Washed microbiota transplantation improves clinical symptoms, gut microbiota, and metabolic profiles in autism spectrum disorder in a twin cohort.

Overview

1. Department of Gastroenterology and Microbiota Medicine, The First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, China
2. Guangdong Provincial Key Laboratory for Research and Evaluation of Pharmaceutical Preparations, Guangzhou, China

Abstract

Objective: Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition characterized by impaired social communication, repetitive behaviors, and restricted interests. Dysregulation of the microbiota–gut–brain axis is closely associated with the pathogenesis of ASD. Washed microbiota transplantation (WMT) has emerged as a promising intervention for ASD, but existing cohort studies lack genetically identical controls, making it difficult to distinguish intervention-related changes from genetic and environmental confounding factors. This twin-paired controlled study adopted a study design that minimizes the influence of genetics and shared environment, to explore the associations of WMT with clinical symptoms, gut microbiota, and metabolic profiles in children with ASD.

Methods: Three pairs of age- and environment-matched twins (one ASD-affected, one typically developing sibling) were enrolled. WMT was administered to the ASD participant in each pair. Fecal samples were collected at baseline and post-intervention. Gut microbiota and metabolic profiles were analyzed using metagenomic sequencing and targeted metabolomics, respectively. Clinical outcomes were evaluated using the Childhood Autism Rating Scale (CARS), Autism Behavior Checklist (ABC), Sleep Disturbance Scale for Children (SDSC), and Bristol Stool Form Scale (BSFS). Relevant observations were carried out to explore potential changing trends.

Results: After WMT, CARS, ABC, SDSC, and BSFS exhibited small numerical directional shifts toward healthier values, but none reached statistical significance. Gut microbial structure and function presented a shifting trend toward the profile of their typically developing twin siblings. Abnormal lipid and energy metabolism indicators showed partial ameliorative trends, and the number of differential metabolites between ASD patients and healthy siblings was markedly reduced. Tyrosine and phenylalanine metabolic pathways, together with Segatella, Negativibacillus, and Sangeribacter, may be associated with incomplete phenotypic changes in this cohort.

Limitations: Although the twin-pair design has high internal validity and can provide strong causal inference evidence for the effect of microbiota transplantation in treating ASD, this study has limitations such as a small sample size, a single-center non-randomized observational design. All findings in this pilot study are merely descriptive trends, and the relevant mechanism analysis only provides correlational clues. A single session of microbiota transplantation failed to fully adjust aromatic amino acid metabolism in ASD children. No definitive causal relationship can be concluded based on the findings of this small-sample pilot study.

Conclusion: Under tightly controlled genetic and environmental conditions, gut microbial dysbiosis presents correlational characteristics with ASD-related phenotypes. WMT was associated with consistent remodeling of gut microbial ecology and partial resolution of metabolic dysregulation in ASD children, with multi-omic signatures converging toward healthy twins. Clinical rating scales only displayed non-significant minor numerical shifts, which cannot be interpreted as evidence of clinical symptom improvement. These initial findings provide exploratory mechanistic clues and phenotypic data supporting WMT as a targeted microbiome intervention approach for ASD, and await further validation through large-scale randomized controlled trials.

Clinical trial registration: Identifier ChiCTR2400091105.

Reproduced under the paper's license (CC BY), from the paper cited above.

Code

The paper links to its data, not to its authors' code: see the Data section.

Tracing map

A tracing map links a paper to the code its authors published: this paper has none, so it has no map.

Data

Data links

• ncbi.nlm.nih.gov/bioproject/1458179, NCBI; found in “Data availability statement”

Data availability statement

The raw sequencing data are publicly available in the NCBI SRA database under BioProject accession PRJNA1458179: http://www.ncbi.nlm.nih.gov/bioproject/1458179. The raw targeted metabolomics data can be found in Supplementary Table 1. Individual anonymized clinical data of all enrolled twin participants (including all baseline and post-intervention scale scores) are presented in Supplementary Table 2. All clinical, metagenomic and metabolomics data can be obtained by reasonable request to the corresponding authors.

Reproduced under the paper's license (CC BY), from the paper cited above.

Versions

The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.

Version 1, 27 September 2026: the first record

Recorded: type, language, journal, volume, pages, dates, 11 authors, 6 keywords, 61 references.

Cite

This paper

Feng, S., Si, X., Lu, C., Gao, Z., Wang, J., Yang, Q., Lu, S., Su, T., Yang, J., He, X., & Wu, L. (2026). Washed microbiota transplantation improves clinical symptoms, gut microbiota, and metabolic profiles in autism spectrum disorder in a twin cohort. Frontiers in microbiology, 17, 1885281. https://doi.org/10.3389/fmicb.2026.1885281

Similar papers

The papers with a page that share the most with this one: the tools found in their code, their categories, datasets, cited references and authors, the rarest counting most.

[1] doi:10.1159/000552681

Microbiota-Based Interventions Differentially Rescue Gut and Social Behavior Phenotypes in <italic>Drosophila</italic> with Kdm5 Deficiency.

Journal: Developmental neuroscience

In common: clinical / translational, 5 references

[2] doi:10.1186/s12866-026-05296-x

Gut bacterial characteristics in children with autism spectrum disorder according to symptom severity: a cross-sectional study.

Journal: BMC microbiology

In common: autism, genetics / omics, 3 references

[3] doi:10.3389/fmicb.2026.1811435

Research on the role of gut microbiota metabolites in autism by multi-omics and network pharmacology.

Journal: Frontiers in microbiology

In common: autism, genetics / omics, 3 references

[4] doi:10.1038/s41380-026-03609-0 [code]

Gut microbiome alterations are sex-dependently associated with brain abnormalities in a mouse model of Neurofibromatosis type I.

Journal: Molecular psychiatry

In common: autism, 3 references

[5] doi:10.3389/fnins.2026.1858005 [code]

Targeted stool metabolomics suggests exploratory catecholamine- and tryptophan-linked metabolic features in autism spectrum disorder.

Journal: Frontiers in neuroscience

In common: autism, clinical / translational, genetics / omics, 2 references

[6] doi:10.1186/s12888-026-08178-8 [code]

Machine learning model to identify gut microbiome-derived metabolites as potential biomarkers of autism spectrum disorder: a pilot study.

Journal: BMC psychiatry

In common: autism, clinical / translational, genetics / omics, 1 reference

[7] doi:10.1242/jeb.252086 [code]

Transcriptional predictors of rescue behaviour in ants.

Journal: The Journal of experimental biology

In common: genetics / omics, 2 references

[8] doi:10.3390/nu18152441 [code]

Clinical Improvement and Taxonomic-Functional Gut Microbiome Remodeling After Six Months of Multi-Strain Synbiotic Supplementation in Mexican Children with Autism Spectrum Disorder.

Journal: Nutrients

In common: autism, clinical / translational, 1 reference

[9] doi:10.3390/genes17080874 [code]

Multi-Omic Analysis of Cerebrospinal Fluid Metabolites in Autism Spectrum Disorder: Biomarker Identification, Metabolic Genetics Insights, and Network Toxicology.

Journal: Genes

In common: autism, genetics / omics, 1 reference

[10] doi:10.1371/journal.pone.0351146

Differential analysis of Short chain fatty acids incubation in autistic organoids based on transcriptome sequencing.

Journal: PloS one

In common: autism, genetics / omics, 1 reference

Contribute

The authors of this paper can claim it, correct its record and validate its tracing map, and the maintainers of its code (its owner, or a public member of its organization) correct what it says of their repository; anyone signed in can ask for its removal. Every request goes to OSCR's own machine, which answers it; your account page follows them.

Request its removal

To ask OSCR to remove this record, the copies of its authors' scripts or its tracing map, use the removal request page: signed in, you say who you are, what to remove and why, then review and confirm the request. Published rules decide every request (how).

Discussion: questions and error reports about this paper and its code, from signed-in readers and its authors. It opens with sign-in.

Reproductions: reports from readers who ran the authors' code: what they reproduced, with which environment, commit and data. It opens with sign-in.

Activity: what happens around this paper: new versions of its record, its map's validation, discussions and reproductions. It opens with sign-in.