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Microglial over-pruning of synapses during development in autism-associated SCN2A-deficient mice and human cerebral organoids

Molecular Psychiatry (2024)Cite this article

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Abstract

Autism spectrum disorder (ASD) is a major neurodevelopmental disorder affecting 1 in 36 children in the United States. While neurons have been the focus of understanding ASD, an altered neuro-immune response in the brain may be closely associated with ASD, and a neuro-immune interaction could play a role in the disease progression. As the resident immune cells of the brain, microglia regulate brain development and homeostasis via core functions including phagocytosis of synapses. While ASD has been traditionally considered a polygenic disorder, recent large-scale human genetic studies have identified SCN2A deficiency as a leading monogenic cause of ASD and intellectual disability. We generated a Scn2a-deficient mouse model, which displays major behavioral and neuronal phenotypes. However, the role of microglia in this disease model is unknown. Here, we reported that Scn2a-deficient mice have impaired learning and memory, accompanied by reduced synaptic transmission and lower spine density in neurons of the hippocampus. Microglia in Scn2a-deficient mice are partially activated, exerting excessive phagocytic pruning of post-synapses related to the complement C3 cascades during selective developmental stages. The ablation of microglia using PLX3397 partially restores synaptic transmission and spine density. To extend our findings from rodents to human cells, we established a microglia-incorporated human cerebral organoid model carrying an SCN2A protein-truncating mutation identified in children with ASD. We found that human microglia display increased elimination of post-synapse in cerebral organoids carrying the SCN2A mutation. Our study establishes a key role of microglia in multi-species autism-associated models of SCN2A deficiency from mouse to human cells.

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Fig. 1: Adult Scn2a-deficient (HOM) mice exhibit impaired learning and memory as well as impaired synaptic functions and structures.
Fig. 2: Hippocampal microglia in Scn2a-deficient (HOM) mice exhibit altered morphology and increased volume of the lysosome (CD68 marker).
Fig. 3: Excessive phagocytic pruning of microglia occurs during development and continues into adulthood in the hippocampus of Scn2a-deficient (HOM) mice.
Fig. 4: Complement components C3 localized to post-synapses (PSD95) in hippocampal CA1 of Scn2a-deficient (HOM) mice during development.
Fig. 5: Microglia depletion during development restores neural transmission and spine density of hippocampal pyramidal neurons of Scn2a-deficient (HOM) mice.
Fig. 6: Increased post-synapse elimination by hiPSC-derived microglia in cortical organoid carrying autism-associated SCN2A-C959X mutation.

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Data availability

The datasets and additional materials can be obtained by contacting the corresponding author (YY) upon request. Supplementary information is accessible on MP’s website.

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Acknowledgements

We thank Dr. William C. Skarnes from Jackson Laboratory Genomic Medicine for providing the reference and edited iPSC lines, and Dr. Adam Kimbrough from Purdue University kindly provided access to Imaris software. This work is partially supported by the Showalter Research Trust, and the Purdue Big Idea Challenge 2.0 on Autism (to YY). The research reported in this publication was also supported by the NINDS of the NIH (R01NS117585 and R01NS123154 to YY). The authors gratefully acknowledge support from the FamilieSCN2A Foundation for the Hodgkin-Huxley Award as well as Action Potential Award. This project was supported in part by the Indiana Spinal Cord & Brain Injury Research Fund and the Indiana CTSI, funded in part by UL1TR002529 from the NIH. The authors thank all members of the Yang laboratory at Purdue University for insightful discussions.

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  1. These authors contributed equally: Jiaxiang Wu, Jingliang Zhang, Xiaoling Chen.

Authors and Affiliations

  1. Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, 47907, USA

    Jiaxiang Wu, Jingliang Zhang, Xiaoling Chen, Kyle Wettschurack, Zhefu Que, Brody A. Deming, Maria I. Olivero-Acosta, Ningren Cui, Muriel Eaton, Yuanrui Zhao, Sophia M. Li, Matthew Suzuki, Ian Chen, Tiange Xiao, Manasi S. Halurkar, Purba Mandal & Yang Yang

  2. Purdue Institute for Integrative Neuroscience, Purdue University, West Lafayette, IN, 47907, USA

    Jiaxiang Wu, Jingliang Zhang, Xiaoling Chen, Kyle Wettschurack, Zhefu Que, Brody A. Deming, Maria I. Olivero-Acosta, Ningren Cui, Muriel Eaton, Yuanrui Zhao, Sophia M. Li, Matthew Suzuki, Ian Chen, Tiange Xiao, Manasi S. Halurkar, Purba Mandal, Wendy A. Koss & Yang Yang

  3. Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN, 47907, USA

    Chongli Yuan

  4. College of Veterinary Medicine, Purdue University, West Lafayette, IN, 47907, USA

    Ranjie Xu

  5. School of Life Sciences, Shanghai University, Shanghai, 200444, China

    Dongshu Du & Fuxue Chen

  6. Department of Neurology, Mayo Clinic, Rochester, MN, 55905, USA

    Long-jun Wu

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Contributions

JW, JZ, and YY conceived and designed the experiments. JW, JZ, XC, KW, ZQ, BAD, MIO-A, NC, ME, YY, YZ, SML, MS, IC, TX, MSH, PM performed the experiments and analysis the data. CY, RX, WAK, DD, FC, and L-JW participated in data analysis and experimental design. YY supervised the project. JW and YY wrote the paper with input from all authors.

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Correspondence to Yang Yang.

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Wu, J., Zhang, J., Chen, X. et al. Microglial over-pruning of synapses during development in autism-associated SCN2A-deficient mice and human cerebral organoids. Mol Psychiatry (2024). https://doi.org/10.1038/s41380-024-02518-4

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