We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies.
- Authors
Aguilar-González, Araceli; González-Correa, Juan Elías; Barriocanal-Casado, Eliana; Ramos-Hernández, Iris; Lerma-Juárez, Miguel A.; Greco, Sara; Rodríguez-Sevilla, Juan José; Molina-Estévez, Francisco Javier; Montalvo-Romeral, Valle; Ronzitti, Giuseppe; Sánchez-Martín, Rosario María; Martín, Francisco; Muñoz, Pilar
- Abstract
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.
- Subjects
GENE therapy; CELL lines; MUSCLE cells; ANIMAL models in research; GLYCOGEN storage disease type II; GLUCOSIDASES
- Publication
International Journal of Molecular Sciences, 2022, Vol 23, Issue 11, p6298
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms23116298