Novel miRNA-based drug CD5-2 reduces liver tumor growth in diethylnitrosamine-treated mice by normalizing tumor vasculature and altering immune infiltrate

Ken Liu; Jinbiao Chen; Yang Zhao; Jade Boland; Ka Ka Ting; Glen Lockwood; Catriona McKenzie; James Kench; Mathew A Vadas; Jennifer R Gamble; Geoffrey W McCaughan

doi:10.3389/fimmu.2023.1245708

Novel miRNA-based drug CD5-2 reduces liver tumor growth in diethylnitrosamine-treated mice by normalizing tumor vasculature and altering immune infiltrate

Front Immunol. 2023 Sep 18:14:1245708. doi: 10.3389/fimmu.2023.1245708. eCollection 2023.

Authors

Ken Liu^{1

2

3}, Jinbiao Chen³, Yang Zhao³, Jade Boland³, Ka Ka Ting³, Glen Lockwood⁴, Catriona McKenzie⁵, James Kench⁵, Mathew A Vadas^{2

3}, Jennifer R Gamble^{2

3}, Geoffrey W McCaughan^{1

2

3}

Affiliations

¹ AW Morrow Gastroenterology and Liver Centre, Royal Prince Alfred Hospital, Sydney, NSW, Australia.
² Faculty of Medicine and Health, University of Sydney, Sydney, NSW, Australia.
³ Centenary Institute, University of Sydney, Sydney, NSW, Australia.
⁴ Biogerontology Group, ANZAC Research Institute, Sydney, NSW, Australia.
⁵ New South Wales Health Pathology, Royal Prince Alfred Hospital, Sydney, NSW, Australia.

Abstract

Introduction: Liver cancers exhibit abnormal (leaky) vasculature, hypoxia and an immunosuppressive microenvironment. Normalization of tumor vasculature is an emerging approach to treat many cancers. Blockmir CD5-2 is a novel oligonucleotide-based inhibitor of the miR-27a interaction with VE-Cadherin, the endothelial-specific cadherin. The combination of a vasoactive medication with inhibition of immune checkpoints such as programmed cell death protein 1 (PD1) has been shown to be effective in treating liver cancer in humans. We aimed to study the effect of CD5-2 combined with checkpoint inhibition (using an antibody against PD1) on liver tumor growth, vasculature and immune infiltrate in the diethylnitrosamine (DEN)-induced liver tumor mouse model.

Methods: We first analyzed human miR-27a and VE-Cadherin expression data from The Cancer Genome Atlas for hepatocellular carcinoma. CD5-2 and/or anti-PD1 antibody were given to the DEN-treated mice from age 7-months until harvest at age 9-months. Tumor and non-tumor liver tissues were analyzed using histology, immunohistochemistry, immunofluorescence and scanning electron microscopy.

Results: Human data showed high miR-27a and low VE-Cadherin were both significantly associated with poorer prognosis. Mice treated with CD5-2 plus anti-PD1 antibody had significantly smaller liver tumors (50% reduction) compared to mice treated with either agent alone, controls, or untreated mice. There was no difference in tumor number. Histologically, tumors in CD5-2-treated mice had less leaky vessels with higher VE-Cadherin expression and less tumor hypoxia compared to non-CD5-2-treated mice. Only tumors in the combination CD5-2 plus anti-PD1 antibody group exhibited a more favorable immune infiltrate (significantly higher CD3+ and CD8+ T cells and lower Ly6G+ neutrophils) compared to tumors from other groups.

Discussion: CD5-2 normalized tumor vasculature and reduced hypoxia in DEN-induced liver tumors. CD5-2 plus anti-PD1 antibody reduced liver tumor size possibly by altering the immune infiltrate to a more immunosupportive one.

Keywords: angiogenesis; hepatocellular carcinoma; hypoxia; immunotherapy; vascular normalization.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Diethylnitrosamine / toxicity
Humans
Hypoxia
Infant
Liver Neoplasms* / chemically induced
Liver Neoplasms* / drug therapy
Liver Neoplasms* / genetics
Mice
MicroRNAs* / genetics
MicroRNAs* / metabolism
Tumor Microenvironment

Substances

Diethylnitrosamine
MicroRNAs