Controlling therapeutic protein expression via inhalation of a butter flavor molecule.
Adrian BertschiBozhidar-Adrian StefanovShuai XueGhislaine Charpin-El HamriAna Palma TeixeiraMartin FusseneggerPublished in: Nucleic acids research (2023)
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.
Keyphrases
- poor prognosis
- blood glucose
- small molecule
- copy number
- binding protein
- genome wide
- induced apoptosis
- wild type
- type diabetes
- transcription factor
- single cell
- cell cycle arrest
- genome wide identification
- protein protein
- gene expression
- long non coding rna
- blood pressure
- dna methylation
- cell death
- adipose tissue
- circulating tumor
- single molecule
- skeletal muscle
- cell proliferation
- physical activity
- drug delivery
- oxidative stress
- signaling pathway
- body mass index
- metabolic syndrome
- genome wide analysis
- high fat diet induced