Editor’s Note: From November 10th to 14th, 2023, the annual international highlight event in the field of hepatology, the 2023 American Association for the Study of Liver Diseases (AASLD) Annual Meeting, is being held in Boston, USA. Dr. Liang Peng’s team from the Third Affiliated Hospital of Sun Yat-sen University has had multiple research results included in the conference. One randomized controlled trial evaluated the safety and effectiveness of tenofovir alafenamide fumarate (TAF) treatment for hepatitis B “healthy” carriers, and preliminary results indicate that TAF is safe for this population, with a rapid decrease in HBV DNA after 24 weeks of antiviral therapy, especially in the HBeAg-negative group where the HBV DNA undetectable rate is high [1].
In addition, the team also reported preliminary research results on the over-inhibition of ANXA4 in chronic hepatitis B to limit HBV replication through MCM2 autophagic degradation [2], as well as the latest discovery that three-dimensional cultured umbilical cord-derived mesenchymal stem cells have therapeutic effects on acute liver failure in mice comparable to traditional two-dimensional cultured cells [3]. “Hepatolgoy Digest” hereby reports on these findings, and the relevant content is shared below.
TAF Treatment for Hepatitis B “Healthy” Carriers: Preliminary Results of a Randomized Controlled Trial Released
Research Background
Currently, guidelines in various countries have gradually expanded indications for antiviral treatment in patients with chronic hepatitis B. However, there is still a lack of high-level evidence in evidence-based medicine for antiviral treatment in “hepatitis B healthy carriers.” Tenofovir alafenamide fumarate (TAF) is a first-line drug recommended for the treatment of chronic hepatitis B in guidelines. This study aimed to evaluate the safety and effectiveness of TAF treatment for hepatitis B “healthy” carriers.
Research Methods
This is a prospective randomized controlled trial that included hepatitis B “healthy” carriers, defined as HBV DNA positive (HBV DNA>20 IU/ml for HBeAg-positive individuals, and HBV DNA 20–2000 IU/ml for HBeAg-negative individuals), alanine aminotransferase (ALT) normal, liver stiffness value <5.8 KPa based on Fibroscan, no signs of cirrhosis, liver cancer, or liver fibrosis on imaging, and exclusion of a family history of cirrhosis or liver cancer. Participants were randomly assigned in a 1:1 ratio to the treatment group (received TAF treatment) and the control group (regular follow-up) to assess the safety and effectiveness of TAF treatment for hepatitis B “healthy” carriers. The detailed study protocol has been published in BMJ Open (BMJ Open. 2021;11: e048410), and the study design is shown in Figure 1.
Research Results
As of May 15, 2023, a total of 122 patients were included in the study, including 61 HBeAg-positive patients (30 received TAF treatment, 31 did not receive treatment) and 61 HBeAg-negative patients (29 received TAF treatment, 32 did not receive treatment). The researchers analyzed subjects who completed the 24-week follow-up (20 cases in the HBeAg-positive TAF group, 17 cases in the control group, 22 cases in the HBeAg-negative TAF group, and 17 cases in the control group).
In both the HBeAg-positive and HBeAg-negative cohorts, the TAF group and the control group had similar baseline characteristics. No serious adverse reactions were observed in any group during the 24-week follow-up, and there were no significant changes in ALT, creatinine, albumin, and Fibroscan.
In the HBeAg-positive cohort, the baseline average levels of HBV DNA in the TAF group and the control group were 7.85 and 8.03 log10 IU/ml, respectively. At 12 and 24 weeks, HBV DNA in the TAF group was significantly lower than in the control group, with an average decrease of 4.24 and 5.24 log10 IU/ml. The undetectable rate of HBV DNA at 24 weeks of treatment was 10% (2/20). In the HBeAg-negative cohort, the baseline average levels of HBV DNA in the TAF group and the control group were 2.74 and 2.60 log10 IU/ml, respectively. At 24 weeks, HBV DNA in the TAF group was significantly lower than in the control group, with an average decrease of 2.5 log10 IU/ml, and the undetectable rate of HBV DNA reached 82% (18/22). Changes in virological indicators compared to baseline at the 24-week follow-up for each group are shown in Figure 2.
Research Conclusion
Preliminary results indicate that TAF treatment in hepatitis B carriers is safe. In hepatitis B carriers receiving TAF antiviral treatment for 24 weeks, there is a rapid decrease in HBV DNA, especially in the HBeAg-negative group where the undetectable rate of HBV DNA is high. In this study, there was not a significant change in HBsAg before and after treatment, but the reduction in the HBeAg-positive group was greater than that in the HBeAg-negative group. For HBeAg-positive patients, HBV pgRNA increased compared to baseline after 24 weeks of TAF treatment, and this phenomenon needs further observation in subsequent treatment.
ANXA4 Limits HBV Replication by Inhibiting Autophagic Degradation of MCM2 in Chronic Hepatitis B – Preliminary Results Revealed!
Research Background
Hepatitis B virus (HBV) is a small enveloped DNA virus that can cause acute and chronic inflammation of the liver. Globally, about 250 million people are infected with chronic hepatitis B (CHB), and each year, over 800,000 deaths occur due to end-stage liver diseases such as cirrhosis and hepatocellular carcinoma caused by chronic hepatitis B. Understanding the mechanisms of HBV replication and transcription is crucial for overcoming HBV. Annexin A4 (ANXA4) is one of the membrane-associated proteins with calcium-regulating and phospholipid-binding properties. Previous studies have suggested that ANXA4 may play a potential role in the diagnosis, prognosis, and treatment of cancer. The latest research, using proteomics, found that ANXA4 protein has a protective role against viral interference.
Research Methods
Clinical samples were collected from patients with hepatitis, liver donors, or healthy individuals. Transcriptome sequencing was performed on CHB liver tissues and HBV-infected cells. Models with the full-length genome of HBV, HepG2.2.15 cells, and HBV-infected HepG2-NTCP cells were established. A mouse model of HBV infection was established using adenovirus.
Research Results
During CHB infection, the expression of ANXA4 increased. Downregulation of ANXA4 promoted HBV replication, worsening liver damage, while overexpression of ANXA4 alleviated liver damage. Mechanistically, the autophagic pathway was activated in the absence of ANXA4, promoting the autophagic degradation of minichromosome maintenance complex component 2 (MCM2). Inhibiting MCM2 activated HBV replication, while overexpression of MCM2 weakened ANXA4 deficiency-induced HBV replication and liver damage. Clinically, the expression of HBV protein was negatively correlated with ANXA4 levels, and CHB patients with higher ANXA4 levels (> 8 ng/ml) were more sensitive to interferon treatment
Research Conclusion
ANXA4 plays a protective role in the process of HBV infection. Under HBV attack, the expression of ANXA4 increases, inhibiting the autophagic degradation of MCM2 to suppress HBV replication and transcription, thereby alleviating liver damage and inhibiting the CHB infection process. ANXA4 also enhances the sensitivity of CHB patients to interferon treatment. ANXA4 is expected to become a new target for the treatment and prognosis assessment of CHB.
Researchers’ Comments
The pathogenicity of HBV is related to the bidirectional interaction between the virus and the host. HBV depends on membrane-associated proteins to bind and enter host cells. Here, we discovered the protective role of ANXA4 as a factor during the HBV infection process. Under the attack of HBV, the expression of ANXA4 increases, inhibiting the autophagic degradation of MCM2, thereby restricting HBV replication and transcription, alleviating liver damage, and suppressing the CHB infection process. Therefore, ANXA4 is expected to become a new target for the treatment and prognosis assessment of CHB
Treatment Effect of Three-Dimensional Cultured Umbilical Cord-Derived Mesenchymal Stem Cells on Acute Liver Failure Mice is not Inferior to Traditional Two-Dimensional Cultured Cells
The mortality rate of acute liver failure (ALF) is high. In existing treatments for ALF, the widespread use of liver transplantation and artificial liver support systems is constrained by various factors. Previous studies have suggested that mesenchymal stem cells (MSCs) may treat ALF through mechanisms such as immune regulation, anti-inflammatory effects, anti-cell apoptosis, and promotion of liver cell regeneration. Compared to two-dimensional culture, three-dimensional culture systems better simulate the real environment of cells in tissues. At the same time, to achieve a sufficient number of MSCs for cell transplantation and tissue engineering applications, three-dimensional culture of MSCs may be a more reliable method. In recent years, several studies have indicated that three-dimensional culture enhances the therapeutic potential of MSCs in disease models such as renal ischemia-reperfusion and ischemic stroke, but the exact effectiveness of three-dimensional cultured MSCs in treating ALF is not clear. This study aims to compare the therapeutic effects of three-dimensional and two-dimensional cultured MSCs in an ALF mouse model.
The study used a bioreactor and a three-dimensional dynamic system composed of microcarriers and traditional two-dimensional culture bottles to expand umbilical cord MSCs to obtain three-dimensional MSCs (3D-MSCs) and two-dimensional MSCs (2D-MSCs). Eight-week-old male C57BL/6J mice were intraperitoneally injected with 250 mg/kg thioacetamide (TAA) to construct the ALF mouse model. Twelve hours after modeling, the three groups were intravenously injected with an equal volume of phosphate-buffered saline (PBS), 1×106 2D-MSCs, or 3D-MSCs.
The results showed that during the cell culture stage, both two-dimensional and three-dimensional cultures could maintain the morphology, multi-directional differentiation ability, and expression of surface molecular markers of MSCs. In terms of cost and energy consumption, taking the harvest of 1×1010 MSCs as an example, the artificial cost, energy consumption, and space occupancy of three-dimensional culture were reduced by 83.3%, 97%, and 80%, respectively, compared to two-dimensional culture.
Compared to the PBS group, the prognosis of ALF mice was significantly improved in the 2D and 3D-MSC groups (PBS vs. 2D, P=0.0124; PBS vs. 3D, P=0.0009), with no significant difference between the 2D and 3D groups (Figure 5).
Compared to the PBS group, the levels of serum aspartate aminotransferase (AST) in ALF mice were significantly reduced in the 2D and 3D-MSC groups (36h: PBS vs. 2D: 8767.14±2457.29 U/L vs. 5673.00±2541.11 U/L, P=0.01; PBS vs. 3D: 8767.14±2457.29 U/L vs. 6289.5±1935.99 U/L, P=0.012; 60h: PBS vs. 2D: 4325.00±1025.30 U/L vs. 966.67±409.18 U/L, P=0.001; PBS vs. 3D: 4325.00±1025.30 U/L vs. 1400.00±1020.94 U/L, P=0.002), with no significant difference between the 2D and 3D groups (Figure 6)
Compared to the PBS group, the necrotic area of the liver in ALF mice was reduced in the 2D and 3D-MSC groups (Figure 7), and the liver necrotic area scores were significantly lower (60h: PBS vs. 2D: 0.71±0.08 vs. 0.43±0.07, P=0.02; PBS vs. 3D: 0.71±0.08 vs. 0.34±0.10, P＜0.000; 84h: PBS vs. 2D: 0.44±0.07 vs. 0.17±0.04, P=0.001; PBS vs. 3D: 0.44±0.07 vs. 0.13±0.12, P=0.007). There was no significant difference between the 2D and 3D groups (Figure 8)
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The results of this study demonstrate that both two-dimensional and three-dimensional cultured MSCs have excellent therapeutic potential in acute liver failure. The therapeutic effect of three-dimensional cultured MSCs on ALF mice is not inferior to that of two-dimensional MSCs.
References:
[1] Luo Q, Xu W, Li X, et al. TENOFOVIR ALAFENAMIDE FUMARATE THERAPY IN HEALTHY HBsAg CARRIERS: PRELIMINARY RESULTS OF A RANDOMIZED CONTROLLED TRIAL. AASLD 2023. Abstract 1442-C.
[2] Yang L, Liu X, Zhen L, et al. ANXA4 RESTRICTS HBV REPLICATION BY INHIBITING AUTOPHAGIC DEGRADATION OF MCM2 IN CHRONIC HEPATITIS B. AASLD 2023. Abstract 43236.
[3] Zhu S, Zhang Z, Peng L. THE THERAPEUTIC CAPACITIES OF THREE-DIMENSIONAL HUMAN UMBILICAL MESENCHYMAL STEM CELLS IN ACUTE LIVER FAILURE MICE. AASLD 2023. Abstract 4321-A.
TAG: AASLD 2023, HBV
