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Updated: Jan 2, 2026
Experimental Melanoma Immunotherapy Model Using Tumor Vaccination with a Hematopoietic Cytokine
Published on: February 24, 2023
NEDD4 Negatively Regulates GITR via Ubiquitination in Immune Microenvironment of Melanoma
Yu Guo1, Lichang Yang1, Shaorong Lei1
1Department of Plastic Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, People's Republic of China.
Introduction:
Melanoma is a common skin cancer that is usually associated with poor clinical outcomes. Recently, the immune checkpoint GITR has been identified as a promising target for immunotherapy of melanoma. In this study, we aimed to investigate the post-translational regulation mechanism of GITR in melanoma.
Methods:
Western blotting was used to evaluate the protein expression of NEDD4, GITR and Foxp3. Real-time PCR (RT-PCR) was performed to determine expression levels of NEDD4, GITR, Foxp3 and IL-2. Cell viability was detected by MTT assay. The ubiquitination of GITR was evaluated by immunoprecipitation. NEDD4 expression data and melanoma survival data were obtained from The Cancer Genome Atlas (TCGA) and cBioPortal databases.
Results:
We demonstrate that E3 ligase NEDD4 binds to GITR and mediates ubiquitination and degradation of GITR. Overexpression of NEDD4 inhibits anti-tumor immunity mediated by T cells against melanoma cells. We also found that the expression of NEDD4 is increased in metastatic melanoma. High NEDD4 expression level is correlated with the poor prognosis of melanoma patients.
Discussion:
In summary, our findings demonstrated that E3 ligase NEDD4 mediates ubiquitination and degradation of GITR and suppresses T-cell-mediated-killings on melanoma cells. Our work highlighted the E3 ligase NEDD4 as a novel prognosis biomarker and therapeutic target for melanoma.
Insights
E3 ligase NEDD4 targets GITR for degradation, suppressing anti-tumor immunity in melanoma. Increased NEDD4 expression correlates with poor prognosis, identifying it as a potential therapeutic target and biomarker for melanoma.
Area of Science:
- Immunology
- Oncology
- Molecular Biology
Background:
- Melanoma is a prevalent skin cancer with poor outcomes.
- Glucocorticoid-induced tumor necrosis factor receptor-related protein (GITR) is a promising immunotherapy target.
- Understanding GITR's regulation is crucial for melanoma treatment.
Purpose of the Study:
- Investigate the post-translational regulation of GITR in melanoma.
- Determine the role of E3 ligase NEDD4 in GITR regulation.
- Evaluate NEDD4 as a prognostic biomarker and therapeutic target.
Main Methods:
- Western blotting and RT-PCR for protein and gene expression analysis (NEDD4, GITR, Foxp3, IL-2).
- MTT assay for cell viability.
- Immunoprecipitation to assess GITR ubiquitination.
- Analysis of TCGA and cBioPortal data for NEDD4 and melanoma survival.
Main Results:
- NEDD4 binds to GITR, mediating its ubiquitination and degradation.
- NEDD4 overexpression inhibits T-cell-mediated anti-tumor immunity against melanoma.
- NEDD4 expression is elevated in metastatic melanoma and linked to poor patient prognosis.
Conclusions:
- NEDD4-mediated GITR degradation suppresses T-cell anti-melanoma activity.
- NEDD4 serves as a novel prognostic biomarker for melanoma.
- NEDD4 represents a potential therapeutic target for melanoma treatment.
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