# NAA20-mediated ACF1 lactylation drives neuroblastoma progression through enhancing GCLM-dependent glutathione synthesis

## TL;DR

This study finds that ACF1 lactylation, driven by NAA20, boosts glutathione production in neuroblastoma cells, promoting tumor growth and suggesting a new treatment target.

## Contribution

The study identifies a novel NAA20-ACF1-GCLM axis that enhances glutathione synthesis and tumor progression in neuroblastoma.

## Key findings

- ACF1 knockdown reduces tumor growth and increases treatment sensitivity in neuroblastoma cells.
- NAA20-mediated ACF1 lactylation activates GCLM transcription, boosting glutathione levels.
- NAA20 and GCLM are upregulated in high-risk neuroblastoma tissues.

## Abstract

Neuroblastoma (NBL) is a pediatric malignancy with poor prognosis in high-risk cases. This study explores the function of albumin conformation factor 1 (ACF1) in NBL progression and delves into the underpinning mechanism. Exome and transcriptome sequencing were applied to analyze ACF1 mutations/expression in NBL tissues versus controls. ACF1 was knocked down in NBL cell lines (KELLY, BE2C, N2a) for in vitro assays (viability, proliferation, migration, apoptosis, therapy sensitivity) or in vivo xenograft/metastasis models with radiation/cisplatin. Mechanisms were probed via RNA-sequencing, chromatin immunoprecipitation, luciferase assays, co-immunoprecipitation, and immunofluorescence assays. Expression patterns and the correlations between ACF1, GCLM, and NAA20 were detected in human NBL tissue microarrays. ACF1 mutations and elevated expression correlated with advanced tumor staging, high-risk factors, and unfavorable prognosis in NBL datasets and TMAs. ACF1 knockdown suppressed NBL cell proliferation, mobility, and in vivo tumor growth/metastasis, while enhancing cisplatin/radiation sensitivity and apoptosis. Mechanistically, ACF1 knockdown reduced GCLM transcription via decreased H3K27ac/H3K4me3/Myc at its promoter, elevating lipid peroxidation and lowering glutathione (GSH) levels. Lactate induced ACF1 lactylation and nuclear translocation, promoted by NAA20 interaction (enhanced by lactate). NAA20 knockdown phenocopied ACF1 effects, rescued by GCLM overexpression. NAA20 and GCLM were upregulated in NBL datasets/TMAs. This study suggests that the NAA20-mediated ACF1 lactylation drives GCLM-dependent GSH synthesis, promoting NBL cell growth and metastasis. Targeting this axis may improve therapy response.

This study uncovers a novel mechanism in which NAA20 promotes ACF1 nuclear translocation through lactylation, which in turn activates transcription of GCLM and enhances the antioxidant machinery of NBL cells, thereby promoting malignancy. As a result, the NAA20-ACF1-GCLM axis may represent a potential therapeutic target for NBL. Although our experiments confirm the direct interaction between NAA20 and ACF1, the precise lactylation sites on ACF1 remain uncharacterized. Further investigations are needed to determine whether NAA20 directly controls ACF1 lactylation or modulates the chromatin environment to achieve this effect.

The online version contains supplementary material available at 10.1007/s10565-026-10154-7.