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A conformation-specific ON-switch for controlling CAR T cells with an orally available drug
Molecular ON-switches in which a chemical compound induces protein–protein interactions can allow cellular function to be controlled with small molecules. ON-switches based on clinically applicable compounds and human proteins would greatly facilitate their therapeutic use. Here, we developed an ON-...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
National Academy of Sciences
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7334647/ https://www.ncbi.nlm.nih.gov/pubmed/32554495 http://dx.doi.org/10.1073/pnas.1911154117 |
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author | Zajc, Charlotte U. Dobersberger, Markus Schaffner, Irene Mlynek, Georg Pühringer, Dominic Salzer, Benjamin Djinović-Carugo, Kristina Steinberger, Peter De Sousa Linhares, Annika Yang, Nicole J. Obinger, Christian Holter, Wolfgang Traxlmayr, Michael W. Lehner, Manfred |
author_facet | Zajc, Charlotte U. Dobersberger, Markus Schaffner, Irene Mlynek, Georg Pühringer, Dominic Salzer, Benjamin Djinović-Carugo, Kristina Steinberger, Peter De Sousa Linhares, Annika Yang, Nicole J. Obinger, Christian Holter, Wolfgang Traxlmayr, Michael W. Lehner, Manfred |
author_sort | Zajc, Charlotte U. |
collection | PubMed |
description | Molecular ON-switches in which a chemical compound induces protein–protein interactions can allow cellular function to be controlled with small molecules. ON-switches based on clinically applicable compounds and human proteins would greatly facilitate their therapeutic use. Here, we developed an ON-switch system in which the human retinol binding protein 4 (hRBP4) of the lipocalin family interacts with engineered hRBP4 binders in a small molecule-dependent manner. Two different protein scaffolds were engineered to bind to hRBP4 when loaded with the orally available small molecule A1120. The crystal structure of an assembled ON-switch shows that the engineered binder specifically recognizes the conformational changes induced by A1120 in two loop regions of hRBP4. We demonstrate that this conformation-specific ON-switch is highly dependent on the presence of A1120, as demonstrated by an ∼500-fold increase in affinity upon addition of the small molecule drug. Furthermore, the ON-switch successfully regulated the activity of primary human CAR T cells in vitro. We anticipate that lipocalin-based ON-switches have the potential to be broadly applied for the safe pharmacological control of cellular therapeutics. |
format | Online Article Text |
id | pubmed-7334647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-73346472020-07-15 A conformation-specific ON-switch for controlling CAR T cells with an orally available drug Zajc, Charlotte U. Dobersberger, Markus Schaffner, Irene Mlynek, Georg Pühringer, Dominic Salzer, Benjamin Djinović-Carugo, Kristina Steinberger, Peter De Sousa Linhares, Annika Yang, Nicole J. Obinger, Christian Holter, Wolfgang Traxlmayr, Michael W. Lehner, Manfred Proc Natl Acad Sci U S A Biological Sciences Molecular ON-switches in which a chemical compound induces protein–protein interactions can allow cellular function to be controlled with small molecules. ON-switches based on clinically applicable compounds and human proteins would greatly facilitate their therapeutic use. Here, we developed an ON-switch system in which the human retinol binding protein 4 (hRBP4) of the lipocalin family interacts with engineered hRBP4 binders in a small molecule-dependent manner. Two different protein scaffolds were engineered to bind to hRBP4 when loaded with the orally available small molecule A1120. The crystal structure of an assembled ON-switch shows that the engineered binder specifically recognizes the conformational changes induced by A1120 in two loop regions of hRBP4. We demonstrate that this conformation-specific ON-switch is highly dependent on the presence of A1120, as demonstrated by an ∼500-fold increase in affinity upon addition of the small molecule drug. Furthermore, the ON-switch successfully regulated the activity of primary human CAR T cells in vitro. We anticipate that lipocalin-based ON-switches have the potential to be broadly applied for the safe pharmacological control of cellular therapeutics. National Academy of Sciences 2020-06-30 2020-06-17 /pmc/articles/PMC7334647/ /pubmed/32554495 http://dx.doi.org/10.1073/pnas.1911154117 Text en Copyright © 2020 the Author(s). Published by PNAS. http://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (http://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Biological Sciences Zajc, Charlotte U. Dobersberger, Markus Schaffner, Irene Mlynek, Georg Pühringer, Dominic Salzer, Benjamin Djinović-Carugo, Kristina Steinberger, Peter De Sousa Linhares, Annika Yang, Nicole J. Obinger, Christian Holter, Wolfgang Traxlmayr, Michael W. Lehner, Manfred A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title | A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title_full | A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title_fullStr | A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title_full_unstemmed | A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title_short | A conformation-specific ON-switch for controlling CAR T cells with an orally available drug |
title_sort | conformation-specific on-switch for controlling car t cells with an orally available drug |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7334647/ https://www.ncbi.nlm.nih.gov/pubmed/32554495 http://dx.doi.org/10.1073/pnas.1911154117 |
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