Biomaterials Research (생체재료학회지)

  • Volume 28
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  • Pages.0043.1-0043.18
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  • 2024
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  • 1226-4601(pISSN)
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  • 2055-7124(eISSN)
Korean Society for Biomaterials (한국생체재료학회)
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Genetically Modified Hepatocytes Targeting Bilirubin and Ammonia Metabolism for the Construction of Bioartificial Liver System

  • Ke Wang (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Yuankui Zhu (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Mengqing Li (School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology) ;
  • Yaxi Yang (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Dianbao Zuo (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Junfeng Sheng (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Xinhai Zhang (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University) ;
  • Wei Wang (Wuhan TOGO Medical Technology Co. Ltd.) ;
  • Ping Zhou (Wuhan TOGO Medical Technology Co. Ltd.) ;
  • Mingqian Feng (College of Biomedicine and Health, College of Life Science and Technology, Huazhong Agricultural University)
  • Received : 2024.03.01
  • Accepted : 2024.05.08
  • Published : 2024.12.01
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Abstract

Acute liver failure (ALF) is a complex syndrome that impairs the liver's function to detoxify bilirubin, ammonia, and other toxic metabolites. Bioartificial liver (BAL) aims to help ALF patients to pass through the urgent period by temporarily undertaking the liver's detoxification functions and promoting the recovery of the injured liver. We genetically modified the hepatocellular cell line HepG2 by stably overexpressing genes encoding UGT1A1, OATP1B1, OTC, ARG1, and CPS1. The resulting SynHeps-II cell line, encapsulated by Cytopore microcarriers, dramatically reduced the serum levels of bilirubin and ammonia, as demonstrated both in vitro using patient plasma and in vivo using ALF animal models. More importantly, we have also completed the 3-dimensional (3D) culturing of cells to meet the demands for industrialized rapid and mass production, and subsequently assembled the plasma-cell contacting BAL (PCC-BAL) system to fulfill the requirements of preclinical experiments. Extracorporeal blood purification of ALF rabbits with SynHeps-II-embedded PCC-BAL saved more than 80% of the animals from rapid death. Mechanistically, SynHeps-II therapy ameliorated liver and brain inflammation caused by high levels of bilirubin and ammonia and promoted liver regeneration by modulating the nuclear factor κB (NF-κB) and signal transducer and activator of transcription 3 (STAT3) pathways. Also, SynHeps-II treatment reduced cerebral infiltration of neutrophils, reduced reactive oxygen species (ROS) levels, and mitigated hepatic encephalopathy. Taken together, SynHeps-II cell-based BAL was promising for the treatment of ALF patients and warrants clinical trials.

Keywords

Acknowledgement

Funding: This work was supported by the National Natural Science Foundation of China (32270992). Author contributions: K.W. performed the experiments, analyzed the data, and drafted the manuscript. Y.Z. and Y.Y. assisted the animal studies. M.L. assisted with immunohistochemistry and immunofluorescence experiments. D.Z. and J.S. assisted in partial in vivo experiments. X.Z. assisted in partial in vitro experiments. W.W. and P.Z. provided partial technical or supervisory support. M.F. designed and supervised the project and finalized the manuscript. All authors read and approved the final manuscript. Competing interests: The authors declare that they have no competing interests.

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