Single-nucleus RNA sequencing reveals HBV-driven metabolic reprogramming and TIMP1-mediated fibrosis in human-liverchimeric mice

Xiaonan Ren¹Cong Wang¹Boyin Qin¹Hua Yang¹Min Wu¹Zhan-qing Zhang¹Wei Lu¹Chao Wang¹Yabin Liu¹Xiaonan Zhang^2*Xiaohui Zhou^1*

• ¹Shanghai Public Health Clinical Center, Fudan University, Shanghai, China
• ²University of Canberra, Canberra, Australia

Hepatitis B virus (HBV) infection remains a leading cause of chronic liver disease, cirrhosis, and hepatocellular carcinoma worldwide. Despite advances in antiviral therapies, the mechanisms underlying HBV-induced metabolic reprogramming and liver fibrosis remain poorly understood. Here, we employed single-nucleus RNA sequencing (snRNA-seq) which is particularly suitable for hepatocytic sequencing to dissect the transcriptional landscape of HBV-infected and uninfected hepatocytes in humanized URG mice (Hu-URG). Chronic HBV infection was successfully established in Hu-URG mice, with progressive increases in serum HBV DNA, HBsAg, and HBeAg levels. snRNA-seq revealed distinct human hepatocyte clusters (clusters 9, 16, 23) characterizing elevated expression of metabolic genes (ALB, UGT2B17, CYP2A6) in HBV-infected cells, while HBV-uninfected cells exhibited upregulation of TIMP1 and pro-fibrotic pathways. Immunofluorescence and histological analyses confirmed that HBV-uninfected hepatocytes (HBsAg -) displayed higher TIMP1 expression and reduced albumin (hALB) levels, correlating with increased collagen deposition in HBV-hu-URG mice. Notably, this TIMP1 + HBsAg - hALB low phenotype was also observed in liver biopsies from chronic HBV patients, underscoring its clinical relevance. Our findings highlight HBV-driven metabolic adaptation and identify TIMP1 as a potential mediator of fibrosis in uninfected hepatocytes, offering novel insights into HBV pathogenesis and therapeutic targeting.