Multiomics analysis reveals the exacerbating effect of constipation on autism-related symptoms in children with autism spectrum disorder.

Constipated ASD children exhibited more severe autism-related symptoms than non-constipated ASD children.

• Participants were classified into constipated and non-constipated groups based on assessment of gastrointestinal and autism-related symptoms.
• Autism-related symptom severity was compared between the two groups using standardized assessment tools.
• The study used 16S rRNA sequencing and non-targeted metabolomics to explore underlying mechanisms.

Constipated ASD children showed alterations in four bacterial taxa compared to non-constipated ASD children.

• The four altered bacterial taxa were the phylum Bacteroidetes, the family Barnesiellaceae, and the genera Alistipes and Bilophila.
• Bacterial profiling was performed using 16S rRNA sequencing.
• Three of the four taxa—Bacteroidetes, Alistipes, and Bilophila—were found to exacerbate the relationship between constipation and autism-related symptoms.

Constipated ASD children showed alterations in 451 metabolites compared to non-constipated ASD children.

• Metabolite profiling was conducted using non-targeted metabolomics.
• A total of 451 metabolites were identified as altered between constipated and non-constipated ASD children.
• Five metabolites derived from the three altered bacterial taxa were significantly associated with autism-related symptoms.

Five specific metabolites derived from Bacteroidetes, Alistipes, and Bilophila were significantly associated with autism-related symptoms.

• The five metabolites identified were chenodeoxycholic acid, palmitic acid, glutaric acid, arachidonic acid, and choline.
• These metabolites were linked to the three bacterial taxa that exacerbated the constipation–autism symptom relationship.
• The association between these metabolites and autism-related symptoms was identified through multiomics integration analysis.

Three bacterial taxa mediated or exacerbated the relationship between constipation and autism-related symptoms.

• The three taxa were Bacteroidetes (phylum level), Alistipes (genus level), and Bilophila (genus level).
• Barnesiellaceae (family level) was altered in constipated ASD children but was not among the three taxa identified as exacerbating the constipation–autism symptom relationship.
• The exacerbating role of these taxa was identified through integration of microbiome and metabolomics data.