RT Journal Article
T1 Isogenic GAA-KO Murine Muscle Cell Lines Mimicking Severe Pompe Mutations as Preclinical Models for the Screening of Potential Gene Therapy Strategies.
A1 Aguilar-González, Araceli
A1 González-Correa, Juan Elías
A1 Barriocanal-Casado, Eliana
A1 Ramos-Hernández, Iris
A1 Lerma-Juárez, Miguel A
A1 Greco, Sara
A1 Rodríguez-Sevilla, Juan José
A1 Molina-Estévez, Francisco Javier
A1 Montalvo-Romeral, Valle
A1 Ronzitti, Giuseppe
A1 Sánchez-Martín, Rosario María
A1 Martín, Francisco
A1 Muñoz, Pilar
K1 CRISPR/Cas9 technology
K1 Pompe disease
K1 adeno-associated virus
K1 cellular disease models
K1 lentiviral vectors
K1 optimised GAA (acid alpha-glucosidase)
AB 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.
YR 2022
FD 2022-06-04
LK http://hdl.handle.net/10668/21164
UL http://hdl.handle.net/10668/21164
LA en
DS RISalud
RD Apr 7, 2025