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Controlling therapeutic protein expression via inhalation of a butter flavor molecule

Precise control of the delivery of therapeutic proteins is critical for gene- and cell-based therapies, and expression should only be switched on in the presence of a specific trigger signal of appropriate magnitude. Focusing on the advantages of delivering the trigger by inhalation, we have develop...

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Autores principales: Bertschi, Adrian, Stefanov, Bozhidar-Adrian, Xue, Shuai, Charpin-El Hamri, Ghislaine, Teixeira, Ana Palma, Fussenegger, Martin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018347/
https://www.ncbi.nlm.nih.gov/pubmed/36625292
http://dx.doi.org/10.1093/nar/gkac1256
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author Bertschi, Adrian
Stefanov, Bozhidar-Adrian
Xue, Shuai
Charpin-El Hamri, Ghislaine
Teixeira, Ana Palma
Fussenegger, Martin
author_facet Bertschi, Adrian
Stefanov, Bozhidar-Adrian
Xue, Shuai
Charpin-El Hamri, Ghislaine
Teixeira, Ana Palma
Fussenegger, Martin
author_sort Bertschi, Adrian
collection PubMed
description Precise control of the delivery of therapeutic proteins is critical for gene- and cell-based therapies, and expression should only be switched on in the presence of a specific trigger signal of appropriate magnitude. Focusing on the advantages of delivering the trigger by inhalation, we have developed a mammalian synthetic gene switch that enables regulation of transgene expression by exposure to the semi-volatile small molecule acetoin, a widely used, FDA-approved food flavor additive. The gene switch capitalizes on the bacterial regulatory protein AcoR fused to a mammalian transactivation domain, which binds to promoter regions with specific DNA sequences in the presence of acetoin and dose-dependently activates expression of downstream transgenes. Wild-type mice implanted with alginate-encapsulated cells transgenic for the acetoin gene switch showed a dose-dependent increase in blood levels of reporter protein in response to either administration of acetoin solution via oral gavage or longer exposure to acetoin aerosol generated by a commercial portable inhaler. Intake of typical acetoin-containing foods, such as butter, lychees and cheese, did not activate transgene expression. As a proof of concept, we show that blood glucose levels were normalized by acetoin aerosol inhalation in type-I diabetic mice implanted with acetoin-responsive insulin-producing cells. Delivery of trigger molecules using portable inhalers may facilitate regular administration of therapeutic proteins via next-generation cell-based therapies to treat chronic diseases for which frequent dosing is required.
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spelling pubmed-100183472023-03-17 Controlling therapeutic protein expression via inhalation of a butter flavor molecule Bertschi, Adrian Stefanov, Bozhidar-Adrian Xue, Shuai Charpin-El Hamri, Ghislaine Teixeira, Ana Palma Fussenegger, Martin Nucleic Acids Res Methods Online Precise control of the delivery of therapeutic proteins is critical for gene- and cell-based therapies, and expression should only be switched on in the presence of a specific trigger signal of appropriate magnitude. Focusing on the advantages of delivering the trigger by inhalation, we have developed a mammalian synthetic gene switch that enables regulation of transgene expression by exposure to the semi-volatile small molecule acetoin, a widely used, FDA-approved food flavor additive. The gene switch capitalizes on the bacterial regulatory protein AcoR fused to a mammalian transactivation domain, which binds to promoter regions with specific DNA sequences in the presence of acetoin and dose-dependently activates expression of downstream transgenes. Wild-type mice implanted with alginate-encapsulated cells transgenic for the acetoin gene switch showed a dose-dependent increase in blood levels of reporter protein in response to either administration of acetoin solution via oral gavage or longer exposure to acetoin aerosol generated by a commercial portable inhaler. Intake of typical acetoin-containing foods, such as butter, lychees and cheese, did not activate transgene expression. As a proof of concept, we show that blood glucose levels were normalized by acetoin aerosol inhalation in type-I diabetic mice implanted with acetoin-responsive insulin-producing cells. Delivery of trigger molecules using portable inhalers may facilitate regular administration of therapeutic proteins via next-generation cell-based therapies to treat chronic diseases for which frequent dosing is required. Oxford University Press 2023-01-10 /pmc/articles/PMC10018347/ /pubmed/36625292 http://dx.doi.org/10.1093/nar/gkac1256 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Bertschi, Adrian
Stefanov, Bozhidar-Adrian
Xue, Shuai
Charpin-El Hamri, Ghislaine
Teixeira, Ana Palma
Fussenegger, Martin
Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title_full Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title_fullStr Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title_full_unstemmed Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title_short Controlling therapeutic protein expression via inhalation of a butter flavor molecule
title_sort controlling therapeutic protein expression via inhalation of a butter flavor molecule
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10018347/
https://www.ncbi.nlm.nih.gov/pubmed/36625292
http://dx.doi.org/10.1093/nar/gkac1256
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