Nitric oxide inhibition ameliorates cortical proteomic changes in the Cntnap2<sup>-/-</sup> and Shank3<sup>Δ4-22</sup> mouse models of autism spectrum disorder.

Overview

1. Institute for Drug Research, School of Pharmacy, Faculty of Medicine, The Hebrew University of Jerusalem,Jerusalem, Israel
2. Rosamund Stone Zander and Hansjoerg Wyss Translational Neuroscience Center, Boston Children’s Hospital, Harvard Medical School, Harvard University,Boston, MA USA

Abstract

Background: Autism spectrum disorder (ASD) is a neurodevelopmental disorder with a strong genetic component, and over a thousand associated genes have been identified, including CNTNAP2 and SHANK3. Our previous work using Cntnap2-/- and Shank3Δ4–22 ASD mouse models implicated dysregulated nitric oxide (NO) signaling in ASD-related behaviors, which were improved by inhibition of neuronal nitric oxide synthase (nNOS) with 7-Nitroindazole (7-NI). However, the molecular mechanisms linking NO signaling to ASD pathology remain poorly defined.

Methods: We performed mass spectrometry-based global proteomic profiling of cortical tissue from both mouse models under baseline conditions and following 7-NI treatment. Systems biology and bioinformatics analyses were used to identify differentially expressed proteins, enriched pathways, and treatment-responsive networks. Cross-model comparisons were performed to assess molecular convergence and overlap with human ASD-risk genes. Behavioral and biochemical assessments were reanalyzed to evaluate ASD-like phenotypes and treatment effects.

Results: Treatment with 7-NI improved ASD-like behavioral deficits in Cntnap2 and Shank3 mutant mice, including increased sociability and reduced anxiety-like behavior. 7-NI was also associated with attenuation of cortical protein alterations across synaptic, neuronal, and metabolic pathways, shifting subsets of dysregulated proteins toward wild-type expression levels. Despite distinct genetic mutations, the two models converged at the protein and pathway levels, including treatment-responsive proteins encoded by high-confidence human ASD risk genes.

Limitations: Analyses were restricted to cortical tissue; additional brain regions may reveal complementary mechanisms. Mass spectrometry may underrepresent low-abundance proteins; larger sample sizes could improve statistical power. Potential off-target effects of 7-NI should also be considered.

Conclusions: These findings show that nNOS inhibition improves ASD-like behaviors and is associated with partial normalization of altered cortical proteins across two genetically distinct ASD mouse models that display convergent molecular changes, including proteins encoded by high-confidence ASD risk genes.

Supplementary Information: The online version contains supplementary material available at 10.1186/s13229-026-00716-1.

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

Datasets cited

• figshare:32096005, at figshare; found in DataCite
• figshare:32096008, at figshare; found in DataCite
• figshare:32096014, at figshare; found in DataCite
• figshare:32096017, at figshare; found in DataCite

Data availability

The dataset(s) supporting the conclusions of this article are included in this article and its Supplementary Files. Please see the section “Sample preparation” in the Methods for a full list of materials available and where they can be accessed. The mass spectrometry proteomics data have been deposited with the ProteomeXchange Consortium via the PRIDE repository under the dataset identifier PXD068474.

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, 29 September 2026: the first record

Recorded: type, language, journal, volume, issue, pages, dates, 4 authors, 10 keywords, 17 MeSH terms, 4 funders, 125 references.

Cite

This paper

Bazbaz, W., Kartawy, M., Khaliulin, I., & Amal, H. (2026). Nitric oxide inhibition ameliorates cortical proteomic changes in the Cntnap2<sup>-/-</sup> and Shank3<sup>Δ4-22</sup> mouse models of autism spectrum disorder. Molecular autism, 17(1), 21. https://doi.org/10.1186/s13229-026-00716-1

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:10.1038/s41593-026-02287-z [code]

Autism subtypes identified using cross-species functional connectivity analyses.

Journal: Nature neuroscience

In common: autism, mouse, 4 references

[2] doi:10.1016/j.isci.2026.117278

Altered striatal long-term potentiation in the eIF4E-TG mouse model of autism spectrum disorder.

Journal: iScience

In common: autism, mouse, cellular / molecular, 2 references

[3] doi:10.1038/s41467-026-74958-1

Acute rapamycin treatment reveals distinct mechanisms of dysfunction in a maternal inflammation mouse model.

Journal: Nature communications

In common: autism, mouse, cellular / molecular, 2 references

[4] doi:10.1126/sciadv.aeb5842 [code]

Regulation of autism-related self-injurious behavior by electrical stimulation of corticostriatal circuits in mice and humans.

Journal: Science advances

In common: autism, mouse, 2 references

[5] doi:10.1038/s41467-026-74320-5 [code]

Spatial architecture of autism pathogenesis reveals mosaic structural disarray during early development.

Journal: Nature communications

In common: autism, genetics / omics, 2 references

[6] doi:10.1038/s41398-026-03952-4 [code]

Perineuronal nets in cerebellar nuclei neurons orchestrate social behaviour via regulation of neuronal activity in circuits innervated by the cerebellum.

Journal: Translational psychiatry

In common: autism, mouse, cellular / molecular, 2 references

[7] doi:10.1126/sciadv.adz3398 [code]

Copper deficiency impairs oligodendrocyte maturation and social behavior via mitophagy and mTOR suppression in ASD.

Journal: Science advances

In common: autism, mouse, cellular / molecular, 2 references

[8] doi:10.1016/j.xgen.2026.101278 [code]

Single-cell profiling of DNA methylation in autism spectrum disorder prefrontal cortex reveals distinct regulatory and aging signatures.

Journal: Cell genomics

In common: autism, genetics / omics, 2 references

[9] doi:10.1101/gr.280394.124 [code]

De novo structural variants in autism spectrum disorder disrupt distal regulatory interactions of neuronal genes.

Journal: Genome research

In common: autism, cellular / molecular, 2 references

[10] doi:10.21203/rs.3.rs-9839901/v1 [code]

Early life ventricular enlargement precedes emergence of autistic traits

Journal: Research Square (preprint)

In common: autism, 2 references

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.