Study of the immune cell microenvironment and transcriptional and miRNA landscape of Kupffer cells in DEN-CCl4 mouse models of liver fibrosis and HCC
(2025)
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Details
- Authors
- Supervisors
- Smets, Françoise
- Abstract
- Hepatocellular carcinoma (HCC) is a classic inflammation-driven cancer that usually develops in the context of chronic liver disease (CLD), progressing from fibrosis to cirrhosis and ultimately to malignancy. To better understand the immune dysregulation that drives this progression, we used a well-established DEN-CCl4-induced mouse model, which closely mimics human HCC due to tumor development in the background of CLD. We first characterized immune alterations across disease stages and spatial compartments by subdividing HCC livers into tumor, invasive margin (IM), and non-tumor tissue (NTT). We observed that this model closely resembles the immunosuppressive subtype of human HCC, characterized by an increase in regulatory T cells (Tregs) and shift in the hepatic macrophage population from Kupffer cell (KC) dominant to infiltrating macrophage dominant composition, and notably a lack of intratumoral CD8⁺ T cell expansion. This immune profile reflects a dysfunctional anti-tumor response. Spatial localization by multiplex immunofluorescence shows that the increased presence of immune cells in the IM region highlights its role as a critical interface where tumor cells interact with the surrounding parenchyma, while the rise in the NTT region indicates significant immune dysregulation in advanced CLD, potentially promoting tumor growth (Chapter 2). Given the importance of Tregs, we aimed to study their transcriptomic and miRNA profiles. However, technical limitations due to the low abundance of hepatic Tregs and poor RNA quality prevented detailed molecular analyses (Annex 2). We therefore shifted focus to KCs, the liver’s resident 8 macrophages which play a central role in regulating hepatic immunity. We observed increased expression of gene sets associated with immune activation, ion and protein transport, carbohydrate metabolism, lipid storage, collagen biosynthesis, extracellular matrix (ECM) remodeling, angiogenesis, epithelial-mesenchymal transition (EMT), hypoxia responses, and several key signaling pathways. Importantly, we found downregulation of miR-29c-3p in KCs from HCC livers, implicating enhanced TGF-β signaling, ECM reorganization, and EMT in tumor progression. Meanwhile, upregulated miRNAs such as miR-214, miR-709, miR-5128, and miR-216-5p were predicted to target genes involved in platelet homeostasis, ion transport, and cGMP signaling, suggesting a role in immune and metabolic dysregulation (Chapter 3). In conclusion, our study delineates the progression of immune alterations from fibrosis to HCC in the DEN-CCl4 model and demonstrates KC reprogramming at the mRNA-miRNA level. This is, to our knowledge, the first study to analyze the integrated transcriptomic and post-transcriptional landscape of KCs during liver disease. Altogether, these findings provide mechanistic insights into the drivers of tumor progression and the central role of KCs in the immunopathology of HCC.
- Affiliations
Citations
Ajith, A. (2025). Study of the immune cell microenvironment and transcriptional and miRNA landscape of Kupffer cells in DEN-CCl4 mouse models of liver fibrosis and HCC.