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

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

Gut. 2025 Jul 11:gutjnl-2025-334813. doi: 10.1136/gutjnl-2025-334813. Online ahead of print.

Authors

Caorui Lin^#^{1

2}, Ying Huang^#², Ning Ran^#^{3

4}, Jie Liu⁵, Linjie Luo², Xin Zhang², Xiaosang Zheng², Zhen Xun², Siyi Xu², Can Liu², Xiaohong Kong⁴, Shiqing Feng⁶, Haiting Mao¹, Qishui Ou⁷

Affiliations

¹ Department of Clinical Laboratory, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China ouqishui@fjmu.edu.cn caoruilin@email.sdu.edu.cn shiqingfeng@sdu.edu.cn maohaiting@sdu.edu.cn.
² Department of Laboratory Medicine, Fujian Key Laboratory of Laboratory Medicine, Fujian Clinical Research Center for Laboratory Medicine of Immunology, Gene Diagnosis Research Center, National Reginal Medical Center, Binhai Campus of the First Affiliated Hospital, The First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, China.
³ Institute of Medical Sciences, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
⁴ Shandong University Orthopedic Research Center, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
⁵ Department of Clinical Laboratory, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
⁶ Shandong University Orthopedic Research Center, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China ouqishui@fjmu.edu.cn caoruilin@email.sdu.edu.cn shiqingfeng@sdu.edu.cn maohaiting@sdu.edu.cn.
⁷ Department of Laboratory Medicine, Fujian Key Laboratory of Laboratory Medicine, Fujian Clinical Research Center for Laboratory Medicine of Immunology, Gene Diagnosis Research Center, National Reginal Medical Center, Binhai Campus of the First Affiliated Hospital, The First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, China ouqishui@fjmu.edu.cn caoruilin@email.sdu.edu.cn shiqingfeng@sdu.edu.cn maohaiting@sdu.edu.cn.

^# Contributed equally.

PMID: 40645766
DOI: 10.1136/gutjnl-2025-334813

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: ANTIVIRAL THERAPY; CHRONIC VIRAL HEPATITIS; DIAGNOSTIC VIROLOGY; HEPATITIS B; NUTRITION.