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Original research
Lineage tracing and single-cell analysis reveal proliferative Prom1+ tumour-propagating cells and their dynamic cellular transition during liver cancer progression
  1. Lei Zhou1,2,
  2. Ken HO Yu1,3,
  3. Tin Lok Wong1,4,
  4. Zhao Zhang5,
  5. Chun Ho Chan1,
  6. Jane HC Loong1,
  7. Noelia Che1,
  8. Hua Jian Yu1,
  9. Kel Vin Tan6,
  10. Man Tong1,2,4,
  11. Elly S Ngan7,
  12. Joshua WK Ho1,3,
  13. Stephanie Ma1,2,4
  1. 1School of Biomedical Sciences, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China
  2. 2Department of Clinical Oncology, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China
  3. 3Laboratory of Data Discovery for Health Limited (D24H), Hong Kong Science Park, Hong Kong SAR, China
  4. 4State Key Laboratory of Liver Research, The University of Hong Kong, Hong Kong SAR, China
  5. 5Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China
  6. 6Department of Diagnostic Radiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China
  7. 7Department of Surgery, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong SAR, China
  1. Correspondence to Dr Stephanie Ma, School of Biomedical Sciences, The University of Hong Kong, Hong Kong, Hong Kong; stefma{at}hku.hk; Dr Joshua WK Ho, School of Biomedical Sciences, The University of Hong Kong, Hong Kong, Hong Kong; jwkho{at}hku.hk

Abstract

Objective Hepatocellular carcinoma (HCC) has high intratumoral heterogeneity, which contributes to therapeutic resistance and tumour recurrence. We previously identified Prominin-1 (PROM1)/CD133 as an important liver cancer stem cell (CSC) marker in human HCC. The aim of this study was to investigate the heterogeneity and properties of Prom1+ cells in HCC in intact mouse models.

Design We established two mouse models representing chronic fibrotic HCC and rapid steatosis-related HCC. We performed lineage tracing post-HCC induction using Prom1C-L/+; Rosa26tdTomato/+ mice, and targeted depletion using Prom1C-L/+; Rosa26DTA/+ mice. Single-cell RNA sequencing (scRNA-seq) was carried out to analyse the transcriptomic profile of traced Prom1+ cells.

Results Prom1 in HCC tumours marks proliferative tumour-propagating cells with CSC-like properties. Lineage tracing demonstrated that these cells display clonal expansion in situ in primary tumours. Labelled Prom1+ cells exhibit increasing tumourigenicity in 3D culture and allotransplantation, as well as potential to form cancers of differential lineages on transplantation. Depletion of Prom1+ cells impedes tumour growth and reduces malignant cancer hallmarks in both HCC models. scRNA-seq analysis highlighted the heterogeneity of Prom1+ HCC cells, which follow a trajectory to the dedifferentiated status with high proliferation and stem cells traits. Conserved gene signature of Prom1 linage predicts poor prognosis in human HCC. The activated oxidant detoxification underlies the protective mechanism of dedifferentiated transition and lineage propagation.

Conclusion Our study combines in vivo lineage tracing and scRNA-seq to reveal the heterogeneity and dynamics of Prom1+ HCC cells, providing insights into the mechanistic role of malignant CSC-like cells in HCC progression.

  • hepatocellular carcinoma
  • stem cells

Data availability statement

Data is available on GEO (GSE181515).

Statistics from Altmetric.com

Data availability statement

Data is available on GEO (GSE181515).

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Footnotes

  • LZ, KHY and TLW contributed equally.

  • Correction notice This article has been corrected since it published Online First. A second corresponding author has been added.

  • Contributors LZ and SM conceived the project, designed the experiments and wrote the manuscript. LZ performed the majority of the experiments including mouse model establishment and characterisation, cell culture, sequencing and data analysis. LZ, KHY and TLW analysed the single-cell sequencing data and provided critical scientific input. TLW helped with manuscript editing. ZZ, CHC, JHL, NC, HJY and MT aided in animal experiments. KVT performed MRI scanning. ESN and JWH provided guidance in single cell sequencing analysis and manuscript editing. SM, JWH and LZ provided funding support. SM and JWH supervised the project.

  • Funding The authors have not declared a specific grant for this research from any funding agency in the public, commercial or not-for-profit sectors.

  • Competing interests None declared.

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

  • Supplemental material This content has been supplied by the author(s). It has not been vetted by BMJ Publishing Group Limited (BMJ) and may not have been peer-reviewed. Any opinions or recommendations discussed are solely those of the author(s) and are not endorsed by BMJ. BMJ disclaims all liability and responsibility arising from any reliance placed on the content. Where the content includes any translated material, BMJ does not warrant the accuracy and reliability of the translations (including but not limited to local regulations, clinical guidelines, terminology, drug names and drug dosages), and is not responsible for any error and/or omissions arising from translation and adaptation or otherwise.

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