CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)

Abstract The neuronal ceroid lipofuscinoses (NCLs) are a group of devastating monogenetic lysosomal disorders that affect children and young adults with no cure or effective treatment currently available. One of the more severe infantile forms of the disease (INCL or CLN1 disease) is due to mutation...

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Main Authors: S. L. Eaton, C. Proudfoot, S. G. Lillico, P. Skehel, R. A. Kline, K. Hamer, N. M. Rzechorzek, E. Clutton, R. Gregson, T. King, C. A. O’Neill, J. D. Cooper, G. Thompson, C. B. Whitelaw, T. M. Wishart
Format: Article
Language:English
Published: Nature Portfolio 2019-07-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-019-45859-9
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author S. L. Eaton
C. Proudfoot
S. G. Lillico
P. Skehel
R. A. Kline
K. Hamer
N. M. Rzechorzek
E. Clutton
R. Gregson
T. King
C. A. O’Neill
J. D. Cooper
G. Thompson
C. B. Whitelaw
T. M. Wishart
author_facet S. L. Eaton
C. Proudfoot
S. G. Lillico
P. Skehel
R. A. Kline
K. Hamer
N. M. Rzechorzek
E. Clutton
R. Gregson
T. King
C. A. O’Neill
J. D. Cooper
G. Thompson
C. B. Whitelaw
T. M. Wishart
author_sort S. L. Eaton
collection DOAJ
description Abstract The neuronal ceroid lipofuscinoses (NCLs) are a group of devastating monogenetic lysosomal disorders that affect children and young adults with no cure or effective treatment currently available. One of the more severe infantile forms of the disease (INCL or CLN1 disease) is due to mutations in the palmitoyl-protein thioesterase 1 (PPT1) gene and severely reduces the child’s lifespan to approximately 9 years of age. In order to better translate the human condition than is possible in mice, we sought to produce a large animal model employing CRISPR/Cas9 gene editing technology. Three PPT1 homozygote sheep were generated by insertion of a disease-causing PPT1 (R151X) human mutation into the orthologous sheep locus. This resulted in a morphological, anatomical and biochemical disease phenotype that closely resembles the human condition. The homozygous sheep were found to have significantly reduced PPT1 enzyme activity and accumulate autofluorescent storage material, as is observed in CLN1 patients. Clinical signs included pronounced behavioral deficits as well as motor deficits and complete loss of vision, with a reduced lifespan of 17 ± 1 months at a humanely defined terminal endpoint. Magnetic resonance imaging (MRI) confirmed a significant decrease in motor cortical volume as well as increased ventricular volume corresponding with observed brain atrophy and a profound reduction in brain mass of 30% at necropsy, similar to alterations observed in human patients. In summary, we have generated the first CRISPR/Cas9 gene edited NCL model. This novel sheep model of CLN1 disease develops biochemical, gross morphological and in vivo brain alterations confirming the efficacy of the targeted modification and potential relevance to the human condition.
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spelling doaj.art-7155c59c589e4aa29b872c1109c35b0c2022-12-21T18:01:45ZengNature PortfolioScientific Reports2045-23222019-07-01911810.1038/s41598-019-45859-9CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)S. L. Eaton0C. Proudfoot1S. G. Lillico2P. Skehel3R. A. Kline4K. Hamer5N. M. Rzechorzek6E. Clutton7R. Gregson8T. King9C. A. O’Neill10J. D. Cooper11G. Thompson12C. B. Whitelaw13T. M. Wishart14The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghCentre for Discovery Brain Science, University of Edinburgh, Hugh Robson BuildingThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghRoyal (Dick) School of Veterinary Studies, The University of EdinburghCentre for Clinical Brain Sciences, University of Edinburgh, Chancellor’s Building, 49 Little France CrescentWellcome Trust Critical Care Laboratory for Large Animals, Roslin Institute, Easter BushWellcome Trust Critical Care Laboratory for Large Animals, Roslin Institute, Easter BushThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghBioMarin Pharmaceutical IncLos Angeles Biomedical Research Institute at Harbor-UCLA Medical Center, David Geffen School of Medicine, UCLACentre for Discovery Brain Science, University of Edinburgh, Hugh Robson BuildingThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghThe Roslin Institute and Royal (Dick) School of Veterinary Studies, University of EdinburghAbstract The neuronal ceroid lipofuscinoses (NCLs) are a group of devastating monogenetic lysosomal disorders that affect children and young adults with no cure or effective treatment currently available. One of the more severe infantile forms of the disease (INCL or CLN1 disease) is due to mutations in the palmitoyl-protein thioesterase 1 (PPT1) gene and severely reduces the child’s lifespan to approximately 9 years of age. In order to better translate the human condition than is possible in mice, we sought to produce a large animal model employing CRISPR/Cas9 gene editing technology. Three PPT1 homozygote sheep were generated by insertion of a disease-causing PPT1 (R151X) human mutation into the orthologous sheep locus. This resulted in a morphological, anatomical and biochemical disease phenotype that closely resembles the human condition. The homozygous sheep were found to have significantly reduced PPT1 enzyme activity and accumulate autofluorescent storage material, as is observed in CLN1 patients. Clinical signs included pronounced behavioral deficits as well as motor deficits and complete loss of vision, with a reduced lifespan of 17 ± 1 months at a humanely defined terminal endpoint. Magnetic resonance imaging (MRI) confirmed a significant decrease in motor cortical volume as well as increased ventricular volume corresponding with observed brain atrophy and a profound reduction in brain mass of 30% at necropsy, similar to alterations observed in human patients. In summary, we have generated the first CRISPR/Cas9 gene edited NCL model. This novel sheep model of CLN1 disease develops biochemical, gross morphological and in vivo brain alterations confirming the efficacy of the targeted modification and potential relevance to the human condition.https://doi.org/10.1038/s41598-019-45859-9
spellingShingle S. L. Eaton
C. Proudfoot
S. G. Lillico
P. Skehel
R. A. Kline
K. Hamer
N. M. Rzechorzek
E. Clutton
R. Gregson
T. King
C. A. O’Neill
J. D. Cooper
G. Thompson
C. B. Whitelaw
T. M. Wishart
CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
Scientific Reports
title CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
title_full CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
title_fullStr CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
title_full_unstemmed CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
title_short CRISPR/Cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis (CLN1 disease)
title_sort crispr cas9 mediated generation of an ovine model for infantile neuronal ceroid lipofuscinosis cln1 disease
url https://doi.org/10.1038/s41598-019-45859-9
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