NEDDylation Regulates CD8+ T-cell Metabolism and Antitumor Immunity

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.

Authors

Institutions

Publication Details

Journal
Cancer Immunology Research
Published
2025-04-22
DOI
https://doi.org/10.1158/2326-6066.cir-24-0127
Citations
4
Primary Topic
Peptidase Inhibition and Analysis
Type
article
Field-Weighted Citation Impact
4.18
Controls
|||
ALL TIME
JAN
FEB
MAR
APR
MAY
JUN
JUL
AUG
SEP
OCT
article

NEDDylation Regulates CD8+ T-cell Metabolism and Antitumor Immunity

Ashwin Woodhoo, Ana García-del Río, Asier Antoñana-Vildosola, Endika Prieto‐Fernández et al.
4 citations
Cancer Immunology Research
Peptidase Inhibition and Analysis
4.18
article

NEDDylation Regulates CD8+ T-cell Metabolism and Antitumor Immunity

Ashwin Woodhoo, Ana García-del Río, Asier Antoñana-Vildosola, Endika Prieto‐Fernández, Asís Palazón, Sebastian Peer, Naroa Goikoetxea‐Usandizaga, Marina Serrano‐Maciá, Samanta Romina Zanetti, Adrián Barreira-Manrique, Gottfried Baier, Thomas Gruber, Félix Elortza, Leire Egia‐Mendikute, Borja Jiménez-Lasheras, Alexandre Bosch, Maria Luz Martínez‐Chantar, Mikel Azkargorta, Ander de Blas, Paloma Velasco-Beltrán, So Young Lee, Lorena Pérez, Jone Etxaniz Díaz de Durana
article en
4 citations

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.

Cancer Immunology ResearchVol. 13(7)
Ikerbasque (ES), Hebron University (PS), Innsbruck Medical University (AT), Universität Innsbruck (AT), Universidade de Santiago de Compostela (ES), Instituto de Salud Carlos III (ES), Vall d'Hebron Institut de Recerca (ES), Center for Research in Molecular Medicine and Chronic Diseases (ES), CIC bioGUNE (ES), Vall d'Hebron Hospital Universitari (ES), Instituto de Investigación Sanitaria de Santiago (ES), Vall d'Hebron Institute of Oncology (ES)
Good health and well-being
Openalex Percentile: Top 6%
Peptidase Inhibition and Analysis
4.18
DOIOpen Access PDFOpenAlex
AI Navigator

Ask Laika to Summarize, Analyze, and Connect papers live on the map.

Summarize Papers & Methodologies

Extract key findings, datasets, and comparative methods across publications.

Benchmark Rankings & Visual Analytics

Rank top research institutions, authors, funders, topics, and journals by Field-Weighted Citation Impact (FWCI) and paper volume with instant charts.

Connect Distant Disciplines

Bridge topological clusters on the map to find hidden collaborative intersections.