Introduction

This study explores potential associations between the gut microbiome and fibrosis in patients with viral hepatitis, using network analysis to reveal correlations between multiple microbes and clinical data. The gut-liver axis plays a crucial role in human health, with weakened tight junctions, increased influx of substances into the liver, and IgA-related immune abnormalities observed in patients with chronic hepatitis and liver cirrhosis. However, the causative bacteria remain elusive. Network analysis can reveal potential biomarkers and therapeutic targets, with the hope of improving patient outcomes.

Aims & Methods

We aimed to elucidate the relationship between fibrosis markers and gut microbiome by constructing networks, analyzing fecal samples and clinical indicators of 145 HCV antibody-positive (HCV group) and 84 HBs antigen-positive (HBV group) patients with chronic liver disease who visited our hospital between June 2016 and April 2021. We identified microbes at the genus level by analyzing DNA from the 16S rRNA region (V3-V4) obtained from fecal samples using the Miseq platform, QIIME2, and Silva database.
Patients were divided into HBV and HCV groups to establish reproducible relationships. We calculated Spearman rank correlation coefficients between relative microbial abundance and clinical data, including BMI, age, albumin , total cholesterol, estimated glomerular filtration rate , AST, ALT,GTP, total bilirubin, hemoglobin, platelet count, Fib4-index, prognostic nutritional index (PNI), and ALBI score. We considered p-values below 0.05 and absolute correlation coefficients above 0.3 as significant correlations, which will help in understanding the complex relationships between fibrosis markers and gut microbiome in patients with viral hepatitis.

Results

Patient backgrounds differed between the HBV group (median age 56.5 years, Fib4-index 1.74, PNI 49.5, ALBI -2.8) and the HCV group (68 years, Fib4-index 3.21, PNI 46.5, ALBI -2.6). We identified 263 microbial genus and 34,453 combinations (₂₆₃C₂). Among these, 1,619 combinations in the HBV group and 4,730 in the HCV group showed significant correlations with over 90% showing positive correlations. There were 586 common combinations between both groups, which were highly likely to be a unique bacterial combination maintained across different patient backgrounds.
We found 85 correlations in the HBV group and 20 in the HCV group in the 3,682 combination between 14 types of clinical data and 263 bacterial genus. Specifically, only Veillonella and Streptococcus demonstrated a positive correlation with Fib4-index and a negative correlation with PNI. Lactobacillus showed a positive correlation with age in common between the HBV and HCV groups . They did not have a direct correlation with age, ALT, AST, or platelet count, suggesting a potential relationship with fibrosis. However, the age-correlated Lactobacillus showed a positive correlation with Veillonella and Streptococcus, indicating a possible secondary relationship. Additionally, Streptococcus, occupying a key position in the network with positive correlations with multiple bacteria, may have a significant influence on these microbes.

Conclusion

We found potential associations between specific gut microbes and fibrosis in viral hepatitis patients. Streptococcus, Veillonella, and Lactobacillus were significant in their relationship with fibrosis markers, with Streptococcus playing a central role in the network. Thhs suggests Streptococcus as a potential biomarker and network analysis as a valuable tool for potential diagnostic and therapeutic applications.

Introduction

Liver cirrhosis is characterized by microbial dysbiosis, intestinal inflammation and a weakened gut barrier and associated with an increased risk of complications. We previously observed a positive influence of L-ornithine-L-aspartate (LOLA) on gut microbiome composition in cirrhosis (1). The current study aimed to investigate the effect of LOLA on microbiome dysbiosis, quality of life, sarcopenia, intestinal permeability, inflammation and metabolite composition.

Aims & Methods

In this monocentric, open-label, phase 4 study, LOLA (18g/day) was administered orally for three months to liver cirrhosis patients with hepatic encephalopathy grades (0-2). We investigated changes in their gut microbiome composition (16s rRNA sequencing, alpha and beta diversity, differential abundance), quality of life (SF36), blood ammonia concentrations, measurements of sarcopenia and frailty (appendicular lean muscle mass, anthropometric measures, liver frailty index, SARC-F), biomarkers of gut permeability, inflammation and sarcopenia as well as metabolic alterations in stool, serum, and urine.

Results

65 patients were enrolled, and 52 patients completed the 3-month study (40.4% female, age 62 (58; 65)). While LOLA reduced the abundance of the genus Romboutsia, it did not significantly change overall microbiome diversity. LOLA led to a significant improvement in the vitality dimension of the SF36 questionnaire (45 (35; 60) to 50 (45; 60), p=0.019) and decreased plasma ammonia levels in patients with elevated baseline ammonia (69.5 (54; 225) to 46.0 (22; 66) µmol/L, p=0.004) Although muscle mass generally declined across the study, this was not observed in patients whose elevated ammonia levels improved. Sarcopenia and frailty scores and the muscle biomarkers myostatin, irisin, fibroblast growth factor 21, insulin-like growth factor 1 were unaffected by LOLA treatment. Diamine oxidase, a marker of gut barrier dysfunction, significantly decreased (from 12.75 U/mL (11.25; 17.45) to 11.15 U/mL (9.9; 15.65), p=0.016), while LPS binding protein, a marker of innate immune response to bacterial translocation, significantly increased (from 17.05 µg/mg (14.7; 19.8) to 17.39 µg/mg (16.2; 21.9), p=0.006). Calprotectin, zonulin and sCD14 did not show significant changes. Metabolomic analysis revealed an increase in serum alanine levels (pFDR=0.014) with additional subtle shifts in lipoprotein and amino acid profiles.

Conclusion

In summary, LOLA was associated with an improvement in vitality, an important patient-reported outcome accompanied by alterations in gut microbial composition and prevented muscle loss in patients with hyperammonaemia. Furthermore, LOLA was associated with improvements in gut permeability and endotoxin handling. The slight increase in alanine observed in the metabolomics analysis may be a consequence of LOLA metabolism. LOLA may be a valuable supportive treatment for enhancing quality of life in cirrhosis by targeting the gut liver axis and may be particularly beneficial in preventing muscle loss in patients with hyperammonaemia.

References

1. Horvath A, Traub J, Aliwa B, Bourgeois B, Madl T, Stadlbauer V. Oral Intake of L-Ornithine-L-Aspartate Is Associated with Distinct Microbiome and Metabolome Changes in Cirrhosis. Nutrients. 2022 Feb 10;14(4):748.

Introduction

Autoimmune hepatitis (AIH) and its variants are uncommon causes of chronic liver disease (CLD) in India [1]. Concomitant metabolic dysfunction-associated steatotic liver disease (MASLD) in patients with autoimmune hepatitis (AIH) is a distinct entity [2]. We aimed to explore the frequency and impact of concomitant MASLD in patients with AIH [3].

Aims & Methods

This is a retrospective study conducted at Sanjay Gandhi Post Graduate Institute of Medical Sciences and King George Medical University, Lucknow, India. Data of patients with AIH who underwent treatment between 2013-2022 were collected from a prospectively filled-up electronic database. Standard criteria were used to diagnose AIH and MASLD [3][4]. Patients presenting with acute severe-AIH, acute-on-chronic-liver-failure, simple steatosis without any metabolic dysfunction, and hepatocellular carcinoma HCC) were excluded.Criteria to diagnose MASLD were applied to patients with AIH. Their demographic, clinical, biochemical, serological parameters, treatment response and long-term outcomes were measured and compared.

Results

Of 175 patients with AIH, 51 were excluded. A total of 122 patients [median age: 46 (34-54) years; 69.6% female] were included in the final analysis. The median follow-up period was 61 (50-76) months. Of the 122 patients, 38 (31%) had concomitant MASLD (AIH-MASLD). Patients withAIH-MASLD had higher baseline liver enzymes [aspertate aminotranferase 226 (121-278) vs 174 (113-225); p= 0.033, alanine aminotranferase: 115 (113-189) vs 97 (47-169); p=0.041]. A higher number of patients with AIH-MASLD presented with advanced fibrosis and decompensation (20.4% vs 10.7%; p=<0.001,30.3% vs 25%; p=0.030, respectively). Following immunosuppression, Patients with MASLD-AIH had attenuated remission rates compared to AIH alone (34.2% vs 77.4%; p=<0.001) [table: 1].

Table: 1
Outcome of patients with AIH and MASLD-AIH

Conclusion

About a third of patients with biopsy-proven AIH have concomitant MASLD. A significant number of patients with AIH-MASLD present with higher liver enzymes, advanced fibrosis, and decompensation compared to AIH alone. Patients with AIH-MASLD respond poorly to steroids and have a lower remission rate. Proper work-up is necessary in patients with steroid non-responsive-AIH to rule out concomitant MASLD.

References

1. Choudhuri G, Somani SK, Baba CS, et al. Autoimmune hepatitis in India: profile of an uncommon disease. BMC Gastroenterol. 2005 Aug 15;5:27. doi: 10.1186/1471-230X-5-27. PMID: 16098234; PMCID: PMC1236920.
2. Zachou K, Azariadis K, Lytvyak E, et al; International Autoimmune Hepatitis Group (IAIHG). Treatment responses and outcomes in patients with autoimmune hepatitis and concomitant features of non-alcoholic fatty liver disease. JHEP Rep. 2023 Apr 22;5(8):100778. doi: 10.1016/j.jhepr.2023.100778. PMID: 37456672; PMCID: PMC10339258.
3. Wang G, Tanaka A, Zhao H, et al. The Asian Pacific Association for the Study of the Liver clinical practice guidance: the diagnosis and management of patients with autoimmune hepatitis. Hepatol Int. 2021 Apr;15(2):223-257. doi: 10.1007/s12072-021-10170-1. Epub 2021 May 4. PMID: 33942203; PMCID: PMC8144150.
4. Eslam M, Newsome PN, Sarin SK, et al. A new definition for metabolic dysfunction-associated fatty liver disease: An international expert consensus statement. J Hepatol. 2020 Jul;73(1):202-209. doi: 10.1016/j.jhep.2020.03.039. Epub 2020 Apr 8. PMID: 32278004.

Introduction

The development of nonalcoholic fatty liver disease (NAFLD) is significantly influenced by genetic and environmental factors. Hepatobiliary diseases are associated with an imbalance of bacterial subpopulations and the associated deleterious effects on the colonized host. Otherwise, more and more evidence points to a close relationship between the composition of the microbiota and the development and progression of NAFLD. The genus Candida and the species Candida albicans could play a central role in the pathogenesis and progression of fatty liver infiltration with further steatohepatitis progression by the impact on the gut-liver axis. For example, rodents colonized with C. albicans develop liver disease without additional stimuli. Fungi might hence possibly exacerbate liver disease in a two-hit model, one hit being alcohol, the Western diet, or a toxin, and another hit being the presence and deleterious impact of fungi.

Aims & Methods

The aim of the study was to examine the prevalence of Candida and comporision of the gut microbiome in patients with NAFLD.

Results

Conclusion

The prevalence of Candida in gut microbiota of patients with NAFLD is significantly higher than in patients without fatty liver infiltration (36% vs. 13%).
The Candida growth can impact TNF-a and TG increasing in patients with NAFLD.
Candida could be associated with F/B index growth in patients with NAFLD.
Such results are potentially interesting for the search of personalized approaches in treatment and prevention the NAFLD. The role of gut microbiome is one of the important parts of the fatty infiltration development pathogenesis.

Introduction

Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly known as nonalcoholic fatty liver disease (NAFLD), is a widespread chronic liver condition closely associated with obesity, insulin resistance, and dyslipidemia. Recent evidence points to gut microbiota dysbiosis as a key player in MASLD pathogenesis, affecting intestinal barrier function and systemic inflammation.

Aims & Methods

The aim was to investigate gut microbiota composition in patients with MASLD and assess its association with disease progression and gut barrier dysfunction. A total of 152 MASLD patients and 150 matched controls without liver pathology were examined. Diagnosis was confirmed by ultrasound and steatometry, alongside assessment of cardiometabolic risk factors. Gut microbiota composition was analyzed using real-time PCR, focusing on Bacteroidetes, Firmicutes, and Actinobacteria. Hydrogen breath testing was used to diagnose small intestinal bacterial overgrowth (SIBO), and the presence of Akkermansia muciniphila was evaluated as a marker of gut barrier integrity.

Results

MASLD patients demonstrated a significant increase in Firmicutes (50.3%) and Actinobacteria (15.3%) proportions compared to controls, with a higher Firmicutes/Bacteroidetes (F/B) ratio. SIBO was detected in 52% of MASLD patients versus 40.5% in controls (p<0.05). The presence of Akkermansia muciniphila was notably lower in MASLD patients (33%) compared to controls (63%), suggesting impaired gut barrier function. Strong positive correlations were observed between Actinobacteria abundance and the F/B index (r=0.75, p<0.05), while Akkermansia muciniphila presence negatively correlated with MASLD development (r=–0.69, p<0.05).

Conclusion

Gut microbiota dysbiosis, characterized by elevated Firmicutes and Actinobacteria and reduced Akkermansia muciniphila, is closely associated with MASLD pathogenesis. The findings highlight the potential of microbiota-targeted interventions for improving gut barrier function and modifying disease progression in MASLD patients.