During the last decade it has been recognized that blood brain barrier (BBB) dysfunction exists in most neurological diseases, including multiple sclerosis, stroke, Alzheimer's and Parkinson's diseases, brain infections and epilepsy. Inflammation plays significant role in BBB injury, secondary to pro-inflammatory factors produced in the brain or blood and leukocyte engagement of brain endothelium. Brain microvascular endothelial cells (BMVEC) are active participants and regulators of inflammatory processes at a site of inflammation. Inflammatory responses in brain endothelium involve hundreds of genes which expression requires fine-tuned regulation. microRNAs (miRNAs) recently emerged as major regulators of gene expression. Very limited information exists about their participation in inflammatory responses in brain endothelial cells. We propose to investigate the role of miRNAs in brain endothelium using well-established in vitro systems for functional studies of BBB and in vivo imaging of brain microvasculature in the model of neuroinflammation. Based on our studies of BBB dysfunction during neuroinflammation, we propose that barrier protection is best achieved when the intervening agents possess anti-inflammatory properties and can stabilize tight junctions as we have shown via inhibition of glycogen synthase kinase (GSK3? To identify a set of relevant miRNAs, we activated primary human BMVEC with the inflammatory stimulus, TNF?identified 137 down-regulated miRNAs and 180 up-regulated miRNAs. In replicate BMVEC, we inhibited GSK3?n BMVEC during TNF?timulation and detected several miRNAs with expression patterns that were regulated by GSK3?The same miRNAs were changed in brain microvessels isolated from mice exposed to TNF?miRNA overexpression prevented secretion of inflammatory mediators in BMVEC. In our proposed study, we will test the overexpression or inhibition of selected miRNAs on BBB tightness and the effects on monocyte-endothelial cell engagement (adhesion/migration). Using bioinformatics, we will identify other targets for miRNAs. Next, we will perform miRNA transfection in vivo and monitor how this will change leukocyte adhesion/migration in an animal model of neuroinflammation. Proposed experiments will provide identification and functional assessment of miRNAs in brain endothelium.

Public Health Relevance

Endothelial dysfunction is perhaps the earliest event in the initiation of vascular disease, caused by inflammation. In this work we propose to use miRNA machinery as a therapeutic tool to prevent deleterious effects of endothelial dysfunction on neuroinflammation. Being able to avoid blood brain barrier from increased permeability would prevent vascular malfunction, which is a most common features of several central nervous system disorders, such as Alzheimer's and Parkinson's diseases, multiple sclerosis, HIV-1-associated neurocognitive disorder, stroke, atherosclerosis and traumatic brain injury.

National Institute of Health (NIH)
National Institute of Neurological Disorders and Stroke (NINDS)
Exploratory/Developmental Grants (R21)
Project #
Application #
Study Section
Brain Injury and Neurovascular Pathologies Study Section (BINP)
Program Officer
Bosetti, Francesca
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Temple University
Schools of Medicine
United States
Zip Code
Persidsky, Yuri; Hill, Jeremy; Zhang, Ming et al. (2016) Dysfunction of brain pericytes in chronic neuroinflammation. J Cereb Blood Flow Metab 36:794-807
Rom, Slava; Zuluaga-Ramirez, Viviana; Reichenbach, Nancy L et al. (2016) PARP inhibition in leukocytes diminishes inflammation via effects on integrins/cytoskeleton and protects the blood-brain barrier. J Neuroinflammation 13:254
Rom, Slava; Zuluaga-Ramirez, Viviana; Dykstra, Holly et al. (2015) Poly(ADP-ribose) polymerase-1 inhibition in brain endothelium protects the blood-brain barrier under physiologic and neuroinflammatory conditions. J Cereb Blood Flow Metab 35:28-36
Zuluaga-Ramirez, Viviana; Rom, Slava; Persidsky, Yuri (2015) Craniula: A cranial window technique for prolonged imaging of brain surface vasculature with simultaneous adjacent intracerebral injection. Fluids Barriers CNS 12:24
Rom, Slava; Reichenbach, Nancy L; Dykstra, Holly et al. (2015) The dual action of poly(ADP-ribose) polymerase -1 (PARP-1) inhibition in HIV-1 infection: HIV-1 LTR inhibition and diminution in Rho GTPase activity. Front Microbiol 6:878
Rom, Slava; Dykstra, Holly; Zuluaga-Ramirez, Viviana et al. (2015) miR-98 and let-7g* protect the blood-brain barrier under neuroinflammatory conditions. J Cereb Blood Flow Metab 35:1957-65
Persidsky, Yuri; Fan, Shongshan; Dykstra, Holly et al. (2015) Activation of Cannabinoid Type Two Receptors (CB2) Diminish Inflammatory Responses in Macrophages and Brain Endothelium. J Neuroimmune Pharmacol 10:302-8