Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies
Pompe disease (PD) is a rare disorder caused by mutations in the acid alpha-glucosidase (GAA) gene. Most gene therapies (GT) partially rely on the cross-correction of unmodified cells through the uptake of the GAA enzyme secreted by corrected cells. In the present study, we generated isogenic murine...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/11/6298 |
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| author | Araceli Aguilar-González Juan Elías González-Correa Eliana Barriocanal-Casado Iris Ramos-Hernández Miguel A. Lerma-Juárez Sara Greco Juan José Rodríguez-Sevilla Francisco Javier Molina-Estévez Valle Montalvo-Romeral Giuseppe Ronzitti Rosario María Sánchez-Martín Francisco Martín Pilar Muñoz |
| author_facet | Araceli Aguilar-González Juan Elías González-Correa Eliana Barriocanal-Casado Iris Ramos-Hernández Miguel A. Lerma-Juárez Sara Greco Juan José Rodríguez-Sevilla Francisco Javier Molina-Estévez Valle Montalvo-Romeral Giuseppe Ronzitti Rosario María Sánchez-Martín Francisco Martín Pilar Muñoz |
| author_sort | Araceli Aguilar-González |
| collection | DOAJ |
| description | Pompe disease (PD) is a rare disorder caused by mutations in the acid alpha-glucosidase (GAA) gene. Most gene therapies (GT) partially rely on the cross-correction of unmodified cells through the uptake of the GAA enzyme secreted by corrected cells. In the present study, we generated isogenic murine GAA-KO cell lines resembling severe mutations from Pompe patients. All of the generated GAA-KO cells lacked GAA activity and presented an increased autophagy and increased glycogen content by means of myotube differentiation as well as the downregulation of mannose 6-phosphate receptors (CI-MPRs), validating them as models for PD. Additionally, different chimeric murine GAA proteins (IFG, IFLG and 2G) were designed with the aim to improve their therapeutic activity. Phenotypic rescue analyses using lentiviral vectors point to IFG chimera as the best candidate in restoring GAA activity, normalising the autophagic marker p62 and surface levels of CI-MPRs. Interestingly, in vivo administration of liver-directed AAVs expressing the chimeras further confirmed the good behaviour of IFG, achieving cross-correction in heart tissue. In summary, we generated different isogenic murine muscle cell lines mimicking the severe PD phenotype, as well as validating their applicability as preclinical models in order to reduce animal experimentation. |
| first_indexed | 2024-03-10T01:12:52Z |
| format | Article |
| id | doaj.art-132e0370e4fc4c1589e3ab86c5f7f940 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:12:52Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-132e0370e4fc4c1589e3ab86c5f7f9402023-11-23T14:13:06ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-06-012311629810.3390/ijms23116298Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy StrategiesAraceli Aguilar-González0Juan Elías González-Correa1Eliana Barriocanal-Casado2Iris Ramos-Hernández3Miguel A. Lerma-Juárez4Sara Greco5Juan José Rodríguez-Sevilla6Francisco Javier Molina-Estévez7Valle Montalvo-Romeral8Giuseppe Ronzitti9Rosario María Sánchez-Martín10Francisco Martín11Pilar Muñoz12GENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainInstituto de Investigación del Hospital Universitario La Paz, IdiPAZ, 28029 Madrid, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGénéthon, Integrare Research Unit UMR_S951, INSERM, Université Paris-Saclay, Univ Evry, 91002 Evry, FranceGénéthon, Integrare Research Unit UMR_S951, INSERM, Université Paris-Saclay, Univ Evry, 91002 Evry, FranceGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainGENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government PTS Granada-Avenida de la Ilustración 114, 18016 Granada, SpainPompe disease (PD) is a rare disorder caused by mutations in the acid alpha-glucosidase (GAA) gene. Most gene therapies (GT) partially rely on the cross-correction of unmodified cells through the uptake of the GAA enzyme secreted by corrected cells. In the present study, we generated isogenic murine GAA-KO cell lines resembling severe mutations from Pompe patients. All of the generated GAA-KO cells lacked GAA activity and presented an increased autophagy and increased glycogen content by means of myotube differentiation as well as the downregulation of mannose 6-phosphate receptors (CI-MPRs), validating them as models for PD. Additionally, different chimeric murine GAA proteins (IFG, IFLG and 2G) were designed with the aim to improve their therapeutic activity. Phenotypic rescue analyses using lentiviral vectors point to IFG chimera as the best candidate in restoring GAA activity, normalising the autophagic marker p62 and surface levels of CI-MPRs. Interestingly, in vivo administration of liver-directed AAVs expressing the chimeras further confirmed the good behaviour of IFG, achieving cross-correction in heart tissue. In summary, we generated different isogenic murine muscle cell lines mimicking the severe PD phenotype, as well as validating their applicability as preclinical models in order to reduce animal experimentation.https://www.mdpi.com/1422-0067/23/11/6298Pompe diseaselentiviral vectorscellular disease modelsoptimised GAA (acid alpha-glucosidase)CRISPR/Cas9 technologyadeno-associated virus |
| spellingShingle | Araceli Aguilar-González Juan Elías González-Correa Eliana Barriocanal-Casado Iris Ramos-Hernández Miguel A. Lerma-Juárez Sara Greco Juan José Rodríguez-Sevilla Francisco Javier Molina-Estévez Valle Montalvo-Romeral Giuseppe Ronzitti Rosario María Sánchez-Martín Francisco Martín Pilar Muñoz Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies International Journal of Molecular Sciences Pompe disease lentiviral vectors cellular disease models optimised GAA (acid alpha-glucosidase) CRISPR/Cas9 technology adeno-associated virus |
| title | Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies |
| title_full | Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies |
| title_fullStr | Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies |
| title_full_unstemmed | Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies |
| title_short | Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies |
| title_sort | isogenic gaa ko murine muscle cell lines mimicking severe pompe mutations as preclinical models for the screening of potential gene therapy strategies |
| topic | Pompe disease lentiviral vectors cellular disease models optimised GAA (acid alpha-glucosidase) CRISPR/Cas9 technology adeno-associated virus |
| url | https://www.mdpi.com/1422-0067/23/11/6298 |
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