Cargando…
Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU
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...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435137/ https://www.ncbi.nlm.nih.gov/pubmed/28520731 http://dx.doi.org/10.1371/journal.pone.0176286 |
_version_ | 1783237176758108160 |
---|---|
author | Durrer, Katherine E. Allen, Michael S. Hunt von Herbing, Ione |
author_facet | Durrer, Katherine E. Allen, Michael S. Hunt von Herbing, Ione |
author_sort | Durrer, Katherine E. |
collection | PubMed |
description | 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 PAH(enu2) (PKU model) mice. Initial trials of two PAH(enu2) 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. |
format | Online Article Text |
id | pubmed-5435137 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-54351372017-05-26 Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU Durrer, Katherine E. Allen, Michael S. Hunt von Herbing, Ione PLoS One Research Article 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 PAH(enu2) (PKU model) mice. Initial trials of two PAH(enu2) 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. Public Library of Science 2017-05-17 /pmc/articles/PMC5435137/ /pubmed/28520731 http://dx.doi.org/10.1371/journal.pone.0176286 Text en © 2017 Durrer et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Durrer, Katherine E. Allen, Michael S. Hunt von Herbing, Ione Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title | Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title_full | Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title_fullStr | Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title_full_unstemmed | Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title_short | Genetically engineered probiotic for the treatment of phenylketonuria (PKU); assessment of a novel treatment in vitro and in the PAH(enu2) mouse model of PKU |
title_sort | genetically engineered probiotic for the treatment of phenylketonuria (pku); assessment of a novel treatment in vitro and in the pah(enu2) mouse model of pku |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5435137/ https://www.ncbi.nlm.nih.gov/pubmed/28520731 http://dx.doi.org/10.1371/journal.pone.0176286 |
work_keys_str_mv | AT durrerkatherinee geneticallyengineeredprobioticforthetreatmentofphenylketonuriapkuassessmentofanoveltreatmentinvitroandinthepahenu2mousemodelofpku AT allenmichaels geneticallyengineeredprobioticforthetreatmentofphenylketonuriapkuassessmentofanoveltreatmentinvitroandinthepahenu2mousemodelofpku AT huntvonherbingione geneticallyengineeredprobioticforthetreatmentofphenylketonuriapkuassessmentofanoveltreatmentinvitroandinthepahenu2mousemodelofpku |