2026-10-10T17:04:07Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34012506 2026-09-28T09:03:57Z category_7 portal_147 item_type_3 month_year_09_2026

Supplemental Material for: Unexpected Limited Immune Potentiation of Common Multikinase Inhibitors Combined with Immune Checkpoint Blockade in HCC: Insights from Murine Models figshare admin karger (2628495) Ying-Chun Shen (10213046) Chia-Lang Hsu (219856) Ching-Ping Yeh (25121311) Wan-Ying Lin (13142126) Yi-Ting Liu (2087746) Ann-Lii Cheng (285432) Medicine Medicine Introduction:Blockade of vascular endothelial growth factor (VEGF) signaling can normalize tumor vasculature and enhance antitumor immunity, as exemplified by atezolizumab plus bevacizumab in hepatocellular carcinoma (HCC). VEGF receptor (VEGFR)-targeting multikinase inhibitors (MKIs) have therefore been considered superior partners for immune checkpoint inhibitors (ICIs) because of their broader inhibition of potentially immune-potentiating kinases. However, most MKI–ICI combinations have shown limited clinical benefit. We hypothesized that MKIs may induce additional biological perturbations beyond VEGFR inhibition that counterbalance immune potentiation.Methods:Murine HCC models (Hepa1-6 and Hep53.4) were treated with a selective anti-VEGFR2 antibody (DC101) or VEGFR-targeting MKIs (cabozantinib, lenvatinib, regorafenib) at low and high doses. Antiangiogenic activity was assessed by CD31⁺ microvessel density (MVD). Transcriptomic remodeling was analyzed by bulk RNA sequencing with gene set variation analysis and principal component analysis. Tumor-infiltrating lymphocytes (TILs) were evaluated for proliferation (Ki67 expression) and interferon-γ (IFN-γ) production. Antitumor efficacy with or without anti–PD-1 was assessed in vivo.Results: All MKIs demonstrated broadly comparable antiangiogenic activity to DC101 (mean CD31+ MVD reduction vs. control: DC101, 27.6%; MKIs: 23.4–42%). Despite this, selective VEGFR2 blockade induced minimal immune perturbation, whereas MKIs induced divergent transcriptomic remodeling across immune, metabolic, stress-response, and proliferation-related pathways. Notably, low-dose cabozantinib and lenvatinib showed reductions in both immune-activating and immune-suppressive gene signatures, suggesting potentially opposing immunologic effects. Functionally, neither DC101 nor MKIs enhanced proliferation or IFN-γ production of tumor-infiltrating T cells. Moreover, MKIs did not improve the antitumor efficacy of anti–PD-1 therapy.Conclusion: Despite similar antiangiogenic effects, MKIs induce broader transcriptomic remodeling without enhancing anti–PD-1 efficacy. These findings suggest that VEGFR-targeting MKIs can induce opposing immune effects beyond VEGF–VEGFR blockade, potentially limiting net enhancement of antitumor immunity. 2026-09-28T09:03:57Z Dataset Dataset 10.6084/m9.figshare.34012506.v1 https://figshare.com/articles/dataset/Supplemental_Material_for_Unexpected_Limited_Immune_Potentiation_of_Common_Multikinase_Inhibitors_Combined_with_Immune_Checkpoint_Blockade_in_HCC_Insights_from_Murine_Models/34012506 CC BY 4.0