Cargando…

AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria

Phenylketonuria (PKU) due to recessively inherited phenylalanine hydroxylase (PAH) deficiency results in hyperphenylalaninemia, which is toxic to the central nervous system. Restriction of dietary phenylalanine intake remains the standard of PKU care and prevents the major neurologic manifestations...

Descripción completa

Detalles Bibliográficos
Autores principales: Richards, Daelyn Y., Winn, Shelley R., Dudley, Sandra, Nygaard, Sean, Mighell, Taylor L., Grompe, Markus, Harding, Cary O.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6962637/
https://www.ncbi.nlm.nih.gov/pubmed/31970201
http://dx.doi.org/10.1016/j.omtm.2019.12.004
_version_ 1783488187081949184
author Richards, Daelyn Y.
Winn, Shelley R.
Dudley, Sandra
Nygaard, Sean
Mighell, Taylor L.
Grompe, Markus
Harding, Cary O.
author_facet Richards, Daelyn Y.
Winn, Shelley R.
Dudley, Sandra
Nygaard, Sean
Mighell, Taylor L.
Grompe, Markus
Harding, Cary O.
author_sort Richards, Daelyn Y.
collection PubMed
description Phenylketonuria (PKU) due to recessively inherited phenylalanine hydroxylase (PAH) deficiency results in hyperphenylalaninemia, which is toxic to the central nervous system. Restriction of dietary phenylalanine intake remains the standard of PKU care and prevents the major neurologic manifestations of the disease, yet shortcomings of dietary therapy remain, including poor adherence to a difficult and unpalatable diet, an increased incidence of neuropsychiatric illness, and imperfect neurocognitive outcomes. Gene therapy for PKU is a promising novel approach to promote lifelong neurological protection while allowing unrestricted dietary phenylalanine intake. In this study, liver-tropic recombinant AAV2/8 vectors were used to deliver CRISPR/Cas9 machinery and facilitate correction of the Pah(enu2) allele by homologous recombination. Additionally, a non-homologous end joining (NHEJ) inhibitor, vanillin, was co-administered with the viral drug to promote homology-directed repair (HDR) with the AAV-provided repair template. This combinatorial drug administration allowed for lifelong, permanent correction of the Pah(enu2) allele in a portion of treated hepatocytes of mice with PKU, yielding partial restoration of liver PAH activity, substantial reduction of blood phenylalanine, and prevention of maternal PKU effects during breeding. This work reveals that CRISPR/Cas9 gene editing is a promising tool for permanent PKU gene editing.
format Online
Article
Text
id pubmed-6962637
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher American Society of Gene & Cell Therapy
record_format MEDLINE/PubMed
spelling pubmed-69626372020-01-22 AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria Richards, Daelyn Y. Winn, Shelley R. Dudley, Sandra Nygaard, Sean Mighell, Taylor L. Grompe, Markus Harding, Cary O. Mol Ther Methods Clin Dev Article Phenylketonuria (PKU) due to recessively inherited phenylalanine hydroxylase (PAH) deficiency results in hyperphenylalaninemia, which is toxic to the central nervous system. Restriction of dietary phenylalanine intake remains the standard of PKU care and prevents the major neurologic manifestations of the disease, yet shortcomings of dietary therapy remain, including poor adherence to a difficult and unpalatable diet, an increased incidence of neuropsychiatric illness, and imperfect neurocognitive outcomes. Gene therapy for PKU is a promising novel approach to promote lifelong neurological protection while allowing unrestricted dietary phenylalanine intake. In this study, liver-tropic recombinant AAV2/8 vectors were used to deliver CRISPR/Cas9 machinery and facilitate correction of the Pah(enu2) allele by homologous recombination. Additionally, a non-homologous end joining (NHEJ) inhibitor, vanillin, was co-administered with the viral drug to promote homology-directed repair (HDR) with the AAV-provided repair template. This combinatorial drug administration allowed for lifelong, permanent correction of the Pah(enu2) allele in a portion of treated hepatocytes of mice with PKU, yielding partial restoration of liver PAH activity, substantial reduction of blood phenylalanine, and prevention of maternal PKU effects during breeding. This work reveals that CRISPR/Cas9 gene editing is a promising tool for permanent PKU gene editing. American Society of Gene & Cell Therapy 2019-12-24 /pmc/articles/PMC6962637/ /pubmed/31970201 http://dx.doi.org/10.1016/j.omtm.2019.12.004 Text en © 2019 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Richards, Daelyn Y.
Winn, Shelley R.
Dudley, Sandra
Nygaard, Sean
Mighell, Taylor L.
Grompe, Markus
Harding, Cary O.
AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title_full AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title_fullStr AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title_full_unstemmed AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title_short AAV-Mediated CRISPR/Cas9 Gene Editing in Murine Phenylketonuria
title_sort aav-mediated crispr/cas9 gene editing in murine phenylketonuria
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6962637/
https://www.ncbi.nlm.nih.gov/pubmed/31970201
http://dx.doi.org/10.1016/j.omtm.2019.12.004
work_keys_str_mv AT richardsdaelyny aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT winnshelleyr aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT dudleysandra aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT nygaardsean aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT mighelltaylorl aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT grompemarkus aavmediatedcrisprcas9geneeditinginmurinephenylketonuria
AT hardingcaryo aavmediatedcrisprcas9geneeditinginmurinephenylketonuria