In Vitro-In Vivo Correlation of Blood–Brain Barrier Permeability of Drugs: A Feasibility Study Towards Development of Prediction Methods for Brain Drug Concentration in Humans

Ito, Ryo; Morio, Hanae; Baba, Tomoyo; Sakaguchi, Yasuyuki; Wakayama, Naomi; Isogai, Ryuto; Yamaura, Yoshiyuki; Komori, Takafumi; Furihata, Tomomi

doi:10.1007/s11095-022-03189-y

Cited by 3 publications

(5 citation statements)

References 28 publications

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“…Even if single bands were detected by Western blotting, the expression level was low, and clear membrane localization was not observed by immunocytochemistry. In a past comparative study, the expression level of Claudin-5 was found to be lower in HBMEC/ci18 than in primary cultured HBMECs [ 42 ]. Claudin-5 is the dominant tight junction protein in BMECs [ 43 , 44 ]; however, recent studies indicated an inverse correlation between Claudin-5 and TJ functionality [ 45 , 46 ].…”

Section: Discussionmentioning

confidence: 99%

Usefulness of a humanized tricellular static transwell blood–brain barrier model as a microphysiological system for drug development applications. - A case study based on the benchmark evaluations of blood-brain barrier microphysiological system

Nakayama-Kitamura

Shigemoto-Mogami

Toyoda³

et al. 2023

Regenerative Therapy

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Section: Discussionmentioning

confidence: 99%

Usefulness of a humanized tricellular static transwell blood–brain barrier model as a microphysiological system for drug development applications. - A case study based on the benchmark evaluations of blood-brain barrier microphysiological system

Nakayama-Kitamura

Shigemoto-Mogami

Toyoda³

et al. 2023

Regenerative Therapy

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“…The limitation of our study lies in the limited number of drug permeability evaluations required for establishing comparisons with in vivo data, making it challenging to predict human permeability values. Additionally, because the in vivo data used were K p,uu,CSF from animals, not represented by K p,uu,Brain , or Q ECF in the human brain [ 19 ], using this study as a tool for predicting real values requires the incorporation of modeling or simulations. Furthermore, the viscosity in the medium we used was relatively low compared to that of human blood (0.035–0.055 dyn·s/cm 2 ), making it a significant factor influencing shear stress [ 45 ].…”

Section: Discussionmentioning

confidence: 99%

“…Subsequently, quantitative real-time polymerase chain reaction (real-time PCR) was conducted on a Step One Plus Real-time PCR System Cycler (Applied Biosystems Co., Waltham, MA, USA) using a Power SYBR Green PCR mix (Applied Biosystems Co., Waltham, MA, USA). GAPDH served as the reference gene for the PCR primer sequences [ 19 , 20 , 21 ], which are detailed in Table S1 .…”

Section: Methodsmentioning

confidence: 99%

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An Improved In Vitro Blood-Brain Barrier Model for the Evaluation of Drug Permeability Using Transwell with Shear Stress

Kim,

Shin,

Lee

et al. 2023

Pharmaceutics

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The development of drugs targeting the central nervous system (CNS) is challenging because of the presence of the Blood-Brain barrier (BBB). Developing physiologically relevant in vitro BBB models for evaluating drug permeability and predicting the activity of drug candidates is crucial. The transwell model is one of the most widely used in vitro BBB models. However, this model has limitations in mimicking in vivo conditions, particularly in the absence of shear stress. This study aimed to overcome the limitations of the transwell model using immortalized human endothelial cells (hCMEC/D3) by developing a novel dish design for an orbital shaker, providing shear stress. During optimization, we assessed cell layer integrity using trans-endothelial electrical resistance measurements and the % diffusion of lucifer yellow. The efflux transporter activity and mRNA expression of junctional proteins (claudin-5, occludin, and VE-cadherin) in the newly optimized model were verified. Additionally, the permeability of 14 compounds was evaluated and compared with published in vivo data. The cell-layer integrity was substantially increased using the newly designed annular shaking-dish model. The results demonstrate that our model provided robust conditions for evaluating the permeability of CNS drug candidates, potentially improving the reliability of in vitro BBB models in drug development.

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“…The GRCh38 (hg38) was used to align the readings of the RNA sequence and quantification of gene expression levels was performed using HTSEQ v0.6.1. A different dataset from the same RNA-seq study was previously published: HBMECs (Ito et al 2022) and HBPCs (Umehara et al 2018).…”

Section: Rna Sequencingmentioning

confidence: 99%

Systemic inflammation modulates the levels of EBI2 and 7α,25-OHC synthesizing (CH25H, CYP7B1) and degrading (HSD3B7) enzymes in brain microvascular cells

Caratis

Karaszewski

Klejbor

et al. 2023

Preprint

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The EBI2 receptor is a major modulator of innate immunity and, together with its ligand, oxysterol 7α,25OHC, has been implicated in several neuroinflammatory and autoimmune diseases. 7alpha,25OHC is synthesised from cholesterol with CH25H and CYP7B1 enzymes and degraded with HSD3B7. The concentration of 7alpha,25OHC in the brain increases in the early phases of the murine model of multiple sclerosis, leading to an enhanced central nervous system (CNS) infiltration with EBI2-expressing lymphocytes. Here, we aimed to investigate whether the enzymes involved in the synthesis and degradation of 7alpha,25OHC are expressed directly in the mouse brain microvascular cells and whether systemic inflammation modulates their levels in these cells. Normal mouse brain capillaries were isolated and immunostained for EBI2, CH25H, CYP7B1 and HSD3B7. Subsequently, mice were challenged with lipopolysaccharide and the mRNA expression in whole brain homogenates was measured. Changes in the receptor and enzyme levels were quantified directly in endothelial cells (ECs), pericytes and astrocytes. The data indicated high levels of EBI2 in the brain microvascular ECs, pericytes and astrocytes with the highest co-localisation in pericytes. CH25H was detected in ECs and astrocytes with low levels in pericytes. CYP7B1 was moderately expressed in ECs, astrocytes and pericytes. The 7alpha,25OHC degrading enzyme HSD3B7 was the least detected in astrocytes. Moreover, the data indicated that systemic inflammation downregulated the mRNA levels of Ebi2 and upregulated Ch25h expression in the whole brain. Specifically in each cell type, EBI2 was not induced in ECs but increased in astrocytes and pericytes. CH25H levels increased in astrocytes and pericytes and CYP7B1 in ECs and astrocytes. The degrading enzyme, HSD3B7, was least affected by systemic inflammation. Taken together, we here demonstrate that EBI2 and the enzymes regulating its ligand levels are differentially expressed in mouse brain microvessels and are highly modulated by systemic inflammation. Upregulated concentration of 7alpha,25OHC in the brain during inflammation may lead to an increased migration of EBI2-expressing immune cells into the CNS during infection or neuroinflammatory disease. Modulation of the EBI2/oxysterol system in the brain or directly in the brain blood vessels may thus provide a new approach to treating neuroinflammatory diseases including multiple sclerosis.

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In Vitro-In Vivo Correlation of Blood–Brain Barrier Permeability of Drugs: A Feasibility Study Towards Development of Prediction Methods for Brain Drug Concentration in Humans

Cited by 3 publications

References 28 publications

Usefulness of a humanized tricellular static transwell blood–brain barrier model as a microphysiological system for drug development applications. - A case study based on the benchmark evaluations of blood-brain barrier microphysiological system

Usefulness of a humanized tricellular static transwell blood–brain barrier model as a microphysiological system for drug development applications. - A case study based on the benchmark evaluations of blood-brain barrier microphysiological system

An Improved In Vitro Blood-Brain Barrier Model for the Evaluation of Drug Permeability Using Transwell with Shear Stress

Systemic inflammation modulates the levels of EBI2 and 7α,25-OHC synthesizing (CH25H, CYP7B1) and degrading (HSD3B7) enzymes in brain microvascular cells

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