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Hevin–calcyon interaction promotes synaptic reorganization after brain injury

Abstract

Hevin, also known as SPARC-like protein 1 (SPARCL1 or SC1), is a synaptogenic protein secreted by astrocytes and modulates the formation of glutamatergic synapses in the developing brain by interacting with synaptic adhesion proteins, such as neurexin and neuroligin. Here, we identified the neuron-specific vesicular protein calcyon as a novel interaction partner of hevin and demonstrated that this interaction played a pivotal role in synaptic reorganization after an injury in the mature brain. Astrocytic hevin was upregulated post-injury in a photothrombotic stroke model. Hevin was fragmented by MMP3 induced during the acute stage of brain injury, and this process was associated with severe gliosis. At the late stage, the functional hevin level was restored as MMP3 expression decreased. The C-terminus of hevin interacted with the N-terminus of calcyon. By using RNAi and binding competitor peptides in an ischemic brain injury model, we showed that this interaction was crucial in synaptic and functional recoveries in the sensory-motor cortex, based on histological and electrophysiological analyses. Regulated expression of hevin and calcyon and interaction between them were confirmed in a mouse model of traumatic brain injury and patients with chronic traumatic encephalopathy. Our study provides direct evidence for the causal relationship between the hevin–calcyon interaction and synaptic reorganization after brain injury. This neuron-glia interaction can be exploited to modulate synaptic reorganization under various neurological conditions.

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Fig. 1: Spatiotemporal expression of hevin and synaptic proteins following brain injury in the sensory-motor cortical region.
Fig. 2: Hevin interacts with calcyon.
Fig. 3: Identification of an interaction domain between hevin and calcyon.
Fig. 4: The role of the hevin–calcyon interaction during synaptic recovery after brain injury.
Fig. 5: The synaptogenic function of hevin is modulated by MMP3 under brain injury conditions.
Fig. 6: Interaction between hevin and calcyon in the pathological brain of traumatic brain injury (TBI) model and chronic traumatic encephalopathy patients.

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Funding

This work was supported by a grant from the Basic Science Research Program through the National Research Foundation (NRF), which is funded by the Korean government (Ministry of Science, ICT and Future Planning, MSIP) (2016M3C7A1904148, 2017M3C7A1043842, and NRF-2020M3E5D9079764).

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Contributions

JHK, EMH, and KS designed the research. JHK and JH conducted animal experiments and acquired the data. HGJ, AK, SJH, YSL, and EMH provided the AAV-shRNA and conducted the Co-IP assay. JHJ and JKL carried out electrophysiological experiments. HSS and HR performed immunohistochemistry and real-time PCR on human brain tissues. JHK, HGJ, and AK analyzed the data. JHK, EMH, and KS drafted the manuscript, with final editing by all the authors. EMH and KS supervised the work.

Corresponding authors

Correspondence to Eun Mi Hwang or Kyoungho Suk.

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The authors declare no competing interests.

Ethical approval

The current study was approved by the appropriate local ethics committee. Neuropathological processing of control and CTE human brain samples was performed according to the procedures previously established by the BUADC and CTE Center. Institutional review board approval for ethical permission was obtained through the BUADC and CTE Center [40, 41]. This study was reviewed by the Boston University School of Medicine Institutional Review Board (Protocol H-28974) and was approved as exempt because the study involved only tissue collected from post-mortem individuals that are not classified as human subjects. Nevertheless, next of kin provided informed consent for participation and brain donation. The study was performed in accordance with the institutional regulatory guidelines and principles of human subject protection in the Declaration of Helsinki. All animal care and handling were approved by the institutional guidelines of Institutional Animal Care and Use Committee at the Korea Institute of Science and Technology (IACUC-2017-056) and Kyungpook National University Animal Care Committee (KNU 2019-90).

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Kim, JH., Jung, HG., Kim, A. et al. Hevin–calcyon interaction promotes synaptic reorganization after brain injury. Cell Death Differ 28, 2571–2588 (2021). https://doi.org/10.1038/s41418-021-00772-5

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