title: High-Throughput Screening for Identification of Blood-Brain Barrier Integrity Enhancers: A Drug Repurposing Opportunity to Rectify Vascular Amyloid Toxicity.
abstract: The blood-brain barrier (BBB) is a dynamic interface that maintains brain homeostasis and protects it from free entry of chemicals, toxins, and drugs. The barrier function of the BBB is maintained mainly by capillary endothelial cells that physically separate brain from blood. Several neurological diseases, such as Alzheimer's disease (AD), are known to disrupt BBB integrity. In this study, a high-throughput screening (HTS) was developed to identify drugs that rectify/protect BBB integrity from vascular amyloid toxicity associated with AD progression. Assessing Lucifer Yellow permeation across in-vitro BBB model composed from mouse brain endothelial cells (bEnd3) grown on 96-well plate inserts was used to screen 1280 compounds of Sigma LOPAC®1280 library for modulators of bEnd3 monolayer integrity. HTS identified 62 compounds as disruptors, and 50 compounds as enhancers of the endothelial barrier integrity. From these 50 enhancers, 7 FDA approved drugs were identified with EC50 values ranging from 0.76-4.56 μM. Of these 7 drugs, 5 were able to protect bEnd3-based BBB model integrity against amyloid toxicity. Furthermore, to test the translational potential to humans, the 7 drugs were tested for their ability to rectify the disruptive effect of Aβ in the human endothelial cell line hCMEC/D3. Only 3 (etodolac, granisetron, and beclomethasone) out of the 5 effective drugs in the bEnd3-based BBB model demonstrated a promising effect to protect the hCMEC/D3-based BBB model integrity. These drugs are compelling candidates for repurposing as therapeutic agents that could rectify dysfunctional BBB associated with AD.
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Hisham
authors.initials: H
authors.lastname: Qosa
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Loqman A
authors.initials: LA
authors.lastname: Mohamed
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Sweilem B
authors.initials: SB
authors.lastname: Al Rihani
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Yazan S
authors.initials: YS
authors.lastname: Batarseh
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Quoc-Viet
authors.initials: QV
authors.lastname: Duong
authors.affiliation: Pennington Biomedical Research Center, Louisiana State University, Baton Rouge, LA, USA.
authors.forename: Jeffrey N
authors.initials: JN
authors.lastname: Keller
authors.affiliation: Department of Basic Pharmaceutical Sciences, School of Pharmacy, University of Louisiana at Monroe, Monroe, LA, USA.
authors.forename: Amal
authors.initials: A
authors.lastname: Kaddoumi
mesh_terms: D016229:Amyloid beta-Peptides / Q000737:chemistry* / Q000633:toxicity*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000187:drug effects*; D001921:Brain / Q000166:cytology; D002461:Cell Line, Transformed; D062507:Claudin-5 / Q000378:metabolism; D018920:Coculture Techniques; D058492:Drug Repositioning / Q000379:methods*; D042783:Endothelial Cells / Q000187:drug effects*; D066298:In Vitro Techniques; D066290:Libraries, Special; D051379:Mice; D008954:Models, Biological; D010446:Peptide Fragments; D010539:Permeability / Q000187:drug effects
chemical_list: D016229:Amyloid beta-Peptides; D062507:Claudin-5; D010446:Peptide Fragments
keywords: Amyloid-β; blood-brain barrier; high-throughput screening; permeability; repurposing