2. Academic Validation
  3. Upregulation of GLT25D1 in Hepatic Stellate Cells Promotes Liver Fibrosis via the TGF-β1/SMAD3 Pathway In Vivo and In vitro

Upregulation of GLT25D1 in Hepatic Stellate Cells Promotes Liver Fibrosis via the TGF-β1/SMAD3 Pathway In Vivo and In vitro

  • J Clin Transl Hepatol. 2023 Feb 28;11(1):1-14. doi: 10.14218/JCTH.2022.00005.
Shiwei Wang 1,  Lingling He 1,  Fan Xiao 2,  Meixin Gao 1,  Herui Wei 1,  Junru Yang 1,  Yang Shu 1,  Fuyang Zhang 1,  Xiaohui Ye 3,  Ping Li 1,  Xiaohua Hao 4,  Xingang Zhou 5,  Hongshan Wei 1 6
Affiliations

Affiliations

  • 1 Department of Gastroenterology, Beijing Ditan Hospital, Capital Medical University, Beijing, China.
  • 2 Institute of Infectious Diseases, Beijing Ditan Hospital, Capital Medical University, Beijing, China.
  • 3 Department of Gastroenterology, Beijing Huaxin Hospital, the First Affiliated Hospital of Tsinghua University, Beijing, China.
  • 4 Beijing Ditan Hospital, Capital Medical University, Beijing, China.
  • 5 Department of Pathology, Beijing Ditan Hospital, Capital Medical University, Beijing, China.
  • 6 Department of Gastroenterology, Peking University Ditan Teaching Hospital, Beijing, China.
PMID: 36406310 DOI: 10.14218/JCTH.2022.00005
Abstract

Background and aims: Collagen β(1-O) galactosyltransferase 25 domain 1 (GLT25D1) is associated with Collagen production and glycosylation, and its knockout in mice results in embryonic death. However, its role in liver fibrosis remains elusive, particularly in hepatic stellate cells (HSCs), the primary collagen-producing cells associated with liver fibrogenesis. Herein, we aimed to elucidate the role of GLT25D1 in HSCs.

Methods: Bile duct ligation (BDL)-induced mouse liver fibrosis models, primary mouse HSCs (mHSCs), and transforming growth factor beta 1 (TGF-β1)-stimulated LX-2 human hepatic stellate cells were used in in vivo and in vitro studies. Stable LX-2 cell lines with either GLT25D1 overexpression or knockdown were established using lentiviral transfection. RNA-seq was performed to investigate the genomic differences. HPLC-MS/MS were used to identify glycosylation sites. Scanning electronic microscopy (SEM) and second-harmonic generation/two-photon excited fluorescence (SHG/TPEF) were used to image Collagen fibril morphology.

Results: GLT25D1 expression was upregulated in nonparenchymal cells in human cirrhotic liver tissues. Meanwhile, its knockdown attenuated Collagen deposition in BDL-induced mouse liver fibrosis and inhibited mHSC activation. GLT25D1 was overexpressed in activated versus quiescence LX-2 cells and regulated in vitro LX-2 cell activation, including proliferation, contraction, and migration. GLT25D1 also significantly increased liver fibrogenic gene and protein expression. GLT25D1 upregulation promoted HSC activation and enhanced Collagen expression through the TGF-β1/SMAD signaling pathway. Mass spectrometry showed that GLT25D1 regulated the glycosylation of Collagen in HSCs, affecting the diameter of Collagen fibers.

Conclusions: Collectively, the upregulation of GLT25D1 in HSCs promoted the progression of liver fibrosis by affecting HSCs activation and Collagen stability.

Keywords

Collagen; Glycosylation; Hepatic stellate cells; Liver fibrosis; TGF-β1.

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