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Therapeutic Role of a Cysteine Precursor, OTC, in Ischemic Stroke Is Mediated by Improved Proteostasis in Mice

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Abstract

Oxidative stress aggravates brain injury following ischemia/reperfusion (I/R). We previously showed that ubiquilin-1 (Ubqln1), a ubiquitin-like protein, improves proteostasis and protects brains against oxidative stress and I/R-induced brain injury. Here, we demonstrate that a small molecule compound, L-2-oxothiazolidine-4-carboxylic acid (OTC) that functions as a precursor of cysteine, upregulated Ubqln1 and protected cells against oxygen-glucose deprivation–induced cell death in neuronal cultures. Further, the administration of OTC either at 1 h prior to ischemia or 3 h after the reperfusion significantly reduced brain infarct injury and improved behavioral outcomes in a stroke model. Administration of OTC also increased glutathione (GSH) level and decreased superoxide production, oxidized protein, and neuroinflammation levels in the penumbral cortex after I/R in the stroke mice. Furthermore, I/R reduced both Ubqln1 and the glutathione S-transferase protein levels, whereas OTC treatment restored both protein levels, which was associated with reduced ubiquitin-conjugated protein level. Interestingly, in the Ubqln1 knockout (KO) mice, OTC treatment showed reduced neuroprotection and increased ubiquitin-conjugated protein level when compared to the similarly treated non-KO mice following I/R, suggesting that OTC-medicated neuroprotection is, at least partially, Ubqln1-dependent. Thus, OTC is a potential therapeutic agent for stroke and possibly for other neurological disorders and its neuroprotection involves enhanced proteostasis.

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Acknowledgments

We would like to thank the Physiology Core Facility at the University of South Dakota (USD) Division of Basic Biomedical Sciences for access to equipment and assistance with data analysis, Mr. Doug Jennewein from the USD-IT Research Computing for help in the database installation and servers operation, and Mrs. Daniela Paez from the USD proteomics Core Facility for assistance in processing and data organization.

New Author Contribution Statement

FQ and EM contributed to some in vitro studies. EC contributed to mass spectrometric analysis of OTC-interacting proteins.

Funding

This study was funded by the National Institute of Neurological Disorders and Stroke under research grant NS088084 and by the National Institute of General Medical Sciences under the grant P20GM103443.

Author information

Correspondence to Hongmin Wang.

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The authors declare that they have no conflict of interest.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed in the study. This article does not contain any studies with human participants performed by any of the authors.

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Liu, Y., Min, J., Feng, S. et al. Therapeutic Role of a Cysteine Precursor, OTC, in Ischemic Stroke Is Mediated by Improved Proteostasis in Mice. Transl. Stroke Res. 11, 147–160 (2020). https://doi.org/10.1007/s12975-019-00707-w

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Keywords

  • Stroke
  • Oxidative stress
  • Proteostasis
  • Cysteine precursor
  • Ubiquilin-1