Dataset Information

MS-based analysis of the role of protein neddylation in the context of T-cell-mediated anti-tumor immunity (2)

ABSTRACT: Neddylation is one type of post-translational modification (PTM) by which the ubiquitin-like molecule NEDD8 is covalently attached to protein substrates, in a process that depends on NAE1 (NEDD8 activating enzyme E1 regulatory subunit). Here we study the role of protein neddylation in the context of T-cell-mediated anti-tumor immunity. Analysis of publicly available single-cell RNA sequencing databases revealed that tumor infiltrating lymphocytes (TILs) upregulate the expression of Nedd8 during their differentiation into effector memory cells. In vitro activation of mouse and human CD8+ T cells drives the upregulation of the neddylation pathway, resulting in an enrichment of neddylated proteins. In vivo tumor challenge assays demonstrated that CD8+ T cells lacking NAE1 (NAE1-KO) have a reduced anti-tumor capacity, and display a less activated phenotype with impaired differentiation into effector T cells. LC MS/MS proteomic analyses conducted on activated NAE1-KO CD8+ T cells and on CD8+ T cells subjected to the treatment of neddylation inhibitor MLN4924 elucidated a pronounced impairment in both glycolytic and oxidative phosphorylation metabolic pathways. In this context, we validated three novel NEDD8 interactors (LDHA, HK1 and ENOA), which are relevant enzymes in the glycolytic pathway. This can clarify the role of neddylation in the metabolic shift to glycolysis exerted by CD8+ T cells during their anti-tumor activity.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Mikel Azkargorta

LAB HEAD: Felix Elortza

PROVIDER: PXD050529 | Pride | 2025-04-15

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	220411_220331A_Borja_KO_1_S1-A1_1_2920.d.rar	Other
	220411_220331A_Borja_KO_2_S1-A2_1_2921.d.rar	Other
	220411_220331A_Borja_KO_3_S1-A3_1_2922.d.rar	Other
	220411_220331A_Borja_KO_4_S1-A4_1_2923.d.rar	Other
	220411_220331A_Borja_MLN_10_S1-A10_1_2929.d.rar	Other

Items per page:

1 - 5 of 13

Similar Datasets

Project description:The heart undergoes significant structural, metabolic, gene expression and functional alterations during the perinatal to postnatal transition. While recent studies have identified multiple epigenetic and transcriptional regulators of cardiac maturation, post-transcriptional mechanisms regulating this process remain poorly understood. Neddylation is a post-translational modification that conjugates a small ubiquitin-like protein, NEDD8, to protein substrates via an E1-E2-E3 enzymatic cascade. The goal of this study was to define the role of neddylation in perinatal cardiac development and cardiac maturation. Neddylation was inhibited in adult mouse hearts by cardiac-specific deletion of NAE1 gene, a regulatory subunit of NEDD8 E1 enzyme, or in neonatal cardiomyocytes (CMs) with a pharmacological neddylation inhibitor, MLN4924. The impact on cardiac transcriptome, metabolism, maturation and function was assessed. Mosaic deletion of NAE1 in ~40% neonatal CMs disrupted aspects of maturation, including transverse-tubule formation, cellular hypertrophy and fetal/adult isoform switching, whereas deletion of NAE1 in over 80% CMs led to rapid development of cardiomyopathy and heart failure. Transcriptome analysis demonstrated an association of metabolic derangement with immature cardiomyocyte signature. Biochemical, ultrastructural and metabolomics analyses confirmed downregulation of fatty acid and oxidative phosphorylation genes, deficits in fatty acid utilization, mitochondrial dysfunction, and significantly altered metabolic profiles in NAE1-deficient hearts or MLN4924-treated neonatal CMs. Mechanistically, we found that HIF1α, a transcription factor known to promote glycolysis and suppress oxidative metabolism, is a putative NEDD8 target. Inhibition of neddylation resulted in HIF1α accumulation and activation, which contributed to diminished fatty acid utilization. Taken together, we conclude that neddylation plays a crucial role in CM maturation and postnatal cardiac development through sustaining the glycolytic to oxidative metabolic switch in perinatal hearts.

Dataset Information

MS-based analysis of the role of protein neddylation in the context of T-cell-mediated anti-tumor immunity (2)

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets