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Exploring ITM2A as a new potential target for brain delivery
BACKGROUND: Integral membrane protein 2A (ITM2A) is a transmembrane protein expressed in a variety of tissues; little is known about its function, particularly in the brain. ITM2A was found to be highly enriched in human brain versus peripheral endothelial cells by transcriptomic and proteomic studi...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8935840/ https://www.ncbi.nlm.nih.gov/pubmed/35313913 http://dx.doi.org/10.1186/s12987-022-00321-3 |
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author | Cegarra, Céline Chaves, C. Déon, C. Do, T. M. Dumas, B. Frenzel, A. Kuhn, P. Roudieres, V. Guillemot, J. C. Lesuisse, D. |
author_facet | Cegarra, Céline Chaves, C. Déon, C. Do, T. M. Dumas, B. Frenzel, A. Kuhn, P. Roudieres, V. Guillemot, J. C. Lesuisse, D. |
author_sort | Cegarra, Céline |
collection | PubMed |
description | BACKGROUND: Integral membrane protein 2A (ITM2A) is a transmembrane protein expressed in a variety of tissues; little is known about its function, particularly in the brain. ITM2A was found to be highly enriched in human brain versus peripheral endothelial cells by transcriptomic and proteomic studies conducted within the European Collaboration on the Optimization of Macromolecular Pharmaceutical (COMPACT) Innovative Medicines Initiative (IMI) consortium. Here, we report the work that was undertaken to determine whether ITM2A could represent a potential target for delivering drugs to the brain. METHODS: A series of ITM2A constructs, cell lines and specific anti-human and mouse ITM2A antibodies were generated. Binding and internalization studies in Human Embryonic Kidney 293 (HEK293) cells overexpressing ITM2A and in brain microvascular endothelial cells from mouse and non-human primate (NHP) were performed with these tools. The best ITM2A antibody was evaluated in an in vitro human blood brain barrier (BBB) model and in an in vivo mouse pharmacokinetic study to investigate its ability to cross the BBB. RESULTS: Antibodies specifically recognizing extracellular parts of ITM2A or tags inserted in its extracellular domain showed selective binding and uptake in ITM2A-overexpressing cells. However, despite high RNA expression in mouse and human microvessels, the ITM2A protein was rapidly downregulated when endothelial cells were grown in culture, probably explaining why transcytosis could not be observed in vitro. An attempt to directly demonstrate in vivo transcytosis in mice was inconclusive, using either a cross-reactive anti-ITM2A antibody or in vivo phage panning of an anti-ITM2A phage library. CONCLUSIONS: The present work describes our efforts to explore the potential of ITM2A as a target mediating transcytosis through the BBB, and highlights the multiple challenges linked to the identification of new brain delivery targets. Our data provide evidence that antibodies against ITM2A are internalized in ITM2A-overexpressing HEK293 cells, and that ITM2A is expressed in brain microvessels, but further investigations will be needed to demonstrate that ITM2A is a potential target for brain delivery. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12987-022-00321-3. |
format | Online Article Text |
id | pubmed-8935840 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-89358402022-03-23 Exploring ITM2A as a new potential target for brain delivery Cegarra, Céline Chaves, C. Déon, C. Do, T. M. Dumas, B. Frenzel, A. Kuhn, P. Roudieres, V. Guillemot, J. C. Lesuisse, D. Fluids Barriers CNS Research BACKGROUND: Integral membrane protein 2A (ITM2A) is a transmembrane protein expressed in a variety of tissues; little is known about its function, particularly in the brain. ITM2A was found to be highly enriched in human brain versus peripheral endothelial cells by transcriptomic and proteomic studies conducted within the European Collaboration on the Optimization of Macromolecular Pharmaceutical (COMPACT) Innovative Medicines Initiative (IMI) consortium. Here, we report the work that was undertaken to determine whether ITM2A could represent a potential target for delivering drugs to the brain. METHODS: A series of ITM2A constructs, cell lines and specific anti-human and mouse ITM2A antibodies were generated. Binding and internalization studies in Human Embryonic Kidney 293 (HEK293) cells overexpressing ITM2A and in brain microvascular endothelial cells from mouse and non-human primate (NHP) were performed with these tools. The best ITM2A antibody was evaluated in an in vitro human blood brain barrier (BBB) model and in an in vivo mouse pharmacokinetic study to investigate its ability to cross the BBB. RESULTS: Antibodies specifically recognizing extracellular parts of ITM2A or tags inserted in its extracellular domain showed selective binding and uptake in ITM2A-overexpressing cells. However, despite high RNA expression in mouse and human microvessels, the ITM2A protein was rapidly downregulated when endothelial cells were grown in culture, probably explaining why transcytosis could not be observed in vitro. An attempt to directly demonstrate in vivo transcytosis in mice was inconclusive, using either a cross-reactive anti-ITM2A antibody or in vivo phage panning of an anti-ITM2A phage library. CONCLUSIONS: The present work describes our efforts to explore the potential of ITM2A as a target mediating transcytosis through the BBB, and highlights the multiple challenges linked to the identification of new brain delivery targets. Our data provide evidence that antibodies against ITM2A are internalized in ITM2A-overexpressing HEK293 cells, and that ITM2A is expressed in brain microvessels, but further investigations will be needed to demonstrate that ITM2A is a potential target for brain delivery. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12987-022-00321-3. BioMed Central 2022-03-21 /pmc/articles/PMC8935840/ /pubmed/35313913 http://dx.doi.org/10.1186/s12987-022-00321-3 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Cegarra, Céline Chaves, C. Déon, C. Do, T. M. Dumas, B. Frenzel, A. Kuhn, P. Roudieres, V. Guillemot, J. C. Lesuisse, D. Exploring ITM2A as a new potential target for brain delivery |
title | Exploring ITM2A as a new potential target for brain delivery |
title_full | Exploring ITM2A as a new potential target for brain delivery |
title_fullStr | Exploring ITM2A as a new potential target for brain delivery |
title_full_unstemmed | Exploring ITM2A as a new potential target for brain delivery |
title_short | Exploring ITM2A as a new potential target for brain delivery |
title_sort | exploring itm2a as a new potential target for brain delivery |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8935840/ https://www.ncbi.nlm.nih.gov/pubmed/35313913 http://dx.doi.org/10.1186/s12987-022-00321-3 |
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