Editor’s Note:

From November 10th to 14th, the annual international event in hepatology—the American Association for the Study of Liver Diseases (AASLD) 2023—was grandly held in Boston, USA. According to incomplete statistics, more than 200 abstracts from Chinese experts and scholars were selected for oral presentations or poster exchanges at this conference, achieving fruitful results. Congratulations are extended here! The Beijing University People’s Hospital and Peking University Hepatology Institute team achieved 1 oral and 9 poster presentations on scientific research in the fields of hepatic encephalopathy, fatty liver, hepatitis C, hepatitis B, and liver cancer. Some research abstracts are shared below, and “Hepatology Digest” takes you to appreciate the team’s excellence.

01 Neutrophil-to-Lymphocyte Ratio Predicts Short- and Long-Term Readmission of Patients with Hepatic Encephalopathy

Abstract ID: 3092-A

Authors: Rui Huang, Lin Zhang

Corresponding Author: Rui Huang

Research Background:

Hepatic encephalopathy (HE) is a critical complication in the late stages of liver disease, indicating poor prognosis. The 30-day and 90-day readmission rates for patients with cirrhosis are 20%-30%, with HE being the most common cause. Multiple factors have been reported as predictors for readmission in HE patients. However, besides liver function indicators, few new serum markers have been found to predict readmission in HE patients. In order to explore simple and effective predictors for short-term and long-term readmission of HE patients, we conducted this retrospective study.

Research Methods:

We conducted a single-center retrospective study, including patients admitted due to HE. The main endpoints were the first liver-related readmission within 30 days, 90 days, and 180 days. Logistic regression analysis and multiple linear regression analysis were applied to identify predictors related to readmission and length of the first hospitalization.

Research Results:

We included 424 patients admitted due to HE, with 24 (5.7%), 63 (14.8%), and 92 (21.7%) patients readmitted within 30 days, 90 days, and 180 days, respectively. Among these patients, 283 (66.7%) were male, with a mean age of 59.9 ± 11.5 years. Alcoholic liver disease (ALD) was present in 120 patients (28.3%). At baseline, 40 (9.4%), 246 (58.0%), 67 (15.8%), and 20 (4.7%) patients had hepatocellular carcinoma (HCC), ascites, variceal bleeding, and spontaneous bacterial peritonitis (SBP), respectively.

Patients readmitted within 180 days were older (62.0 ± 9.6 vs. 59.3 ± 12.0, P=0.025), had a higher proportion of uninsured individuals [25 (27.2%) vs. 52 (15.7%), P=0.010], a higher proportion of ALD [34 (37.0%) vs. 86 (25.9%), P=0.027], a higher proportion of chronic kidney disease (CKD) [21 (22.8%) vs. 39 (11.7%), P=0.007], a higher proportion of HCC (15 (16.3%) vs. 25 (7.5%), P=0.012), and a higher proportion of ascites (65 (70.7%) vs. 181 (54.5%), P=0.004). Reasons for admission, in addition to HE, included volume-related (247 cases, 58.3%) and other complications (177 cases, 41.7%). The average length of the first hospitalization for all patients was 17.9 ± 12.1 days.

HE was the most common cause of readmission, with 15 (62.5%), 30 (47.6%), and 48 (52.1%) patients readmitted within 30 days, 90 days, and 180 days, respectively. In addition, compared to patients not readmitted within 30 days, 90 days, or 180 days, readmitted patients had a higher neutrophil-to-lymphocyte ratio (NLR) at the time of first discharge (8.17 ± 12.1 vs. 3.95 ± 4.96; 7.17 ± 9.72 vs. 3.67 ± 4.44; 6.80 ± 8.62 vs. 3.47 ± 4.28, P<0.001). Patients readmitted within 30 days had the longest length of hospital stay during the first admission, significantly higher than patients not readmitted within 30 days (24.8 ± 19.2 vs. 17.7 ± 11.90, P<0.001).

Logistic regression analysis showed significant predictors of readmission, including lack of insurance, ALD, ascites, Model for End-Stage Liver Disease (MELD) score, and NLR at the time of first discharge. Age and HCC were also significant predictors of readmission within 90 days and 180 days (Table 1). However, there were gender differences in predictors of readmission. In males, in addition to HCC, ascites, MELD score, and NLR at the time of discharge were significant predictors of short-term and long-term readmission. Interestingly, in females, ALD was not a significant predictor of readmission, while hypertension was significantly associated with readmission within 180 days. Significant predictors of the length of the first hospitalization included variceal bleeding at the time of first admission (P=0.006), CKD (P=0.003), and MELD score at the time of discharge (P=0.024).

Research Conclusions:

The Neutrophil-to-Lymphocyte Ratio (NLR) at the time of discharge is a significant predictor for both short-term and long-term readmission of patients with Hepatic Encephalopathy (HE). Chronic Kidney Disease (CKD) is a significant factor influencing the length of the first hospitalization in HE patients.

Table 1. Predictors of readmission at 30 days, 90 days, and 180 days.

Corresponding Author’s Brief Introduction:

Associate Chief Physician, Department of Hepatology, Peking University People’s Hospital, and Peking University Hepatology Institute.

02 Sustained Virologic Response Improved the Long-term Health-Related Quality of Life In Patients With Chronic Hepatic C: A Prospective National Study In China

Abstract ID: 1856-A

Authors: Rui Huang, Huiying Rao

Corresponding Author: Huiying Rao

Research Background:

Chronic hepatitis C virus (HCV) infection significantly impacts the Health-Related Quality of Life (HRQoL) of patients. While it is known that Sustained Virologic Response (SVR) helps improve HRQoL, there is a lack of long-term follow-up prospective studies in real-world settings. We conducted a nationwide multicenter prospective study to investigate trends in HRQoL changes among HCV-infected individuals and assess the long-term impact of antiviral treatment.

Research Methods:

We conducted a prospective observational study in 28 hospitals nationwide (as part of the CCgnos study), targeting adult patients diagnosed with chronic HCV infection. Data on patients’ demographic, clinical characteristics, and EQ-5D questionnaire responses were collected. Generalized Estimating Equation (GEE) models were applied to assess the relationship between these variables and changes in HRQoL over time.

Research Results:

The study included 456 patients, with a median age of 46.5 (36.5–57.0) years, 262 (57.5%) were male, and 44 (9.6%) had cirrhosis. The primary HCV genotypes were genotype 1 (46.1%) and genotype 2 (25.4%). The baseline log HCV RNA was 6.10 (5.34, 6.55) IU/mL. Among 335 patients receiving antiviral treatment (interferon and ribavirin therapy), 73.5% achieved SVR24 by the end of follow-up. The baseline EQ-5D index and EQ-VAS scores were 0.916 ± 0.208 and 80.6 ± 13.0, respectively.

At baseline, there was no significant difference in EQ-5D index between patients who achieved SVR and those who did not (0.959 ± 0.077 & 0.913 ± 0.215, P=0.241). From the first year onwards, the SVR group had significantly higher EQ-5D index and EQ-5D VAS scores than the non-SVR group. However, by the fifth year, these inter-group differences were no longer significant. The EQ-5D index and EQ-VAS scores increased each year, with the proportion of patients reporting moderate to severe problems in any of the five dimensions of EQ-5D gradually decreasing over time. Figure 1 shows the number of patients reporting moderate to severe problems in each dimension of EQ-5D during the follow-up period.

Stratifying by SVR status, differences over time became significant between patients who achieved SVR and those who did not, especially in the MO and UA dimensions [Fifth year: MO: 8 (4.8%) and 8 (18.6%), P=0.032; UA: 4 (2.4%) and 7 (16.3%), P=0.021]. Additionally, the proportion of patients reporting pain or discomfort was significantly higher in the non-SVR group than the SVR group [First year: 21 (12.2%) and 39 (23.9%), P=0.005; Second year: 20 (9.9%) and 39 (20.2%), P=0.000; Third year: 29 (15.3%) and 28 (25.0%), P=0.047; Fourth year: 19 (10.9%) and 18 (22.8%), P=0.027].

GEE model analysis was used to identify predictors of the longitudinal EQ-5D index and EQ-VAS in HCV-infected individuals. Univariate analysis showed that gender, age, residence, marital status, education level, income, cirrhosis, Beck Depression score, and follow-up duration were significantly associated with the EQ-5D index. Furthermore, achieving SVR24 was positively correlated with the EQ-5D index [β=0.049, 95% CI: 0.033–0.065, P=0.000]. Gender, age, residence, marital status, education level, income, cirrhosis, genotype, and follow-up duration were significant predictors of EQ-VAS. At the same time, SVR24 was a strong predictor of EQ-VAS over time [β=5.933, 95% CI (4.827–7.040), P=0.000].

Multivariate analysis revealed that age, income, and SVR24 [β=0.040, 95% CI: 0.023–0.057, P=0.000] were significant predictors of the EQ-5D index, while gender, age, genotype, and SVR24 [β=5.333, 95% CI: 4.204–6.462, P=0.000] were associated with changes in EQ-VAS over time.

Research Conclusions:

Our study demonstrates that achieving viral clearance through antiviral treatment can improve the long-term Health-Related Quality of Life (HRQoL) of HCV patients. Specific demographic factors, along with the presence of cirrhosis and genotype, also significantly influence the long-term changes in patient life quality.

03 Identification of Novel Hub Genes that may Predict the Risk of Liver Fibrosis Progressing to Hepatocellular Carcinoma In Non-Alcoholic Fatty Liver Disease

Abstract ID: 2053-A First Author: Baiyi Liu Corresponding Author: Huiying Rao

Research Background:

Non-Alcoholic Fatty Liver Disease (NAFLD) is the most common chronic liver disease worldwide, including non-alcoholic fatty liver, non-alcoholic steatohepatitis, liver fibrosis, and cirrhosis. If left untreated, NAFLD fibrosis can progress to Hepatocellular Carcinoma (HCC). In recent years, with the continuous increase in the number of NAFLD patients, NAFLD is increasingly considered a contributing factor to HCC, with liver fibrosis being a critical stage. Currently, there are many diagnostic methods for NAFLD, including non-invasive and invasive assessments such as ultrasound, MRE, FIB-4, FAST score, and liver tissue biopsy.

Although these clinical examinations provide effective assistance in diagnosing NAFLD and its various stages, they cannot predict the likelihood of NAFLD progressing to HCC. With the development of biomedicine, bioinformatics has gained increasing attention for detecting specific biomarkers for various diseases and analyzing the molecular mechanisms of disease occurrence and progression. However, there are currently few biomarkers that can predict the risk of NAFLD progressing to HCC. Therefore, it is necessary to explore relevant biomarkers to address this gap.

Research Methods:

To screen for molecular biomarkers predicting the progression of NAFLD to HCC, we first downloaded RNA-seq data from the GEO database for NAFLD with fibrosis and NAFLD-related HCC to identify differentially expressed genes. Next, to explore the potential functions and key genes for risk prediction of differentially expressed genes, we conducted GO and KEGG enrichment analyses and protein-protein interaction analysis on the intersection genes of the two sets of differentially expressed genes. We further identified key genes predicting the risk of NAFLD progressing to HCC. Subsequently, we performed Lasso Cox regression analysis on the key genes to determine the gene model most suitable for predicting the progression of NAFLD with liver fibrosis to HCC.

To evaluate the performance of the gene model, we used two additional sets of liver tissue sequencing data for validation. The first set included sequencing data for NAFLD with liver fibrosis and NAFLD-related HCC, and the second set included sequencing data for NAFLD with liver fibrosis and NASH-related HCC. We then used ROC curves to show the evaluation results of the model performance on these two datasets. Finally, using the gene model, we divided NAFLD patients into high-risk and low-risk groups for HCC and performed GSEA enrichment analysis and tumor immune infiltration analysis on the sequencing data of the two groups of patients’ liver tissues.

Research Results:

In the dataset of NAFLD with liver fibrosis, 559 differentially expressed genes were identified, and in the dataset of NAFLD-related HCC, 2164 differentially expressed genes were identified, with 88 genes in common (Figure 2A). GO and KEGG enrichment analysis showed that the intersection genes were closely associated with the positive regulation of reactive oxygen response, stress-activated MAPK cascade, apoptosis signaling pathway, and IL-17 signaling pathway (Figure 2B~C). The protein-protein interaction network identified 7 genes as key genes, including TIPM1, COL1A1, CXCL9, FOS, VWF, CD79A, and VCAN, and these 7 key genes were used to construct the gene model (Figure 2D).

The results of the gene model calculations showed that the Area Under the ROC Curve (AUC) for risk prediction in the dataset of NAFLD-related HCC was 0.893, and in the dataset of NASH-related HCC, the AUC for risk prediction was 0.751 (Figure 2E~F). In the TCGA dataset, the AUC for risk prediction was 0.572, and the overall survival of the high-risk group for HCC was significantly shorter than that of the low-risk group (Figure 2G~H). GSEA enrichment analysis showed that the PPAR signaling pathway, insulin signaling pathway, and fatty acid metabolism were significantly upregulated in the high-risk group, while negative regulation of the apoptosis signaling pathway was significantly upregulated in the low-risk group (Figure 2I~L). Tumor immune infiltration analysis showed that CXCL9 was significantly positively correlated with M1 macrophages in the high-risk group, and VCAN was significantly negatively correlated with M2 macrophages (Figure 2M).

Research Conclusions:

Through bioinformatics research methods, this study identified TIPM1, COL1A1, CXCL9, FOS, VWF, CD79A, and VCAN as potential biomarkers for predicting the risk of NAFLD progressing to HCC. These 7 genes may also play a crucial role in mediating the potential molecular mechanisms of NAFLD progression to HCC.

Research Conclusions:

This study, employing bioinformatics research methods, identified TIPM1, COL1A1, CXCL9, FOS, VWF, CD79A, and VCAN as potential biomarkers for predicting the risk of NAFLD progressing to HCC. These seven genes may also play a crucial role in mediating the potential molecular mechanisms of NAFLD progression to HCC.

Corresponding Author’s Brief Introduction:

Chief Physician, Professor, Doctoral Supervisor. Director of Peking University Hepatology Institute, Deputy Director of Clinical Trials Institution at Peking University People’s Hospital.

04 Drp1 regulates GSDMD mediated mitochondrial dysfunction and hepatocyte pyroptosis promoting the occurrence of alcoholic hepatitis

Abstract ID: 3539-C Corresponding Author: Yandi Xie

Research Background:

In recent years, the global incidence of alcohol-related diseases has been on the rise. Alcohol-induced liver toxicity and damage can lead to the occurrence of alcoholic liver disease. Alcoholic hepatitis (AH) is a common form of alcoholic liver disease with a high short-term mortality rate and currently lacks effective treatment methods. Exploring the pathogenesis of alcoholic hepatitis is crucial for finding new targeted treatment methods. In the study of the pathogenesis of alcoholic hepatitis, the mechanisms and influences of Gasdermin D (GSDMD) activation are not yet clear. In the current research, we explored whether dynamin-related protein 1 (Drp1) regulates GSDMD-mediated mitochondrial dysfunction and hepatocyte pyroptosis, promoting the occurrence of alcoholic hepatitis.

Research Methods:

We created a mouse model of alcoholic hepatitis, evaluated the degree of liver damage by HE staining of liver tissues from alcoholic hepatitis mice, and measured serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), and triglycerides (TG) in alcoholic hepatitis mice. We assessed hepatocyte pyroptosis by detecting levels of interleukin (IL)-1β, IL-18, inflammatory proteins, and hepatocyte death. We evaluated mitochondrial function by measuring mitochondrial DNA (mtDNA) levels, reactive oxygen species (ROS) generation, mitochondrial membrane potential, ATP content, levels of mitochondrial function-related proteins, and changes in mitochondrial morphology.

Research Results:

Compared to normal mice, HE staining of liver tissues from alcoholic hepatitis mice showed cytoplasmic vacuolation and lipid degeneration of hepatocytes, focal necrosis with neutrophil infiltration, and sinusoidal congestion, indicating more severe liver damage in alcoholic hepatitis mice. Serum levels of AST, ALT, TC, and TG were significantly increased in alcoholic hepatitis mice. Serum levels of IL-1β and IL-18 were significantly increased in alcoholic hepatitis mice. Increased expression of GSDMD-N, NLRP3, and Caspase-11 proteins in the liver tissues of alcoholic hepatitis mice suggested GSDMD activation. Downregulating GSDMD could alleviate alcohol-induced hepatocyte pyroptosis. Mitochondrial dysfunction in hepatocytes caused by alcohol was observed in the liver tissues of alcoholic hepatitis mice, and this phenomenon was improved when GSDMD was downregulated. Improving mitochondrial function also suppressed alcohol-induced hepatocyte pyroptosis. Knocking out GSDMD or Drp1 improved alcoholic hepatitis liver damage, accompanied by a reduction in hepatocyte pyroptosis.

Research Conclusions:

In the process of alcohol-induced liver inflammation and damage, Drp1 regulates GSDMD-mediated mitochondrial dysfunction and hepatocyte pyroptosis. These findings may lay the foundation for the development of new treatments for alcoholic hepatitis.

Corresponding Author’s Brief Introduction:

Medical Doctor, Associate Chief Physician

Department of Liver Diseases, Peking University People’s Hospital, Peking University Hepatology Institute.