oxygen Therapy Requirements in People with Idiopathic Pulmonary Fibrosis. A Retrospective Study

Introduction. Exertional desaturation (ED) is common in advanced idiopathic pulmonary fibrosis (IPF) stages. Ambulatory O2 therapy could increase physical activity by preventing ED in people with IPF. Objective. This study aimed to assess ED and ambulatory O2 requirements in people with IPF using a protocol that involved up to four 6-minute walking tests (6MWT). Method. An observational study of a dynamic retrospective cohort from a high-complexity hospital was conducted. The ambulatory O2 requirement assessment protocol involves performing up to four 6MWT depending on ED. All participants performed the baseline test (no additional O2). If ED (SpO2 < 90%) was observed, up to three additional 6MWTs were performed with two, four, and six O2 liters/minute until ED was avoided. Results. Twenty-eight patients (16 female; mean age 73 years) were referred for assessment of ambulatory O2 requirements. Twenty-three (82%) had ED during baseline 6MWT. Twenty-two patients performed the 6MWT with two liters/minute of O2, ten performed a third 6MWT with four liters/minute of O2, and seven with six liters/minute of O2. The six participants who performed all four 6MWTs significantly increased their walking distance by 56.33 meters (SD 36.45) compared to the baseline (p = 0.001). Four of the seven patients (57.14%) who performed the last 6MWT had ED despite O2 supplementation. Conclusion. The prevalence of ED during baseline 6MWT was high. Some participants even experienced ED with six liters/minute of supplemental O2. Despite this, walking distance increased more than the learning effect.

Non-radiomics imaging (US-CEUS) features and clinical text features: correlation with microvascular invasion and tumor grading in hepatocellular carcinoma.

To predict microvascular invasion (MVI) status and tumor grading of hepatocellular carcinoma (HCC) by evaluating preoperative non-radiomics ultrasound and contrast-enhanced ultrasound (US-CEUS) features and determine the influences of MVI/tumor grading on the category of CEUS LI-RADS for HCC. A total of 506 HCC patients who underwent preoperative US-CEUS examinations from 8 hospitals between July 2020 and June 2023 were enrolled. According to the MVI status, all the patients were classified, and HCC differentiation was assessed by using Edmondson-Steiner (ES) grading: MVI-negative (M0) and low-grade ES (GI/II) (MN-L, n = 297) and MVI-positive (M1/M2) and/or high-grade ES (GIII/IV) (MP-H, n = 209). Stratified analysis was performed based on fibrosis stage and tumor size. The results proved that MN-L HCC was more frequently classified into the LR-5 category (p = 0.034), while MP-H HCC was more frequently classified into the LR-TIV (p = 0.010). The heterogeneously arterial phase hyperenhancement (APHE) is significantly correlated with MVI(+)/high grade-ES (p = 0.003). Compared with MN-L HCC, the onset of washout was earlier, washout rate was higher, and tumor-invasion border was larger (all p < 0.01) in MP-H HCC. In addition, fibrosis stage and tumor size significantly influenced the onset of washout and washout rate of HCC (all p < 0.01). The tumor-invasion border was only positively correlated with tumor size (p < 0.001) rather than fibrosis stage (p > 0.05). MVI status and tumor grading influence the classification of LR-5 and LR-TIV. Heterogeneous APHE, higher washout rate, earlier onset of washout (≤65s), larger tumor-invasion border (≥3mm) and higher alpha fetoprotein level indicate the presence of MVI and/or high-grade ES.

Morphological Changes of Liver Among Post-Fontan Surgery Patients

Abstract Purpose Liver screening and longitudinal study of Fontan Associated Liver Diseases (FALD) is essential to identifying hepatomegaly and how hepatomegaly relates to various stages of liver fibrosis. In this study, we investigated longitudinal liver shape changes and liver stiffness in a cohort of patients with Fontan Associated Liver Disease. Methods We used 170 image volumes of 40 Fontan stage 3 completion patients. We also used 65 computed tomography images of healthy individuals from three datasets for comparison. Thirteen radiomic shape features of Fontan patients and individuals with a healthy liver were extracted and analyzed longitudinally. We studied correlations among features, liver spleen ratio, and liver stiffness with shape features. Results The enlargement of the liver, along with all shape features, was observed in all post-surgery intervals related to hepatomegaly and fibrosis. The shape features of healthy individuals and Fontan cases differ significantly in the longitudinal analysis and in the liver-spleen ratio. There is a positive correlation among body mass index, body surface area, age, Fontan surgery years, and liver stiffness. Conclusion The changes in shape features between Fontan patients and healthy subjects are statistically significant, which shows the relation for hepatomegaly and liver fibrosis. Accurate delineation of these features with artificial intelligence-based segmentation could serve as a valuable adjunct for the clinical follow-up of Fontan patients.

Utility of AI digital pathology as an aid for pathologists scoring fibrosis in MASH

Background & AimsIntra and inter-pathologist variability poses a significant challenge in metabolic dysfunction-associated steatohepatitis (MASH) biopsy evaluation, leading to suboptimal selection of patients and confounded assessment of histological response in clinical trials. We evaluated the utility of an artificial intelligence (AI) digital pathology (DP) platform to aid pathologists improve the reliability of fibrosis staging. MethodsA total of 120 digitized histology slides from two trials (NCT03517540, NCT03912532) were analysed by four expert hepatopathologists, with and without AI-assistance in a randomized, cross-over design. We utilized the HistoIndex AI DP platform, consisting of unstained second harmonic generation/two photon excitation fluorescence (SHG/TPEF) images and AI quantitative fibrosis (qF) values. ResultsAI-assistance significantly improved inter-pathologist kappa for fibrosis (F)-staging, particularly for early fibrosis (F0-F2), with reduced variance around the median reads. Intra-pathologist kappa was unchanged. AI-assistance increased pathologist concordance for identifying clinical trial inclusion subjects (F2-F3) from 45% to 71%, exclusion subjects (F0/F1/F4) from 38% to 55%, and evaluation of fibrosis response to treatment from 49% to 61%. SHG/TPEF images, qFibrosis continuous values, and qF-stage were considered useful by at least 3 out of 4 pathologists in 83%, 55%, and 38% cases, respectively. In the context of a clinical trial, the increase in inter-pathologist concordance in this study is modeled to result in a ∼25% reduction in the potential need for adjudication as well as a ∼50% increase in the study power. ConclusionsThe use of AI DP enhances inter-rater reliability of fibrosis staging for MASH. This indicates that the SHG/TPEF-based AI DP tool is useful for assisting pathologists in assessing fibrosis, thereby enhancing clinical trial efficiency and reliability of fibrosis readouts in response to treatments.

Heterogeneous development of liver fibrosis in chronic hepatitis C patients; assessment by extracellular volume fraction map generated from routine clinical CT data

ObjectivesTo clarify the heterogeneous development of liver fibrosis in patients with chronic hepatitis C (CHC) using extracellular volume fraction (ECV) map obtained from routine clinical CT data. MethodsBetween November 2012 and July 2020, patients with CHC were retrospectively recruited who had undergone four-phase CT and MR elastography (MRE) within one year. Patients were divided into 4 grades to represent different cirrhotic/fibrotic stage, using two different methods; one based on liver stiffness measured by MRE (MRE model), and the other by mALBI grades (mALBI model). Liver was anatomically divided into 16 sections, namely peripheral and central areas of each segment. ECV map was generated according to the previously reported method, and ECV was measured for the 16 sections. Estimated pathological fibrosis grade was assigned for each section based on the previously reported data. ResultsThere were 150 patients available. In each anatomical section, ECV significantly increases as cirrhotic /fibrotic stage progresses. The peripheral areas of segments 4,5 and 8 were the earliest to show F2 or F3-equivalent ECV (p < 0.05), followed by central areas or other segments. The central areas of segments 6 and 7 were the last to be involved by fibrosis both in MRE and mALBI models, finally almost all sections showing F4-equivalent ECV at the end stage fibrosis. ConclusionFibrosis starts at the peripheral areas of segments 4, 5, and 8, and spreads towards other parts of the liver, with the central areas of segments 6 and 7 being the last, in patients with CHC.

Inhaled exogenous thymosin beta 4 suppresses bleomycin-induced pulmonary fibrosis in mice via TGF-β1 signalling pathway.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and fibrotic interstitial lung disease. The two drugs indicated for IPF have limited efficacy and there is an urgent need to develop new drugs. Thymosin β4 (Tβ4) is a natural endogenous repair factor whose antifibrotic effects have been reported. This study aimed to evaluate the effect of exogenous recombinant human thymosin beta 4 (rhTβ4) on pulmonary fibrosis. Pulmonary fibrosis was induced in mice with bleomycin, and rhTβ4 was administrated by nebulization following three strategies: early dosing, mid-term dosing, and late dosing. The rhTβ4 efficacy was assessed by hydroxyproline, lung function, and lung histopathology. In vitro, the effects of rhTβ4 on fibroblast and lung epithelial cell phenotypes, as well as the TGF-β1 pathway, were evaluated. Aerosol administration of rhTβ4 could alleviate bleomycin-induced pulmonary fibrosis in mice at different stages of fibrosis. Studies conducted in vitro suggested that rhTβ4 could suppress lung fibroblasts from proliferating, migrating, and activation via regulating the TGF-β1 signalling pathway. In vitro, rhTβ4 also inhibited the epithelial-mesenchymal transition-like process of pulmonary epithelial cells. This study suggests that nebulized rhTβ4 is a potential treatment for IPF.

Evolution of characteristics of MASLD with and without diabetes: a meta-analysis of placebo arms

BackgroundWe explored the changes in metabolic dysfunction-associated steatotic liver disease (MASLD) severity over time by analyzing data from the placebo arms of randomized controlled trials (RCTs), focusing on the presence of diabetes.MethodsRCTs on MASLD that included a placebo arm were identified using a systematic search of the literature. Primary outcomes were changes in hepatic steatosis and fibrosis.ResultsThe meta-analysis included 8 RCTs involving 386 patients without diabetes and 24 RCTs involving 637 patients with diabetes. The pooled estimate of mean change in steatosis grade was − 0.1 in patients without diabetes, and − 0.37 in patients with diabetes (P = 0.066). The mean change in fibrosis stage was 0.05 in patients without diabetes, and − 0.03 in patients with diabetes (P = 0.359). The mean change in nonalcoholic fatty liver disease activity score was − 0.55 in patients without diabetes, and − 1.50 in patients with diabetes (P = 0.100). The mean change in ALT and AST were significantly larger in patients without diabetes compared to those with diabetes (P < 0.05).ConclusionPlacebo treatment had a greater effect in improving liver steatosis in patients with diabetes compared to those without. These findings highlight the importance of tailored treatment strategies in MASLD, particularly considering diabetes status.

Manganese-based type I collagen-targeting MRI probe for in vivo imaging of liver fibrosis.

Liver fibrosis is a common pathway shared by all forms of progressive chronic liver disease. There is an unmet clinical need for noninvasive imaging tools to diagnose and stage fibrosis, which presently relies heavily on percutaneous liver biopsy. Here we explored the feasibility of using a novel type I collagen-targeted manganese (Mn)-based MRI probe, Mn-CBP20, for liver fibrosis imaging. In vitro characterization of Mn-CBP20 demonstrated its high binding affinity for human collagen (Kd = 9.6 µM), high T1-relaxivity (48.9 mM-1s-1 at 1.4T and 27°C), and kinetic inertness to Mn release under forcing conditions. We demonstrated MRI using Mn-CBP20 performs comparably to previously reported gadolinium-based type I collagen-targeted probe EP-3533 in a mouse model of carbon tetrachloride-induced liver fibrosis, and further demonstrate efficacy to detect fibrosis in a diet-induced mouse model of metabolically-associated steatohepatitis. Biodistribution studies using the Mn-CBP20 radio-labeled with the positron-emitting 52Mn isotope demonstrate efficient clearance of Mn-CBP20 primarily via renal excretion. Mn-CBP20 represents a promising candidate that merits further evaluation and development for molecular imaging of liver fibrosis.

Predictors of Atherosclerotic Cardiovascular Disease Events in Patients with Metabolic Dysfunction-associated Steatotic Liver Disease.

Objective Metabolic dysfunction-associated steatotic liver disease (MASLD) is a chronic liver disease associated with metabolic comorbidities. However, the risk factors for atherosclerotic cardiovascular disease (ASCVD) in these patients remain unclear. Therefore, this study investigated predictors of ASCVD in patients with MASLD. Methods This single-center retrospective study examined 372 patients with MASLD ≥40 years old with liver biopsies and available Hisayama scores (median follow-up, 7.4 years; range, 0.6-22 years). We compared baseline characteristics, liver histology (stage, lobular inflammation, steatosis, and hepatocellular ballooning), fibrosis-4 (FIB-4) index (<1.3/1.3-2.66/≥2.67), and laboratory data between patients with and without ASCVD. A predictive model for the onset of ASCVD based on the Hisayama score (low/intermediate/high) and ASCVD incidence was evaluated according to the liver fibrosis stage and FIB-4 index. Results ASCVD incidence was 11.1/1,000 person-years, with cumulative incidences of 4.4%, 9.0%, 14%, and 32% at 5, 10, 15, and 20 years, respectively. Regarding the incidence of ASCVD, the liver fibrosis stage and an FIB-4 index ≥2.67 were not significant predictors, but type IV collagen 7S was a significant predictor. The incidence of ASCVD was higher in the intermediate- and high-risk Hisayama score groups than in the low-risk group. In the multivariate Cox proportional hazards model, the Hisayama score and type IV collagen 7S predicted the incidence of ASCVD more accurately than an FIB-4 index ≥2.67. Conclusions The Hisayama score predicted ASCVD risk in patients with MASLD. These findings will help predict and improve the prognosis of MASLD.

How far are we? Assessing progress in hepatitis C response towards the WHO 2030 elimination goals by the civil society monitoring in 25 European countries, period 2020 to 2023

BackgroundWith the advent of direct acting antivirals (DAAs) the World Health Organisation (WHO) adopted global strategy to eliminate hepatitis C virus (HCV) infection by 2030. In Europe, people who inject drugs (PWID) account for the majority of new cases, however testing and treatment remain suboptimal. The aim was to monitor progress in HCV policy and cascade-of-care for PWID, led by the civil society organisations (CSO) that provide harm reduction services for PWID across Europe.MethodsIn period 2020–2023, CSOs representing focal points of Correlation-European Harm Reduction Network were annually invited to complete online questionnaire on use/impact of HCV test-and-treat guidelines for PWID, availability/functioning of continuum-of-care, and role/limitations of harm reduction services for PWID. A retrospective longitudinal analysis of responses to questions answered each year by the same respondents was performed, and a comparison among the studied years was made.ResultsTwenty-five CSOs from cities in 25 European countries were included and responded to 25 questions. Between 2020 and 2023, there was positive trend in number of HCV treatment guidelines, separate guidelines for PWID, and their positive impact on acess to testing/treatment (24/25, 5/25, and 16/25 in 2023, respectively). DAAs were available in all countries, predominantly prescribed by specialist physicians only (slight increase at primary care), with restrictions including active drug use, stage of liver fibrosis or/and reimbursement policies (2/25, 4/25, and 3/25 in 2023, respectively). A decrease in HCV testing sites was noted. Treatment was consistently most common at clinical settings, however an increase outside the specialist settings was detected, particularly in prisons (12/25 and 15/25 in 2020–2021, respectively). Comparing 2022–2023, number of HCV-testing services increased in many cities with positive dynamic in nearly all the settings; increase in treatment at harm reduction services/community centres was noted (6/25 to 8/25, respectively). Between 2020 and 2023 the frequency of various limitations to CSOs addressing HCV was oscillating, presenting an increase between 2022 and 2023 (9/25 to 14/25, respectively).ConclusionThe overall progress towards WHO HCV elimination goals across Europe remains insufficient, most probably also due to the influence of Covid-19 pandemic. Further improvements are needed, also by including CSOs for PWID in continuum-of-care services, and in monitoring progress.

Association between advanced fibrosis and epigenetic age acceleration among individuals with MASLD.

The biological process of aging plays an important role in the progress of liver fibrosis. However, epidemiological evidence about the associations between advanced fibrosis and epigenetic age acceleration (EAA) among individuals with metabolic dysfunction-associated steatotic liver disease (MASLD) is limited. We utilized publicly available DNA methylation data (GSE180474) for our analysis. Five EAA measures were calculated in this study, including IEAA, PhenoAA, GrimAA, DunedinPACE, and DNAmTLAA. Separate linear regression models were conducted to explore the associations between different fibrosis grades and each measure of EAA. A total of 325 participants were included in this study, with a mean (± SD) age of 48.56 ± 11.50years. Of these participants, 64.6% with no fibrosis, 16.9% with bridging fibrosis, 11.1% with incomplete cirrhosis, and 7.4% with cirrhosis. After adjusting for demographics and medication status, MASLD individuals with advanced fibrosis were associated with a 5% increase in the pace of aging (DunedinPACE, β = 0.05, 95% CI: 0.03-0.07) and a 10% decrease in DNAmTLAA (β = -0.10, 95% CI: -0.13 to -0.07) compared those without fibrosis. Similarly, higher stages of fibrosis were associated with an increased pace of aging (DunedinPACE, β = 0.02, 95% CI: 0.01-0.03, Ptrend < 0.001) and decreased DNAmTLAA (β = -0.05, 95% CI: -0.07 to -0.04, Ptrend < 0.001). However, no significant association was found between advanced fibrosis and IEAA, PhenoAA, and GrimAA. Our findings suggest that advanced fibrosis was associated with an accelerated pace of aging, as measured by the third-generation EA measure DunedinPACE, and shorter telomere length, captured by DNAmTLAA, among individuals with MASLD. This finding has potential prognostic implications and suggests EAA may serve as a surrogate marker of therapeutic efficacy in MASLD.

Application of the Chinese Visceral Adiposity Index in the Diagnosis of Hepatic Steatosis and Liver Fibrosis: A Cross-Sectional Study

Background: Visceral fat exerts a significant impact on the histopathology of metabolic dysfunction-associated steatotic liver disease (MASLD). Hence, we evaluated the application of the Chinese Visceral Adiposity Index (CVAI), a novel visceral fat marker, in predicting and diagnosing hepatic steatosis and liver fibrosis. Methods: After excluding individuals with liver disease or significant alcohol consumption, a total of 273 patients (57.9% males) aged over 18 were eligible for this cross-sectional study. Chinese Visceral Adiposity Index was calculated by age, Body Mass Index (BMI), waist circumference )WC(, triglyceride, and high-density lipoprotein cholesterol (HDL-C). Hepatic steatosis and fibrosis were measured by elastography. A P-value of less than 0.05 was considered significant. Results: After adjusting for age, gender, metabolic diseases, and visceral obesity indices, each unit increase in CVAI elevated the odds of hepatic steatosis and liver fibrosis by 3.3% (P = 0.002) and 2.9% (P = 0.001), respectively. According to the analysis of variance (ANOVA), the grades of liver steatosis had a great impact on CVAI (P &lt; 0.001), and CVAI well-discriminated grades of liver steatosis. Furthermore, the stages of liver fibrosis greatly impacted CVAI (P &lt; 0.001), but it cannot distinguish the stages of liver fibrosis. The Area Under the Receiver Operating Characteristic Curve (AUROC) for S ≥ S1 was 0.800 (0.747 to 0.846), and for F ≥ F2 was 0.810 (0.759 to 0.855). Conclusions: We concluded that CVAI is a strong predictor and accurate diagnostic factor for liver steatosis and significant fibrosis; thus, it could be a tool for screening individuals at risk of liver steatosis and the irreversible complications of fibrosis.

Artificial Intelligence and Image Analysis-Assisted Diagnosis for Fibrosis Stage of Metabolic Dysfunction-Associated Steatotic Liver Disease Using Ultrasonography: A Pilot Study.

Elastography increased the diagnostic accuracy of liver fibrosis. However, several challenges persist, including the widespread utilization of equipment, difficulties in measuring certain cases, and the influence of viscosity factors. A rough surface and a blunted hepatic margin have long been acknowledged as valuable characteristics indicative of hepatic fibrosis. The objective of this study was to conduct an image analysis and quantitative assessment of the contour of the sagittal section of the left lobe of the liver. Between February and October 2020, 486 consecutive outpatients underwent ultrasound examinations at our hospital. A total of 214 images were manually annotated by delineating the liver contour to create annotation images. U-Net was employed for liver segmentation, with the dataset divided into training (n = 128), testing (n = 42), and validation (n = 44) subsets. Additionally, 43 Metabolic Dysfunction Associated Steatotic Liver Disease (MASLD) cases with pathology data from between 2015 and 2020 were included. Segmentation was performed using the program developed in the first step. Subsequently, shape analysis was conducted using ImageJ. Liver segmentation exhibited high accuracy, as indicated by Dice loss of 0.044, Intersection over Union of 0.935, and an F score of 0.966. The accuracy of the classification of the liver surface as smooth or rough via ResNet 50 was 84.6%. Image analysis showed MinFeret and Minor correlated with liver fibrosis stage (p = 0.046, 0.036, respectively). Sensitivity, specificity, and AUROC of Minor for ≥F3 were 0.571, 0.862, and 0.722, respectively, and F4 were 1, 0.600, and 0.825, respectively. Deep learning segmentation of the sagittal cross-sectional contour of the left lobe of the liver demonstrated commendable accuracy. The roughness of the liver surface was correctly judged by artificial intelligence. Image analysis showed the thickness of the left lobe inversely correlated with liver fibrosis stage.

CD206+ macrophages are relevant non-invasive imaging biomarkers and therapeutic targets in experimental lung fibrosis

BackgroundInterstitial lung diseases (ILDs) include a large number of diseases associated with progressive pulmonary fibrosis (PPF), including idiopathic pulmonary fibrosis (IPF). Despite the rarity of each of the fibrotic ILDs...

Clinical- and Cost-Effectiveness of Liver Disease Staging in Hepatitis C Virus Infection: A Microsimulation Study.

Liver disease assessment is a key aspect of chronic hepatitis C virus (HCV) infection pre-treatment evaluation but guidelines differ on the optimal testing modality given trade-offs in availability and accuracy. We compared clinical outcomes and cost-effectiveness of common fibrosis staging strategies. We simulated adults with chronic HCV receiving care at US health centers through a lifetime microsimulation across five strategies: (1) no staging or treatment (comparator), (2) indirect serum biomarker testing (Fibrosis-4 index [FIB-4]) only, (3) transient elastography (TE) only, (4) staged approach: FIB-4 for all, TE only for intermediate FIB-4 scores (1.45-3.25), and (5) both tests for all. Outcomes included infections cured, cirrhosis cases, liver-related deaths, costs, quality-adjusted life years (QALYs), and incremental cost-effectiveness ratios (ICERs). We used literature-informed loss to follow-up (LTFU) rates and 2021 Medicaid perspective and costs. FIB-4 alone generated the best clinical outcomes: 87.7% cured, 8.7% developed cirrhosis, and 4.6% had liver-related deaths. TE strategies cured 58.5%-76.6%, 16.8%-29.4% developed cirrhosis, and 11.6%-22.6% had liver-related deaths. All TE strategies yielded worse clinical outcomes at higher costs per QALY than FIB-4 only, which had an ICER of $12 869 per QALY gained compared with no staging or treatment. LTFU drove these findings: TE strategies were only cost-effective with no LTFU. In a point-of-care HCV test-and-treat scenario, treatment without any staging was most clinically and cost-effective. FIB-4 staging alone resulted in optimal clinical outcomes and was cost-effective. Treatment for chronic HCV should not be delayed while awaiting fibrosis staging with TE.

Non-invasive testing in metabolic dysfunction-associated steatotic liver disease.

Metabolic dysfunction-associated steatotic liver disease (MASLD), previously referred to as non-alcoholic fatty liver disease (NAFLD), is a leading cause of chronic liver disease, affecting up to 30% of the global population. MASLD is strongly associated with metabolic risk factors such as obesity and type 2 diabetes, and can progress to advanced stages including cirrhosis and hepatocellular carcinoma. Early diagnosis and accurate staging of fibrosis are critical in managing the disease and preventing complications. While liver biopsy has long been considered the gold standard for assessing fibrosis, it is invasive and carries associated risks. In response, non-invasive tests (NITs) have emerged as essential alternatives for the diagnosis and monitoring of MASLD. Key methods include blood-based biomarkers such as the Fibrosis-4 (FIB-4) score, NAFLD Fibrosis Score (NFS), and Enhanced Liver Fibrosis (ELF) test, as well as imaging modalities like vibration-controlled transient elastography (VCTE) and magnetic resonance elastography (MRE). These tests provide safer, more accessible methods for identifying liver fibrosis and guiding clinical management. They are integral in assessing disease severity, guiding treatment decisions, and monitoring disease progression, particularly in light of emerging therapies. NITs have become increasingly recommended by clinical guidelines as they reduce the need for invasive procedures like liver biopsy, improving patient care and outcomes. In conclusion, non-invasive testing plays a crucial role in the effective management of MASLD, offering reliable alternatives for diagnosis and monitoring while minimizing risks associated with traditional invasive methods.

Mathematical model of liver cirrhosis formation during morphological and molecular-genetic preclinical studies

BACKGROUND: Currently, researchers describe challenges in developing new treatments for fibrosis and cirrhosis: poor quality of preclinical models, insufficient trial duration, and lack of markers of therapeutic response. A separate task is to standardize the process of liver cirrhosis formation in preclinical trials, which is necessary to obtain accurate quantitative estimates in a short timeframe. AIM: This study aimed to develop a mathematical model for the formation of liver cirrhosis during preclinical trials. MATERIALS AND METHODS: Liver fibrosis and cirrhosis were induced in male Wistar rats using freshly prepared thioacetamide solution for 17 weeks. The area of connective tissue was determined as a percentage of the image area. The area of interlobular veins was measured in µm2. The numbers of cells expressing the FAP marker and the α-SMA marker were counted. The level of mRNA expression of the Vegfa and Yap1 genes was assessed by real-time polymerase chain reaction. A mathematical model for classifying observations into stages was constructed using multiple logistic regression with stepwise selection of predictors, followed by calculation of sensitivity, specificity, and area under the curve with a 95% confidence interval based on ROC analysis. RESULTS: As a result of the analysis, a mathematical model of liver cirrhosis formation was developed. The model is based on the values of two indicators: FAP+ cells and Yap1 mRNA and demonstrated good quality. The resulting value of the area under the ROC curve of 0.883 suggests good results for classifying cases. CONCLUSIONS: The mathematical model makes it possible to differentiate the stage of liver cirrhosis from the stage of fibrosis during preclinical studies. It provides a foundation for studying the pathogenesis of liver fibrosis and cirrhosis, identifying new potential molecular targets for antifibrotic therapy, and reducing the number of expensive, labor-intensive laboratory tests.

Significant Within-Individual Variability in VCTE Liver Stiffness Measurements at Two Intercostal Spaces in Subjects with MASLD: Implications for Evaluating Improvement in Liver Fibrosis After Weight-Loss or Liver-Directed Therapy.

Studies have compared the group-averages of liver stiffness measures (LSMs) from multiple rib spaces by vibration-controlled transient elastography (VCTE) to stage liver fibrosis. No previous study has assessed within-individual liver stiffness variation from two rib spaces in individuals with metabolic-dysfunction associated steatotic liver disease (MASLD). We evaluated within-individual LSM variation according to body weight classification and its clinical implication. From October 2019 to March 2024, VCTE was performed on MASLD patients or those at high risk, in accordance with FibroScan guidelines. The LSMs were categorized into stages: <5 kPa (stage 0), 5-7.99 kPa (stage 1), 8-9.99 kPa (stage 2), 10-13.99 kPa (stage 3), and 14+ kPa (stage 4). Measurements with 10 values and IQR/median ≤ 0.30 were included, using SPSS V25.0 for analysis. Among 1107 subjects (age 54.4 ± 13.9 years, 56.9% female), 7.7% were normal weight, 20.7% overweight, 28.9% class 1 obesity, 21.3% class 2 obesity, and 21.2% class 3 obesity. Significant within-individual variation was noted: 67% (0-2 kPa) variation, 23.4% (2.1-6 kPa), and 10% (≥6.1 kPa). Class 3 obese individuals had the maximum variation. Comparing the group-average of LSM at each ICS site showed that 95% of individuals were within one fibrosis stage. While LSM group-averages at different rib sites provides reliable fibrosis staging, significant within-individual variability exists especially in class 3 obesity. This should be considered when serial LSM assessments are used to assess medical therapeutic efficacy.

Liver transcriptome analysis reveals PSC-attributed gene set associated with fibrosis progression

BackgroundPrimary Sclerosing Cholangitis (PSC) is a chronic heterogenous cholangiopathy with unknown etiology where chronic inflammation of the bile ducts leads to multifocal biliary strictures and biliary fibrosis with consecutive cirrhosis development. We here aimed to identify a PSC-specific gene signature associated with biliary fibrosis development. MethodsWe performed RNA-sequencing of 47 liver biopsies from people with PSC (n = 16), primary biliary cholangitis (PBC, n = 15), and metabolic dysfunction-associated steatotic liver disease (MASLD, n = 16) with different fibrosis stages to identify a PSC-specific gene signature associated with biliary fibrosis progression. For validation, we compared an external transcriptome data set of liver biopsies from people with PSC (n = 73) with different fibrosis stages (DOI: 10.1002/hep.31488; baseline samples from NCT01672853). ResultsDifferential gene expression analysis of the liver transcriptome from PSC patients with advanced versus early fibrosis revealed 431 genes associated with fibrosis development. Of those, 367 were identified as PSC-associated when compared to PBC or MASLD. Validation against an external data set of 73 liver biopsies from PSC patients with different fibrosis stages led to a condensed set of 150 (out of 367) DEGs. Cell type specificity assignment of those genes by using published single cell RNA-seq data revealed genetic disease drivers expressed by cholangiocytes (e.g. CXCL1, SPP1), fibroblasts, innate and adaptive immune cells while deconvolution along fibrosis progression of the PSC, PBC, MASLD samples highlighted an early involvement of macrophage- and neutrophil-associated genes in PSC fibrosis. ConclusionWe reveal a PSC-attributed gene signature associated with biliary fibrosis development that may enable the identification of potential new biomarkers and therapeutic targets in PSC-related fibrogenesis. IMPACT AND IMPLICATIONSPrimary sclerosing cholangitis (PSC) is an inflammatory liver disease that is characterized by multifocal inflammation of bile ducts and subsequent biliary fibrosis. Herein, we identify a PSC-specific gene set of biliary fibrosis progression attributing to a uniquely complex milieu of different cell types, including innate and adaptive immune cells while neutrophils and macrophages showed an earlier involvement in fibrosis initiation in PSC in contrast to PBC and MASLD. Thus, our unbiased approach lays an important groundwork for further mechanistic studies for research into PSC-specific fibrosis.