Genetically engineered probiotic for the treatment of phenylketonuria (PKU); Assessment of a novel treatment in vitro and in the PAHenu2 mouse model of PKU

Katherine E. Durrer, Michael S. Allen, Ione Hunt Von Herbing

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Phenylketonuria (PKU) is a genetic disease characterized by the inability to convert dietary phenylalanine to tyrosine by phenylalanine hydroxylase. Given the importance of gut microbes in digestion, a genetically engineered microbe could potentially degrade some ingested phenylalanine from the diet prior to absorption. To test this, a phenylalanine lyase gene from Anabaena variabilis (AvPAL) was codon-optimized and cloned into a shuttle vector for expression in Lactobacillus reuteri 100-23C (pHENOMMenal). Functional expression of AvPAL was determined in vitro, and subsequently tested in vivo in homozygous PAHenu2 (PKU model) mice. Initial trials of two PAHenu2 homozygous (PKU) mice defined conditions for freeze-drying and delivery of bacteria. Animals showed reduced blood phe within three to four days of treatment with pHENOMMenal probiotic, and blood phe concentrations remained significantly reduced (P < 0.0005) compared to untreated controls during the course of experiments. Although pHENOMMenal probiotic could be cultured from fecal samples at four months post treatment, it could no longer be cultivated from feces at eight months post treatment, indicating eventual loss of the microbe from the gut. Preliminary screens during experimentation found no immune response to AvPAL. Collectively these studies provide data for the use of a genetically engineered probiotic as a potential treatment for PKU.

Original languageEnglish
Article numbere0176286
JournalPLoS ONE
Volume12
Issue number5
DOIs
StatePublished - May 2017

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phenylketonuria
Phenylketonurias
Probiotics
Phenylalanine
probiotics
animal models
phenylalanine
Blood
phenylalanine 4-monooxygenase
microorganisms
Phenylalanine Hydroxylase
Genetic Vectors
Lyases
Anabaena variabilis
digestive system
Lactobacillus reuteri
Nutrition
genetic vectors
Inborn Genetic Diseases
Freeze Drying

Cite this

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title = "Genetically engineered probiotic for the treatment of phenylketonuria (PKU); Assessment of a novel treatment in vitro and in the PAHenu2 mouse model of PKU",
abstract = "Phenylketonuria (PKU) is a genetic disease characterized by the inability to convert dietary phenylalanine to tyrosine by phenylalanine hydroxylase. Given the importance of gut microbes in digestion, a genetically engineered microbe could potentially degrade some ingested phenylalanine from the diet prior to absorption. To test this, a phenylalanine lyase gene from Anabaena variabilis (AvPAL) was codon-optimized and cloned into a shuttle vector for expression in Lactobacillus reuteri 100-23C (pHENOMMenal). Functional expression of AvPAL was determined in vitro, and subsequently tested in vivo in homozygous PAHenu2 (PKU model) mice. Initial trials of two PAHenu2 homozygous (PKU) mice defined conditions for freeze-drying and delivery of bacteria. Animals showed reduced blood phe within three to four days of treatment with pHENOMMenal probiotic, and blood phe concentrations remained significantly reduced (P < 0.0005) compared to untreated controls during the course of experiments. Although pHENOMMenal probiotic could be cultured from fecal samples at four months post treatment, it could no longer be cultivated from feces at eight months post treatment, indicating eventual loss of the microbe from the gut. Preliminary screens during experimentation found no immune response to AvPAL. Collectively these studies provide data for the use of a genetically engineered probiotic as a potential treatment for PKU.",
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Genetically engineered probiotic for the treatment of phenylketonuria (PKU); Assessment of a novel treatment in vitro and in the PAHenu2 mouse model of PKU. / Durrer, Katherine E.; Allen, Michael S.; Von Herbing, Ione Hunt.

In: PLoS ONE, Vol. 12, No. 5, e0176286, 05.2017.

Research output: Contribution to journalArticle

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