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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.

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Author notes

  1. Yanying Liu and Jia-Wei Min contributed equally to this work.

Authors and Affiliations

  1. Division of Basic Biomedical Sciences and Center for Brain and Behavior Research, Sanford School of Medicine, University of South Dakota, Vermillion, SD, 57069, USA

    Yanying Liu, Shelley Feng, Kalpana Subedi, Emily Mammenga, Eduardo Callegari & Hongmin Wang

  2. Department of Neurology, McGovern Medical School, University of Texas Health Science Center at Houston, MSE R334, 6431 Fannin St, Houston, TX, 77030, USA

    Jia-Wei Min

  3. Eppley Institute for Research in Cancer, University of Nebraska Medical Center, Omaha, NE, 68198, USA

    Fangfang Qiao

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Correspondence to Hongmin Wang.

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Liu, Y., Min, JW., 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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