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 (<i>GBA1</i>). Besides causing GD, <i>GBA1</i> mutations constitute the main genetic risk factor for developing Parkinson’s disease. The molecular basi...
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| Format: | Article |
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MDPI AG
2020-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/9/3268 |
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| author | Eleonora Pavan Maximiliano Ormazabal Paolo Peruzzo Emilio Vaena Paula Rozenfeld Andrea Dardis |
| author_facet | Eleonora Pavan Maximiliano Ormazabal Paolo Peruzzo Emilio Vaena Paula Rozenfeld Andrea Dardis |
| author_sort | Eleonora Pavan |
| collection | DOAJ |
| description | Gaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the acid β-glucosidase gene (<i>GBA1</i>). Besides causing GD, <i>GBA1</i> 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 <i>GBA1</i> 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 (<i>GBA1</i> 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 <i>GBA1</i> mutant cells showed that the mutant enzyme was retained in the ER and subjected to proteasomal degradation, triggering unfolded protein response (UPR). U87 <i>GBA1</i> 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. |
| first_indexed | 2024-03-10T20:02:01Z |
| format | Article |
| id | doaj.art-f71202a490a64bd483dd3b17be28cb0e |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T20:02:01Z |
| publishDate | 2020-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f71202a490a64bd483dd3b17be28cb0e2023-11-19T23:32:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-05-01219326810.3390/ijms21093268CRISPR/Cas9 Editing for Gaucher Disease ModellingEleonora Pavan0Maximiliano Ormazabal1Paolo Peruzzo2Emilio Vaena3Paula Rozenfeld4Andrea Dardis5Regional Coordinator Centre for Rare Diseases, Academic Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, Academic Hospital of Udine, 33100 Udine, ItalyRegional Coordinator Centre for Rare Diseases, Academic Hospital of Udine, 33100 Udine, ItalyDepartamento de Ciencias Biológicas, IIFP, Universidad Nacional de La Plata, CONICET, Facultad de Ciencias Exactas Bv120 Nro.1489, La Plata 1900, ArgentinaDepartamento de Ciencias Biológicas, IIFP, Universidad Nacional de La Plata, CONICET, Facultad de Ciencias Exactas Bv120 Nro.1489, La Plata 1900, ArgentinaRegional Coordinator Centre for Rare Diseases, Academic Hospital of Udine, 33100 Udine, ItalyGaucher disease (GD) is an autosomal recessive lysosomal storage disorder caused by mutations in the acid β-glucosidase gene (<i>GBA1</i>). Besides causing GD, <i>GBA1</i> 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 <i>GBA1</i> 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 (<i>GBA1</i> 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 <i>GBA1</i> mutant cells showed that the mutant enzyme was retained in the ER and subjected to proteasomal degradation, triggering unfolded protein response (UPR). U87 <i>GBA1</i> 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.https://www.mdpi.com/1422-0067/21/9/3268Gaucher diseasecellular modelacid β-glucosidaseCRISPR/Cas9unfolded protein responseneuroinflammation |
| spellingShingle | Eleonora Pavan Maximiliano Ormazabal Paolo Peruzzo Emilio Vaena Paula Rozenfeld Andrea Dardis CRISPR/Cas9 Editing for Gaucher Disease Modelling International Journal of Molecular Sciences Gaucher disease cellular model acid β-glucosidase CRISPR/Cas9 unfolded protein response neuroinflammation |
| title | 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_short | CRISPR/Cas9 Editing for Gaucher Disease Modelling |
| title_sort | crispr cas9 editing for gaucher disease modelling |
| topic | Gaucher disease cellular model acid β-glucosidase CRISPR/Cas9 unfolded protein response neuroinflammation |
| url | https://www.mdpi.com/1422-0067/21/9/3268 |
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