The tumour microenvironment shapes innate lymphoid cells in patients with hepatocellular carcinoma

Gut. 2022 Jun;71(6):1161-1175. doi: 10.1136/gutjnl-2021-325288. Epub 2021 Aug 2.


Objective: Hepatocellular carcinoma (HCC) represents a typical inflammation-associated cancer. Tissue resident innate lymphoid cells (ILCs) have been suggested to control tumour surveillance. Here, we studied how the local cytokine milieu controls ILCs in HCC.

Design: We performed bulk RNA sequencing of HCC tissue as well as flow cytometry and single-cell RNA sequencing of enriched ILCs from non-tumour liver, margin and tumour core derived from 48 patients with HCC. Simultaneous measurement of protein and RNA expression at the single-cell level (AbSeq) identified precise signatures of ILC subgroups. In vitro culturing of ILCs was used to validate findings from in silico analysis. Analysis of RNA-sequencing data from large HCC cohorts allowed stratification and survival analysis based on transcriptomic signatures.

Results: RNA sequencing of tumour, non-tumour and margin identified tumour-dependent gradients, which were associated with poor survival and control of ILC plasticity. Single-cell RNA sequencing and flow cytometry of ILCs from HCC livers identified natural killer (NK)-like cells in the non-tumour tissue, losing their cytotoxic profile as they transitioned into tumour ILC1 and NK-like-ILC3 cells. Tumour ILC composition was mediated by cytokine gradients that directed ILC plasticity towards activated tumour ILC2s. This was liver-specific and not seen in ILCs from peripheral blood mononuclear cells. Patients with high ILC2/ILC1 ratio expressed interleukin-33 in the tumour that promoted ILC2 generation, which was associated with better survival.

Conclusion: Our results suggest that the tumour cytokine milieu controls ILC composition and HCC outcome. Specific changes of cytokines modify ILC composition in the tumour by inducing plasticity and alter ILC function.

Keywords: hepatocellular carcinoma; immune response; immunology; immunoregulation; liver immunology.

MeSH terms

  • Carcinoma, Hepatocellular* / metabolism
  • Cytokines / metabolism
  • Humans
  • Immunity, Innate
  • Killer Cells, Natural / metabolism
  • Leukocytes, Mononuclear
  • Liver Neoplasms* / metabolism
  • Lymphocytes
  • RNA / metabolism
  • Tumor Microenvironment


  • Cytokines
  • RNA