@article{2894219f647940a5960a471a979eca6a,
title = "Reversible blood-brain barrier opening utilizing the membrane active peptide melittin in vitro and in vivo",
abstract = "The blood-brain barrier (BBB) tightly controls entry of molecules and cells into the brain, restricting the delivery of therapeutics. Blood-brain barrier opening (BBBO) utilizes reversible disruption of cell-cell junctions between brain microvascular endothelial cells to enable transient entry into the brain. Here, we demonstrate that melittin, a membrane active peptide present in bee venom, supports transient BBBO. From endothelial and neuronal viability studies, we first identify the accessible concentration range for BBBO. We then use a tissue-engineered model of the human BBB to optimize dosing and elucidate the mechanism of opening. Melittin and other membrane active variants transiently increase paracellular permeability via disruption of cell-cell junctions that result in transient focal leaks. To validate the results from the tissue-engineered model, we then demonstrate that transient BBBO can be reproduced in a mouse model. We identify a minimum clinically effective intra-arterial dose of 3 μM min melittin, which is reversible within one day and neurologically safe. Melittin-induced BBBO represents a novel technology for delivery of therapeutics into the brain.",
keywords = "Blood-brain barrier, Drug delivery, Melittin, Peptide, Tissue engineering",
author = "Linville, {Raleigh M.} and Alexander Komin and Xiaoyan Lan and DeStefano, {Jackson G.} and Chengyan Chu and Guanshu Liu and Piotr Walczak and Kalina Hristova and Searson, {Peter C.}",
note = "Funding Information: This work was supported by NIH / NINDS ( R01NS106008 , R01NS09111 , R01NS102675 ), DTRA ( HDTRA1-15-1-0046 ), and NSF ( DMR 1709892 ). RML acknowledges a National Science Foundation Graduate Research Fellowship under Grant No. DGE1746891 , AK acknowledges support from a Kirschstein-NRSA Individual Predoctoral Fellowship (F31) under award number NINDS 1F31NS101875 , JGD acknowledges support from the Nanotechnology for Cancer Research training program. We also acknowledge Dr. William Wimley (Tulane University) for useful discussions and materials, and Matt Sklar, Gabrielle Grifno, Alanna Farrell, and Erin Gallagher for assistance with cell culture and microfabrication. Funding Information: Corning transwell supports (Cat. no. 3470) were coated with 25 μg mL−1 fibronectin and 50 μg mL−1 collagen IV overnight at 37 °C. iBMECs were seeded on transwells in BBB induction medium at 106 cells cm−2. On day 1 after seeding, the medium was changed to the BBB maintenance medium after one wash with the maintenance medium. On day 2, TEER measurements were performed using an EVOM-2 and STX-100 electrodes (World Precision Instruments, Sarasota, FL). Monolayers with TEER >1500 Ω cm2 were used for the permeability experiments. All TEER measurements are reported after subtracting the value for a blank transwell correcting for the membrane area. For transwell permeability measurements, cell monolayers were washed with BBB maintenance medium, followed by the addition of 500 kDa dextran-fluorescein (1.2 μM) to the apical side of the transwell with or without 5 μM melittin. The transwells were incubated, while rocking, for 90 min at 37 °C, 5% CO2. After the permeability experiments, the TEER across monolayers was measured. Dextran fluorescence in the basolateral and apical chambers was measured using the Synergy H4 plate reader (Biotek Instruments Inc., Winooski, VT) and was converted to a dextran concentration using dextran-fluorescein standard curves. Dextran concentrations in the basolateral chambers were less than 10% of the input concentration (average = 0.1%), confirming that transport was in the linear regime [44]. The apparent permeability (Papp) was calculated from:This work was supported by NIH/NINDS (R01NS106008, R01NS09111, R01NS102675), DTRA (HDTRA1-15-1-0046), and NSF (DMR 1709892). RML acknowledges a National Science Foundation Graduate Research Fellowship under Grant No. DGE1746891, AK acknowledges support from a Kirschstein-NRSA Individual Predoctoral Fellowship (F31) under award number NINDS 1F31NS101875, JGD acknowledges support from the Nanotechnology for Cancer Research training program. We also acknowledge Dr. William Wimley (Tulane University) for useful discussions and materials, and Matt Sklar, Gabrielle Grifno, Alanna Farrell, and Erin Gallagher for assistance with cell culture and microfabrication. Publisher Copyright: {\textcopyright} 2021",
year = "2021",
month = aug,
doi = "10.1016/j.biomaterials.2021.120942",
language = "English (US)",
volume = "275",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier BV",
}