Proteomic signature of endothelial dysfunction identified in the serum of acute ischemic stroke patients by the iTRAQ-based LC-MS approach

Rakesh Sharma, Harsha Gowda, Sandip Chavan, Jayshree Advani, Dhanashree Kelkar, G. S Sameer Kumar, Mitali Bhattacharjee, Raghothama Chaerkady, T. S Keshava Prasad, Akhilesh Pandey, Dindagur Nagaraja, Rita Christopher

Research output: Contribution to journalArticle

Abstract

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

Original languageEnglish (US)
Pages (from-to)2466-2479
Number of pages14
JournalJournal of Proteome Research
Volume14
Issue number6
DOIs
StatePublished - Jun 5 2015

Fingerprint

Proteomics
Stroke
Biomarkers
Serum
Proteins
Blood
Imaging techniques
Cerebrovascular Disorders
Mass spectrometry
Tandem Mass Spectrometry
Hemostasis
Blood-Brain Barrier
Blood Proteins
Early Diagnosis
Enzyme-Linked Immunosorbent Assay
Inflammation

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Proteomic signature of endothelial dysfunction identified in the serum of acute ischemic stroke patients by the iTRAQ-based LC-MS approach. / Sharma, Rakesh; Gowda, Harsha; Chavan, Sandip; Advani, Jayshree; Kelkar, Dhanashree; Kumar, G. S Sameer; Bhattacharjee, Mitali; Chaerkady, Raghothama; Prasad, T. S Keshava; Pandey, Akhilesh; Nagaraja, Dindagur; Christopher, Rita.

In: Journal of Proteome Research, Vol. 14, No. 6, 05.06.2015, p. 2466-2479.

Research output: Contribution to journalArticle

Sharma, R, Gowda, H, Chavan, S, Advani, J, Kelkar, D, Kumar, GSS, Bhattacharjee, M, Chaerkady, R, Prasad, TSK, Pandey, A, Nagaraja, D & Christopher, R 2015, 'Proteomic signature of endothelial dysfunction identified in the serum of acute ischemic stroke patients by the iTRAQ-based LC-MS approach', Journal of Proteome Research, vol. 14, no. 6, pp. 2466-2479. https://doi.org/10.1021/pr501324n
Sharma, Rakesh ; Gowda, Harsha ; Chavan, Sandip ; Advani, Jayshree ; Kelkar, Dhanashree ; Kumar, G. S Sameer ; Bhattacharjee, Mitali ; Chaerkady, Raghothama ; Prasad, T. S Keshava ; Pandey, Akhilesh ; Nagaraja, Dindagur ; Christopher, Rita. / Proteomic signature of endothelial dysfunction identified in the serum of acute ischemic stroke patients by the iTRAQ-based LC-MS approach. In: Journal of Proteome Research. 2015 ; Vol. 14, No. 6. pp. 2466-2479.
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abstract = "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",
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AU - Advani, Jayshree

AU - Kelkar, Dhanashree

AU - Kumar, G. S Sameer

AU - Bhattacharjee, Mitali

AU - Chaerkady, Raghothama

AU - Prasad, T. S Keshava

AU - Pandey, Akhilesh

AU - Nagaraja, Dindagur

AU - Christopher, Rita

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N2 - 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

AB - 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

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