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Hepatology
Original article
Glial cell line-derived neurotrophic factor (GDNF) mediates hepatic stellate cell activation via ALK5/Smad signalling
  1. Le Tao1,2,
  2. Wenting Ma2,
  3. Liu Wu2,
  4. Mingyi Xu3,
  5. Yanqin Yang4,
  6. Wei Zhang1,
  7. Wenjun Sha5,
  8. Hongshan Li6,
  9. Jianrong Xu7,
  10. Rilu Feng8,
  11. Dongying Xue2,
  12. Jie Zhang2,
  13. Steven Dooley8,
  14. Ekihiro Seki9,
  15. Ping Liu10,11,
  16. Cheng Liu1,2,12
  1. 1 Central Laboratory, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China
  2. 2 Laboratory of Liver Disease, Department of Infectious Disease, PutuoHospital , Shanghai University of Traditional Chinese Medicine, Shanghai, China
  3. 3 Department of Gastroenterology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
  4. 4 Department of Pathology, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China
  5. 5 Departmentof Endocrinology and Metabolism, PutuoHospital , Shanghai University of Traditional Chinese Medicine, Shanghai, China
  6. 6 Department of Hepatology, HwaMei Hospital, University of Chinese Academy of Sciences, Ningbo, China
  7. 7 Department of Pharmacology and Chemical Biology, Faculty of Basic Medicine, Shanghai Jiao Tong University School of Medicine, Shanghai, China
  8. 8 Department of Medicine II, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany
  9. 9 Division of Digestive and Liver Diseases, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California, USA
  10. 10 Shanghai University of Traditional Chinese Medicine, Shanghai, China
  11. 11 Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China
  12. 12 Shanghai Putuo Central School of Clinical Medicine, Anhui Medical University, Shanghai, China
  1. Correspondence to Steven Dooley, Department of Medicine II, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany; steven.dooley{at}medma.uni-heidelberg.de, Ekihiro Seki, Department of Medicine, Cedars-Sinai Medical Center, Los Angeles, California, USA; Ekihiro.Seki{at}cshs.org, Ping Liu, Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China; liuliver{at}vip.sina.com and Cheng Liu, Central Laboratory and Laboratory of liver disease, Department of Infectious Disease, Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200062, China; liucheng0082010{at}163.com

Abstract

Objective Although glial cell line-derived neurotrophic factor (GDNF) is a member of the transforming growth factor-β superfamily, its function in liver fibrosis has rarely been studied. Here, we investigated the role of GDNF in hepatic stellate cell (HSC) activation and liver fibrosis in humans and mice.

Design GDNF expression was examined in liver biopsies and sera from patients with liver fibrosis. The functional role of GDNF in liver fibrosis was examined in mice with adenoviral delivery of the GDNF gene, GDNF sgRNA CRISPR/Cas9 and the administration of GDNF-blocking antibodies. GDNF was examined on HSC activation using human and mouse primary HSCs. The binding of activin receptor-like kinase 5 (ALK5) to GDNF was determined using surface plasmon resonance (SPR), molecular docking, mutagenesis and co-immunoprecipitation.

Results GDNF mRNA and protein levels are significantly upregulated in patients with stage F4 fibrosis. Serum GDNF content correlates positively with α-smooth muscle actin (α-SMA) and Col1A1 mRNA in human fibrotic livers. Mice with overexpressed GDNF display aggravated liver fibrosis, while mice with silenced GDNF expression or signalling inhibition by GDNF-blocking antibodies have reduced fibrosis and HSC activation. GDNF is confined mainly to HSCs and contributes to HSC activation through ALK5 at His39 and Asp76 and through downstream signalling via Smad2/3, but not through GDNF family receptor alpha-1 (GFRα1). GDNF, ALK5 and α-SMA colocalise in human and mouse HSCs, as demonstrated by confocal microscopy.

Conclusions GDNF promotes HSC activation and liver fibrosis through ALK5/Smad signalling. Inhibition of GDNF could be a novel therapeutic strategy to combat liver fibrosis.

  • hepatic fibrosis
  • chronic liver disease
  • hepatic stellate cell

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/.

https://doi.org/10.1136/gutjnl-2018-317872

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    Footnotes

    • LT, WM, LW and MX contributed equally.

    • Contributors CL, SD, PL and ES conceived and designed the experiments. LT, WM, LW, WS, MX, HL, RF, WZ and JX carried out experiments. YY, DX, JZ, MX and helped with the human liver biopsy specimen. ES assisted with data analysis. CL and SD wrote the manuscript, which was read, edited and approved by all authors.

    • Funding This work was supported by The National Natural Science Foundation of China (no. 81673788 (to CL), 81873136 (to CL), 81530101 (to PL), 81803232 (to WM), 81803898 (to LT), 81570547 (to MX), 81770597 (to MX), 81704027 (to WS), 81873109 (to HL)). The National Institutes of Health Grant (no. R01DK085252, R01AA027036 (to ES)) and Winnick Research award from Cedars-Sinai Medical Center (to ES). The Federal Ministry of Education and Research grant ‘LiSyM’ (to SD).

    • Competing interests None declared.

    • Ethics approval Human samples and study protocol were approved by the Clinical Ethics Committee of Putuo Hospital, Shanghai University of Traditional Chinese Medicine and Shanghai General Hospital, Jiao Tong University School of Medicine.

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

    • Correction notice This article has been corrected since it published Online First. Figure 8 has been corrected.

    • Patient consent for publication Obtained.