Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments

<b>Background/Objectives</b>: The low permeability of the blood-brain barrier (BBB) represents a significant challenge to effective systemic chemotherapy for primary and metastatic brain cancers. Kinin receptors play a crucial role in modulating BBB permeability, and their agonist analog...

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Main Authors: Carolina Batista, João Victor Roza Cruz, Michele Siqueira, João Bosco Pesquero, Joice Stipursky, Fabio de Almeida Mendes
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/18/4/591
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author Carolina Batista
João Victor Roza Cruz
Michele Siqueira
João Bosco Pesquero
Joice Stipursky
Fabio de Almeida Mendes
author_facet Carolina Batista
João Victor Roza Cruz
Michele Siqueira
João Bosco Pesquero
Joice Stipursky
Fabio de Almeida Mendes
author_sort Carolina Batista
collection DOAJ
description <b>Background/Objectives</b>: The low permeability of the blood-brain barrier (BBB) represents a significant challenge to effective systemic chemotherapy for primary and metastatic brain cancers. Kinin receptors play a crucial role in modulating BBB permeability, and their agonist analogs have been explored in preclinical animal models to enhance drug delivery to the brain. In this study, we investigated whether des-Arg<sup>9</sup>-bradykinin (DBK), a physiological agonist of kinin B<sub>1</sub> receptor (B1R), acts as a brain drug delivery adjuvant by promoting the transient opening of the BBB. <b>Methods</b>: Human brain microvascular endothelial cells (HBMECs) were treated with DBK in the culture medium and in conditioned media from glioblastoma cell lines, namely T98G (CMT98G) and U87MG (CMU87). Immunofluorescence, RT-qPCR, in-cell Western assay, and proximity ligation assay (PLA) were performed to analyze BBB components, kinin receptors and TLR4, a receptor associated with the kinin pathway and inflammation. The effect of DBK on enhancing paracellular molecule transport was evaluated using Evans blue dye (EB) quantification in a cell culture insert assay and in an in vivo model, where mice with and without brain tumors were treated with DBK. To assess the functional impact of the transient BBB opening induced by DBK, the chemotherapeutic drug doxorubicin (DOX) was administered. <b>Results</b>: Treatment with DBK facilitates the presence of EB in the brain parenchyma by transiently disrupting the BBB, as further evidenced by the increased paracellular passage of the dye in an in vitro assay. B1R activation by DBK induces transient BBB opening lasting less than 48 h, enhancing the bioavailability of the DOX within the brain parenchyma and glioma tumor mass. The interaction between B1R and TLR4 is disrupted by the secreted factors released by glioblastoma cells, as conditioned media from T98G and U87 reduce TLR4 staining in endothelial cells without affecting B1R expression. <b>Conclusions</b>: These results further support the potential of B1R activation as a strategy to enhance targeted drug delivery to the brain.
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spelling doaj-art-6fbc99bdb0b54c57bc92e768b2672e802025-08-20T03:13:48ZengMDPI AGPharmaceuticals1424-82472025-04-0118459110.3390/ph18040591Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma EnvironmentsCarolina Batista0João Victor Roza Cruz1Michele Siqueira2João Bosco Pesquero3Joice Stipursky4Fabio de Almeida Mendes5Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, BrazilInstituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, BrazilInstituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, BrazilDepartamento de Biofísica, Universidade Federal de São Paulo, São Paulo 04039-032, SP, BrazilInstituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, BrazilInstituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-902, RJ, Brazil<b>Background/Objectives</b>: The low permeability of the blood-brain barrier (BBB) represents a significant challenge to effective systemic chemotherapy for primary and metastatic brain cancers. Kinin receptors play a crucial role in modulating BBB permeability, and their agonist analogs have been explored in preclinical animal models to enhance drug delivery to the brain. In this study, we investigated whether des-Arg<sup>9</sup>-bradykinin (DBK), a physiological agonist of kinin B<sub>1</sub> receptor (B1R), acts as a brain drug delivery adjuvant by promoting the transient opening of the BBB. <b>Methods</b>: Human brain microvascular endothelial cells (HBMECs) were treated with DBK in the culture medium and in conditioned media from glioblastoma cell lines, namely T98G (CMT98G) and U87MG (CMU87). Immunofluorescence, RT-qPCR, in-cell Western assay, and proximity ligation assay (PLA) were performed to analyze BBB components, kinin receptors and TLR4, a receptor associated with the kinin pathway and inflammation. The effect of DBK on enhancing paracellular molecule transport was evaluated using Evans blue dye (EB) quantification in a cell culture insert assay and in an in vivo model, where mice with and without brain tumors were treated with DBK. To assess the functional impact of the transient BBB opening induced by DBK, the chemotherapeutic drug doxorubicin (DOX) was administered. <b>Results</b>: Treatment with DBK facilitates the presence of EB in the brain parenchyma by transiently disrupting the BBB, as further evidenced by the increased paracellular passage of the dye in an in vitro assay. B1R activation by DBK induces transient BBB opening lasting less than 48 h, enhancing the bioavailability of the DOX within the brain parenchyma and glioma tumor mass. The interaction between B1R and TLR4 is disrupted by the secreted factors released by glioblastoma cells, as conditioned media from T98G and U87 reduce TLR4 staining in endothelial cells without affecting B1R expression. <b>Conclusions</b>: These results further support the potential of B1R activation as a strategy to enhance targeted drug delivery to the brain.https://www.mdpi.com/1424-8247/18/4/591kinin B<sub>1</sub> receptordes-Arg<sup>9</sup>-bradykininblood-brain barrierendothelial cellsglioblastoma
spellingShingle Carolina Batista
João Victor Roza Cruz
Michele Siqueira
João Bosco Pesquero
Joice Stipursky
Fabio de Almeida Mendes
Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
Pharmaceuticals
kinin B<sub>1</sub> receptor
des-Arg<sup>9</sup>-bradykinin
blood-brain barrier
endothelial cells
glioblastoma
title Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
title_full Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
title_fullStr Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
title_full_unstemmed Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
title_short Kinin B<sub>1</sub> Receptor Agonist Enhances Blood-Brain Barrier Permeability in Healthy and Glioblastoma Environments
title_sort kinin b sub 1 sub receptor agonist enhances blood brain barrier permeability in healthy and glioblastoma environments
topic kinin B<sub>1</sub> receptor
des-Arg<sup>9</sup>-bradykinin
blood-brain barrier
endothelial cells
glioblastoma
url https://www.mdpi.com/1424-8247/18/4/591
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