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Cited by 8Pluripotent Stem Cells ResearchLiver physiology and pathologyDevelopmental Biology and Gene Regulation

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

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Zhang, Shule, Liu, Linghong, Li, Xianyu, Zhou, Tiancheng, Shi, Qing, Li, Dong, Ju, Xiuli·2024·Stem Cell Research & Therapy
DOI 10.1186/s13287-024-04101-8PMC PMC11657659
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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.

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Authors
Zhang, S., Liu, L., Li, X., Zhou, T., Shi, Q., Li, D., & Ju, X.
Affiliations
  • Qilu Hospital of Shandong University (CN)
  • China Academy of Chinese Medical Sciences (CN)
  • Chinese Academy of Sciences (CN)
Corresponding author
  • Li, Dong
  • Ju, Xiuli
ORCID
Venue
Stem Cell Research & Therapy
Publisher
Springer Science and Business Media LLC
Published
2024-12-18
Volume
15
Issue
1
Pages
478-478
ISSN
1757-6512 (ISSN-L)
DOI
10.1186/s13287-024-04101-8
PMC
PMC11657659
OpenAlex
W4405516687
Cited by
8
References
36
Citations by year
2026: 5 · 2025: 3
FWCI
1.09
Type
journal article
Language
en
Open access
Licence
Locations
  • Stem Cell Research & Therapy (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Natural Science Foundation of Shandong Province (ZR2023MH079, ZR2024ZD23)
  • Jinan Science and Technology Bureau (JNKCJN202201)
  • Clinical Research Center of Shandong University (2020SDUCRCA010)
  • Shandong Provincial Hospital for Maternal and Child Health, Ministry of Health Key Laboratory of Reproductive Regulation Technologies (2023005)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
Keywords
hepatic organoids, embryonic stem cells, liver development, vitamin D receptor, 1,25-dihydroxyvitamin D3, hepatoblasts, glycolytic metabolism, metabolic flux, HIF-1 signaling, MAPK signaling, PI3K/Akt signaling, transcription factors, 2-deoxyglucose, organoid differentiation
MeSH terms
Cell Differentiation, Humans, Liver, Organoids, Vitamin D, Receptors, Calcitriol, Proteomics, Embryonic Stem Cells, Transcriptome

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