Stages Of Nonalcoholic Steatohepatitis Research Articles

Correction of the clinical course of non-alcoholic steatohepatitis and diabetic kidney disease in patients with type 2 diabetes

Background. The relevance of finding optimal methods to treat patients with a comorbid non-alcoholic steatohepatitis (NASH) that developed against the background of type 2 diabetes mellitus (T2DM) is due to the fact that these diseases have a number of common cause-and-effect mechanisms, and if diabetic kidney disease (DKD) develops, also mutual burden mechanisms. The purpose of the study was to find out the possible influence of a combination of metformin, rosuvastatin, essential phospholipids and quercetin on the clinical course of non-alcoholic steatohepatitis, diabetic kidney disease, type 2 diabetes mellitus, as well as on the state of the blood lipids, parameters of carbohydrate metabolism compensation, the degree of insulin resistance, which are factors for the progression of NASH and diabetic kidney disease. Materials and methods. Studies were conducted on the dynamics of treatment in 60 patients with NASH, T2DM and DKD stage I–III: 48 (80.0 %) of them had mild NASH, and 12 (20.0 %) had moderate NASH. A comorbid disease in 100 % of patients was moderate type 2 diabetes: 15 (25.0 %) people were diagnosed with diabetes in the stage of compensation, 45 (75.0 %) had subcompensated disease. Results. The positive effect of quercetin was noted by us in relation to the content of low-density lipoprotein cholesterol in the blood that was increased by 1.8 times (p < 0.05) before the treatment: a decrease after it was 1.7 times (p < 0.05) in group 2 and 1.3 times (p < 0.05) in group 1. Comprehensive therapy with the inclusion of quercetin contributed to a probable increase in anti-atherogenic high-density lipoprotein (by 1.3 times, p < 0.05) with the normalization of the indicator after the treatment, while traditional therapy in this contingent did not lead to any probable changes. Conclusions. The combination therapy for type 2 diabetes mellitus and NASH with the addition of quercetin contributed to the elimination of the main clinical and laboratory symptoms of NASH exacerbation, a probable reduction in the liver inflammation (a decrease in markers of cytolysis, mesenchymal inflammation), reversal of hepatic steatosis due to the optimization of cholesterol and triacylglycerols in the blood, a probable increase in high-density lipoproteins, normalization of glycemia, reduction of insulinemia, a decrease in the degree of insulin resistance. The effectiveness of treatment for DKD was also increased: the rate of proteinuria and the degree of hypercreatinemia decreased, and the glomerular filtration rate increased.

Diagnostic accuracy of FibroScan-AST (FAST) score, non-alcoholic fatty liver fibrosis score (NFS), FibroScan, and liver fibrosis index (FIB-4) for identifying fibrotic non-alcoholic steatohepatitis in patients with chronic hepatitis B with metabolic dysfunction-associated fatty liver disease

Objective To evaluate the diagnostic value of the FibroScan-AST (FAST) score, non-alcoholic fatty liver fibrosis score (NFS), FibroScan, and liver fibrosis index (FIB-4) for identifying fibrotic non-alcoholic steatohepatitis (NASH) in patients with chronic hepatitis B (CHB) with metabolic dysfunction-associated fatty liver disease (MAFLD). Methods All patients with CHB and MAFLD who underwent liver biopsy at the Zhenjiang Third Hospital affiliated with Jiangsu University between August 2010 and December 2022 were included in the analysis. The diagnostic accuracy of FAST, NFS, FibroScan, and FIB-4 for diagnosing NASH and liver fibrosis were evaluated based on the area under the receiver-operating characteristic curve (AUC). Results A total of 156 patients with CHB combined with MAFLD were included, including 69 with NASH and fibrosis stage 2 or higher (NASH+F ≥ 2), and 16 with NASH and cirrhosis (NASH+F4). The AUC of FAST, NFS, liver stiffness measurement (LSM), and FIB-4 for diagnosing NASH+F ≥ 2 was 0.739 (p < 0.001), 0.643 (p = 0.006), 0.754 (p < 0.001), and 0.665 (p = 0.003), respectively. The specificity of FAST, NFS, LSM, and FIB-4 was 67%, 51.8%, 78.6% and 76.8%, respectively, and the sensitivity was 75%, 78.6%, 67.9%, and 53.6%, respectively. No significant differences were found between groups. The AUC of FAST, NFS, LSM, and FIB-4 for diagnosing NASH+F4 was 0.650 (p = 0.038), 0.725 (p = 0.001), 0.851 (p < 0.001), and 0.560 (p = 0.533), respectively. The specificity of the FAST, NFS, LSM, and FIB-4 was 55.9%, 50.0%, 71.6%, and 75.5%, respectively and the sensitivity was 80.0%, 100%, 100%, and 50.0%, respectively. The differences between AUCs of FIB-4 and FAST compared with LSM were 0.291 and 0.201, respectively (p < 0.05). Conclusion In patients with CHB combined with MAFLD, FAST did not have better accuracy than NFS and FIB-4 for predicting fibrotic NASH, whereas LSM had better accuracy than FAST and FIB-4.

Dynamic Liver Chip Based on Well-Coupled Microfluidics: An Accurate NASH Model for Drug Evaluation.

The convenient liver model in vitro recapitulating the hepatic functions, metabolism, and even steatohepatitis to perform the accurate drug evaluation is still challenging because of the unattainable hominine physiological microenvironment in vitro. Here, the progressed stages of nonalcoholic steatohepatitis (NASH) disease were precisely modeled to accurately evaluate the performance of antilipemic based on the dynamic liver chip adopting the well-coupled microfluidics, which well recapitulated the normal and steatohepatitis of liver in vitro. In brief, the mild nutrient flow and sufficient oxygen supply for parenchymal liver cells could be well supplied through the endothelial cells layer that mimicked the real physiological barrier of endothelium, while the loading of drugs might be obtained by directly adding drug into the running nutrient flow to mimic the intravenously administrable. The progressed degree of steatohepatitis could be directly reflected by the amount of intramyocellular lipid content (IMLC) of the HepG2 cell hepatocyte layer in wells that were induced by different concentrations of free fatty acids (FFA). To prove the concept of the liver chip in drug evaluation, an accurate assessment of the performance of firsocostat, the acetyl-CoA carboxylase (ACC) inhibitor of hepatic mitochondria of hepatocytes, was carried out. The subtle time dependence of firsocostat treatment to different progressed stages of NASH was clearly figured out. Therefore, we prospect the liver chip that adopted well-coupled microfluidics could be an accurate and standard liver model in vitro to carry out the antilipemic evaluation and screening, which significantly enlightens the drug evaluation by liver on chip in vitro.

Association between triglyceride glucose-body mass index and the staging of non-alcoholic steatohepatitis and fibrosis in patients with non-alcoholic fatty liver disease

Objective The objective of this study was to thoroughly investigate the clinical value of triglyceride glucose-body mass index (TyG-BMI) in patients diagnosed with non-alcoholic fatty liver disease (NAFLD). Specifically, we aimed to determine its association with non-alcoholic steatohepatitis (NASH) and the progression of liver fibrosis. Methods The study included 393 patients diagnosed with NAFLD after liver biopsy. The patients were divided into two distinct cohorts: a training cohort (N = 320) and a validation cohort (N = 73). The training cohort was further divided into four groups based on TyG-BMI quartiles. The clinical characteristics of the patients in each group were compared in detail, and the association between TyG-BMI and NASH, NAFLD Activity Score (NAS) ≥ 4, at-risk NASH, significant fibrosis, advanced fibrosis, and cirrhosis was analyzed using multiple models. Additionally, we generated receiver operating characteristic (ROC) curves to evaluate the predictive ability of TyG-BMI for NASH and fibrosis staging in patients with NAFLD. Results Patients with higher TyG-BMI values had a significantly higher prevalence of NASH, NAS ≥ 4, at-risk NASH, significant fibrosis, advanced fibrosis, and cirrhosis (all p < .05). TyG-BMI was an independent predictor of these diseases in both unadjusted and adjusted models (all p < .05). ROC curve analysis further revealed the excellent performance of TyG-BMI in predicting NASH, NAS ≥ 4, at-risk NASH, significant fibrosis, advanced fibrosis, and cirrhosis. The validation cohort yielded analogous results. Furthermore, we constructed three multivariate models of TyG-BMI in conjunction with elastography metrics, which demonstrated elevated diagnostic AUC values of 0.782, 0.792, 0.794, 0.785, 0.834, and 0.845, respectively. Conclusion This study confirms a significant association between insulin resistance and NAFLD, including at-risk NASH and fibrosis staging, as assessed using the TyG-BMI index. TyG-BMI and its associated multivariate models can be valuable noninvasive indicators for NAFLD diagnosis, risk stratification, and disease course monitoring.

Cardiovascular risk in US adults with nonalcoholic steatohepatitis (NASH) vs. matched non-NASH controls, National Health and Nutrition Examination Survey, 2017-2020.

NASH is considered a contributor to atherosclerotic cardiovascular disease (ASCVD) risk; however, its contribution beyond traditional risk factors for CVD, particularly diabetes, is less clearly understood. This study aimed to quantify the cardiovascular-event risk associated with NASH, independent of diabetes status. A cross-sectional analysis was conducted using the 2017-2020 NHANES pre-pandemic cycle. NASH was defined based on presence of steatosis without other causes of liver disease, and FibroScan+AST score from vibration-controlled transient elastography (VCTE). Significant fibrosis (stages F2-F4) was identified by liver stiffness measurement from VCTE. Predicted primary CV-event risk was estimated using both the Pooled Cohort Equations (PCE) and the Framingham Risk Score (FRS). NASH patients were matched with non-NASH controls on age, sex, race/ethnicity, and diabetes status. Weighted logistic regression was conducted, modeling elevated predicted CV risk (binary) as the dependent variable and indicators for NASH / fibrosis stages as independent variables. A sample of 125 NASH patients was matched with 2585 controls. NASH with significant fibrosis was associated with elevated predicted 10-year CV risk, although this association was only statistically significant in PCE analyses (odds ratio and 95% CI 2.34 [1.25, 4.36]). Analyses restricting to ages <65 years showed similar results, with associations of greater magnitude. Independent of diabetes, a significant association was observed between NASH with significant liver fibrosis and predicted primary CV-event risk in US adults, particularly for those <65. These findings suggest the importance of accounting for NASH and liver-fibrosis stage in predicting CV-event risk.

Analysis of Metabolic Changes in Endogenous Metabolites and Diagnostic Biomarkers for Various Diseases Using Liquid Chromatography and Mass Spectrometry.

Analysis of endogenous metabolites in various diseases is useful for searching diagnostic biomarkers and elucidating the molecular mechanisms of pathophysiology. The author and collaborators have developed some LC/tandem mass spectrometry (LC/MS/MS) methods for metabolites and applied them to disease-related samples. First, we identified urinary conjugated cholesterol metabolites and serum N-palmitoyl-O-phosphocholine serine as useful biomarkers for Niemann-Pick disease type C (NPC). For the purpose of intraoperative diagnosis of glioma patients, we developed the LC/MS/MS analysis methods for 2-hydroxyglutaric acid or cystine and found that they could be good differential biomarkers. For renal cell carcinoma, we searched for various biomarkers for early diagnosis, malignancy evaluation and recurrence prediction by global metabolome analysis and targeted LC/MS/MS analysis. In pathological analysis, we developed a simultaneous LC/MS/MS analysis method for 13 steroid hormones and applied it to NPC cells, we found 6 types of reductions in NPC model cells. For non-alcoholic steatohepatitis (NASH), model mice were prepared with special diet and plasma bile acids were measured, and as a result, hydrophilic bile acids were significantly increased. In addition, we developed an LC/MS/MS method for 17 sterols and analyzed liver cholesterol metabolites and found a decrease in phytosterols and cholesterol synthetic markers and an increase in non-enzymatic oxidative sterols in the pre-onset stage of NASH. We will continue to challenge themselves to add value to clinical practice based on cutting-edge analytical chemistry methodology.

Serum miR‑29 is increased in mice with early liver fibrosis.

Non-alcoholic steatohepatitis (NASH) is a fatty liver disease that is not caused by alcohol consumption and is characterized by fatty degeneration, inflammation and hepatocellular damage. Therefore, predicting future fibrosis is critical in the early stages of NASH to prevent disease progression. The present study examined histological changes in the liver as well as microRNA (miR/miRNA) expression changes in the liver and serum of NASH mice model to identify potential biomarker candidates that could predict early fibrosis. This study used 6-week-old C57BL/6NJcl male mice and fed the control with a standard solid diet (CE-2) for breeding and propagation and NASH groups with a high-fat diet [choline-deficient high-fat and 0.1% (w/v) methionine supplemented diet], respectively. Agilent Technologies miRNA microarray was used to investigate microRNA expression in the liver and serum. Hematoxylin and eosin staining of the livers of the NASH group mice during the second week of feeding revealed fatty degeneration, balloon-like degeneration and inflammatory cell infiltration, confirming that the mice were in a state of NASH. The livers of the NASH group mice at 6 weeks of feeding showed fibrosis. Microarray analysis revealed that miRNAs were upregulated and 47 miRNAs were downregulated in the liver of the NASH group. Pathway analysis using OmicsNet predicted miR-29 to target collagen genes. Furthermore, miR-29 was downregulated in the livers of NASH-induced mice but upregulated in serum. These findings suggested that lower miR-29 expression in NASH-induced liver would increase collagen expression and fibrosis. Early liver fibrosis suggests that miR-29 leaks from the liver into the bloodstream, and elevated serum miR-29 levels may be a predictive biomarker for early liver fibrosis.

Bofutsushosan (Fangfengtongshengsan) improves early stages of NASH via the gut–liver axis in diabetes-induced NASH model mice

BackgroundThe mechanisms underlying non-alcoholic fatty liver disease (NAFLD) are not fully understood, but the gut microbiota is deeply involved. Because the Japanese traditional medicine bofutsushosan (BTS; Fangfengtongshengsan in Chinese) is known to improve obesity-induced liver injury and increase gut Akkermansia muciniphila, we evaluated its effects in a mouse model of diabetes-induced nonalcoholic steatohepatitis (NASH). MethodsTwo-day-old male C57BL/6J mice were injected subcutaneously with streptozotocin and fed a high-fat diet (HFD) from 4 weeks. Between 5 and 8 weeks, HFD was supplemented with 5% BTS (BTS) or not supplemented (CONT), and plasma, liver, and stool samples were collected and analyzed. ResultsThe NAFLD activity score (NAS), which was consistent with early steatohepatitis in CONT mice, was significantly lower in BTS mice (P = 0.018). Improvement of steatosis contributed most to decreasing NAS. Relative abundances of probiotic Akkermansia muciniphila and Bifidobacterium pseudolongum were significantly higher in BTS mice (both P < 0.05). Positive correlations were observed among NAS, steatosis, ballooning, and body weight change. Liver macrophage markers correlated with some microbes in BTS mice. ConclusionsBTS administration suppressed liver steatosis and improved the gut microbiome, which correlated with some liver factors. BTS may be a promising treatment for NAFLD by maintaining the gut–liver axis.

Probiotic supplementation for 24 weeks in patients with non-alcoholic steatohepatitis: the PROBILIVER randomized clinical trial.

Considering the increasing prevalence of non-alcoholic steatohepatitis (NASH) and treatment gaps, this study aimed to evaluate the effect of probiotic supplementation on liver function markers, nutritional status, and clinical parameters. This double-blind, randomized clinical trial (ClinicalTrials.gov ID: NCT0346782) included adult outpatients with biopsy-proven NASH. The intervention consisted of 24 weeks of supplementation with the probiotic mix Lactobacillus acidophilus (1 × 109 CFU) + Lactobacillus rhamnosus (1 × 109 CFU) + Lactobacillus paracasei (1 × 109 CFU) + Bifidobacterium lactis (1 × 109 CFU), or placebo, twice a day. The following parameters were evaluated: demographic and clinical data, transient elastography (FibroScan), liver enzymes, NAFLD fibrosis score, fatty liver index, laboratory assessment, serum concentration of toll-like receptor-4 (sTLR-4) and cytokeratin 18 (CK-18), anthropometric data, dietary intake, and physical activity. Regarding data analysis, the comparison between the groups was based on the delta of the difference of each variable analyzed (value at the end of treatment minus the baseline value) using the t-test for independent samples or the Mann-Whitney U-test. Forty-four patients with NASH completed the trial (51.4 ± 11.6 years). At baseline, 87% of participants had a mild liver fibrosis degree on biopsy, normal values of liver enzymes, transient elastography values consistent with grade 1 fibrosis in both groups, increased waist circumference (WC), a BMI of 30.97 kg/m2, and 76% presented with metabolic syndrome (MetS). After the intervention, no differences were observed between the probiotic and placebo groups in terms of MetS, WC, BMI scores, or liver enzyme levels (p > 0.05 for all). The elastography values remained consistent with grade 1 fibrosis in both groups. Although CK-18 was reduced in both groups, a larger effect size was noted in the probiotic group (D = 1.336). sTLR-4 was also reduced in both groups, with no difference between groups (p = 0.885). Intervention with probiotics in the early stages of NASH demonstrated no significant change in hepatic and clinical parameters. ClinicalTrials.gov, identifier NCT0346782.

Longitudinal Magnetic Resonance Imaging with ROS-Responsive Bilirubin Nanoparticles Enables Monitoring of Nonalcoholic Steatohepatitis Progression to Cirrhosis.

Despite the vital importance of monitoring the progression of nonalcoholic fatty liver disease (NAFLD) and its progressive form, nonalcoholic steatohepatitis (NASH), an efficient imaging modality that is readily available at hospitals is currently lacking. Here, a new magnetic-resonance-imaging (MRI)-based imaging modality is presented that allows for efficient and longitudinal monitoring of NAFLD and NASH progression. The imaging modality uses manganese-ion (Mn2+)-chelated bilirubin nanoparticles (Mn@BRNPs) as a reactive-oxygen-species (ROS)-responsive MRI imaging probe. Longitudinal T1-weighted MR imaging of NASH model mice is performed after injecting Mn@BRNPs intravenously. The MR signal enhancement in the liver relative to muscle gradually increases up to 8 weeks of NASH progression, but decreases significantly as NASH progresses to the cirrhosis-like stage at weeks 10 and 12. A new dual input pseudo-three-compartment model is developed to provide information on NASH stage with a single MRI scan. It is also demonstrated that the ROS-responsive Mn@BRNPs can be used to monitor the efficacy of potential anti-NASH drugs with conventional MRI. The findings suggest that the ROS-responsive Mn@BRNPs have the potential to serve as an efficient MRI contrast for monitoring NASH progression and its transition to the cirrhosis-like stage.

Lipidomic signatures discriminate subtle hepatic changes in the progression of porcine nonalcoholic steatohepatitis.

Recently, the development of nonalcoholic steatohepatitis (NASH) in common strains of pigs has been achieved using a diet high in saturated fat, fructose, cholesterol, and cholate and deficient in choline and methionine. The aim of the present work was to characterize the hepatic and plasma lipidomic changes that accompany the progression of NASH and its reversal by switching pigs back to a chow diet. One month of this extreme steatotic diet was sufficient to induce porcine NASH. The lipidomic platform using liquid chromatography-mass spectrometry analyzed 467 lipid species. Seven hepatic phospholipids [PC(30:0), PC(32:0), PC(33:0), PC(33:1), PC(34:0), PC(34:3) and PC(36:2)] significantly discriminated the time of dietary exposure, and PC(30:0), PC(33:0), PC(33:1) and PC(34:0) showed rapid adaptation in the reversion period. Three transcripts (CS, MAT1A, and SPP1) showed significant changes associated with hepatic triglycerides and PC(33:0). Plasma lipidomics revealed that these species [FA 16:0, FA 18:0, LPC(17:1), PA(40:5), PC(37:1), TG(45:0), TG(47:2) and TG(51:0)] were able to discriminate the time of dietary exposure. Among them, FA 16:0, FA 18:0, LPC(17:1) and PA(40:5) changed the trend in the reversion phase. Plasma LDL-cholesterol and IL12P40 were good parameters to study the progression of NASH, but their capacity was surpassed by hepatic [PC(33:0), PC(33:1), and PC(34:0)] or plasma lipid [FA 16:0, FA 18:0, and LPC(17:1)] species. Taken together, these lipid species can be used as biomarkers of metabolic changes in the progression and regression of NASH in this model. The lipid changes suggest that the development of NASH also affects peripheral lipid metabolism.NEW & NOTEWORTHY A NASH stage was obtained in crossbred pigs. Hepatic [PC(33:0), PC(33:1) and PC(34:0)] or plasma [FA 16:0, FA 18:0 and LPC(17:1)] species were sensitive parameters to detect subtle changes in development and regression of nonalcoholic steatohepatitis (NASH). These findings may delineate the liquid biopsy to detect subtle changes in progression or in treatments. Furthermore, phospholipid changes according to the insult-inducing NASH may play an important role in accepting or rejecting fatty livers in transplantation.

Estimating utility values for non-alcoholic steatohepatitis health states: a discrete choice experiment.

Background: This study estimated utility values for non-alcoholic steatohepatitis (NASH). Previous studies have assumed that health-related quality of life does not vary between the early stages of NASH. Materials & Methods: Discrete choice experiment (DCE) surveys estimated the value of avoiding fibrosis progression. Patients also completed the EQ-5D-5L. Marginal rates of substitution estimated utility change associated with fibrosis progression. Results: DCE surveys were completed by the UK general public (n=520) and patients with NASH (n=154). The utility decline between fibrosis stages F1 and F4 decompensated was between -0.521 to -0.646 (depending on method). Conclusion: Three methods were used to estimate utilities for NASH, each one showed sensitivity to advancing fibrosis, including in the early stages, which is often considered asymptomatic.

Elucidating Cuproptosis-Associated Genes in the Progression from Nash to HCC Using Bulk and Single-Cell RNA Sequencing Analyses and Experimental Validation.

Background and Objectives: Non-alcoholic steatohepatitis (NASH) is a significant risk factor for hepatocellular carcinoma (HCC) development. Timely treatment during the NASH stage is essential to minimize the possibility of disease progression to HCC. Cuproptosis is a newly identified form of cellular death that could impact the progression of various diseases and cancers. Materials and Methods: Transcriptome and single-cell sequencing datasets were utilized to investigate the role of cuproptosis-related genes (CRGs) in NASH progression to HCC. FDX1, LIPT1, and PDHP were identified as CRGs in NASH patients, and FDX1, DBT, GCSH, SLC31A1, and DLAT were identified as CRGs in patients with NASH progressing to HCC. FDX1 was found to play a significant role in both NASH patients and patients with NASH progressing to HCC. This study constructed cuproptosis-related clusters (CRCs) using the Nonnegative Matrix Factorization algorithm, and they were linked to fatty acid metabolism and the PPAR signaling pathway in both NASH CRCs and HCC CRCs. The Weighted Correlation Network Analysis algorithm identified CRP, CRC, TAT, CXCL10, and ACTA1 as highly relevant genes in NASH CRCs and HCC CRCs. The expression of FDX1 was validated in both mouse models and human NASH samples. Results: The investigation highlights FDX1 as a pivotal CRG in both NASH and NASH progression to HCC. The comprehensive characterization of CRGs sheds light on their potential biofunctional importance in the context of NASH and HCC. Our experimental results show that FDX1 expression was significantly increased in NASH patients. Conclusions: The present study identified key CRGs, revealing their potential impact on NASH and HCC. Meanwhile, targeting FDX1 may prevent the progression of NASH to HCC.

Diagnostic performance of circulating biomarkers for non-alcoholic steatohepatitis.

There are no approved diagnostic biomarkers for at-risk non-alcoholic steatohepatitis (NASH), defined by the presence of NASH, high histological activity and fibrosis stage ≥2, which is associated with higher incidence of liver-related events and mortality. FNIH-NIMBLE is a multi-stakeholder project to support regulatory approval of NASH-related biomarkers. The diagnostic performance of five blood-based panels was evaluated in an observational (NASH CRN DB2) cohort (n = 1,073) with full spectrum of non-alcoholic fatty liver disease (NAFLD). The panels were intended to diagnose at-risk NASH (NIS4), presence of NASH (OWLiver) or fibrosis stages >2, >3 or 4 (enhanced liver fibrosis (ELF) test, PROC3 and FibroMeter VCTE). The prespecified performance metric was an area under the receiver operating characteristic curve (AUROC) ≥0.7 and superiority over alanine aminotransferase for disease activity and the FIB-4 test for fibrosis severity. Multiple biomarkers met these metrics. NIS4 had an AUROC of 0.81 (95% confidence interval: 0.78-0.84) for at-risk NASH. The AUROCs of the ELF test, PROC3 and FibroMeterVCTE for clinically significant fibrosis (≥stage 2), advanced fibrosis (≥stage 3) or cirrhosis (stage 4), respectively, were all ≥0.8. ELF and FibroMeter VCTE outperformed FIB-4 for all fibrosis endpoints. These data represent a milestone toward qualification of several biomarker panels for at-risk NASH and also fibrosis severity in individuals with NAFLD.

The glycemic profile in patients with non-alcoholic steatohepatitis and type 2 diabetes depending on diabetic kidney disease

Background. State of carbohydrate metabolism and severity of insulin resistance in the comorbid course of non-alcoholic steatohepatitis (NASH) and diabetic kidney disease (DKD) in patients with type 2 diabetes mellitus (DM2) is due to the cascade of possible transformation of NASH into liver cirrhosis. The purpose is to study the interaction of changes in glucose and insulin homeostasis, the degree of insulin resistance and insulin sensitivity, the degree of hemoglobin glycosylation on the clinical course of NASH associated with DM depending on the presence of DKD and its stage. Materials and methods. One hundred and eight patients with NASH and comorbid DM2 were examined. The average age of patients was 58.2 ± 6.1 years. There were 63 women (58.3 %) and 45 men (41.7 %). Depending on the presence of DKD, 4 groups of patients were formed, who were randomized by age, sex, activity of cytolytic syndrome. The comparison group consisted of 30 healthy individuals of the appropriate age and sex. The degree of hepatic steatosis and its nature were determined using SteatoTest, ASH and NASH-Test kits (BioPredictive, France). The stage of liver fibrosis was determined using FibroTest (BioPredictive, France), a set of markers for quantitative biochemical evaluation of fibrosis. Results. In patients with NASH, DM2 and DKD stage I–II, we found a significant decrease in serum albumin by 9.0 % (p &lt; 0.05); glomerular filtration rate (GFR) and urine albumin, on the contrary, increased significantly, by 1.5 times (p &lt; 0.05) compared to those in the control group, which indicates the phenomenon of hyperfiltration and is specific to the initial stage of DKD. When NASH is combined with DM2 and DKD stage III, a significant decrease in serum albumin by 1.2 times (p &lt; 0.05) is reported; GFR and albuminuria were significantly increased, by 1.4 and 11.7 times (p &lt; 0.05), respectively, compared to the control group. In patients with NASH, DM2 and DKD stage IV, we found a significant decrease in serum albumin by 1.4 times (p &lt; 0.05), it was significantly increased by 30.2 times (p &lt; 0.05) compared to the indicator in the control group, and the GFR, on the contrary, was significantly reduced by 1.7 times (p &lt; 0.05), which indicates the progression of DKD. Conclusions. Disorders of glucose homeostasis due to insulin resistance are one of the probable risk factors for the progression of non-alcoholic steatohepatitis and type 2 diabetes mellitus in the presence of stage I–IV diabetic kidney disease.

Current Clinical Trial Status and Future Prospects of PPAR-Targeted Drugs for Treating Nonalcoholic Fatty Liver Disease.

The number of patients with nonalcoholic fatty liver disease (NAFLD)/nonalcoholic steatohepatitis (NASH) is increasing globally and is raising serious concerns regarding the increasing medical and economic burden incurred for their treatment. The progression of NASH to more severe conditions such as cirrhosis and hepatocellular carcinoma requires liver transplantation to avoid death. Therefore, therapeutic intervention is required in the NASH stage, although no therapeutic drugs are currently available for this. Several anti-NASH candidate drugs have been developed that enable treatment via the modulation of distinct signaling cascades and include a series of drugs targeting peroxisome proliferator-activated receptor (PPAR) subtypes (PPARα/δ/γ) that are considered to be attractive because they can regulate both systemic lipid metabolism and inflammation. Multiple PPAR dual/pan agonists have been developed but only a few of them have been evaluated in clinical trials for NAFLD/NASH. Herein, we review the current clinical trial status and future prospects of PPAR-targeted drugs for treating NAFLD/NASH. In addition, we summarize our recent findings on the binding modes and the potencies/efficacies of several candidate PPAR dual/pan agonists to estimate their therapeutic potentials against NASH. Considering that the development of numerous PPAR dual/pan agonists has been abandoned because of their serious side effects, we also propose a repositioning of the already approved, safety-proven PPAR-targeted drugs against NAFLD/NASH.

Hepatic regulatory T-cells promote colorectal liver metastasis in NASH

Abstract Background: Non-alcoholic steatohepatitis (NASH), a progressive inflammatory form of NAFLD, underlies an extremely versatile and dynamic inflammatory microenvironment along with aberrant metabolism and ongoing liver regeneration. Regulatory T cells (Tregs), as essential immune regulatory cells, are critical for the inhibition of tumor-promoting inflammation and cancer cell escape. Our recent study found that Tregs were selectively increased in both mouse models and patients with NAFLD, which promotes us to study the role of Tregs in NASH-HCC. However, the role of Tregs in the development of hepatic metastasis in NASH liver remains unknown. Methods: A Western diet mouse model and the STAM mouse model were utilized for NASH development. MC38 murine colorectal tumor cells were injected via portal vein at 12 weeks of feeding or orthotopically injected into cecum subserosa. Tumor development will be determined 3 or 4 weeks after injection. Depletion of Tregs by FoxP3-DTR mice or CD25 antibody treatment. Transcriptomic profiling of leucocytes in hepatic immune microenvironment was determined by scRNA-seq. Results: There is a positive correlation between increased Tregs and hepatic tumor growth in NASH liver. Depleting Tregs inhibits the development of hepatic metastasis in NASH liver compared with control liver. NASH development led to a significant transcriptomic shift in hepatic immune cells, such as Kupffer cells, Tregs and MDSCs. Blocking the interaction of Tregs and PMN-MDSCs in the TME prevents tumor growth in the NASH liver. Conclusions: Tregs can suppress immunosurveillance in the premalignant stages of NASH. Therapies targeting Tregs could offer a potential strategy for preventing hepatic metastasis in patients with NASH. RO1GM137203

Influence of non-alcoholic steatohepatitis on the renal functional status in patients with type 2 diabetes and diabetic kidney disease

Background. Hepatic steatosis and nonalcoholic steatohepatitis (NASH) are considered the most common liver pathologies, which in developed countries is observed in 20–30% of the adult population. Patients with diabetes mellitus (DM) often develop non-specific kidney lesions such as asymptomatic bacteriuria, pyelonephritis, kidney carbuncle, kidney abscess, kidney tuberculosis, necrotizing papillitis, or papillary necrosis, which significantly complicate the course of diabetic nephropathy (DN). DN accounts for 40% of complications in type 1 DM and 5–15% among patients with type 2 DM. The purpose of the study is to clarify the impact of non-alcoholic steatohepatitis on the renal functional status in patients with type 2 diabetes and diabetic kidney disease (DКD). Materials and methods. One hundred and sixty patients with NASH with comorbid type 2 DM of moderate severity and DКD stages I–IV were examined. A prospective study was conducted, which involved 160 patients and 30 practically healthy individuals. In particular, 25 patients with NASH and type 2 diabetes (group 1), 20 people with NASH and class I obesity (group 2), 70 patients with NASH stage I–IV (group 3), of whom 31 patients (44.3%) had DКD stage I–II (group 3a), 20 (28.6%) stage III (group 3b), 19 (27.1%) had DKD stage IV (group 3c). Results. In patients with NASH, type 2 DM, and DKD stage I–IV, hypoalbuminemia was found to be probably higher compared to that of patients with type 2 DM, DКD stage I–IV and without NASH that indicates the influence of NASH on the level of albuminemia; higher blood level of creatinine and urea; lower glomerular filtration rate, which confirms the negative effect of NASH on the course of type 2 DM with DКD. In the presence of NASH, patients with DКD had a higher level of albuminuria, urinary creatinine, albumin-creatinine ratio than if it’s absent. Conclusions. Indicators of the renal functional state in patients with NASH and type 2 DM varied depending on the presence of DКD and its stage. As the stage of DКD increased, the content of albumin in the blood decreased, the levels of creatinine and urea increased. The result of the detected changes was an increase in glomerular filtration rate, which indicates the phenomenon of hyperfiltration, which is specific for the initial stages of DКD.

A phase 2, adaptive randomized, double-blind, placebo-controlled, multicenter, 52-week study of HM15211 in patients with biopsy-confirmed non-alcoholic steatohepatitis – Study design and rationale of HM-TRIA-201 study

Non-alcoholic steatohepatitis (NASH) is a multifactorial disease with an increasing prevalence worldwide due to the obesity pandemic. HM15211 (efocipegtrutide), a novel, long-acting glucagon-like peptide-1/glucagon/glucose-dependent insulinotropic polypeptide triple incretin agonist has shown promising efficacy in in vitro, preclinical rodent models of NASH and phase 1 studies with manageable toxicity. Though liver biopsy is recommended for grading and staging of NASH, its invasive nature necessitates innovative approaches in clinical trials that decrease the burden of patients otherwise subjected to this invasive procedure. We report an innovative study design of phase 2 study of HM15211. MethodsHM-TRIA-201 is a multicenter, randomized, double-blind, 52-week, placebo-controlled, parallel-group adaptive design study of 217 patients with biopsy-proven NASH. The primary endpoint is the proportion of patients with complete resolution of steatohepatitis (defined as Non-alcoholic fatty liver disease Activity Score of 0–1 for inflammation, 0 for ballooning, and any other value for steatosis) on overall histopathological reading and no worsening of liver fibrosis on NASH Clinical Research Network fibrosis score. An interim analysis is planned after 15 patients/group complete 26 weeks of treatment, after which one HM15211 dose group will be discontinued based on safety and efficacy risk-to-benefit analysis; patients of the dropped dosing arm will be re-randomized into 2 remaining HM15211 groups. ConclusionThe adaptive design study of HM15211 minimizes the number of patients to be exposed to a liver biopsy while optimizing the sample size of patients exposed to safe and effective doses of HM15211 to inform ideal dose for further clinical development in NASH.

The state of the cytokine profile in pregnant women with non-alcoholic fatty liver disease at the stage of non-alcoholic steatohepatitis with varying degrees of comorbid obesity under the influence of the developed complex therapy program

The aim of the study: to evaluate the cytokine profile state in pregnant women with non-alcoholic fatty liver disease (NAFLD) at the stage of non-alcoholic steatohepatitis (NASH) with varying degrees of obesity under the influence of the developed complex therapy program. Material and methods. We examined 197 pregnant women with NAFLD at the stage of NASH in combination with obesity. The main group I consisted of 98 pregnant women with NAFLD at the stage of NASH with varying degrees of obesity, who were divided into 3 subgroups depending on body mass index (BMI). Among them, 26 pregnant women with BMI of 25.0–29.9 kg/m2 were included in IA group, 48 pregnant women with BMI of 30.0–34.9 kg/m2 were included in IB group, and 24 pregnant women with BMI of 35.0–39.9 kg/m2 – in IC group. All pregnant women in the main group were prescribed complex therapy including vitamin E at a dose of 400 IU/day, ursodeoxycholic acid (UDCA) at a dose of 15 mg/kg/day, and L-carnitine at a dose of 3 g per day. The comparison group consisted of 69 women with NAFLD at the stage of NASH and abdominal obesity, who corresponded to subgroups of the main group (IIA – 23 patients, IIB – 25 women, IIC – 21 pregnant women) and received basic therapy. The control group consisted of 30 healthy women. To evaluate the cytokine profile, levels of IL-1β, IL-6, IL-10 and TNF-α were determined by ELISPOT. Results. Analysis of the cytokine profile in women with NASH and obesity showed the presence of systemic inflammation links in the examined groups, which was manifested by increased levels of pro-inflammatory and decreased levels of anti-inflammatory interleukins in blood serum of pregnant women. A prescription of the complex treatment contributed to a decreased activity of the inflammatory response, which was manifested by an improvement in the levels of cytokine profile indicators. Conclusions. NASH during pregnancy is accompanied by significant changes in the cytokine profile. The prescription of complex therapy in the form of vitamin E, UDCA and L-carnitine is effective in the treatment of pregnant women with NAFLD at the stage of NASH due to cumulative and potentiating effects, reducing manifestations of systemic inflammation by normalizing the level of cytokines.