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PER2 binding to HSP90 enhances immune response against oral squamous cell carcinoma by inhibiting IKK/NF-κB pathway
Zhiwei Zhang1, Deping Sun2, Hong Tang1
1Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Background:
Programmed death-ligand 1 (PD-L1) contributes to the immune escape of tumor cells and is a critical target for antitumor immunotherapy. However, the molecular mechanisms regulating PD-L1 expression remain unclear, hindering the development of effective therapies. Here we investigate the role and molecular mechanism of the core clock gene Period2 (PER2) in regulating PD-L1 expression and its role in the combination therapy of oral squamous cell carcinoma (OSCC).
Methods:
Quantitative real-time PCR, western blotting or immunohistochemistry to detect expression of PER2 and PD-L1 in OSCC tissues and cells. Overexpression and knockdown of PER2 detects the function of PER2. Bioinformatics, immunoprecipitation, GST pull-down, CHX chase assay and western blot and strip to detect the mechanism of PER2 regulation for PD-L1. A humanized immune reconstitution subcutaneous xenograft mouse model was established to investigate the combination therapy efficacy.
Results:
In OSCC tissues and cells, PER2 expression was reduced and PD-L1 expression was increased, the expression of PER2 was significantly negatively correlated with PD-L1. In vitro and in vivo experiments demonstrated that PER2 inhibited PD-L1 expression and enhanced T-cell-mediated OSCC cell killing by suppressing the IKK/NF-κB pathway. Mechanistically, PER2 binds to heat shock protein 90 (HSP90) through the PAS1 domain and reduces the interaction of HSP90 with inhibitors of kappa B kinase (IKKs), promoting the ubiquitination of IKKα/β and p65 nuclear translocation to inhibit IKK/NF-κB pathway, thereby suppressing PD-L1 expression. In humanized immune reconstitution subcutaneous xenograft mouse model, it was demonstrated that PER2 targeting combined with anti-PD-L1 treatment improved the inhibition of OSCC growth by promoting CD8+ T-cell infiltration into the tumor.
Conclusions:
Our findings reveal the role and mechanism of PD-L1 regulation by PER2 and support the potential clinical application of PER2 targeting in combination with anti-PD-L1 in OSCC immunotherapy.
Insights
The core clock gene Period2 (PER2) suppresses programmed death-ligand 1 (PD-L1) in oral squamous cell carcinoma (OSCC). Targeting PER2 with anti-PD-L1 immunotherapy enhances anti-tumor effects by promoting T-cell infiltration.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
- Chronobiology
Background:
- Programmed death-ligand 1 (PD-L1) promotes tumor immune escape and is a key immunotherapy target.
- Mechanisms regulating PD-L1 expression are not fully understood, limiting therapeutic development.
- Oral squamous cell carcinoma (OSCC) is a significant health concern where immune escape is a challenge.
Purpose of the Study:
- Investigate the role of the core clock gene Period2 (PER2) in regulating PD-L1 expression in OSCC.
- Elucidate the molecular mechanism by which PER2 influences PD-L1.
- Evaluate the efficacy of combining PER2 targeting with anti-PD-L1 therapy for OSCC.
Main Methods:
- Quantitative real-time PCR, western blotting, and immunohistochemistry were used to assess PER2 and PD-L1 expression in OSCC.
- In vitro and in vivo experiments involved PER2 overexpression and knockdown.
- Mechanism studies included bioinformatics, immunoprecipitation, GST pull-down, CHX chase assays, and western blotting.
- A humanized immune reconstitution subcutaneous xenograft mouse model was utilized.
Main Results:
- PER2 expression was decreased, while PD-L1 expression was increased in OSCC tissues and cells, showing a negative correlation.
- PER2 inhibited PD-L1 expression and enhanced T-cell-mediated OSCC cell killing by suppressing the IKK/NF-κB pathway.
- PER2 binds HSP90, reducing HSP90-IKK interaction, promoting IKK ubiquitination, and inhibiting NF-κB signaling, thus suppressing PD-L1.
- Combination therapy of PER2 targeting and anti-PD-L1 improved OSCC growth inhibition and CD8+ T-cell infiltration in a mouse model.
Conclusions:
- PER2 plays a crucial role in regulating PD-L1 expression in OSCC.
- The findings elucidate the molecular mechanism involving PER2, HSP90, and the IKK/NF-κB pathway.
- PER2 targeting combined with anti-PD-L1 immunotherapy shows potential for clinical application in OSCC treatment.
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