Therapeutic inhibition of HBsAg and HBV cccDNA through a novel phased combination treatment: glycine and interferon-α

Caorui Lin; Ying Huang; Ning Ran; Jie Liu; Linjie Luo; Xin Zhang; Xiaosang Zheng; Zhen Xun; Siyi Xu; Can Liu; Xiaohong Kong; Shiqing Feng; Haiting Mao; Qishui Ou

doi:10.1136/gutjnl-2025-334813

. 2025 Jul 11;74(12):e334813. doi: 10.1136/gutjnl-2025-334813

Therapeutic inhibition of HBsAg and HBV cccDNA through a novel phased combination treatment: glycine and interferon-α

Caorui Lin ^1,^2,^*,⁰, Ying Huang ^2,⁰, Ning Ran ^3,^4,⁰, Jie Liu ¹, Linjie Luo ², Xin Zhang ², Xiaosang Zheng ², Zhen Xun ², Siyi Xu ², Can Liu ², Xiaohong Kong ⁴, Shiqing Feng ^4,^*, Haiting Mao ^1,^*, Qishui Ou ^2,^✉

PMCID: PMC12703313 PMID: 40645766

Abstract

Background

The elimination of HBV covalently closed circular DNA (cccDNA) remains a critical hurdle for chronic hepatitis B (CHB) management.

Objective

In this investigation, we examined the efficacy of glycine administration and its potential enhancement in interferon-α (IFN-α) antiviral efficacy to stimulate hepatocyte proliferation to mitigate cccDNA levels.

Design

The study cohort comprised 89 healthy individuals and 496 HBV-infected patients, with subgroups of 30 and 42 participants receiving randomised nucleos(t)ide analogue (NA) and PegIFN-α treatments, respectively. Glycine concentrations were quantified via liquid chromatography‒tandem mass spectrometry, and its diagnostic potential was assessed via receiver operating characteristic curve analysis. The therapeutic impact of glycine was evaluated in various HBV-infected cell lines and murine models via various methodologies including transcriptomic sequencing, metabolomics sequencing, flow cytometry, immunofluorescence and in situ hybridisation.

Results

Elevated serum glycine levels with a robust positive correlation with serum alanine aminotransferase levels (R=0.7650) were observed in HBV-infected patients relative to healthy controls. The area under the curve for differentiating patients with HBeAg-expressing CHB from healthy controls was 0.9701. Glycine supplementation diminished HBV cccDNA levels by approximately 50% by promoting hepatocyte proliferation. Glycine is metabolised into a one-carbon unit, activating mTORC1 signalling via glycine transporter-1. Furthermore, glycine ameliorates hepatic inflammation by inhibiting the nuclear factor-kappa B signalling pathway through glycine receptors. Combination therapy with IFN-α effectively suppressed HBV replication, achieving a 60% reduction in HBsAg levels and sustained viral suppression in mice.

Conclusion

Glycine has the potential to reduce HBV cccDNA levels by stimulating hepatocyte proliferation. The phased administration of glycine and IFN-α significantly enhances its therapeutic efficacy. These findings suggest a novel and promising strategy for the treatment of CHB.

Keywords: HEPATITIS B, CHRONIC VIRAL HEPATITIS, DIAGNOSTIC VIROLOGY, ANTIVIRAL THERAPY, NUTRITION

WHAT IS ALREADY KNOWN ON THIS TOPIC

The therapeutic clearance of HBV covalently closed circular DNA (cccDNA) remains a challenge, which hinders the development of effective treatments for chronic hepatitis B (CHB).
Hepatocyte proliferation can lead to the dilution and potential elimination of HBV cccDNA because cccDNA is not faithfully copied to daughter cells during division.

WHAT THIS STUDY ADDS

Serum glycine levels are positively correlated with alanine aminotransferase levels and may serve as a potential biomarker for hepatic injury in HBV infection.
Glycine administration inhibits HBV cccDNA expression and enhances hepatocyte proliferation.Glycine administration triggers hepatocyte regeneration through activation of mTORC1 signalling and its metabolism into one-carbon units via glycine transporter-1 activity.
Glycine improves hepatic inflammation by blocking the nuclear factor-kappa B signalling pathway through glycine receptors.
Phased combination therapy of glycine and interferon-α (IFN-α) elicits therapeutic HBV replication inhibition.

HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY

Our findings reveal that glycine is an effective cccDNA inhibitor in HBV infection. Phased administration of glycine and IFN-α significantly elicits therapeutic levels of HBV replication suppression and thus opens a fundamentally novel avenue to achieve sustained viral suppression in patients with CHB.

Introduction

Chronic HBV infection poses a significant health burden globally, affecting approximately 254 million people worldwide.¹ Despite advances in antiviral therapies, a clinical cure for HBV remains elusive.² Current treatment strategies, including the administration of nucleos(t)ide analogues (NAs) and interferon-α (IFN-α), have shown limited efficacy in completely eradicating the virus.³ While the administration of NAs effectively suppresses viral replication, it fails to eliminate the viral reservoir. The administration of IFN-α, known for its immunomodulatory and antiviral properties, has a sustained response (SR) in only 30–40% of patients.^{4 5} The primary obstacle to effective treatment is the persistence of viral covalently closed circular DNA (cccDNA) in the nuclei of infected hepatocytes. This stable viral reservoir serves as a template for HBV replication and is resistant to current antiviral agents, thus maintaining chronic infection and contributing to the risk of cirrhosis and hepatocellular carcinoma.⁶,⁸ The challenge of eradicating viral cccDNA underscores the need for novel therapeutic strategies that can effectively target this viral form.^{9 10}

Recent research has focused on developing methods to inhibit cccDNA and promote its clearance from infected hepatocytes.^{10 11} Several approaches are currently under investigation, including direct targeting of cccDNA, interference with cccDNA formation, epigenetic modulation of cccDNA and gene editing technologies.¹²,¹⁴ Another promising strategy involves promoting hepatocyte proliferation, which can lead to the dilution and potential elimination of cccDNA.^{15 16} Studies have demonstrated that during hepatocyte division, cccDNA is not accurately copied to daughter cells, resulting in a reduction in the number of cccDNA copies per cell.^{17 18} This natural mechanism of cccDNA clearance through cell proliferation has sparked interest in the development of therapies that can safely stimulate hepatocyte turnover.^{10 19} Combining such approaches with current antiviral treatments may offer a more effective path towards achieving functional cures in patients with chronic HBV.

Glycine, a non-essential amino acid, has emerged as a versatile therapeutic agent across various pathological conditions.^{20 21} In our previous studies, we demonstrated that glycine administration promoted muscle regeneration in Duchenne muscular dystrophy model mice.²² Recent studies have highlighted the therapeutic benefits of glycine in treating liver diseases. Glycine can attenuate experimental non-alcoholic fatty liver disease by stimulating hepatic fatty acid β-oxidation and glutathione synthesis.²³ Moreover, glycine has anti-inflammatory and antiapoptotic effects, thereby protecting hepatocytes from damage and reducing liver fibrosis.²⁴,²⁷ Notably, earlier studies also demonstrated that glycine administration may enhance liver transplantation in injured rats.²⁸,³⁰ Given its ability to stimulate cell proliferation and its hepatoprotective properties, glycine has intriguing therapeutic potential and could be explored as an adjunct to existing antiviral therapies for patients with chronic HBV infection.

Here, we showed that serum glycine levels were significantly correlated with alanine aminotransferase (ALT) levels and highlighted their diagnostic value across various stages of natural HBV infection. Glycine supplementation resulted in enhanced HBV replication inhibition in vitro and in vivo. The efficacy of glycine can be attributed to its ability to suppress HBV cccDNA expression and increase hepatocyte regeneration induction, which in turn is mediated by an elevated one-carbon unit pool and the activation of the mammalian target of rapamycin complex 1 (mTORC1) signalling pathway via glycine transporter-1. Importantly, the phased administration of glycine and IFN-α significantly increased the therapeutic levels of HBsAg and HBV cccDNA suppression in HBV-infected mice. These findings provide insight into the role of glycine in the inhibition of HBV cccDNA and the crucial therapeutic effects of combined treatment with IFN-α to achieve sustained viral suppression in patients with chronic hepatitis B (CHB).

Materials and methods

Materials and methods used in this study are described in the online supplemental materials and methods.

Results

Serum glycine levels increase on HBV infection in humans

To examine the alterations in amino acid abundance in the circulation of HBV-infected patients, we used liquid chromatography‒tandem mass spectrometry (LC‒MS/MS) to analyse the serum amino acid composition and abundance in patients with different phases of chronic HBV infection who were naïve to treatment and in healthy controls (HCs) according to the flow chart of the patient selection process (online supplemental figure 1), and the characteristics of the patients are depicted in online supplemental table 1. As shown in figure 1A, HBV infection altered amino acid metabolism and composition; this, in particular, affected the levels of glycine (figure 1B), glutamate (online supplemental figure 2A) and glutamine (online supplemental figure 2B). Interestingly, we found that serum glycine levels were significantly different between patients in the IA (immune reactive) phase and HCs (figure 1C) or those in the immune tolerant (IT) phase (figure 1D). To determine the change in serum glycine levels during HBV infection, we collected serum from 64 HCs and 304 patients in different HBV infection phases, as depicted in online supplemental figure 1 and table 2. The LC‒MS/MS results revealed that circulating glycine levels were significantly greater in patients with HBV infection than in HCs, especially in patients in the IA phase (approximately twofold) (figure 1E). Interestingly, increased extracellular and decreased intracellular glycine levels were observed in HBV-infected cells, including HepG2-NTCP cells infected with HBV (online supplemental figure 2C), HepAD38 cells not treated with doxycycline (an inhibitor of HBV production) (online supplemental figure 2D) and HepG2.2.15 cells (online supplemental figure 2E).

Consistent with the in vitro data, we also detected significantly greater serum glycine levels in C57BL/6 mice subjected to rAAV8-mediated HBV replication than in wild-type C57BL/6 mice (online supplemental figure 2F and figure 1F). We found that glycine levels were significantly lower in the livers of HBV-infected mice than in those of normal controls (figure 1G). In addition, we analysed the correlations between serum glycine levels and HBV markers or serum ALT (a marker of liver damage) levels in HBV-infected patients and HCs, and the results revealed that glycine levels were significantly positively correlated with the ALT levels (correlation coefficient 0.7650) (figure 1H) and not with HBV markers (online supplemental figure 2G). These findings suggest that serum glycine level may serve as a novel indicator related to liver injury rather than a direct correlate of HBV replication activity. Furthermore, we used a receiver operating characteristic (ROC) curve to analyse the diagnostic value of serum glycine levels in patients in the IT phase compared with those in the IA phase and in HCs compared with those in the IA phase. The area under the ROC curve (AUC) in the IT versus IA groups was 0.9336 (figure 1I), and the AUC between the HC and IA groups was 0.9701 (figure 1J). Strikingly, we observed that serum glycine levels were significantly lower after treatment than before treatment in a cohort of patients with CHB who were receiving clinical treatment (figure 1K and online supplemental table 3). The above results revealed a significant increase in circulating glycine levels and a decrease in intrahepatic glycine levels in HBV-infected individuals, suggesting a potential role for glycine in the pathogenesis of CHB. Moreover, our findings suggest that serum glycine level may serve as a prognostic biomarker for liver injury in CHB.

Glycine inhibits HBV infection in vitro and in vivo

On the basis of the changes in glycine levels during HBV infection, we sought to investigate the role of glycine in HBV replication. First, we performed dose‒response assays using varying concentrations of glycine for 48 hours in HepG2-NTCP cells that had been infected with HBV for 5 days. Unexpectedly, compared with glycine depletion, the administration of high concentrations of glycine significantly inhibited HBV infection (0.4 mM glycine represents a normal culture medium level of glycine). This inhibition was indicated by a marked reduction in the levels of HBsAg (figure 2A), HBeAg (figure 2B), pgRNA (figure 2C) and HBV DNA (figure 2D); the most pronounced effect was observed with 1.2 mM glycine, resulting in an HBeAg inhibition rate near 70%. Notably, there was also a significant downregulation of HBx (Hepatitis B virus X protein) expression, accompanied by an unexpected increase in the number of cells present after treatment with 1.2 mM glycine (figure 2E and online supplemental figure 3A). Additionally, we investigated other HBV-expressing cell lines, including HepG2.2.15 cells and HepAD38 cells. As shown in online supplemental figure 3B,C, substantial inhibition of HBV replication was detected in HBV-expressing cells treated with 1.2 mM glycine, particularly based on the suppression of HBeAg levels.

To further evaluate the therapeutic potential of glycine, we administered glycine to a C57BL/6 mouse model of HBV infection mediated by rAAV8-HBV delivery (figure 2F and online supplemental table 4). Our findings revealed that glycine treatment significantly elevated both the serum (online supplemental figure 3D) and hepatic glycine concentrations (figure 2G). Consistent with the in vitro data, HBV replication was markedly suppressed by glycine treatment (figure 2H–J), especially HBeAg and HBV DNA expression. These results collectively indicate that glycine has the capacity to inhibit HBV replication, suggesting its potential as a therapeutic agent in the management of HBV infection.

Glycine enhances liver regeneration and contributes to total cccDNA reduction

Building on the observations regarding the effect of glycine on the number of cells presented in figure 2E, we hypothesise that glycine plays a pivotal role in cell viability. To test this hypothesis, we treated wild-type C57BL/6 mice with glycine. The results revealed that glycine administration facilitated an increase in body weight in wild-type mice (online supplemental figure 4A), whereas the liver to body weight ratio remained unchanged (online supplemental figure 4B). In contrast, we noted a concomitant increase in both body weight (figure 3A) and the liver to body weight ratio (figure 3B) in the HBV-infected mice, suggesting that glycine treatment predominantly enhances the hepatic mass during HBV infection. A marked increase in the population of dividing hepatocytes was detected via pathological examination of liver tissues (figure 3C,D). Therefore, we conducted experiments to further assess the effects of glycine administration on cell proliferation. As depicted in online supplemental figure 4C–E, glycine significantly promoted the proliferation and viability of HL-7702 cells (a normal human hepatocyte line). Additionally, 5′-bromo-2′-deoxyuridine (BrdU) incorporation assays demonstrated a notable increase in BrdU-positive cell counts in HBV-infected livers treated with glycine (figure 3E,F). These findings suggest that glycine can promote the proliferation of HBV-infected hepatocytes, which was confirmed by increased numbers of ethynyl deoxyuridine-positive cells (online supplemental figure 4F) and enhanced colony formation (online supplemental figure 4G) in HBV-infected HepG2-NTCP cells treated with glycine.

Integrating these findings with those from recent research,¹⁵ we propose that glycine inhibits the expression of HBV cccDNA by stimulating hepatocyte proliferation, thereby suppressing HBV replication. As expected, we detected a significant reduction in HBV cccDNA levels with glycine treatment in HBV-infected HepG2-NTCP cells (online supplemental figure 4H), with an inhibition rate of 50% in HBV-infected or HBV-expressing cell lines (figure 3G). A marked downregulation of the HBV receptor sodium taurocholate cotransporting polypeptide (NTCP)³¹ was observed (figure 3H), which corroborates existing findings regarding the decreased expression of NTCP in newly formed hepatocytes.³² Moreover, we normalised HBV replication levels relative to cell counts to accurately assess viral activity on a per-cell basis in HBV-infected HepG2-NTCP (figure 3I), HepAD38 (figure 3J) and HepG2.2.15 cells (figure 3K). The results indicated that glycine treatment significantly suppressed HBV replication, as evidenced by 70–85% reductions per cell in HBeAg levels and by 60–80% reductions per cell in HBsAg levels across cell lines. Notably, glycine treatment exhibited limited antiviral efficacy in primary human hepatocytes (maintained in a differentiated, non-proliferative state, online supplemental figure 4I,J). This contrast to its robust activity in proliferating hepatocyte models strongly supports that glycine’s antiviral activity is mitosis dependent, requiring active hepatocyte division to dilute cccDNA. To further substantiate this, we used in situ hybridisation to detect cccDNA expression in HBV-infected liver tissues (hepatocyte nuclear factor-4α was used as a hepatocyte identity marker), and the results revealed that glycine promoted liver regeneration and significantly reduced cccDNA levels in nuclei in generated hepatocytes (figure 3L,M). Collectively, these results suggest that glycine reduces cccDNA levels by promoting hepatocyte regeneration, in turn inhibiting HBV replication.

Glycine induces liver regeneration by activating the AKT-mTORC1 signalling pathway, resulting in HBV cccDNA reduction and antiviral efﬁcacy

To investigate the mechanism of action of glycine, we performed RNA-seq on HepG2.2.15 cells following a 48-hour glycine treatment. As shown in figure 4A, glycine administration led to significant upregulation of 284 genes and downregulation of 50 genes. Gene set enrichment analysis (GSEA) revealed that the genes with upregulated expression were markedly enriched in pathways associated with the cell cycle and cell division (figure 4B). Subsequent cell cycle analysis of HBV-infected HepG2-NTCP cells revealed that glycine treatment resulted in an increased proportion of cells in the S phase and G2 phase (figure 4C–E), concomitantly increasing the expression of key genes related to cell proliferation, including cyclin-dependent kinase 1 (CDK1), cyclin D1 (CCND1) and proliferating cell nuclear antigen (PCNA) genes, as validated by quantitative real-time reverse transcriptase PCR (RT-PCR) (figure 4F). These findings suggest that glycine administration effectively promotes hepatocyte proliferation.

Moreover, KEGG (Kyoto Encyclopedia of Genes and Genomes) (online supplemental figure 5A) and GSEA analyses (figure 4G) demonstrated that glycine administration facilitated the activation of the PI3K-AKT signalling pathway, as evidenced by the increased phosphorylation of AKT (figure 4H). Strikingly, the phosphorylation levels of mTOR, a hallmark of speciﬁc activation by AKT; the phosphorylation levels of ribosomal protein S6 kinase 1 (p-S6K1); the phosphorylation levels of ribosomal protein S6 (p-S6); and the phosphorylation levels of eukaryotic initiation factor 4E-binding protein 1 (p-4EBP1), key downstream targets of mTORC1, were elevated in HBV-infected HepG2-NTCP cells (figure 4I,J) and HepG2.2.15 cells (online supplemental figure 5B) treated with glycine compared with those in the untreated controls. Blockade of mTORC1 activation via PP242, a small-molecule protein kinase inhibitor targeting the ATP-binding site of mTOR,³³ in HBV-infected HepG2-NTCP cells (figure 4K,L) and in HepG2.2.15 cells (online supplemental figure 5C,D) abolished the enhancement potentiated by glycine, as cell numbers (figure 4M and online supplemental figure 5E) and cell viability (figure 4N and online supplemental figure 5F) significantly declined. The antiviral efficacy was also negated, as HBV replication (figure 4O,P and online supplemental figure 5G,H) dramatically increased in HBV-infected cells treated with glycine coadministered with PP242, confirming the relevance of mTORC1 to the functionality of glycine. Additionally, pharmacological inhibition of AKT with LY294002³⁴ significantly attenuated glycine-induced AKT phosphorylation and downstream mTORC1 activation (online supplemental figure 5I), thereby reversing glycine’s suppression of HBV replication (online supplemental figure 5J). These results indicate that glycine administration promotes AKT-mTORC1 activation, which in turn enhances hepatocyte proliferation and antiviral efficacy.

Glycine increases liver regeneration by boosting the one-carbon unit pool via glycine transporter-1

To understand the role of glycine in promoting liver regeneration and to determine whether active transport of glycine is critical for its functionality, we evaluated the expression of glycine transporter-1 (GlyT1), the predominant glycine transporter,³⁵ in HBV-infected HepG2-NTCP cells or HepAD38 cells treated with glycine. As shown in figure 5A and online supplemental figure 6A, upregulation of GlyT1 expression was observed in cells treated with glycine compared with cells treated with a lower dose of glycine or in glycine-depleted cells. We subsequently blocked GlyT1 expression with bitopertin (BP), a specific GlyT1 inhibitor,³⁶ in glycine-treated HBV-infected cells. Strikingly, cell proliferation was completely compromised in the presence of BP (figure 5B and online supplemental figure 6B), as the activation of the mTORC1 signalling pathway significantly declined in cells coadministered with glycine and BP (figure 5C and online supplemental figure 6C). The antiviral efficacy was consistently negated when BP was coadministered with glycine, as HBsAg, HBeAg, HBV DNA, pgRNA and cccDNA levels signiﬁcantly decreased (figure 5D and online supplemental figure 6D), indicating that active transport is crucial for the functionality of glycine.

Additionally, we harvested liver tissues from HBV-infected mice following glycine treatment for comprehensive metabolomic analysis via LC–MS/MS. As shown in online supplemental figure 6E, glycine treatment significantly increased the abundance of 51 metabolites and decreased the abundance of 45 metabolites compared with those in the glycine-depleted group. Notably, the metabolites with upregulated abundance were predominantly enriched in nucleotides and their metabolites, methylenetetrahydrofolate, carbohydrates and their metabolites, and others (figure 5E). KEGG pathway analysis revealed significant clustering related to one-carbon metabolism (figure 5F), with the major metabolites characterised as one-carbon group donors or carriers (figure 5G). To validate this, we inhibited the activity of dihydrofolate reductase (DHFR) using methotrexate (MTX), a DHFR competitive antagonist,^{37 38} to validate the role of glycine metabolism in HBV replication. Compared with those in the glycine-treated cells, the number and viability of the cells in the MTX-treated, glycine and MTX combination-treated and control groups were significantly lower (figure 5H,I and online supplemental figure 6F). In parallel with reduced cell proliferation, a dramatic increase in HBsAg, HBeAg, HBV DNA, pgRNA and cccDNA levels was observed in HBV-infected cells coadministered with glycine and MTX compared with those administered glycine alone (figure 5J and online supplemental figure 6G). These results suggest that glycine contributes to liver regeneration and antiviral efficacy by replenishing one-carbon units and promoting mTORC1 activation (figure 5K).

Glycine administration triggers glycine receptor signalling to improve hepatic inflammation

Following the decrease in HBV replication, we detected notable improvements in the pathological morphology of liver tissues (figure 6A), alongside a significant reduction in serum ALT level (figure 6B). Immunohistochemical analysis revealed a marked decrease in F4/80-positive cells (macrophages) within the liver (figure 6C). The above data suggest that glycine treatment effectively ameliorated the hepatocytic necrosis and inflammatory infiltration associated with CHB infection. We determined via GSEA that glycine administration significantly activated calcium ion transport channels (figure 6D). Moreover, high-expression gene profiling (online supplemental figure 7A) and RT-PCR validation (figure 6E) revealed that glycine administration significantly upregulated the expression of several genes related to calcium ion transport, including solute carrier family 26 member 9 (SLC26A9),³⁹ ATPase sarcoplasmic/endoplasmic reticulum Ca²⁺ transporting 3 (ATP2A3)⁴⁰ and transmembrane channel-like 5 (TMC5) genes.⁴¹ Further intracellular calcium ion measurements in HBV-infected HepG2-NTCP cells indicated that glycine treatment significantly reduced the intracellular calcium ion concentration (figure 6F).

Glycine receptors are reportedly involved in the exchange of calcium and chloride ions,^{20 42} and we hypothesise that glycine administration may attenuate cytosolic calcium ion levels via glycine receptors, thereby exerting anti-inflammatory effects during HBV infection. Compared with those in the control group, a notable decline in the elevated ROS (Reactive Oxygen Species) levels (figure 6G) caused by HBV infection was observed in the glycine treatment group. Western blot analyses confirmed that glycine treatment enhanced the expression of glycine receptors (figure 6H) and suppressed the activation of the nuclear factor-kappa B (NF-κB) signalling pathway (figure 6I). To confirm that glycine administration enables glycine receptors to transport calcium ions, we knocked down the expression of glycine receptors via siRNA in HepG2-NTCP cells (online supplemental figure 7B). As depicted in online supplemental figure 7C,D, this intervention did not affect cell proliferation or glycine-induced activation of the mTORC1 signalling pathway. However, glycine-induced calcium ion reduction (figure 6J), a decrease in ROS levels (figure 6K) and NF-κB activation (figure 6L) were substantially blocked by the administration of siRNA targeting the glycine receptor in HBV-infected HepG2-NTCP cells compared with those in cells subjected to glycine treatment. Taken together, these results prove that glycine administration may inhibit the increase in intracellular calcium ion levels via activation of glycine receptors, which results in the suppression of NF-κB signalling to ameliorate hepatic inflammation and necrosis during HBV infection (figure 6M).

Glycine enhances IFN-α antiviral efficacy through a phased administration strategy

Given the limited effect of glycine on HBsAg inhibition, we wanted to investigate whether the administration of IFN-α, an anti-HBV drug used in clinical therapy for patients with CHB with a markedly high-serum HBsAg clearance rate,^{43 44} can augment glycine potency in a model of HBV infection. IFN-α was administered to a 5-week-old C3H mouse model of HBV infection mediated by rAAV8-HBV delivery. This HBV-infected mouse model closely recapitulates the clinical manifestations of patients with viral hepatitis and shows high levels of persistent HBV cccDNA.⁴⁵,⁴⁷ These mice were treated with IFN-α at a dose of 50 µg/kg twice a week^{46 48} for 3 weeks, and glycine was supplemented via the diet daily (online supplemental figure 8A). Unfortunately, we noted an increase in the serum HBsAg level in the combination therapy group compared with that in the IFN-α alone group, which was similar to that in the glycine group (online supplemental figure 8B), whereas a significant reduction in the serum HBeAg (online supplemental figure 8C) and HBV DNA levels (online supplemental figure 8D) was observed in the combination therapy group compared with that in the untreated control group, suggesting that the IFN-α treatment in the combination therapy group may not have fulfilled its expected therapeutic role. Notably, we detected that the baseline serum glycine levels in the IFN-α SR group were significantly lower than those in the non-response group (figure 7A) in a cohort of patients with CHB receiving PegIFN-α therapy (online supplemental table 5). These findings imply that IFN-α administration exerts its antiviral effects primarily under low-serum glycine conditions. Consequently, we devised a novel treatment protocol for IFN-α/glycine (IG) combination therapy: oral supplementation of 5% glycine for three consecutive days per week for 3 weeks, a previously used dosing regimen,⁴⁹ alongside IFN-α injection twice a week for 3 weeks prior to a glycine-containing diet (figure 7B).

Notably, the IG group presented significant decreases in the levels of serum HBV DNA (figure 7C), HBeAg (figure 7D) and HBsAg (figure 7E) during the treatment period; in particular, the inhibition rate of HBsAg reached 60% 1 week after the last treatment compared with that of the groups receiving IFN-α alone and glycine alone and the untreated controls. Additionally, the expression of intrahepatic cccDNA and pgRNA was significantly decreased (figure 7F), which was consistent with the obviously reduced areas of HBsAg and HBcAg expression (figure 7G,H) in the livers of IG-treated mice, providing robust evidence for the antiviral efficacy of IG combination therapy. Importantly, the number of monocyte infiltration and necrotic cells (online supplemental figure 8E) and the hepatic fibrosis areas (figure 7I,J) were substantially decreased in the livers of the HBV-infected mice treated with IG compared with those of the other groups of treated mice. Concordantly, as a result of treatment with IG, the serum levels of enzymes related to liver function, specifically ALT (figure 7K) and aspartate aminotransferase (figure 7L), which are highly abundant in the liver, were significantly decreased in mice. A marked increase in the level of albumin (figure 7M), a protein secreted from the liver that reflects synthetic function, was observed. The above data indicate improved hepatic function in the IG-treated mice. There were no morphological alterations in the kidneys (online supplemental figure 8F) and no differences in the levels of creatinine (online supplemental figure 8G) and uric acid (online supplemental figure 8H), which are biochemical parameters of the kidney, between the IG-treated and the other groups of treated mice. These data suggest that this combination therapy for phased incorporation of IFN-α and glycine exerts therapeutic antiviral effects and restores liver function without any detectable toxicity.

Discussion

Glycine is vital for mammalian nutrition and metabolism and is known to possess a broad array of beneficial effects against various injuries and diseases.⁵⁰ In this study, we provide novel insight into the role of glycine as a potential therapeutic agent for the treatment of patients with CHB, particularly in combination with IFN-α therapy. Our findings indicate that the serum glycine level can serve as a biomarker capable of distinguishing the natural history of HBV infection and is positively associated with serum ALT level in humans. Intriguingly, exogenous glycine treatment can inhibit HBV replication by nearly 50% according to HBV cccDNA levels both in vitro and in vivo. Dissection of glycine functionality unveils that glycine enhances liver regeneration, which in turn leads to diluted or even depleted HBV cccDNA expression and subsequent HBV suppression. Mechanistically, glycine promotes liver regeneration by boosting the one-carbon unit pool and triggering activation of the mTORC1 signalling pathway via glycine transporter-1. Moreover, glycine ameliorates hepatic inflammation by restraining NF-κB activation via glycine receptors. Importantly, the incorporation of IFN-α via phased intraperitoneal injection further elicits therapeutic levels of HBV inhibition and significantly reduces HBsAg expression by up to 60%, prevents pathological progression and improves liver function in HBV-infected C3H mice. This study demonstrates the potential of a novel therapeutic strategy that combines oral glycine supplementation to induce liver regeneration to deplete HBV cccDNA and IFN-α administration to inhibit HBsAg expression. This approach has been shown to result in sustained viral suppression for HBV in C3H mice and may extend its applicability to patients with CHB, thereby offering a promising new option for the clinical use of glycine.

There are currently no adequate detection methods or host-derived indices, apart from liver biopsy, that can accurately diagnose the specific stage of HBV infection progression.^{2 4} In our prior research, we showed that serum MOTS-c⁵¹ and ATP⁵² levels can be used to diagnose the progression of CHB. Here, we show that serum glycine levels significantly increase with the progression of HBV infection, effectively distinguishing healthy individuals from HBeAg-positive patients with CHB (AUC of 0.9701). Strikingly, our data demonstrate that serum glycine levels significantly decrease in patients with CHB following antiviral therapy, suggesting the utility of glycine as a reliable prognostic marker for monitoring CHB treatment progress. Surprisingly, we observed a strong positive correlation between serum glycine and ALT levels and a modest inverse correlation between glycine and HBV replication markers. These findings indicate that serum glycine levels primarily serve as a biomarker of hepatic injury rather than a direct correlate of HBV replication activity. Further histological and biochemical analyses of liver biopsies from patients at different HBV infection stages are essential to validate this hypothesis. In future studies, we will aim to combine serum glycine levels with other biomarkers for joint diagnostic purposes, which could provide a new diagnostic method to potentially replace or reduce the need for liver biopsy.

Glycine is a conditionally essential amino acid; therefore, it is extensively used as a food additive.⁵³ Currently, there are no approved therapies that directly target and eliminate cccDNA. Efforts to develop curative therapies for HBV are focused on strategies to either silence cccDNA transcription or promote its degradation.⁵⁴ One promising approach involves promoting hepatocyte turnover and proliferation, as the division of infected hepatocytes can lead to the dilution and potential loss of cccDNA.¹⁵ Notably, a previous study reported that cell regeneration can inhibit HBV DNA synthesis in individual hepatocytes.⁵⁵ Our study demonstrates that glycine administration achieves ~50% suppression of HBV markers (HBeAg, HBV DNA, pgRNA) and cccDNA in vitro and in vivo through a unique mitosis-dependent dilution mechanism. Therapeutic efficacy of glycine correlates with its ability to promote hepatocyte proliferation, a process that dilutes cccDNA copies during mitotic segregation, which resulted in proliferation-driven cccDNA clearance instead of direct genome editing. Although glycine levels were elevated in HBV-infected patients and mice serum, the intracellular glycine levels in HBV-infected liver tissue and cell lines were decreased, implying a paradoxical glycine imbalance in HBV infection. Intracellular depletion of glycine may result from HBV-induced membrane damage and pathological efflux, as evidenced by the strong serum glycine-ALT correlation (R=0.7650, p<0.0001). Exogenous glycine can restore hepatocyte homeostasis via (1) glycine transporter-1 upregulation to enhance uptake and (2) extracellular pool saturation to reverse efflux gradients, thereby replenishing intracellular reserves, which are essential for antiviral functions. However, the efficacy of glycine in clearing cccDNA is cell context dependent: while sustained cccDNA reduction occurred in HepAD38 cells (cccDNA dependent), in HepG2.2.15 cells (integrated DNA dominant), rapid HBsAg/HBV DNA rebound post-glycine withdrawal was observed (data not shown). This transient suppression was aligned with the persistent integration-driven transcription. Overall, our findings imply that glycine is targeted in cccDNA-rich microenvironments, particularly during early infection phases to reduce cccDNA production.

Clinical translational challenges arise from glycine’s dual role as a mitotic driver and metabolic regulator. Although hepatocarcinogenic signals in mice (normal serum AFP (Alpha-fetoprotein); data not shown) or increased HBV-integration frequency in vitro (Alu-PCR; data not shown) were not observed, long-term glycine treatment limited the proliferative capacity of hepatocytes in both cultured HepAD38 cell line and treated HBV-infected mouse model (10-week glycine administration; data not shown). These findings indicate that optimising the minimal effective dose to balance cccDNA dilution and hepatocyte replicative capacity is extremely necessary in the following clinical translational research. Notably, glycine treatment could reduce NTCP expression, a finding initially interpreted through the lens of HBV entry inhibition. However, NTCP also serves as the primary transporter for conjugated bile acids, and its downregulation could theoretically impair hepatic bile acid clearance, leading to systemic accumulation. Chronic glycine administration in clinical settings warrants rigorous monitoring of bile acid profiles, particularly in patients with pre-existing cholestatic conditions. Additionally, the limited effect of glycine on HBsAg levels may be attributed to its insufficient impact on integrated HBV DNA, the main source of HBsAg^{56 57}; this requires further exploration. Collectively, these findings delineate the therapeutic roles of glycine as a transient adjunct to reduce cccDNA burden. More administration methods, such as pulsatile dose-based administration, should be developed to mitigate proliferative exhaustion and off-target metabolic effects of glycine. From this study, we propose glycine as a phase I-ready adjunct therapy through pulsatile dosing and combined with RNA interference (targeting integrated DNA transcripts) or immune modulators (eg, IFN-α) to achieve functional cure in the clinic. Furthermore, long-term safety in patients with cirrhosis and cccDNA clearance kinetics using single-cell genomic approaches should be validated in future studies.

Mechanistically, glycine appears to exert its effects through dual pathways. It enters hepatocytes via specific transporter-1, where it is metabolised into one-carbon units, thereby activating the mTORC1 signalling pathway to promote cell proliferation. Simultaneously, glycine interacts with glycine receptors to inhibit calcium influx, suppressing NF-κB signalling and alleviating hepatic inflammation. This dual mechanism impedes HBV replication and ameliorates liver inflammation, offering a comprehensive therapeutic approach. Previous studies have demonstrated the utility of glycine in supplements for liver transplantation in patients,⁵⁸ and our findings further support its potential application in CHB treatment.

Although IFN-α administration has been explored for treating patients with CHB in the clinic, its overall effect is limited.⁵⁹ Given the limitations of the effect of glycine on HBsAg levels, we explored IFN-α combination therapy. Unexpectedly, the initial results showed that simultaneous administration of both agents did not yield the desired increase in the effect of IFN-α on HBsAg suppression. Furthermore, a clinical treatment cohort analysis revealed that patients with superior IFN-α treatment responses had lower baseline serum glycine levels. This insight led us to develop a staggered treatment regimen in which glycine administration was adjusted three times per week. This innovative regimen significantly reduced HBsAg and cccDNA expression, resulting in more than 60% suppression of HBsAg. This approach mitigates the drawbacks of high-frequency glycine monotherapy and offers a viable pathway towards sustained viral suppression and potential complete eradication of HBV cccDNA in patients with CHB. Considering the affordability and nutritional role of glycine, future research will focus on clinical trials involving patients with CHB to further validate the effects of this combined therapy. Additionally, we aimed to elucidate the mechanisms underlying the poor IFN-α response in patients with high-baseline glycine levels, potentially identifying new therapeutic targets.

In summary, we demonstrate that glycine inhibits HBV replication by diluting the reservoir of cccDNA through enhancing hepatocyte regeneration. Phased administration of glycine and IFN-α significantly elicits therapeutic levels of HBsAg and HBV cccDNA suppression and thus opens a fundamentally novel avenue to achieve sustained viral suppression in patients with CHB. Moreover, our ﬁndings unveil the role of glycine as a biomarker for monitoring the progression of HBV infection.

Supplementary material

online supplemental file 1

gutjnl-74-12-s001.doc^{(15.4MB, doc)}

DOI: 10.1136/gutjnl-2025-334813

online supplemental file 2

gutjnl-74-12-s002.pptx^{(5MB, pptx)}

DOI: 10.1136/gutjnl-2025-334813

Acknowledgements

We thank all the staff and patients from The First Affiliated Hospital of Fujian Medical University and The Second Hospital of Shandong University for the provision of the samples used in this study. We thank Professor Juan Chen and her team (Chongqing Medical University) for providing assistance in performing the Southern blot experiments.

Footnotes

Funding: This work was supported by the National Natural Science Foundation of China (Grant Nos 82202597 and 82030063), the Postdoctoral Fellowship Program (Grade B) of China Postdoctoral Science Foundation (CPSF) (Grant No GZB20240404), the Taishan Scholars Program of Shandong Province (tsqn202408353) and the Major Special Project of Fujian Provincial Health Technology Project (2024ZD01003).

Provenance and peer review: Not commissioned; externally peer reviewed.

Patient consent for publication: Not applicable.

Ethics approval: This study involves human participants and was approved by the Ethics Committee of The First Affiliated Hospital of Fujian Medical University (Approval No MRCTA, ECFAH of FMU (2022) 278). The study adhered to the principles outlined in the 1975 Declaration of Helsinki. Participants gave informed consent to participate in the study before taking part.

Patient and public involvement: Patients and/or the public were involved in the design, or conduct, or reporting, or dissemination plans of this research. Refer to the Methods section for further details.

Data availability statement

Data are available upon reasonable request.

References

1.World Health Organization . Global hepatitis report 2024 action for access in low- and middle-income countries. World Health Organization; 2024. [Google Scholar]
2.WHO . Guidelines for the prevention, diagnosis, care and treatment for people with chronic hepatitis B infection. World Health Organization; 2024. [PubMed] [Google Scholar]
3.Cui F, Blach S, Manzengo Mingiedi C, et al. Global reporting of progress towards elimination of hepatitis B and hepatitis C. Lancet Gastroenterol Hepatol. 2023;8:332–42. doi: 10.1016/S2468-1253(22)00386-7. [DOI] [PubMed] [Google Scholar]
4.Kramvis A, Chang K-M, Dandri M, et al. A roadmap for serum biomarkers for hepatitis B virus: current status and future outlook. Nat Rev Gastroenterol Hepatol. 2022;19:727–45. doi: 10.1038/s41575-022-00649-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
5.Lok AS, Zoulim F, Dusheiko G, et al. Hepatitis B cure: From discovery to regulatory approval. J Hepatol. 2017;67:847–61. doi: 10.1016/j.jhep.2017.05.008. [DOI] [PubMed] [Google Scholar]
6.Lucifora J, Xia Y, Reisinger F, et al. Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA. Science. 2014;343:1221–8. doi: 10.1126/science.1243462. [DOI] [PMC free article] [PubMed] [Google Scholar]
7.Nassal M. HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic hepatitis B. Gut. 2015;64:1972–84. doi: 10.1136/gutjnl-2015-309809. [DOI] [PubMed] [Google Scholar]
8.Seeger C, Mason WS. Molecular biology of hepatitis B virus infection. Virology (Auckl) 2015;479–480:672–86. doi: 10.1016/j.virol.2015.02.031. [DOI] [PMC free article] [PubMed] [Google Scholar]
9.Revill PA, Chisari FV, Block JM, et al. A global scientific strategy to cure hepatitis B. Lancet Gastroenterol Hepatol. 2019;4:545–58. doi: 10.1016/S2468-1253(19)30119-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
10.Fung S, Choi HSJ, Gehring A, et al. Getting to HBV cure: The promising paths forward. Hepatology. 2022;76:233–50. doi: 10.1002/hep.32314. [DOI] [PubMed] [Google Scholar]
11.Levrero M, Pollicino T, Petersen J, et al. Control of cccDNA function in hepatitis B virus infection. J Hepatol. 2009;51:581–92. doi: 10.1016/j.jhep.2009.05.022. [DOI] [PubMed] [Google Scholar]
12.Belloni L, Pollicino T, De Nicola F, et al. Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function. Proc Natl Acad Sci U S A. 2009;106:19975–9. doi: 10.1073/pnas.0908365106. [DOI] [PMC free article] [PubMed] [Google Scholar]
13.Yang HC, Kao JH. Persistence of hepatitis B virus covalently closed circular DNA in hepatocytes: molecular mechanisms and clinical significance. Emerg Microbes Infect. 2014;3:e64. doi: 10.1038/emi.2014.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
14.Wu K, He M, Mao B, et al. Enhanced delivery of CRISPR/Cas9 system based on biomimetic nanoparticles for hepatitis B virus therapy. J Control Release. 2024;374:293–311. doi: 10.1016/j.jconrel.2024.08.019. [DOI] [PubMed] [Google Scholar]
15.Allweiss L, Volz T, Giersch K, et al. Proliferation of primary human hepatocytes and prevention of hepatitis B virus reinfection efficiently deplete nuclear cccDNA in vivo. Gut. 2018;67:542–52. doi: 10.1136/gutjnl-2016-312162. [DOI] [PubMed] [Google Scholar]
16.Chang J, Guo F, Zhao X, et al. Therapeutic strategies for a functional cure of chronic hepatitis B virus infection. Acta Pharm Sin B. 2014;4:248–57. doi: 10.1016/j.apsb.2014.05.002. [DOI] [PMC free article] [PubMed] [Google Scholar]
17.Lutgehetmann M, Volz T, Köpke A, et al. In vivo proliferation of hepadnavirus-infected hepatocytes induces loss of covalently closed circular DNA in mice. Hepatology. 2010;52:16–24. doi: 10.1002/hep.23611. [DOI] [PubMed] [Google Scholar]
18.Köck J, Rösler C, Zhang J-J, et al. Generation of covalently closed circular DNA of hepatitis B viruses via intracellular recycling is regulated in a virus specific manner. PLoS Pathog. 2010;6:e1001082. doi: 10.1371/journal.ppat.1001082. [DOI] [PMC free article] [PubMed] [Google Scholar]
19.Dandri M, Petersen J. Mechanism of Hepatitis B Virus Persistence in Hepatocytes and Its Carcinogenic Potential. Clin Infect Dis. 2016;62 Suppl 4:S281–8. doi: 10.1093/cid/ciw023. [DOI] [PMC free article] [PubMed] [Google Scholar]
20.Wheeler MD, Ikejema K, Enomoto N, et al. Glycine: a new anti-inflammatory immunonutrient. Cell Mol Life Sci. 1999;56:843–56. doi: 10.1007/s000180050030. [DOI] [PMC free article] [PubMed] [Google Scholar]
21.Zhong Z, Wheeler MD, Li X, et al. L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. Curr Opin Clin Nutr Metab Care. 2003;6:229–40. doi: 10.1097/01.mco.0000058609.19236.a4. [DOI] [PubMed] [Google Scholar]
22.Lin C, Han G, Ning H, et al. Glycine Enhances Satellite Cell Proliferation, Cell Transplantation, and Oligonucleotide Efficacy in Dystrophic Muscle. Mol Ther. 2020;28:1339–58. doi: 10.1016/j.ymthe.2020.03.003. [DOI] [PMC free article] [PubMed] [Google Scholar]
23.Rom O, Liu Y, Liu Z, et al. Glycine-based treatment ameliorates NAFLD by modulating fatty acid oxidation, glutathione synthesis, and the gut microbiome. Sci Transl Med. 2020;12:eaaz2841. doi: 10.1126/scitranslmed.aaz2841. [DOI] [PMC free article] [PubMed] [Google Scholar]
24.Qu P, Rom O, Li K, et al. DT-109 ameliorates nonalcoholic steatohepatitis in nonhuman primates. Cell Metab. 2023;35:742–57. doi: 10.1016/j.cmet.2023.03.013. [DOI] [PubMed] [Google Scholar]
25.Ghrayeb A, Finney AC, Agranovich B, et al. Serine synthesis via reversed SHMT2 activity drives glycine depletion and acetaminophen hepatotoxicity in MASLD. Cell Metab. 2024;36:116–29. doi: 10.1016/j.cmet.2023.12.013. [DOI] [PMC free article] [PubMed] [Google Scholar]
26.Wang Z, Zhang J, Wang L, et al. Glycine mitigates renal oxidative stress by suppressing Nox4 expression in rats with streptozotocin-induced diabetes. J Pharmacol Sci. 2018;137:387–94. doi: 10.1016/j.jphs.2018.08.005. [DOI] [PubMed] [Google Scholar]
27.Al-Saeedi M, Liang R, Schultze DP, et al. Glycine protects partial liver grafts from Kupfer cell-dependent ischemia–reperfusion injury without negative efect on regeneration. Amino Acids. 2019;51:903–11. doi: 10.1007/S00726-019-02722-5/TABLES/3. [DOI] [PubMed] [Google Scholar]
28.Schemmer P, Bradford BU, Rose ML, et al. Intravenous glycine improves survival in rat liver transplantation. Am J Physiol. 1999;276:G924–32. doi: 10.1152/ajpgi.1999.276.4.G924. [DOI] [PubMed] [Google Scholar]
29.Bachmann S, Peng XX, Currin RT, et al. Glycine in Carolina rinse solution reduces reperfusion injury, improves graft function, and increases graft survival after rat liver transplantation. Transplant Proc. 1995;27:741–2. [PubMed] [Google Scholar]
30.Benko T, Frede S, Gu Y, et al. Glycine pretreatment ameliorates liver injury after partial hepatectomy in the rat. J Invest Surg. 2010;23:12–20. doi: 10.3109/08941930903469466. [DOI] [PubMed] [Google Scholar]
31.Yan H, Zhong G, Xu G, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012;3:e00049. doi: 10.7554/eLife.00049. [DOI] [PMC free article] [PubMed] [Google Scholar]
32.Yan Y, Allweiss L, Yang D, et al. Down-regulation of cell membrane localized NTCP expression in proliferating hepatocytes prevents hepatitis B virus infection. Emerg Microbes Infect. 2019;8:879–94. doi: 10.1080/22221751.2019.1625728. [DOI] [PMC free article] [PubMed] [Google Scholar]
33.Zeng Z, Shi YX, Tsao T, et al. Targeting of mTORC1/2 by the mTOR kinase inhibitor PP242 induces apoptosis in AML cells under conditions mimicking the bone marrow microenvironment. Blood. 2012;120:2679–89. doi: 10.1182/blood-2011-11-393934. [DOI] [PMC free article] [PubMed] [Google Scholar]
34.Bar J, Lukaschuk N, Zalcenstein A, et al. The PI3K inhibitor LY294002 prevents p53 induction by DNA damage and attenuates chemotherapy-induced apoptosis. Cell Death Differ. 2005;12:1578–87. doi: 10.1038/sj.cdd.4401677. [DOI] [PubMed] [Google Scholar]
35.Harsing Jr. L, Juranyi Z, Gacsalyi I, et al. Glycine transporter type-1 and its inhibitors. Curr Med Chem. 2006;13:1017–44. doi: 10.2174/092986706776360932. [DOI] [PubMed] [Google Scholar]
36.Martin-Facklam M, Pizzagalli F, Zhou Y, et al. Glycine transporter type 1 occupancy by bitopertin: a positron emission tomography study in healthy volunteers. Neuropsychopharmacology. 2013;38:504–12. doi: 10.1038/npp.2012.212. [DOI] [PMC free article] [PubMed] [Google Scholar]
37.Stover PJ. One-carbon metabolism-genome interactions in folate-associated pathologies. J Nutr. 2009;139:2402–5. doi: 10.3945/jn.109.113670. [DOI] [PMC free article] [PubMed] [Google Scholar]
38.Albrecht LV, Bui MH, De Robertis EM. Canonical Wnt is inhibited by targeting one-carbon metabolism through methotrexate or methionine deprivation. Proc Natl Acad Sci USA. 2019;116:2987–95. doi: 10.1073/pnas.1820161116. [DOI] [PMC free article] [PubMed] [Google Scholar]
39.Soleimani M. SLC26 Cl-/HCO3- exchangers in the kidney: roles in health and disease. Kidney Int. 2013;84:657–66. doi: 10.1038/ki.2013.138. [DOI] [PMC free article] [PubMed] [Google Scholar]
40.Feng M, Elaïb Z, Borgel D, et al. NAADP/SERCA3-Dependent Ca2+ Stores Pathway Specifically Controls Early Autocrine ADP Secretion Potentiating Platelet Activation. Circ Res. 2020;127:e166–83. doi: 10.1161/CIRCRESAHA.119.316090. [DOI] [PubMed] [Google Scholar]
41.Ebrahim S, Ballesteros A, Zheng WS, et al. Transmembrane channel-like 4 and 5 proteins at microvillar tips are potential ion channels and lipid scramblases. bioRxiv. 2024 doi: 10.1101/2024.08.22.609173. Preprint. [DOI]
42.Qu W, Ikejima K, Zhong Z, et al. Glycine blocks the increase in intracellular free Ca2+ due to vasoactive mediators in hepatic parenchymal cells. Am J Physiol Gastrointest Liver Physiol. 2002;283:G1249–56. doi: 10.1152/ajpgi.00197.2002. [DOI] [PubMed] [Google Scholar]
43.Cao Z, Liu Y, Ma L, et al. A potent hepatitis B surface antigen response in subjects with inactive hepatitis B surface antigen carrier treated with pegylated-interferon alpha. Hepatology. 2017;66:1058–66. doi: 10.1002/hep.29213. [DOI] [PubMed] [Google Scholar]
44.Wong GLH, Gane E, Lok ASF. How to achieve functional cure of HBV: Stopping NUCs, adding interferon or new drug development? J Hepatol. 2022;76:1249–62. doi: 10.1016/j.jhep.2021.11.024. [DOI] [PubMed] [Google Scholar]
45.Li H, Yan L, Shi Y, et al. Hepatitis infection B virus: molecular virology to antiviral drugs. 2020. [Google Scholar]
46.Peng X-H, Ren X-N, Chen L-X, et al. High persistence rate of hepatitis B virus in a hydrodynamic injection-based transfection model in C3H/HeN mice. World J Gastroenterol. 2015;21:3527–36. doi: 10.3748/wjg.v21.i12.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]
47.Yan Z, Zeng J, Yu Y, et al. HBVcircle: A novel tool to investigate hepatitis B virus covalently closed circular DNA. J Hepatol. 2017;66:1149–57. doi: 10.1016/j.jhep.2017.02.004. [DOI] [PubMed] [Google Scholar]
48.Yu S-S, Tang R-C, Zhang A, et al. Deacetylase Sirtuin 1 mitigates type I IFN- and type II IFN-induced signaling and antiviral immunity. J Virol. 2024;98:e00088-24. doi: 10.1128/jvi.00088-24. [DOI] [PMC free article] [PubMed] [Google Scholar]
49.Lin C, Han G, Jia L, et al. Cardio-respiratory and phenotypic rescue of dystrophin/utrophin-deficient mice by combination therapy. EMBO Rep. 2022;23:e53955. doi: 10.15252/embr.202153955. [DOI] [PMC free article] [PubMed] [Google Scholar]
50.Razak MA, Begum PS, Viswanath B, et al. Multifarious Beneficial Effect of Nonessential Amino Acid, Glycine: A Review. Oxid Med Cell Longev. 2017;2017:1716701. doi: 10.1155/2017/1716701. [DOI] [PMC free article] [PubMed] [Google Scholar]
51.Lin C, Luo L, Xun Z, et al. Novel function of MOTS-c in mitochondrial remodelling contributes to its antiviral role during HBV infection. Gut. 2024;73:338–49. doi: 10.1136/gutjnl-2023-330389. [DOI] [PubMed] [Google Scholar]
52.Lin C, Huang Y, Luo L, et al. Adenosine Triphosphate in Serum as a Promising Biomarker for Differential Diagnosis of Hepatitis B Disease Progression. Front Immunol. 2022;13:1–8. doi: 10.3389/fimmu.2022.927761. [DOI] [PMC free article] [PubMed] [Google Scholar]
53.Wang W, Wu Z, Dai Z, et al. Glycine metabolism in animals and humans: implications for nutrition and health. Amino Acids. 2013;45:463–77. doi: 10.1007/s00726-013-1493-1. [DOI] [PubMed] [Google Scholar]
54.Xia Y, Guo H. Hepatitis B virus cccDNA: Formation, regulation and therapeutic potential. Antiviral Res. 2020;180:104824. doi: 10.1016/j.antiviral.2020.104824. [DOI] [PMC free article] [PubMed] [Google Scholar]
55.Ozer A, Khaoustov VI, Mearns M, et al. Effect of hepatocyte proliferation and cellular DNA synthesis on hepatitis B virus replication. Gastroenterology. 1996;110:1519–28. doi: 10.1053/gast.1996.v110.pm8613059. [DOI] [PubMed] [Google Scholar]
56.Rehermann B, Ferrari C, Pasquinelli C, et al. The hepatitis B virus persists for decades after patients’ recovery from acute viral hepatitis despite active maintenance of a cytotoxic T-lymphocyte response. Nat Med. 1996;2:1104–8. doi: 10.1038/nm1096-1104. [DOI] [PubMed] [Google Scholar]
57.Wooddell CI, Yuen M-F, Chan HL-Y, et al. RNAi-based treatment of chronically infected patients and chimpanzees reveals that integrated hepatitis B virus DNA is a source of HBsAg. Sci Transl Med. 2017;9:eaan0241. doi: 10.1126/scitranslmed.aan0241. [DOI] [PMC free article] [PubMed] [Google Scholar]
58.Luntz SP, Unnebrink K, Seibert-Grafe M, et al. HEGPOL: randomized, placebo controlled, multicenter, double-blind clinical trial to investigate hepatoprotective effects of glycine in the postoperative phase of liver transplantation [ISRCTN69350312] BMC Surg. 2005;5:18. doi: 10.1186/1471-2482-5-18. [DOI] [PMC free article] [PubMed] [Google Scholar]
59.Marcellin P, Ahn SH, Ma X, et al. Combination of Tenofovir Disoproxil Fumarate and Peginterferon α-2a Increases Loss of Hepatitis B Surface Antigen in Patients With Chronic Hepatitis B. Gastroenterology. 2016;150:134–44. doi: 10.1053/j.gastro.2015.09.043. [DOI] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

online supplemental file 1

gutjnl-74-12-s001.doc^{(15.4MB, doc)}

DOI: 10.1136/gutjnl-2025-334813

online supplemental file 2

gutjnl-74-12-s002.pptx^{(5MB, pptx)}

DOI: 10.1136/gutjnl-2025-334813

Data Availability Statement

Data are available upon reasonable request.

[R1] 1.World Health Organization . Global hepatitis report 2024 action for access in low- and middle-income countries. World Health Organization; 2024. [Google Scholar]

[R2] 2.WHO . Guidelines for the prevention, diagnosis, care and treatment for people with chronic hepatitis B infection. World Health Organization; 2024. [PubMed] [Google Scholar]

[R3] 3.Cui F, Blach S, Manzengo Mingiedi C, et al. Global reporting of progress towards elimination of hepatitis B and hepatitis C. Lancet Gastroenterol Hepatol. 2023;8:332–42. doi: 10.1016/S2468-1253(22)00386-7. [DOI] [PubMed] [Google Scholar]

[R4] 4.Kramvis A, Chang K-M, Dandri M, et al. A roadmap for serum biomarkers for hepatitis B virus: current status and future outlook. Nat Rev Gastroenterol Hepatol. 2022;19:727–45. doi: 10.1038/s41575-022-00649-z. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R5] 5.Lok AS, Zoulim F, Dusheiko G, et al. Hepatitis B cure: From discovery to regulatory approval. J Hepatol. 2017;67:847–61. doi: 10.1016/j.jhep.2017.05.008. [DOI] [PubMed] [Google Scholar]

[R6] 6.Lucifora J, Xia Y, Reisinger F, et al. Specific and nonhepatotoxic degradation of nuclear hepatitis B virus cccDNA. Science. 2014;343:1221–8. doi: 10.1126/science.1243462. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R7] 7.Nassal M. HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic hepatitis B. Gut. 2015;64:1972–84. doi: 10.1136/gutjnl-2015-309809. [DOI] [PubMed] [Google Scholar]

[R8] 8.Seeger C, Mason WS. Molecular biology of hepatitis B virus infection. Virology (Auckl) 2015;479–480:672–86. doi: 10.1016/j.virol.2015.02.031. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R9] 9.Revill PA, Chisari FV, Block JM, et al. A global scientific strategy to cure hepatitis B. Lancet Gastroenterol Hepatol. 2019;4:545–58. doi: 10.1016/S2468-1253(19)30119-0. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R10] 10.Fung S, Choi HSJ, Gehring A, et al. Getting to HBV cure: The promising paths forward. Hepatology. 2022;76:233–50. doi: 10.1002/hep.32314. [DOI] [PubMed] [Google Scholar]

[R11] 11.Levrero M, Pollicino T, Petersen J, et al. Control of cccDNA function in hepatitis B virus infection. J Hepatol. 2009;51:581–92. doi: 10.1016/j.jhep.2009.05.022. [DOI] [PubMed] [Google Scholar]

[R12] 12.Belloni L, Pollicino T, De Nicola F, et al. Nuclear HBx binds the HBV minichromosome and modifies the epigenetic regulation of cccDNA function. Proc Natl Acad Sci U S A. 2009;106:19975–9. doi: 10.1073/pnas.0908365106. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R13] 13.Yang HC, Kao JH. Persistence of hepatitis B virus covalently closed circular DNA in hepatocytes: molecular mechanisms and clinical significance. Emerg Microbes Infect. 2014;3:e64. doi: 10.1038/emi.2014.64. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R14] 14.Wu K, He M, Mao B, et al. Enhanced delivery of CRISPR/Cas9 system based on biomimetic nanoparticles for hepatitis B virus therapy. J Control Release. 2024;374:293–311. doi: 10.1016/j.jconrel.2024.08.019. [DOI] [PubMed] [Google Scholar]

[R15] 15.Allweiss L, Volz T, Giersch K, et al. Proliferation of primary human hepatocytes and prevention of hepatitis B virus reinfection efficiently deplete nuclear cccDNA in vivo. Gut. 2018;67:542–52. doi: 10.1136/gutjnl-2016-312162. [DOI] [PubMed] [Google Scholar]

[R16] 16.Chang J, Guo F, Zhao X, et al. Therapeutic strategies for a functional cure of chronic hepatitis B virus infection. Acta Pharm Sin B. 2014;4:248–57. doi: 10.1016/j.apsb.2014.05.002. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R17] 17.Lutgehetmann M, Volz T, Köpke A, et al. In vivo proliferation of hepadnavirus-infected hepatocytes induces loss of covalently closed circular DNA in mice. Hepatology. 2010;52:16–24. doi: 10.1002/hep.23611. [DOI] [PubMed] [Google Scholar]

[R18] 18.Köck J, Rösler C, Zhang J-J, et al. Generation of covalently closed circular DNA of hepatitis B viruses via intracellular recycling is regulated in a virus specific manner. PLoS Pathog. 2010;6:e1001082. doi: 10.1371/journal.ppat.1001082. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R19] 19.Dandri M, Petersen J. Mechanism of Hepatitis B Virus Persistence in Hepatocytes and Its Carcinogenic Potential. Clin Infect Dis. 2016;62 Suppl 4:S281–8. doi: 10.1093/cid/ciw023. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R20] 20.Wheeler MD, Ikejema K, Enomoto N, et al. Glycine: a new anti-inflammatory immunonutrient. Cell Mol Life Sci. 1999;56:843–56. doi: 10.1007/s000180050030. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R21] 21.Zhong Z, Wheeler MD, Li X, et al. L-Glycine: a novel antiinflammatory, immunomodulatory, and cytoprotective agent. Curr Opin Clin Nutr Metab Care. 2003;6:229–40. doi: 10.1097/01.mco.0000058609.19236.a4. [DOI] [PubMed] [Google Scholar]

[R22] 22.Lin C, Han G, Ning H, et al. Glycine Enhances Satellite Cell Proliferation, Cell Transplantation, and Oligonucleotide Efficacy in Dystrophic Muscle. Mol Ther. 2020;28:1339–58. doi: 10.1016/j.ymthe.2020.03.003. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R23] 23.Rom O, Liu Y, Liu Z, et al. Glycine-based treatment ameliorates NAFLD by modulating fatty acid oxidation, glutathione synthesis, and the gut microbiome. Sci Transl Med. 2020;12:eaaz2841. doi: 10.1126/scitranslmed.aaz2841. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R24] 24.Qu P, Rom O, Li K, et al. DT-109 ameliorates nonalcoholic steatohepatitis in nonhuman primates. Cell Metab. 2023;35:742–57. doi: 10.1016/j.cmet.2023.03.013. [DOI] [PubMed] [Google Scholar]

[R25] 25.Ghrayeb A, Finney AC, Agranovich B, et al. Serine synthesis via reversed SHMT2 activity drives glycine depletion and acetaminophen hepatotoxicity in MASLD. Cell Metab. 2024;36:116–29. doi: 10.1016/j.cmet.2023.12.013. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R26] 26.Wang Z, Zhang J, Wang L, et al. Glycine mitigates renal oxidative stress by suppressing Nox4 expression in rats with streptozotocin-induced diabetes. J Pharmacol Sci. 2018;137:387–94. doi: 10.1016/j.jphs.2018.08.005. [DOI] [PubMed] [Google Scholar]

[R27] 27.Al-Saeedi M, Liang R, Schultze DP, et al. Glycine protects partial liver grafts from Kupfer cell-dependent ischemia–reperfusion injury without negative efect on regeneration. Amino Acids. 2019;51:903–11. doi: 10.1007/S00726-019-02722-5/TABLES/3. [DOI] [PubMed] [Google Scholar]

[R28] 28.Schemmer P, Bradford BU, Rose ML, et al. Intravenous glycine improves survival in rat liver transplantation. Am J Physiol. 1999;276:G924–32. doi: 10.1152/ajpgi.1999.276.4.G924. [DOI] [PubMed] [Google Scholar]

[R29] 29.Bachmann S, Peng XX, Currin RT, et al. Glycine in Carolina rinse solution reduces reperfusion injury, improves graft function, and increases graft survival after rat liver transplantation. Transplant Proc. 1995;27:741–2. [PubMed] [Google Scholar]

[R30] 30.Benko T, Frede S, Gu Y, et al. Glycine pretreatment ameliorates liver injury after partial hepatectomy in the rat. J Invest Surg. 2010;23:12–20. doi: 10.3109/08941930903469466. [DOI] [PubMed] [Google Scholar]

[R31] 31.Yan H, Zhong G, Xu G, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. Elife. 2012;3:e00049. doi: 10.7554/eLife.00049. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R32] 32.Yan Y, Allweiss L, Yang D, et al. Down-regulation of cell membrane localized NTCP expression in proliferating hepatocytes prevents hepatitis B virus infection. Emerg Microbes Infect. 2019;8:879–94. doi: 10.1080/22221751.2019.1625728. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R33] 33.Zeng Z, Shi YX, Tsao T, et al. Targeting of mTORC1/2 by the mTOR kinase inhibitor PP242 induces apoptosis in AML cells under conditions mimicking the bone marrow microenvironment. Blood. 2012;120:2679–89. doi: 10.1182/blood-2011-11-393934. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R34] 34.Bar J, Lukaschuk N, Zalcenstein A, et al. The PI3K inhibitor LY294002 prevents p53 induction by DNA damage and attenuates chemotherapy-induced apoptosis. Cell Death Differ. 2005;12:1578–87. doi: 10.1038/sj.cdd.4401677. [DOI] [PubMed] [Google Scholar]

[R35] 35.Harsing Jr. L, Juranyi Z, Gacsalyi I, et al. Glycine transporter type-1 and its inhibitors. Curr Med Chem. 2006;13:1017–44. doi: 10.2174/092986706776360932. [DOI] [PubMed] [Google Scholar]

[R36] 36.Martin-Facklam M, Pizzagalli F, Zhou Y, et al. Glycine transporter type 1 occupancy by bitopertin: a positron emission tomography study in healthy volunteers. Neuropsychopharmacology. 2013;38:504–12. doi: 10.1038/npp.2012.212. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R37] 37.Stover PJ. One-carbon metabolism-genome interactions in folate-associated pathologies. J Nutr. 2009;139:2402–5. doi: 10.3945/jn.109.113670. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R38] 38.Albrecht LV, Bui MH, De Robertis EM. Canonical Wnt is inhibited by targeting one-carbon metabolism through methotrexate or methionine deprivation. Proc Natl Acad Sci USA. 2019;116:2987–95. doi: 10.1073/pnas.1820161116. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R39] 39.Soleimani M. SLC26 Cl-/HCO3- exchangers in the kidney: roles in health and disease. Kidney Int. 2013;84:657–66. doi: 10.1038/ki.2013.138. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R40] 40.Feng M, Elaïb Z, Borgel D, et al. NAADP/SERCA3-Dependent Ca2+ Stores Pathway Specifically Controls Early Autocrine ADP Secretion Potentiating Platelet Activation. Circ Res. 2020;127:e166–83. doi: 10.1161/CIRCRESAHA.119.316090. [DOI] [PubMed] [Google Scholar]

[R41] 41.Ebrahim S, Ballesteros A, Zheng WS, et al. Transmembrane channel-like 4 and 5 proteins at microvillar tips are potential ion channels and lipid scramblases. bioRxiv. 2024 doi: 10.1101/2024.08.22.609173. Preprint. [DOI]

[R42] 42.Qu W, Ikejima K, Zhong Z, et al. Glycine blocks the increase in intracellular free Ca2+ due to vasoactive mediators in hepatic parenchymal cells. Am J Physiol Gastrointest Liver Physiol. 2002;283:G1249–56. doi: 10.1152/ajpgi.00197.2002. [DOI] [PubMed] [Google Scholar]

[R43] 43.Cao Z, Liu Y, Ma L, et al. A potent hepatitis B surface antigen response in subjects with inactive hepatitis B surface antigen carrier treated with pegylated-interferon alpha. Hepatology. 2017;66:1058–66. doi: 10.1002/hep.29213. [DOI] [PubMed] [Google Scholar]

[R44] 44.Wong GLH, Gane E, Lok ASF. How to achieve functional cure of HBV: Stopping NUCs, adding interferon or new drug development? J Hepatol. 2022;76:1249–62. doi: 10.1016/j.jhep.2021.11.024. [DOI] [PubMed] [Google Scholar]

[R45] 45.Li H, Yan L, Shi Y, et al. Hepatitis infection B virus: molecular virology to antiviral drugs. 2020. [Google Scholar]

[R46] 46.Peng X-H, Ren X-N, Chen L-X, et al. High persistence rate of hepatitis B virus in a hydrodynamic injection-based transfection model in C3H/HeN mice. World J Gastroenterol. 2015;21:3527–36. doi: 10.3748/wjg.v21.i12.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R47] 47.Yan Z, Zeng J, Yu Y, et al. HBVcircle: A novel tool to investigate hepatitis B virus covalently closed circular DNA. J Hepatol. 2017;66:1149–57. doi: 10.1016/j.jhep.2017.02.004. [DOI] [PubMed] [Google Scholar]

[R48] 48.Yu S-S, Tang R-C, Zhang A, et al. Deacetylase Sirtuin 1 mitigates type I IFN- and type II IFN-induced signaling and antiviral immunity. J Virol. 2024;98:e00088-24. doi: 10.1128/jvi.00088-24. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R49] 49.Lin C, Han G, Jia L, et al. Cardio-respiratory and phenotypic rescue of dystrophin/utrophin-deficient mice by combination therapy. EMBO Rep. 2022;23:e53955. doi: 10.15252/embr.202153955. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R50] 50.Razak MA, Begum PS, Viswanath B, et al. Multifarious Beneficial Effect of Nonessential Amino Acid, Glycine: A Review. Oxid Med Cell Longev. 2017;2017:1716701. doi: 10.1155/2017/1716701. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R51] 51.Lin C, Luo L, Xun Z, et al. Novel function of MOTS-c in mitochondrial remodelling contributes to its antiviral role during HBV infection. Gut. 2024;73:338–49. doi: 10.1136/gutjnl-2023-330389. [DOI] [PubMed] [Google Scholar]

[R52] 52.Lin C, Huang Y, Luo L, et al. Adenosine Triphosphate in Serum as a Promising Biomarker for Differential Diagnosis of Hepatitis B Disease Progression. Front Immunol. 2022;13:1–8. doi: 10.3389/fimmu.2022.927761. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R53] 53.Wang W, Wu Z, Dai Z, et al. Glycine metabolism in animals and humans: implications for nutrition and health. Amino Acids. 2013;45:463–77. doi: 10.1007/s00726-013-1493-1. [DOI] [PubMed] [Google Scholar]

[R54] 54.Xia Y, Guo H. Hepatitis B virus cccDNA: Formation, regulation and therapeutic potential. Antiviral Res. 2020;180:104824. doi: 10.1016/j.antiviral.2020.104824. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R55] 55.Ozer A, Khaoustov VI, Mearns M, et al. Effect of hepatocyte proliferation and cellular DNA synthesis on hepatitis B virus replication. Gastroenterology. 1996;110:1519–28. doi: 10.1053/gast.1996.v110.pm8613059. [DOI] [PubMed] [Google Scholar]

[R56] 56.Rehermann B, Ferrari C, Pasquinelli C, et al. The hepatitis B virus persists for decades after patients’ recovery from acute viral hepatitis despite active maintenance of a cytotoxic T-lymphocyte response. Nat Med. 1996;2:1104–8. doi: 10.1038/nm1096-1104. [DOI] [PubMed] [Google Scholar]

[R57] 57.Wooddell CI, Yuen M-F, Chan HL-Y, et al. RNAi-based treatment of chronically infected patients and chimpanzees reveals that integrated hepatitis B virus DNA is a source of HBsAg. Sci Transl Med. 2017;9:eaan0241. doi: 10.1126/scitranslmed.aan0241. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R58] 58.Luntz SP, Unnebrink K, Seibert-Grafe M, et al. HEGPOL: randomized, placebo controlled, multicenter, double-blind clinical trial to investigate hepatoprotective effects of glycine in the postoperative phase of liver transplantation [ISRCTN69350312] BMC Surg. 2005;5:18. doi: 10.1186/1471-2482-5-18. [DOI] [PMC free article] [PubMed] [Google Scholar]

[R59] 59.Marcellin P, Ahn SH, Ma X, et al. Combination of Tenofovir Disoproxil Fumarate and Peginterferon α-2a Increases Loss of Hepatitis B Surface Antigen in Patients With Chronic Hepatitis B. Gastroenterology. 2016;150:134–44. doi: 10.1053/j.gastro.2015.09.043. [DOI] [PubMed] [Google Scholar]

PERMALINK

Therapeutic inhibition of HBsAg and HBV cccDNA through a novel phased combination treatment: glycine and interferon-α

Caorui Lin

Ying Huang

Ning Ran

Jie Liu

Linjie Luo

Xin Zhang

Xiaosang Zheng

Zhen Xun

Siyi Xu

Can Liu

Xiaohong Kong

Shiqing Feng

Haiting Mao

Qishui Ou

Abstract

Background

Objective

Design

Results

Conclusion

WHAT IS ALREADY KNOWN ON THIS TOPIC

WHAT THIS STUDY ADDS

HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY

Introduction

Materials and methods

Results

Serum glycine levels increase on HBV infection in humans

Glycine inhibits HBV infection in vitro and in vivo

Glycine enhances liver regeneration and contributes to total cccDNA reduction

Glycine induces liver regeneration by activating the AKT-mTORC1 signalling pathway, resulting in HBV cccDNA reduction and antiviral efﬁcacy

Glycine increases liver regeneration by boosting the one-carbon unit pool via glycine transporter-1

Glycine administration triggers glycine receptor signalling to improve hepatic inflammation

Glycine enhances IFN-α antiviral efficacy through a phased administration strategy

Discussion

Supplementary material

Acknowledgements

Footnotes

Data availability statement

References

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

Similar articles

Cited by other articles

Links to NCBI Databases