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Perlecan Improves Blood Spinal Cord Barrier Repair Through the Integrin β1/ROCK/MLC Pathway After Spinal Cord Injury

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Abstract

Spinal cord injury (SCI) can lead to the destruction of the blood-spinal cord barrier (BSCB), causing various inflammatory cytokines, neutrophils, and macrophages to infiltrate the lesion area, resulting in secondary injury. Basement membranes (BMs) are maintained by all types of cells in the BSCB and contribute to BSCB maintenance. Perlecan is an important constituent of vascular BMs, maintaining vascular integrity and neuroprotection. However, it is not clear whether Perlecan is involved in BSCB repair after SCI. In this study, we found that Perlecan was specifically expressed in the BMs in the spinal cord and underwent degradation/remodeling after SCI. Subsequently, a CRISPR/Cas9-based SAM system was used to overexpress Perlecan in the injured spinal cord, resulting in significantly enhanced locomotor recovery and neural regeneration. Overexpression of Perlecan reduced BSCB permeability along with the neuroinflammatory response. Interestingly, Perlecan inhibited stress fiber formation by interacting with integrin β1 and inhibiting downstream ROCK/MLC signaling, resulting in reduced tight junctions (TJs) disassembly and improved BSCB integrity. Furthermore, the integrin receptor antagonist GRGDSP abolished the effects of Perlecan overexpression on BSCB permeability and TJs integrity. Overall, our findings suggest that Perlecan reduces BSCB permeability and the neuroinflammatory response by interacting with integrin β1 and inhibiting the downstream ROCK/MLC pathway to promote neurological recovery after SCI.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Home for Researchers editorial team (www.home-for-researchers.com) for language editing service.

Funding

This work is supported by the Natural Science Foundation of Guangdong Province (2017A030312009, 2017A030313111), the National Natural Science Foundation (82071386, 81974329, 81672140), Key Research & Development Program of Guangzhou Regenerative Medicine and Health Guangdong Laboratory (2018GZR110104008), and Research Grant of Guangdong Province Key Laboratory of Psychiatric Disorders (N201904).

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  1. Changnan Xie and Yihan Wang contributed equally to this work.

Authors and Affiliations

  1. Department of Spinal Surgery, Orthopedic Medical Center, Zhujiang Hospital, Southern Medical University, Guangzhou, 510280, China

    Changnan Xie, Yihan Wang, Jinfeng Wang, Yizhou Xu, Jiasong Guo & Lixin Zhu

  2. Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China

    Yizhou Xu & Jiasong Guo

  3. Department of Rehabilitation Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, 510280, China

    Haining Liu

  4. Guangdong Provincial Key Laboratory of Construction and Detection in Tissue Engineering, Southern Medical University, Guangzhou, 510515, China

    Jiasong Guo

  5. Bioland Laboratory (Guangzhou Regenerative Medicine and Health Guangdong Laboratory), Guangzhou, 510700, China

    Jiasong Guo

  6. Guangdong-Hong Kong-Macao Greater Bay Area Center for Brain Science and Brain-Inspired Intelligence, Guangdong Province Key Laboratory of Psychiatric Disorders, Key Laboratory of Mental Health of the Ministry of Education, Guangzhou, 510515, China

    Jiasong Guo

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Contributions

All authors contributed to the study conception and design. Lixin Zhu and Jiasong Guo designed the study. Material preparation, data collection and analysis were performed by Changnan Xie, Yihan Wang, Jinfeng Wang, Yizhou Xu and Haining Liu. The first draft of the manuscript was written by Changnan Xie and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Jiasong Guo or Lixin Zhu.

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All animal experiments were performed in accordance with Institutional Animal Care and Use committee guidelines and approved protocols at the Southern Medical University (ethics Number: LAEC-2022–064).

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Supplemental Fig. 1

The expression effect of Perlecan by SAM system in injured spinal cords A and B Real time PCR analysis showed the relative mRNA level of Perlecan in injured spinal cords from SCI, LV-GFP and LV-Perlecan groups at 7 and 14 days after SCI (n = 3 animal per group). C Immunostaining of Perlecan in spinal cords from SCI, LV-GFP and LV-Perlecan groups at 14 days after SCI. White dashed lines indicated the injury sites. LC indicated lesion core. Quantitative data in (A and B) were analyzed using one-way ANOVA with Dunnett’s multiple comparison test. **p<0.01, compared with LV-Perlecan group. Data were mean ± SEM. Scale bars, 20 μm (PNG 3.26 MB)

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Xie, C., Wang, Y., Wang, J. et al. Perlecan Improves Blood Spinal Cord Barrier Repair Through the Integrin β1/ROCK/MLC Pathway After Spinal Cord Injury. Mol Neurobiol 60, 51–67 (2023). https://doi.org/10.1007/s12035-022-03041-9

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