Journal Article

N6-methyladenosine-modified circIGF2BP3 inhibits CD8+ T-cell responses to facilitate tumor immune evasion by promoting the deubiquitination of PD-L1 in non-small cell lung cancer

Liu, Zhenchuan, Wang, Tingting, Zhou, Yongxin, She, Yunlang, Wu, Kaiqing, Chen, Chang, Li, Lei, Dong, Chenglai, Gu, Shaorui

2021

Overview

Background An in-depth understanding of immune evasion mechanisms in tumors is crucial to overcome resistance and enable innovative advances in immunotherapy. Circular RNAs (circRNAs) have been implicated in cancer progression. However, much remains unknown regarding whether circRNAs impact immune escape in non-small-cell lung carcinoma (NSCLC). Methods We performed bioinformatics analysis to profile and identify the circRNAs mediating immune evasion in NSCLC. A luciferase reporter assay, RNA immunoprecipitation (RIP), RNA pulldown assays and fluorescence in situ hybridization were performed to identify the interactions among circIGF2BP3, miR-328-3p, miR-3173-5p and plakophilin 3 (PKP3). In vitro T cell-mediated killing assays and in vivo syngeneic mouse models were used to investigate the functional roles of circIGF2BP3 and its downstream target PKP3 in antitumor immunity in NSCLC. The molecular mechanism of PKP3-induced PD-L1 upregulation was explored by immunoprecipitation, RIP, and ubiquitination assays. Results We demonstrated that circIGF2BP3 (hsa_circ_0079587) expression was increased in NSCLC and negatively correlated with CD8 + T cell infiltration. Functionally, elevated circIGF2BP3 inactivated cocultured T cells in vitro and compromised antitumor immunity in an immunocompetent mouse model, and this effect was dependent on CD8 + T cells. Mechanistically, METTL3 mediates the N 6 -methyladenosine (m 6 A) modification of circIGF2BP3 and promotes its circularization in a manner dependent on the m 6 A reader protein YTHDC1. circIGF2BP3 competitively upregulates PKP3 expression by sponging miR-328-3p and miR-3173-5p to compromise the cancer immune response. Furthermore, PKP3 engages with the RNA-binding protein FXR1 to stabilize OTUB1 mRNA, and OTUB1 elevates PD-L1 abundance by facilitating its deubiquitination. Tumor PD-L1 deletion completely blocked the impact of the circIGF2BP3/PKP3 axis on the CD8 + T cell response. The inhibition of circIGF2BP3/PKP3 enhanced the treatment efficacy of anti-PD-1 therapy in a Lewis lung carcinoma mouse model. Collectively, the PKP3/PD-L1 signature and the infiltrating CD8 + T cell status stratified NSCLC patients into different risk groups. Conclusion Our results reveal the function of circIGF2BP3 in causing immune escape from CD8 + T cell-mediated killing through a decrease in PD-L1 ubiquitination and subsequent proteasomal degradation by stabilizing OTUB1 mRNA in a PKP3-dependent manner. This work sheds light on a novel mechanism of PD-L1 regulation in NSCLC and provides a rationale to enhance the efficacy of anti-PD-1 treatment in NSCLC.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Analysis / Animal models / Binding sites / Bioinformatics / Biomedical and Life Sciences / Biomedicine / Cancer Research / CD8 antigen / Circular RNA / CRISPR / Development and progression / Disease susceptibility / Fluorescence in situ hybridization / Gene expression / Immune escape / Immune evasion / Immune response / Immunoprecipitation / Immunotherapy / Lung cancer / Lung cancer, Non-small cell / Lung carcinoma / Lymphocytes T / Metastases / Metastasis / mRNA / Mutagenesis / Mutation / N6-methyladenosine / Non-small cell lung carcinoma / Oncology / PD-1 protein / PD-L1 / PD-L1 protein / Plakophilin 3 / Plasmids / Proteasomes / Protein binding / Risk groups / RNA / RNA-binding protein / Small cell lung carcinoma / T cells / Tumors / Ubiquitin / Ubiquitination