2. Academic Validation
  3. Single-cell transcriptomics reveals liver developmental trajectory during lineage reprogramming of human induced hepatocyte-like cells

Single-cell transcriptomics reveals liver developmental trajectory during lineage reprogramming of human induced hepatocyte-like cells

  • Cell Mol Life Sci. 2025 Apr 6;82(1):139. doi: 10.1007/s00018-025-05677-x.
Nan Jiang # 1 2,  Guangya Li # 3,  Sen Luo # 1 2,  Xi Kong 1 2,  Shigang Yin 1 2,  Jianhua Peng 1 2,  Yong Jiang 1 2,  Wei Tao 4,  Cheng Li 5,  Huangfan Xie 6 7,  Hongkui Deng 8 9,  Bingqing Xie 10 11
Affiliations

Affiliations

  • 1 Laboratory of Neurological Diseases and Brain Function, the Affiliated Hospital, Southwest Medical University, Luzhou, China.
  • 2 Institute of Epigenetics and Brain Science, Southwest Medical University, Luzhou, China.
  • 3 MOE Engineering Research Center of Regenerative Medicine, School of Basic Medical Sciences, State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, the MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
  • 4 Key Laboratory of Cell Proliferation and Differentiation, School of Life Sciences, Peking University, Beijing, China.
  • 5 School of Life Sciences, Center for Bioinformatics, Center for Statistical Science, Peking University, Beijing, China.
  • 6 Laboratory of Neurological Diseases and Brain Function, the Affiliated Hospital, Southwest Medical University, Luzhou, China. xie1991@swmu.edu.cn.
  • 7 Institute of Epigenetics and Brain Science, Southwest Medical University, Luzhou, China. xie1991@swmu.edu.cn.
  • 8 MOE Engineering Research Center of Regenerative Medicine, School of Basic Medical Sciences, State Key Laboratory of Natural and Biomimetic Drugs, Peking University Health Science Center, the MOE Key Laboratory of Cell Proliferation and Differentiation, College of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China. hongkui_deng@pku.edu.cn.
  • 9 Changping Laboratory, Beijing, China. hongkui_deng@pku.edu.cn.
  • 10 Laboratory of Neurological Diseases and Brain Function, the Affiliated Hospital, Southwest Medical University, Luzhou, China. bingqingxie@swmu.edu.cn.
  • 11 Institute of Epigenetics and Brain Science, Southwest Medical University, Luzhou, China. bingqingxie@swmu.edu.cn.
  • # Contributed equally.
PMID: 40188417 DOI: 10.1007/s00018-025-05677-x
Abstract

Hepatocytes are crucial for drug screening, disease modeling, and clinical transplantation, yet generating functional hepatocytes in vitro is challenging due to the difficulty of establishing their authentic gene regulatory networks (GRNs). We have previously developed a two-step lineage reprogramming strategy to generate functionally competent human induced hepatocytes (hiHeps), providing an effective model for studying the establishment of hepatocyte-specific GRNs. In this study, we utilized high-throughput single-cell RNA Sequencing (scRNA-seq) to explore the cell-fate transition and the establishment of hepatocyte-specific GRNs involved in the two-step reprogramming process. Our findings revealed that the late stage of the reprogramming process mimics the natural trajectory of liver development, exhibiting similar transcriptional waves of developmental genes. CD24 and DLK1 were identified as surface markers enriching two distinct hepatic progenitor populations respectively. Lipid metabolism emerged as a key enhancer of hiHeps maturation. Furthermore, transcription factors HNF4A and HHEX were identified as pivotal gatekeepers directing cell fate decisions between hepatocytes and intestinal cells. Collectively, this study provides valuable insights into the establishment of hepatocyte-specific GRNs during hiHeps induction at single-cell resolution, facilitating more efficient production of functional hepatocytes for therapeutic applications.

Keywords

Gene regulatory networks; Human induced hepatocytes; Lineage reprogramming; Lipid metabolism; Single-cell RNA sequencing.

Figures
Products