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J Proteome Res. 2015 Jun 5;14(6):2466-79. doi: 10.1021/pr501324n. Epub 2015 Apr 24.

Proteomic Signature of Endothelial Dysfunction Identified in the Serum of Acute Ischemic Stroke Patients by the iTRAQ-Based LC-MS Approach.

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†Department of Neurochemistry, National Institute of Mental Health and Neuro Sciences, Bangalore 560029, India.
▲Department of Biochemistry, Faculty of Medicine, The University of Hong Kong, Hong Kong.
●Department of Biology and Chemistry, City University of Hong Kong, Hong Kong.
‡Institute of Bioinformatics, International Technology Park, Bangalore 560 066, India.
§Manipal University, Manipal 576 104, India.
○Department of Neurology, Dharwad Institute of Mental Health and Neuro Sciences, Dharwad 580001, India.


Acute ischemic stroke (AIS) is a devastating cerebrovascular disorder that leads to permanent physical and neurological disabilities in adults worldwide. Proteins associated with stroke pathogenesis may appear in the serum of AIS patients due to blood-brain barrier dysfunction, thus permitting the development of blood-based biomarkers for early diagnosis of stroke. These biomarkers could perhaps be an adjunct to the existing imaging modalities and aid in better management and therapeutic intervention during the course of the disease. For this exploratory study, a combination of multiplexed isobaric tagging using iTRAQ reagents and high resolution tandem mass spectrometry was used to identify differentially expressed proteins in serum samples from AIS patients. The quantitative proteomic analysis of serum from both AIS and control subjects revealed 389 high confidence protein identifications and their relative levels. Among them, 60 proteins showed a ≥1.5-fold change in the AIS subjects. We verified the altered serum levels of candidate proteins such as vWF, ADAMTS13, S100A7, and DLG4 through ELISA, and the results also corroborate with the experimental findings. vWF and ADAMTS13 are key players that regulate blood hemostasis, and their altered concentration may contribute to endothelial dysfunction. S100A7 is a novel candidate protein identified in this study that is also known to mediate inflammation, endothelial proliferation, and angiogenesis. The current study provided a potential and novel biomarker panel that may in turn provide diagnostic aid to the existing imaging modalities for the rapid diagnosis of ischemic stroke.


LTQ-Orbitrap mass spectrometer; diagnostic marker; hemostasis; high density lipoprotein (HDL); inflammation; ischemic stroke; low density lipoprotein (LDL); thrombus

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