2. Academic Validation
  3. Transcriptomic and proteomic sequencing unveils the role of vitamin D and metabolic flux shifts in the induction of human hepatic organoids

Transcriptomic and proteomic sequencing unveils the role of vitamin D and metabolic flux shifts in the induction of human hepatic organoids

  • Stem Cell Res Ther. 2024 Dec 18;15(1):478. doi: 10.1186/s13287-024-04101-8.
Shule Zhang 1,  Linghong Liu 2,  Xianyu Li 3,  Tiancheng Zhou 4,  Qing Shi 2,  Dong Li 5,  Xiuli Ju 6 7
Affiliations

Affiliations

  • 1 Department of Pediatrics, Qilu Hospital of Shandong University, 107 Wenhua Xilu, Jinan, 250012, Shandong, P.R. China.
  • 2 Cryomedicine Laboratory, Qilu Hospital of Shandong University, 107 Wenhua Xilu, Jinan, 250012, China.
  • 3 Experimental Research Center, Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment of Major Disease, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
  • 4 Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangdong-Hong Kong Joint Laboratory for Stem Cell and Regenerative Medicine, Center for Cell Lineage and Cell Therapy, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China.
  • 5 Cryomedicine Laboratory, Qilu Hospital of Shandong University, 107 Wenhua Xilu, Jinan, 250012, China. lidong73@sdu.edu.cn.
  • 6 Department of Pediatrics, Qilu Hospital of Shandong University, 107 Wenhua Xilu, Jinan, 250012, Shandong, P.R. China. jxlqlyy@163.com.
  • 7 Cryomedicine Laboratory, Qilu Hospital of Shandong University, 107 Wenhua Xilu, Jinan, 250012, China. jxlqlyy@163.com.
PMID: 39696644 DOI: 10.1186/s13287-024-04101-8
Abstract

Background: Hepatic organoids (HOs), validated through comparative Sequencing with human liver tissues, are reliable models for liver research. Comprehensive transcriptomic and proteomic Sequencing of HOs throughout their induction period will enhance the platform's utility, aiding in the elucidation of liver development's molecular mechanisms.

Methods: We developed hepatic organoids (HOs) from embryonic stem cells (ESCs) through a de novo induction protocol, mimicking the stages of fetal liver development: ESCs to definitive endoderm (DE), then to foregut (FG), hepatoblasts (HB), and finally to HOs stage 1 (HO1), culminating in self-organizing HOs stage 2 (HO2) via dissociation and re-inoculation. The successful establishment of HOs was validated by immunofluorescence staining and RT-qPCR for specific markers. Comprehensive transcriptomic and proteomic Sequencing and analysis were conducted on FG, HB, HO1, and HO2.

Results: Our data suggest that several transcription factors (TFs) activated during the HB stage share overlapping target genes with the vitamin D receptor (VDR). Calcitriol, a direct activator of VDR, notably facilitated the FG to HB stage transition by activating VDR and enhancing key TFs, thereby promoting hepatic progenitor cell maturation. Furthermore, our findings revealed a significant transition towards glycolytic energy metabolism at the HO2 stage, characterized by increased glycolytic flux and reduced Oxidative Phosphorylation. Inhibition of glycolysis using 2-deoxy-D-glucose (2-DG) led to suppressed growth and differentiation at the HO2 stage. Analysis of signaling pathways indicated upregulation of the HIF-1 pathway, which is associated with glycolysis activation, as well as the MAPK and PI3K-AKT pathways, which regulate HIF-1α protein translation.

Conclusions: We elucidated a pivotal role for calcitriol in facilitating the transition from FG to HB by activating VDR and augmenting the expression of critical transcription factors (TFs). Besides, our research underscores a shift in metabolic pathways toward glycolytic energy metabolism in HO2 organoids. Overall, our multiomics approach reveals the intricate molecular regulation during the development of HOs.

Keywords

Calcitriol; Glycolysis; Hepatic organoids; Hepatocyte differentiation; Vitamin D receptor.

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