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Proteomic signature of endothelial dysfunction identified in serum of acute ischemic stroke patients by iTRAQ-based LC-MS approach Rakesh Sharma, Harsha Gowda, Sandip Chavan, Jayshree Advani, Dhanashree S Kelkar, Sameer S Ghantasala Kumar, Mitali Bhattacharjee, Raghothama Chaerkady, T S Keshava Prasad, Akhilesh Pandey, Nagaraja Dindagur, and Rita Christopher J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/pr501324n • Publication Date (Web): 25 Mar 2015 Downloaded from http://pubs.acs.org on April 7, 2015
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Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
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Journal of Proteome Research
Proteomic signature of endothelial dysfunction identified in serum of acute ischemic stroke patients by iTRAQ-based LC-MS approach
Rakesh Sharma1, Harsha Gowda2, Sandip Chavan2,3, Jayshree Advani2,3, Dhanashree Kelkar2, G. S. Sameer Kumar2, Mitali Bhattacharjee2 , Raghothama Chaerkady2,3, T. S. Keshava Prasad2,3, Akhilesh Pandey2,4,5,6,7, Dindagur Nagaraja8* and Rita Christopher1*
1
Department of Neurochemistry, National Institute of Mental Health and Neuro Sciences,
Bangalore 560029, India 2
Institute of Bioinformatics, International Technology Park, Bangalore 560 066, India
3
Manipal University, Manipal 576 104, India
4
McKusick-Nathans Institute of Genetic Medicine and 5Departments of Biological Chemistry,
6
Pathology and 7Oncology, Johns Hopkins University School of Medicine, Baltimore, MD
21205, USA 8
Department of Neurology, Dharwad Institute of Mental Health and Neuro Sciences, Dharwad
580001, India
KEYWORDS Hemostasis, thrombus, inflammation, LTQ-Orbitrap mass spectrometer, diagnostic marker, ischemic stroke. 1
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ABSTRACT Acute ischemic stroke (AIS) is a devastating cerebrovascular disorder which 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 development of blood-based biomarkers for early diagnosis of stroke, which perhaps could be an adjunct to the existing imaging modalities to 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. Quantitative proteomic analysis of serum from AIS and control subjects revealed 389 high confidence protein identifications and their relative levels. Among them, 60 proteins showed ≥1.5 fold change in the AIS. We verified altered serum levels of candidate proteins such as vWF, ADAMTS13, S100A7 and DLG4, through ELISA and their 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 protein identified in this study also known to mediate inflammation, endothelial proliferation and angiogenesis. Current study provided a potential and novel biomarker panel, which may provide diagnostic aid to existing imaging modalities for rapid diagnosis of ischemic stroke
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Journal of Proteome Research
INTRODUCTION Stroke is one of the leading causes of morbidity and mortality in both developed and developing countries. In spite of it being a major public health concern, effective disease management strategies for treatment of acute ischemic stroke (AIS) is limited. Thrombolytic therapy with intravenous tissue plasminogen activator, should be administered within the limited therapeutic time window of 4.5 hours post-stroke and hence only
