Abstract
The objective response rate of immune checkpoint blockade (ICB) in hepatocellular carcinoma (HCC) with anti PD-L1/PD-1 therapy is low. Discovering the signaling pathways regulating PD-L1 might help to improve ICB response rates. Here, we investigate transcription factors IRF-1 and IRF-2 signaling pathways regulating PD-L1 in HCC cells. In vivo studies show that IRF-1 and PD-L1 mRNA expression in human HCC tumors are significantly repressed compared with noncancerous background liver. IRF-1, IRF-2, and PD-L1 mRNA expression correlated positively in HCC tumors. Increased IRF-1 mRNA expression was observed in patients with well-differentiated or early stage HCC tumors. In vitro studies show that IFN-γ induces PD-L1 mRNA and protein expression through upregulation of IRF-1 in mouse and human HCC cells. IRF-1, IRF-2, and PD-L1 mRNA expression is upregulated in murine HCC by co-culture with effector T cells from spleen cells incubated with anti-CD3/CD28 antibodies. IRF-2 over-expression down-regulates IFN-γ induced PD-L1 promoter activity and protein levels in a dose-dependent manner. We identify two IRF-1 response elements (IRE1/IRE2) in the upstream 5′-flanking region of the CD274 (PD-L1) gene promoter. Site-directed mutagenesis shows both IRE1 and IRE2 are functional in transfection promoter assays. IRF-1 traditionally functions as tumor suppressor gene. However, these novel findings show a complex role for IRF-1 which upregulates PD-L1 in the inflammatory tumor microenvironment. IRF-1 antagonizes IRF-2 for binding to the IRE promoter element in PD-L1 which gives new insight to the regulation of PD-L1/PD-1 pathways in HCC ICB therapy.
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Abbreviations
- ATR:
-
Ataxia telangiectasia and Rad3-related protein
- cGAS:
-
Cyclic guanosine monophosphate (GMP)–adenosine monophosphate (AMP) synthase
- CHEK-1:
-
Check point kinase 1
- CTLA-4:
-
Cytotoxic T-lymphocyte-associated protein 4
- HCC:
-
Hepatocellular carcinoma
- ICB:
-
Immune checkpoint blockade
- IFN-γ:
-
Interferon-γ
- IRE:
-
IRF response element
- IRF-1:
-
Interferon regulatory factor 1
- IRF-2:
-
Interferon regulatory factor 2
- JAK:
-
Janus kinase
- MARK:
-
Mitogen-activated protein kinase
- PD-L1:
-
Programmed death-ligand 1
- PD-1:
-
Programmed death protein 1
- PI3K:
-
Phosphatidylinositol 3-kinase
- RLU:
-
Relative luciferase unit
- STAT:
-
Signal transducer and activator of transcription
- STING:
-
Stimulator of interferon gene
- TCGA:
-
The Cancer Genome Atlas
- TNM:
-
Tumor node metastasis classification
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Funding
DAG was supported by funding from National Institute of Health (HHSN276201200017C and P30DK120531-01). YY was supported by funding from Guangxi Natural Science Foundation (2017GXNSFAA198014) and Guangxi High Education Institute Science Foundation (2017KY0099).
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YY and DAG designed the research. YY, LZ, QD, BY, and DAG performed research and also analyzed the data. YY and DAG wrote the paper. All authors edited and approved the submission of this work.
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All individuals provided written informed consent and human tissues were obtained in accordance with the University of Pittsburgh Institutional Review Board (IRB) approved protocol (No. MOD08010372/PRO08010372).
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Yan, Y., Zheng, L., Du, Q. et al. Interferon regulatory factor 1 (IRF-1) and IRF-2 regulate PD-L1 expression in hepatocellular carcinoma (HCC) cells. Cancer Immunol Immunother 69, 1891–1903 (2020). https://doi.org/10.1007/s00262-020-02586-9
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DOI: https://doi.org/10.1007/s00262-020-02586-9