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Proteomics Reveals Plasma Biomarkers for Ischemic Stroke Related to the Coagulation Cascade

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Abstract

Stroke has a high incidence rate and often leads to permanent disability, particularly if it is not treated promptly. However, no blood biomarkers for early diagnosis are available to date. Therefore, we sought to detect stroke-specific blood biomarkers by identifying proteins associated with the underlying coagulation mechanism, which accounts for more than 80% of all stroke cases. Protein profiling was performed using blood samples from 16 healthy controls and 18 patients who suffered a stroke as the discovery set. We identified upregulated proteins (> 1.5-fold change and p value < 0.05) in patients who suffered a stroke relative to the corresponding levels in healthy controls by nano-liquid chromatography-tandem mass spectrometry using data-independent acquisition based on sequential window acquisition of all theoretical mass spectra, which was developed to improve the consistency and accuracy of candidate proteins. Pathway analysis confirmed that the upregulated proteins were mainly involved in blood coagulation. Among these, we selected prothrombin, plasminogen, fibrinogen alpha-chain, and histidine-rich glycoprotein as candidate biomarkers. Multiple reaction monitoring analysis was performed on a validation set of 61 serum samples (31 healthy controls and 30 stroke patients) to assess the diagnostic value of the candidate biomarkers. All four proteins showed higher expression levels in patients with stroke than in healthy controls. The areas under the receiver operating characteristic curve were greater than 0.9, confirming their clinical value. These four blood coagulation proteins may help in diagnosing stroke more accurately and quickly.

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Funding

This research was supported by the Bio & Medical Technology Development Program of the NRF funded by the Korean government, MSIP (Grant No. 2016M3A9B694241).

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Author notes

  1. Jiyeong Lee and Sora Mun contributed equally to this work.

Authors and Affiliations

  1. Department of Biomedical Laboratory Science, College of Health Sciences, Eulji University, Daejeon, Republic of Korea

    Jiyeong Lee

  2. Department of Biomedical Laboratory Science, College of Health Sciences, Eulji University, Seongnam, Republic of Korea

    Sora Mun, Arum Park, Doojin Kim, Yoo-Jin Lee, Hyo-Jin Kim, Hyebin Choi, Miji Shin, Yeon Tae Chun & Hee-Gyoo Kang

  3. Department of Senior Healthcare, BK21 Plus Program, Graduate School, Eulji University, Seongnam, Republic of Korea

    Sora Mun, Yoo-Jin Lee, Hyo-Jin Kim, Hyebin Choi, Miji Shin & Hee-Gyoo Kang

  4. Department of Neurology, Eulji University Hospital, School of Medicine, Eulji University, Daejeon, Republic of Korea

    Soo Joo Lee & Jae Guk Kim

  5. Seongnam Senior Industry Innovation Center, Eulji University, Seongnam, Republic of Korea

    Hee-Gyoo Kang

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  1. Jiyeong Lee
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Contributions

J.L. and S.M. wrote the manuscript, created the figures and the table, and generated proteomic data. A.P., D.K., Y.J.L., H.J.K., H.C., M.S., S.J., J.G., and Y.T.J. performed sample preparation and generated proteomic data. H.G.K. conceived the study and was in charge of overall direction and planning. All the authors discussed the results and have approved the manuscript for publishing.

Corresponding author

Correspondence to Hee-Gyoo Kang.

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The authors declare that they have no competing interests.

Ethics Approval

All samples were obtained from Eulji University, Daejeon, Korea. This study was approved by the local institutional review board (EMC 2016-03-019).

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Lee, J., Mun, S., Park, A. et al. Proteomics Reveals Plasma Biomarkers for Ischemic Stroke Related to the Coagulation Cascade. J Mol Neurosci 70, 1321–1331 (2020). https://doi.org/10.1007/s12031-020-01545-4

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  • DOI: https://doi.org/10.1007/s12031-020-01545-4

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