What in-depth analyses can be performed on the differential metabolites screened by non-targeted metabolomics? After pathway enrichment, can we trace back to find key enzymes or genes from the metabolic pathways?
The differential metabolites identified through untargeted metabolomics can be further analyzed through the following steps:
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Validation of differential metabolites: Verify the accuracy and abundance changes of the screening results through targeted metabolomics.
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Functional annotation and classification: Use databases such as KEGG and HMDB to annotate the metabolites and determine their metabolic pathways.
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Metabolic pathway enrichment analysis: Use tools like KEGG Pathway and MetaboAnalyst to perform enrichment analysis of differential metabolites and identify significantly altered metabolic pathways.
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Reverse deduce key enzymes or genes: Use enrichment analysis to deduce key enzymes or genes that might regulate these metabolites, and further confirm with bioinformatics tools.
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Multi-omics integration analysis: Combine transcriptomics and proteomics data to further verify the molecular mechanisms behind metabolic changes and confirm the roles of genes and enzymes.
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Functional validation experiments: Validate the specific roles of these genes or enzymes in metabolic regulation through functional experiments such as gene knockout, gene overexpression, or enzyme activity assays.
In untargeted metabolomics, the differential metabolites identified can be deeply studied through methods such as validation, enrichment analysis, reverse deduction of key enzymes and genes, and multi-omics integration. After metabolic pathway enrichment, key enzymes can be deduced using bioinformatics tools, and candidate genes and enzymes can be further confirmed using transcriptomics and proteomics data; finally, functional experiments can verify the specific roles of these genes or enzymes in metabolic regulation.
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