Intermittent fasting inhibits Tp53-driven glioma through gut microbiota-mediated methionine-m6A regulation.

Intermittent fasting significantly inhibits GBM progression in Tp53 subtype but not in Cdkn2a subtype mouse models.

• GBM was classified into CDKN2A subtype and TP53 subtype for comparison of IF efficacy.
• IF showed significant inhibitory effects on GBM progression in mice with the Tp53 GBM model.
• The inhibitory effect of IF was not significant in the Cdkn2a GBM model.
• This finding supports a genotype-based hypothesis for the therapeutic effects of IF on tumors.

The efficacy of intermittent fasting is primarily mediated by alterations in gut microbiota composition.

• Multi-omics sequencing was performed in the IF-responsive Tp53 GBM mouse model.
• Systematic biological analysis and rescue experiments were conducted to demonstrate gut microbiota's role.
• Gut microbiota alterations subsequently modulated the production of the microbial metabolite methionine sulfoxide.
• The microbiome was one of several omics layers analyzed, alongside spatial transcriptome, spatial metabolome, single-cell transcriptome, single-cell RNA methylation, and metabolome.

Methionine sulfoxide, a microbial metabolite altered by IF, regulates m6A RNA modification to suppress GBM progression.

• Methionine sulfoxide was identified as the key microbial metabolite mediating IF's anti-tumor effects.
• Methionine sulfoxide regulation of m6A modification was demonstrated through single-cell RNA methylation profiling.
• m6A modification changes led to inhibition of the TGF-β signaling pathway.
• Suppression of TGF-β signaling resulted in inhibition of GBM progression.

A comprehensive multi-omics molecular profiling was performed to delineate the mechanisms of IF in the Tp53 GBM mouse model.

• The multi-omics approach included spatial transcriptome, spatial metabolome, single-cell transcriptome, single-cell RNA methylation, metabolome, and microbiome analyses.
• This approach was applied specifically in the IF-responsive Tp53 GBM mouse model.
• The analysis provided a comprehensive molecular profiling of IF's effects.
• Findings from multi-omics were validated through rescue experiments in the same mouse model.

The TGF-β signaling pathway is suppressed downstream of methionine sulfoxide-mediated m6A modification changes induced by IF.

• Methionine sulfoxide acts as a regulatory metabolite connecting gut microbiota changes to epigenetic RNA modification.
• m6A modification changes caused by methionine sulfoxide inhibit the TGF-β signaling pathway.
• TGF-β pathway suppression was identified as the proximal molecular mechanism resulting in GBM inhibition.
• This mechanistic chain links dietary intervention (IF) to RNA modification-related molecular mechanisms.