Microbiome-derived short-chain fatty acids and tryptophan metabolites in children with autism spectrum disorder : a stool–urine multi-omics analysis

Osredkar, Joško (author), Fabjan, Teja (author), Godnov, Uroš (author), Jekovec-Vrhovšek, Maja (author), Osredkar, Damjan (author), Finderle, Petra (author), Kumer, Kristina (author), Zorec, Maša (author), Fanedl, Lijana (author), Avguštin, Gorazd (author)

Autism spectrum disorder (ASD) has been associated with alterations in the gut microbiota and its metabolites, particularly short-chain fatty acids (SCFAs) and microbiota-derived tryptophan catabolites, which may influence neurodevelopment through immune and epigenetic mechanisms. We investigated whether stool SCFAs and tryptophan-pathway metabolites differ between children with ASD and typically developing controls, and whether these metabolites associate with ASD severity and systemic biochemical signatures. In this cross-sectional study, we analyzed stool samples from 229 children (160 with ASD, 69 controls) with complete SCFA and tryptophan-metabolite data, while urine metabolomics data were available for a subset and were used for exploratory stool–urine integration analyses. Children with ASD and controls were similar in age, but the ASD group had a higher proportion of males. Absolute concentrations of individual SCFAs, total SCFAs, and derived indices were broadly comparable between groups; nominal differences in propionate/acetate ratio and caproate did not remain significant after false discovery rate correction. Similarly, stool tryptophan-pathway metabolites reported as ng/a.u. based on the NanoDrop-derived proxy (tryptophan, kynurenine, indole-3-acetic, indole-3-lactic, indole-3-propionic, indole-3-aldehyde, N-acetyl-tryptophan, serotonin, melatonin, tryptamine) and functional ratios (kynurenine/tryptophan, indole-derived/tryptophan, serotonin/tryptophan) showed no robust ASD–control differences; N-acetyl-tryptophan was nominally higher in ASD but did not survive multiple-testing correction. In the ASD subgroup with available Childhood Autism Rating Scale (CARS) data (n = 34), SCFA and tryptophan indices showed only weak, non-significant correlations with global ASD severity. In contrast, correlation analyses revealed two coherent metabolic modules, i.e., an SCFA block with very strong internal correlations among individual SCFAs and total SCFAs and a tryptophan block with strong correlations between metabolites and their normalized ratios, while cross-module correlations were modest. These results indicate that stool SCFA and microbiota-derived tryptophan profiles do not robustly distinguish ASD from controls in this cohort, but they form stable metabolic modules compatible with microbiome–epigenome frameworks.

• Language: English
• Keywords: autism spectrum disorder, gut microbiota, short-chain fatty acids, tryptophan metabolism, indole metabolites, kynurenine pathway, epigenetics, microbiome–epigenome interaction, metabolomics, pediatrics
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy; FMF - Faculty of Mathematics and Physics; MF - Faculty of Medicine; BF - Biotechnical Faculty
• Publication status: Published
• Publication version: Version of Record
• Year: 2026
• Number of pages: 20 str.
• Numbering: Vol. 27, iss. 9, art. 3988
• PID: 20.500.12556/RUL-185277
• UDC: 616.896+577.21
• ISSN on article: 1422-0067
• DOI: 10.3390/ijms27093988
• COBISS.SI-ID: 276866051
• Publication date in RUL: 30.07.2026

Record is a part of a journal

• Title: International journal of molecular sciences
• Shortened title: Int. j. mol. sci.
• Publisher: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: črevesna mikrobiota, kratkoverižne maščobne kisline, presnova triptofana, indolni metaboliti, kinureninska pot, epigenetika, interakcija med mikrobiomom in epigenomom, metabolomika, pediatrija