The emerging concept of the neurovascular unit emphasizes that cell-cell interactions between vascular and parenchymal compartments mediate an integrative response to injury. Thus, endothelial cell dysfunction may be an upstream trigger for brain damage after stroke. For this R21 Project, we propose to investigate the Proteomic Profiles of Human Brain Endothelial Cells in Hypoxia, to broadly characterize the role of soluble and cytosolic mediators in this in vitro stroke model. Building on our encouraging preliminary findings of novel proteinases and substrates found in oxidative stress, we hypothesize that: In comparison to controls, human endothelial cell cultures exposed to hypoxic insult will have consistent and identifiable soluble-protein and substrate expression profiles, indicating relative up- or down-regulation of both known and unknown factors important in cell-cell and cell-matrix signaling. We propose a three-part study in human brain endothelial cell culture media and lysates, to investigate the mechanism of hypoxia-reoxygenation.
Specific Aim 1 : Using validated techniques of quantitative proteomics technology, examine the protein expression profile of human endothelial cell culture media pre and post hypoxia-reoxygenation.
Specific Aim 2 : Examine the down-stream substrates repertoire of human endothelial cell culture media pre and post hypoxia-reoxygenation, in order to obtain relative quantification of the substrate repertoire characteristic of hypoxic injury. Secondary Aims: Extend Aim 1 and 2 to study cell lysates. Through quantitative proteomic analysis of hypoxia-reoxygenation in human brain endothelial cells, an important component of the neurovascular unit, we hope to provide a screening tool for the discovery of novel proteins in ischemic models, and provide venues for the discovery of new therapeutic targets.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Exploratory/Developmental Grants (R21)
Project #
1R21NS052498-01
Application #
6959803
Study Section
Clinical Neuroscience and Disease Study Section (CND)
Program Officer
Jacobs, Tom P
Project Start
2005-08-05
Project End
2007-06-30
Budget Start
2005-08-05
Budget End
2006-06-30
Support Year
1
Fiscal Year
2005
Total Cost
$202,344
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Deng, Wenjun; Cao, Jing; Chen, Lei et al. (2018) Plasma Glycoproteomic Study of Therapeutic Hypothermia Reveals Novel Markers Predicting Neurologic Outcome Post-cardiac Arrest. Transl Stroke Res 9:64-73
Ning, Mingming; Lo, Eng H; Ning, Pei-Chen et al. (2013) The brain's heart - therapeutic opportunities for patent foramen ovale (PFO) and neurovascular disease. Pharmacol Ther 139:111-23
Chou, Sherry H-Y; Feske, Steven K; Atherton, Juli et al. (2012) Early elevation of serum tumor necrosis factor-? is associated with poor outcome in subarachnoid hemorrhage. J Investig Med 60:1054-8
Chou, S H-Y; Feske, S K; Simmons, S L et al. (2011) Elevated peripheral neutrophils and matrix metalloproteinase 9 as biomarkers of functional outcome following subarachnoid hemorrhage. Transl Stroke Res 2:600-7
Ning, Mingming; Sarracino, David A; Kho, Alvin T et al. (2011) Proteomic temporal profile of human brain endothelium after oxidative stress. Stroke 42:37-43
Chou, Sherry H-Y; Lee, Po-Shun; Konigsberg, Rachael G et al. (2011) Plasma-type gelsolin is decreased in human blood and cerebrospinal fluid after subarachnoid hemorrhage. Stroke 42:3624-7
Ning, Mingming; Sarracino, David A; Buonanno, Ferdinando S et al. (2010) Proteomic Protease Substrate Profiling of tPA Treatment in Acute Ischemic Stroke Patients: A Step Toward Individualizing Thrombolytic Therapy at the Bedside. Transl Stroke Res 1:268-275
Ning, Mingming; Lo, Eng H (2010) Opportunities and Challenges in Omics. Transl Stroke Res 1:233-237
Xing, Changhong; Lee, Sunryung; Kim, Woo Jean et al. (2009) Neurovascular effects of CD47 signaling: promotion of cell death, inflammation, and suppression of angiogenesis in brain endothelial cells in vitro. J Neurosci Res 87:2571-7
Ning, Mingming; Wang, Xiaoying; Lo, Eng H (2009) Reperfusion injury after stroke: neurovascular proteases and the blood-brain barrier. Handb Clin Neurol 92:117-36

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