Optimization of osmotic blood-brain barrier opening to enable intravital microscopy studies on drug delivery in mouse cortex

Chu, Chengyan; Jabłońska, Anna; Lesniak, Wojciech G.; Thomas, Adrian; Lan, Xiaoyan; Linville, Raleigh M.; Shen, Li; Searson, Peter C.; Liu, Guanshu; Pearl, Monica S.; Pomper, Martin G.; Janowski, Mirosław; Magnus, Tim; Walczak, Piotr

doi:10.1016/j.jconrel.2019.11.019

Cited by 39 publications

(32 citation statements)

References 45 publications

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“…Finally the antagonistic roles of MF and MT stabilization in the pathological changes of BBB permeability have been observed in in vivo models. Intracellular protein nanoparticle-induced osmotic potential, a crucial tension factor, could play an important role in the regulation of nonselective BBB permeability, in accordance with previous studies that hyperosmotic stimuli could result effectively in blood-brain barrier opening (OBBBO) or disruption [51,52] .…”

Section: Discussionsupporting

confidence: 87%

Protein Nanoparticle Osmotic Pressure Modifies Nonselective Permeability of the Blood Brain Barrier by Increasing Membrane Fluidity

Chen

Wang

et al. 2020

Preprint

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BackgroundIntracellular tension plays a crucial role in the destruction of the blood brain barrier (BBB) in response to lesion stimuli. Tight junction structure could be primarily affected by tension activity. In this study, we aimed to determine the effects of extracellular BBB damage on intracellular tension activity, and elucidate the mechanism underlying the effects of intracellular protein nanoparticle-dependent osmotic pressure on BBB permeability.MethodsThe intracellular tension for tight junction proteins occludin and ZO1 were evaluated using the fluorescence resonance energy transfer (FRET)-based tension probes and cpstFRET analysis. The efficiency of the probes was examined by acceptor photobleaching FRET (FRET-AB) analysis. The changes in mobility ratios of the transmembrane protein occludin were evaluated via the fluorescence recovery after photobleaching (FRAP) test. The cytoplasmic osmotic pressure (OP) was measured using the Osmomat 3000 Freezing Point Osmometer and 050 Membrane Osmometer. The count rate of cytoplasmic nanoparticles was detected by Nanosight NS300. The activation of cofilin and stathmin was examined by Western blot analysis. The BBB permeability in vivo was determined via investigating the changes of Evans Blue (EB) injected into the SD rats. The tight junction formation was assessed by the measurement of transendothelial electrical resistance (TEER). Intracellular calcium or chloride ions were measured using Fluo-4 AM or MQAE dyes. ResultsBBB lesions were accompanied by changes in occludin/ZO1 tension. Increases in intracellular osmotic pressure were involved in alteration of BBB permeability, possibly through the depolymerization of microfilaments or microtubules and mass production of protein nanoparticles according to the Donnan effect. Recovery of protein nanoparticle osmotic pressure could effectively reverse the effects of changes in occludin/ZO1 tension and BBB lesions. Outward tension of intracellular osmotic potential also caused upregulation of membrane fluidity, which promoted nonselective drug influx.ConclusionsOur results suggest a crucial mechanical mechanism underlying BBB lesions, and protein nanoparticle osmotic pressure could be a novel therapeutic target for BBB lesion-related brain diseases. Our results also provide a basis for further studies on the regulation of intracellular tension activity and its effect on the permeability of the BBB, and development of novel drugs that cross the blood-brain barrier.

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Section: Discussionsupporting

confidence: 87%

Protein Nanoparticle Osmotic Pressure Modifies Nonselective Permeability of the Blood Brain Barrier by Increasing Membrane Fluidity

Chen

Wang

et al. 2020

Preprint

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“…Intracellular protein nanoparticle-related osmotic potential, a crucial tension factor, could play an important role in the regulation of nonselective BBB permeability, in accordance with previous studies that alteration of osmotic potential across membrane could result effectively in blood-brain barrier opening (OBBBO) or disruption. [49][50][51] The transmembrane protein occludin and cytoplasmic attachment protein ZO1 are the main structural proteins of tight junctions (TJs) and have different tension distributions. Microfilament and microtubule pulling tensions could cause occludin/ZO1 tension.…”

Section: Discussionmentioning

confidence: 99%

Protein Nanoparticle-Related Osmotic Pressure Modifies Nonselective Permeability of the Blood–Brain Barrier by Increasing Membrane Fluidity

Chen

Wang

et al. 2021

IJN

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“…To explore the optimal concentration of melittin for blood-brain barrier opening (BBBO), 1 Therefore, even with IA injection, the concentration window for mannitol-induced BBBO is extremely limited and small decreases in concentration resulting from mixing with the blood due to collateral circulation can affect reproducibility and territories of BBBO [47,48]. In contrast, the solubility of melittin is about 350 µM, more than 70-fold larger than the concentrations for causing irreversible damage (5 µM), providing a broad window for establishing reversible and non-cytotoxic conditions.…”

Section: Intraarterial Injection Of Melittin In Micementioning

confidence: 99%

“…To address this challenge and improve the predictability of the territory of BBBO we have developed a technique based on interventional MRI where injection of the BBB-opening agent is preceded by dynamic contrast-enhanced MRI during IA infusion of a contrast agent (Fig. 7D), as previously described [48,49]. Intra-arterial infusion of saline supplemented with 50 mM gadolinium with dynamic T2-weighted MR imaging facilitates detection of hypointense signal from the brain region selectively perfused by the microcatheter.…”

Section: Profile Of Melittin-induced Bbbo In a Mouse Modelmentioning

confidence: 99%

Reversible blood-brain barrier opening utilizing the membrane active peptide melittin in vitro and in vivo

Linville

Komin

Lan

et al. 2021

Preprint

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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. Development of BBBO techniques has been hindered by a lack of physiological models for in vitro study. Here, we utilize an in vitro tissue-engineered microvessel model to demonstrate that melittin, a membrane active peptide present in bee venom, supports BBBO. From endothelial and neuronal viability studies, we 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. In mice, we demonstrate 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 platform for delivery of therapeutics into the brain.

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Optimization of osmotic blood-brain barrier opening to enable intravital microscopy studies on drug delivery in mouse cortex

Cited by 39 publications

References 45 publications

Protein Nanoparticle Osmotic Pressure Modifies Nonselective Permeability of the Blood Brain Barrier by Increasing Membrane Fluidity

Protein Nanoparticle Osmotic Pressure Modifies Nonselective Permeability of the Blood Brain Barrier by Increasing Membrane Fluidity

Protein Nanoparticle-Related Osmotic Pressure Modifies Nonselective Permeability of the Blood–Brain Barrier by Increasing Membrane Fluidity

Reversible blood-brain barrier opening utilizing the membrane active peptide melittin in vitro and in vivo

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