CRISPR/Cas9 Editing for Gaucher Disease Modelling

Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the acid β-glucosidase gene (GBA1). Besides causing GD, GBA1 mutations constitute the main genetic risk factor for developing Parkinson’s disease. The molecular basis of neurological manifestations in GD...

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Autores principales: Pavan, Eleonora, Ormazabal, Maximiliano Emanuel, Peruzzo, Paolo, Vaena, Emilio, Rozenfeld, Paula Adriana, Dardis, Andrea
Formato: Articulo
Lenguaje:Inglés
Publicado: 2020
Materias:
Biología
Gaucher disease
Cellular model
Acid β-glucosidase
Crispr/cas9
Unfolded protein response
Neuroinflammation
Α-synuclein
High-throughput drug screenings
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/107275
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC7246564&blobtype=pdf
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-107275
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Biología
Gaucher disease
Cellular model
Acid β-glucosidase
Crispr/cas9
Unfolded protein response
Neuroinflammation
Α-synuclein
High-throughput drug screenings
spellingShingle Biología
Gaucher disease
Cellular model
Acid β-glucosidase
Crispr/cas9
Unfolded protein response
Neuroinflammation
Α-synuclein
High-throughput drug screenings
Pavan, Eleonora
Ormazabal, Maximiliano Emanuel
Peruzzo, Paolo
Vaena, Emilio
Rozenfeld, Paula Adriana
Dardis, Andrea
CRISPR/Cas9 Editing for Gaucher Disease Modelling
topic_facet Biología
Gaucher disease
Cellular model
Acid β-glucosidase
Crispr/cas9
Unfolded protein response
Neuroinflammation
Α-synuclein
High-throughput drug screenings
description Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the acid β-glucosidase gene (GBA1). Besides causing GD, GBA1 mutations constitute the main genetic risk factor for developing Parkinson’s disease. The molecular basis of neurological manifestations in GD remain elusive. However, neuroinflammation has been proposed as a key player in this process. We exploited CRISPR/Cas9 technology to edit GBA1 in the human monocytic THP-1 cell line to develop an isogenic GD model of monocytes and in glioblastoma U87 cell lines to generate an isogenic GD model of glial cells. Both edited (GBA1 mutant) cell lines presented low levels of mutant acid β-glucosidase expression, less than 1% of residual activity and massive accumulation of substrate. Moreover, U87 GBA1 mutant cells showed that the mutant enzyme was retained in the ER and subjected to proteasomal degradation, triggering unfolded protein response (UPR). U87 GBA1 mutant cells displayed an increased production of interleukin-1β, both with and without inflammosome activation, α-syn accumulation and a higher rate of cell death in comparison with wild-type cells. In conclusion, we developed reliable, isogenic, and easy-to-handle cellular models of GD obtained from commercially accessible cells to be employed in GD pathophysiology studies and high-throughput drug screenings.
format Articulo
Articulo
author Pavan, Eleonora
Ormazabal, Maximiliano Emanuel
Peruzzo, Paolo
Vaena, Emilio
Rozenfeld, Paula Adriana
Dardis, Andrea
author_facet Pavan, Eleonora
Ormazabal, Maximiliano Emanuel
Peruzzo, Paolo
Vaena, Emilio
Rozenfeld, Paula Adriana
Dardis, Andrea
author_sort Pavan, Eleonora
title CRISPR/Cas9 Editing for Gaucher Disease Modelling
title_short CRISPR/Cas9 Editing for Gaucher Disease Modelling
title_full CRISPR/Cas9 Editing for Gaucher Disease Modelling
title_fullStr CRISPR/Cas9 Editing for Gaucher Disease Modelling
title_full_unstemmed CRISPR/Cas9 Editing for Gaucher Disease Modelling
title_sort crispr/cas9 editing for gaucher disease modelling
publishDate 2020
url http://sedici.unlp.edu.ar/handle/10915/107275
http://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC7246564&blobtype=pdf
work_keys_str_mv AT pavaneleonora crisprcas9editingforgaucherdiseasemodelling
AT ormazabalmaximilianoemanuel crisprcas9editingforgaucherdiseasemodelling
AT peruzzopaolo crisprcas9editingforgaucherdiseasemodelling
AT vaenaemilio crisprcas9editingforgaucherdiseasemodelling
AT rozenfeldpaulaadriana crisprcas9editingforgaucherdiseasemodelling
AT dardisandrea crisprcas9editingforgaucherdiseasemodelling
bdutipo_str Repositorios
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