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  • Clinical Research Article
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Integrated metabolomics and proteomics analysis in children with cerebral palsy exposed to botulinum toxin-A

Pediatric Research (2025)Cite this article

Abstract

Background

We previously examined plasma metabolic changes before and after botulinum toxin-A injections of cerebral palsy (CP) children and showed that the glycine, serine and threonine metabolism may play a key role in neuritogenesis. This study analysed untargeted metabolomics combined with proteomics of plasma to discussed which substances are meaningfully changed, to what extent they affect the effects of action.

Methods

Blood samples were collected from 91 children with spastic CP at 4 time points: pre-injection (T1), 1 month post-injection (T2), 3 months post-injection (T3) and 6 months post-injection (T4). Differentially changed metabolites and proteins were selected, and co-expression pathways were constructed to explore the key molecular processes.

Results

A total of 674 proteins and 354 metabolites were identified. The differential metabolites were mainly involved in the linoleic acid metabolism, beta−Alanine metabolism, citrate cycle, pyruvate metabolism and glycolysis or gluconeogenesis. Differential proteins were primarily associated with glucose metabolism, lipid metabolism, immune and inflammation responses. Co-expression pathways showed that ECM-receptor interaction, complement and coagulation cascades, glycolysis or gluconeogenesis, pyruvate metabolism, and linoleic acid metabolism were the main pathways.

Conslusions

Our results indicated the botulinum toxin-A predominantly activated the glucose metabolism, lipid metabolism, and immune and inflammation responses, and energy metabolism changed significantly in this process.

Trial registration details

ChiCTR2000033800, Research on the mechanism of botulinum toxin relieving spasticity in children with cerebral palsy. Approval No. 202023041. Registered 13 June 2020, http://www.chictr.org.cn/showproj.html?proj=52267.

Impact statement

  • This is the first study that combined dynamic metabolomics and proteomics analysis to investigate the molecular changes in children with spastic cerebral palsy after botulinum toxin-A injections, which might provide a theoretical reference for the further subsequent study for targets to increase the efficacy and prolong the duration of botulinum toxin-A, and would be a valuable resource for the metabolomics and proteomics field in this group.

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Fig. 1: Principal component analysis and orthogonal partial least squares-discriminant analysis of different time points.
Fig. 2: Comprehensive metabolomics analysis of plasma samples from different time points.
Fig. 3: Interaction network for differentially expressed proteins.
Fig. 4: Proteomic profile shift of plasma samples from different time points.
Fig. 5: Integrated pathway analysis of differential metabolites and proteins.

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Data availability

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

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Acknowledgements

The authors gratefully acknowledge the extraordinary generosity of the study participants and the support from their caregivers. And we would like to thank Dr. Luo Jie for polishing the language of the article. The study was funded by the Featured Clinical Technique of Guangzhou (2023C-TS59), Guangzhou Municipal Science and Technology Project (2024A03J01274, 2023A03J0919), and Plan on enhancing scientific research in Guangzhou Medical University (GMUCR2024-02020).

Author information

Author notes

  1. These authors contributed equally: Zhaofang Chen, Tingting Peng Sr., Mengru Zhong, Yage Zhang.

Authors and Affiliations

  1. Department of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, Guangzhou, China

    Zhaofang Chen, Tingting Peng Sr., Mengru Zhong, Yage Zhang, Yuan Zhang, Qingfen Hou, Tingting Peng Jr., Xubo Yang, Hongyu Zhou, Liru Liu, Mingshan Han, Hongmei Tang, Lu He, Jinling Li & Kaishou Xu

  2. Department of Sport Rehabilitation, Shanghai University of Sport, Shanghai, Shanghai, China

    Yuan Zhang

  3. Department of Sports and Health, Guangzhou Sport University, Guangzhou, Guangzhou, China

    Qingfen Hou

  4. Genechem Biotechnology Co., Ltd, Shanghai, Shanghai, China

    Huiran Niu

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  1. Zhaofang Chen
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Contributions

K.S.X. contributed to conception and design, and final approval of the version to be published; Z.F.C. and T.T.P.a. wrote this manuscript and performed data analysis; M.R.Z., Y.G.Z. and H.R.N. carried acquisition of data; X.B.Y., Y.Z., T.T.P.b., Q.F.H., H.Y.Z., L.R.L. and M.S.H. carried out sample collection and revised the paper. H.M.T., L.H. and J.L.L. participated in participants recruitment.

Corresponding author

Correspondence to Kaishou Xu.

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The authors declared no competing interests.

Ethics statement

This study was approved by the Health Research Ethics Committee of Guangzhou Women and Children’s Medical Centre (Approval No. 202023041). Informed consent was obtained from all participants in the study.

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Chen, Z., Peng, T., Zhong, M. et al. Integrated metabolomics and proteomics analysis in children with cerebral palsy exposed to botulinum toxin-A. Pediatr Res (2025). https://doi.org/10.1038/s41390-025-04038-5

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