2. Academic Validation
  3. NEDDylation Regulates CD8+ T-cell Metabolism and Antitumor Immunity

NEDDylation Regulates CD8+ T-cell Metabolism and Antitumor Immunity

  • Cancer Immunol Res. 2025 Jul 2;13(7):1004-1021. doi: 10.1158/2326-6066.CIR-24-0127.
Borja Jiménez-Lasheras # 1,  Paloma Velasco-Beltrán # 1,  Leire Egia-Mendikute 1,  Lorena Pérez-Gutiérrez 1,  So Young Lee 1,  Ander de Blas 1,  Ana García-Del Río 1,  Samanta Romina Zanetti 1,  Asier Antoñana-Vildosola 1,  Adrián Barreira-Manrique 1,  Alexandre Bosch 1,  Jone Etxaniz-Díaz de Durana 1,  Endika Prieto-Fernández 2,  Marina Serrano-Maciá 3 4,  Naroa Goikoetxea-Usandizaga 3 4,  Mikel Azkargorta 5,  Félix Elortza 5,  Thomas Gruber 6,  Sebastian Peer 6 7,  Gottfried Baier 6,  Ashwin Woodhoo 8,  María Luz Martínez-Chantar 3 4,  Asis Palazon 1 9
Affiliations

Affiliations

  • 1 Cancer Immunology and Immunotherapy Laboratory, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.
  • 2 Tumor Immunology and Immunotherapy Laboratory, Vall d'Hebron Institute of Oncology (VHIO), Vall d'Hebron Barcelona Hospital Campus, Barcelona, Spain.
  • 3 Liver Disease Laboratory, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Derio, Spain.
  • 4 CIBERehd, Instituto de Salud Carlos III, Madrid, Spain.
  • 5 Proteomics Platform, Center for Cooperative Research in Biosciences (CIC bioGUNE), Basque Research and Technology Alliance (BRTA), Carlos III Networked Proteomics Platform (ProteoRed-ISCIII) Derio, Spain.
  • 6 Department for Genetics, Institute of Cell Genetics, Medical University Innsbruck, Innsbruck, Austria.
  • 7 Internal Medicine V, Medical University Innsbruck, Innsbruck, Austria.
  • 8 Gene Regulatory Control in Disease Laboratory, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Instituto de Investigación Sanitaria de Santiago de Compostela (IDIS), University of Santiago de Compostela, A Coruña, Spain.
  • 9 Ikerbasque, Basque Foundation for Science, Bilbao, Spain.
  • # Contributed equally.
PMID: 40261130 DOI: 10.1158/2326-6066.CIR-24-0127
Abstract

NEDDylation is a posttranslational modification whereby the ubiquitin-like molecule NEDD8 is attached to protein substrates in a process dependent on NEDD8-activating Enzyme regulatory subunit (NAE1). NEDDylation is emerging as a regulator of Cancer biology, but its precise role in antitumor immunity has not been thoroughly characterized. In this study, we examine the impact of NEDDylation in CD8+ T cell-mediated antitumor responses. Analysis of publicly available single-cell RNA Sequencing databases revealed that CD8+ tumor-infiltrating lymphocytes showed increased expression of NEDD8 during their differentiation into effector memory cells. In vitro activation of mouse and human CD8+ T cells drove the upregulation of the NEDDylation enzymatic pathway, resulting in an enrichment of NEDDylated proteins. In vivo tumor challenge assays demonstrated that CD8+ T cells lacking NAE1 exhibited reduced antitumor capability and a less activated phenotype with compromised differentiation into effector cells. Upregulating NEDDylation by knocking out deNEDDylase sentrin-specific protease 8 increased the in vitro cytotoxic capability of CD8+ CAR T cells. In addition, LC MS/MS proteomic analyses of NAE1-deficient CD8+ T cells and CD8+ T cells treated with the NEDDylation inhibitor MLN4924 showed a pronounced impairment in metabolic pathways, including glycolysis and Oxidative Phosphorylation. In this context, we validated Lactate Dehydrogenase A, α-enolase, and Hexokinase 1, which are relevant glycolytic Enzymes, as NEDD8 targets. In line with this, NEDDylation-deficient CD8+ T cells demonstrated reduced transcription, protein expression, and enzymatic activity of Lactate Dehydrogenase. In summary, we uncover NEDDylation as a critical regulator of CD8+ T cell-mediated antitumor immunity.

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