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Updated: Nov 28, 2025
An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis
Published on: September 12, 2019
Regorafenib combined with PD1 blockade increases CD8 T-cell infiltration by inducing CXCL10 expression in
Kohei Shigeta1, Aya Matsui1, Hiroto Kikuchi1
1Edwin L. Steele Laboratories for Tumor Biology, Department of Radiation Oncology, Massachusetts General Hospital, Boston, Massachusetts, USA.
Background And Purpose:
Combining inhibitors of vascular endothelial growth factor and the programmed cell death protein 1 (PD1) pathway has shown efficacy in multiple cancers, but the disease-specific and agent-specific mechanisms of benefit remain unclear. We examined the efficacy and defined the mechanisms of benefit when combining regorafenib (a multikinase antivascular endothelial growth factor receptor inhibitor) with PD1 blockade in murine hepatocellular carcinoma (HCC) models.
Basic Procedures:
We used orthotopic models of HCC in mice with liver damage to test the effects of regorafenib-dosed orally at 5, 10 or 20 mg/kg daily-combined with anti-PD1 antibodies (10 mg/kg intraperitoneally thrice weekly). We evaluated the effects of therapy on tumor vasculature and immune microenvironment using immunofluorescence, flow cytometry, RNA-sequencing, ELISA and pharmacokinetic/pharmacodynamic studies in mice and in tissue and blood samples from patients with cancer.
Main Findings:
Regorafenib/anti-PD1 combination therapy increased survival compared with regofarenib or anti-PD1 alone in a regorafenib dose-dependent manner. Combination therapy increased regorafenib uptake into the tumor tissues by normalizing the HCC vasculature and increasing CD8 T-cell infiltration and activation at an intermediate regorafenib dose. The efficacy of regorafenib/anti-PD1 therapy was compromised in mice lacking functional T cells (Rag1-deficient mice). Regorafenib treatment increased the transcription and protein expression of CXCL10-a ligand for CXCR3 expressed on tumor-infiltrating lymphocytes-in murine HCC and in blood of patients with HCC. Using Cxcr3-deficient mice, we demonstrate that CXCR3 mediated the increased intratumoral CD8 T-cell infiltration and the added survival benefit when regorafenib was combined with anti-PD1 therapy.
Principal Conclusions:
Judicious regorafenib/anti-PD1 combination therapy can inhibit tumor growth and increase survival by normalizing tumor vasculature and increasing intratumoral CXCR3+CD8 T-cell infiltration through elevated CXCL10 expression in HCC cells.
Insights
Combining regorafenib and PD1 blockade enhances survival in liver cancer by normalizing tumor vasculature and increasing CD8 T-cell infiltration. This combination therapy, driven by CXCL10 and CXCR3, offers a promising strategy for hepatocellular carcinoma treatment.
Area of Science:
- Immunology
- Oncology
- Pharmacology
Background:
- Vascular endothelial growth factor (VEGF) and programmed cell death protein 1 (PD1) pathway inhibitors show efficacy in various cancers.
- Mechanisms of combined VEGF and PD1 inhibition remain unclear for specific cancers and agents.
Purpose of the Study:
- To examine the efficacy of combining regorafenib (VEGF inhibitor) with PD1 blockade.
- To define the mechanisms of benefit in murine hepatocellular carcinoma (HCC) models.
Main Methods:
- Orthotopic HCC mouse models with liver damage were used.
- Combination therapy involved regorafenib and anti-PD1 antibodies.
- Tumor vasculature, immune microenvironment, and molecular markers were analyzed using various techniques.
Main Results:
- Combination therapy significantly increased survival in a dose-dependent manner.
- Regorafenib normalized HCC vasculature, increasing CD8 T-cell infiltration and activation.
- CXCL10 upregulation and CXCR3-mediated T-cell infiltration were crucial for efficacy.
Conclusions:
- Regorafenib/anti-PD1 combination therapy inhibits tumor growth and improves survival in HCC.
- Tumor vasculature normalization and enhanced intratumoral CXCR3+CD8 T-cell infiltration via CXCL10 are key mechanisms.
