Abstract
Autism spectrum disorder (ASD) is a complex neurodevelopmental condition with limited treatment options, where neuroinflammation and gut microbiota dysbiosis are emerging as interconnected therapeutic targets. This study evaluated the therapeutic potential of non-gene-edited human chemically induced pluripotent stem cell-derived neural stem cells (hCiPSC-NSCs) in a prenatal valproic acid (VPA)-induced rat model of ASD, using a dual-pathway administration strategy (intravenous systemic delivery combined with an intracerebroventricular boost). The treatment significantly ameliorated core ASD-like behaviors, including improved sociability (increased stranger interaction time, P < 0.0001), reduced repetitive behaviors (decreased marble-burying, P < 0.0001; and self-grooming, P < 0.05), and enhanced spatial memory (shorter escape latency in the Morris water maze, P < 0.01). At the mechanistic level, hCiPSC-NSCs attenuated neuroinflammation (suppressed IL-1β, IL-6, and TNF-α; elevated IL-10, all P < 0.0001), reduced oxidative stress (restored GSH and SOD, decreased MDA and NO), diminished microglial activation in the hippocampus and cortex, and restored synaptic ultrastructure by replenishing synaptic vesicles. Furthermore, 16S rRNA sequencing revealed a rebalancing of the gut microbiota, characterized by a reduced Firmicutes/Bacteroidota ratio, enrichment of beneficial taxa like Bacteroidota and Alloprevotella, suppression of pathobionts such as Desulfovibrionales, and partial restoration of microbial diversity. These findings demonstrate that non-gene-edited hCiPSC-NSCs can simultaneously address neural pathophysiology and gut ecosystem disruption in ASD, highlighting their potential as a gut-brain axis-targeting therapy for neurodevelopmental disorders.
Highlights
First non-gene-edited neural stem cells reversing ASD phenotypes;
Dual-route delivery rescues neuroinflammation & synaptic deficits;
Gut microbiota rebalancing (Alloprevotella ↑ , Desulfovibrionales ↓ );
Concurrent restoration of gut-brain axis and ultrastructure;
Multi-system recovery: behavior, immunity, microbiome.
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Data availability
All datasets generated and analyzed during this study are presented in this published article. The raw 16S rRNA sequencing data generated in this study have been deposited in the Genome Sequence Archive (GSA) at the National Genomics Data Center, China National Center for Bioinformation / Beijing Institute of Genomics, Chinese Academy of Sciences, under accession number GSA：CRA038413.
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Acknowledgements
This work was supported by grants from the Enterprise Joint Fund Project of Hunan Provincial Natural Science Foundation, 2024JJ9097; and the GDAS Project of Science and Technology Development (2022GDASZH-2022010110, 2022GDASZH-2022030603-01, 2023GDASZH-2023030602);The horizontal research projects of the Institute of Biological and Medical Engineering, Guangdong Academy of Sciences (Project Leader: Caixia Wu; Grant numbers: 0525270024; 0525270018; 0525270015; 0525270016; 0525270011; 0525270012; 0525270010; 0525270009; 0524270061; 0524270037; 0524270014; 0524270015; 0524270002; 0524270001; 0523270054; 0523270051; 0523270053; 0523270049; 0523270048; 0523270046; 0523270047; 0523270050; 0523270052; 0522270050; 0522270049; 0522270051; 0522270043; 0522270045; 0522270044).
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ZML, CXW and ZYL conceived and designed the study; ZML, CXW performed the experiments; ZML and CXW analyzed the data; ZML and CXW visualized the figures; ZML and CXW wrote the manuscript draft; and ZML, CXW, XJL, HW, MM, RB and ZYL revised the manuscript and supervised the study. All the authors contributed to the article and approved the submitted version.
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(1)Animal Ethics declaration: All animal experiments were conducted in compliance with the ARRIVE guidelines and approved by the Institutional Animal Care and Use Committee (IACUC) of [Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences] for the project ‘ Establishment of autism models in SD rats and C57 mice and stem cell therapy ‘ (IACUC: No. 2023081). Final approval was granted on [November 20, 2023]. (2) Statement of Life Sciences and Medical Ethics: All procedures followed the guidelines of the National Health and Medical Research Council of China and received approval (GIBH-LMEC2024-091-01(AL)) from the Committee on Life Sciences and Medical Ethics of Guangzhou Institutes of Biomedicine and Health, CAS for the project [Research on the Mechanism of stem Cell Therapy in the mouse Autism Model Induced by VPA] on [August 15, 2024].
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Liu, Z., Wu, C., Li, X. et al. Non-gene-edited neural stem cells reverse neuroinflammation and microbiota dysbiosis in a sprague-dawley rat model of autism spectrum disorder. Transl Psychiatry (2026). https://doi.org/10.1038/s41398-026-03841-w
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DOI: https://doi.org/10.1038/s41398-026-03841-w
