Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays
Abstract Background The pathways that control protein transport across the blood–brain barrier (BBB) remain poorly characterized. Despite great advances in recapitulating the human BBB in vitro, current models are not suitable for systematic analysis of the molecular mechanisms of antibody transport...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | Fluids and Barriers of the CNS |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12987-021-00276-x |
| _version_ | 1818882195904790528 |
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| author | Claire Simonneau Martina Duschmalé Alina Gavrilov Nathalie Brandenberg Sylke Hoehnel Camilla Ceroni Evodie Lassalle Elena Kassianidou Hendrik Knoetgen Jens Niewoehner Roberto Villaseñor |
| author_facet | Claire Simonneau Martina Duschmalé Alina Gavrilov Nathalie Brandenberg Sylke Hoehnel Camilla Ceroni Evodie Lassalle Elena Kassianidou Hendrik Knoetgen Jens Niewoehner Roberto Villaseñor |
| author_sort | Claire Simonneau |
| collection | DOAJ |
| description | Abstract Background The pathways that control protein transport across the blood–brain barrier (BBB) remain poorly characterized. Despite great advances in recapitulating the human BBB in vitro, current models are not suitable for systematic analysis of the molecular mechanisms of antibody transport. The gaps in our mechanistic understanding of antibody transcytosis hinder new therapeutic delivery strategy development. Methods We applied a novel bioengineering approach to generate human BBB organoids by the self-assembly of astrocytes, pericytes and brain endothelial cells with unprecedented throughput and reproducibility using micro patterned hydrogels. We designed a semi-automated and scalable imaging assay to measure receptor-mediated transcytosis of antibodies. Finally, we developed a workflow to use CRISPR/Cas9 gene editing in BBB organoid arrays to knock out regulators of endocytosis specifically in brain endothelial cells in order to dissect the molecular mechanisms of receptor-mediated transcytosis. Results BBB organoid arrays allowed the simultaneous growth of more than 3000 homogenous organoids per individual experiment in a highly reproducible manner. BBB organoid arrays showed low permeability to macromolecules and prevented transport of human non-targeting antibodies. In contrast, a monovalent antibody targeting the human transferrin receptor underwent dose- and time-dependent transcytosis in organoids. Using CRISPR/Cas9 gene editing in BBB organoid arrays, we showed that clathrin, but not caveolin, is required for transferrin receptor-dependent transcytosis. Conclusions Human BBB organoid arrays are a robust high-throughput platform that can be used to discover new mechanisms of receptor-mediated antibody transcytosis. The implementation of this platform during early stages of drug discovery can accelerate the development of new brain delivery technologies. |
| first_indexed | 2024-12-19T15:13:54Z |
| format | Article |
| id | doaj.art-0bec5e6650484393b2bce695057360c1 |
| institution | Directory Open Access Journal |
| issn | 2045-8118 |
| language | English |
| last_indexed | 2024-12-19T15:13:54Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Fluids and Barriers of the CNS |
| spelling | doaj.art-0bec5e6650484393b2bce695057360c12022-12-21T20:16:13ZengBMCFluids and Barriers of the CNS2045-81182021-09-0118111710.1186/s12987-021-00276-xInvestigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arraysClaire Simonneau0Martina Duschmalé1Alina Gavrilov2Nathalie Brandenberg3Sylke Hoehnel4Camilla Ceroni5Evodie Lassalle6Elena Kassianidou7Hendrik Knoetgen8Jens Niewoehner9Roberto Villaseñor10Roche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselRoche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselRoche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselSUN bioscience, EPFL Innovation ParkSUN bioscience, EPFL Innovation ParkSUN bioscience, EPFL Innovation ParkRoche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselRoche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselRoche Pharma Research and Early Development (pRED), Therapeutic Modalities, Roche Innovation Center MunichRoche Pharma Research and Early Development (pRED), Therapeutic Modalities, Roche Innovation Center MunichRoche Pharma Research and Early Development (pRED), Pharmaceutical Sciences, Roche Innovation Center BaselAbstract Background The pathways that control protein transport across the blood–brain barrier (BBB) remain poorly characterized. Despite great advances in recapitulating the human BBB in vitro, current models are not suitable for systematic analysis of the molecular mechanisms of antibody transport. The gaps in our mechanistic understanding of antibody transcytosis hinder new therapeutic delivery strategy development. Methods We applied a novel bioengineering approach to generate human BBB organoids by the self-assembly of astrocytes, pericytes and brain endothelial cells with unprecedented throughput and reproducibility using micro patterned hydrogels. We designed a semi-automated and scalable imaging assay to measure receptor-mediated transcytosis of antibodies. Finally, we developed a workflow to use CRISPR/Cas9 gene editing in BBB organoid arrays to knock out regulators of endocytosis specifically in brain endothelial cells in order to dissect the molecular mechanisms of receptor-mediated transcytosis. Results BBB organoid arrays allowed the simultaneous growth of more than 3000 homogenous organoids per individual experiment in a highly reproducible manner. BBB organoid arrays showed low permeability to macromolecules and prevented transport of human non-targeting antibodies. In contrast, a monovalent antibody targeting the human transferrin receptor underwent dose- and time-dependent transcytosis in organoids. Using CRISPR/Cas9 gene editing in BBB organoid arrays, we showed that clathrin, but not caveolin, is required for transferrin receptor-dependent transcytosis. Conclusions Human BBB organoid arrays are a robust high-throughput platform that can be used to discover new mechanisms of receptor-mediated antibody transcytosis. The implementation of this platform during early stages of drug discovery can accelerate the development of new brain delivery technologies.https://doi.org/10.1186/s12987-021-00276-xBlood–brain barrierOrganoidsReceptor-mediated transcytosisHigh-throughput in vitro model |
| spellingShingle | Claire Simonneau Martina Duschmalé Alina Gavrilov Nathalie Brandenberg Sylke Hoehnel Camilla Ceroni Evodie Lassalle Elena Kassianidou Hendrik Knoetgen Jens Niewoehner Roberto Villaseñor Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays Fluids and Barriers of the CNS Blood–brain barrier Organoids Receptor-mediated transcytosis High-throughput in vitro model |
| title | Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays |
| title_full | Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays |
| title_fullStr | Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays |
| title_full_unstemmed | Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays |
| title_short | Investigating receptor-mediated antibody transcytosis using blood–brain barrier organoid arrays |
| title_sort | investigating receptor mediated antibody transcytosis using blood brain barrier organoid arrays |
| topic | Blood–brain barrier Organoids Receptor-mediated transcytosis High-throughput in vitro model |
| url | https://doi.org/10.1186/s12987-021-00276-x |
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