Journal: Molecular Therapy. Methods & Clinical Development
Article Title: FAP-CAR-T cells reduce dystrophic muscle fibrosis, improving adeno-associated virus gene transfer efficacy
doi: 10.1016/j.omtm.2025.101545
Figure Lengend Snippet: Combined therapy: Gene transfer efficiency (A) Schematic representation of the in vivo protocol. Two-month-old D2.Mdx mice were injected or not with two consecutive doses (1 × 10 6 ) of control T cells or of FAP-CAR-T cells, and after 2 weeks of treatment the mice were injected with a single intravenous dose (5 × 10 12 VG/kg) of rAAV9-microdystrophin (μDystrophin or μDys). Tissues were collected 6 weeks after the AAV injection, 4 animals were included per group and statistical analyses used one-way ANOVA tests with p values indicated above the compared conditions. (B) Quantification of VGCN with ddPCR in TA, EDL, and DIA in DBA2-MDX mice. (C) Gene expression analysis of microdystrophin in TA, EDL, and DIA of DBA2-MDX mice. Results are shown as a histogram of relative abundance of gene expression over P0 normalizer gene. (D) Quantification of microdystrophin expression after capillary western blot with DYS-B antibody in TA, EDL, and DIA at 6 weeks post-injection in mice. Values are represented as normalized area under the curve (AUC). (E) Capillary western blot of protein lysates of TA at 6 weeks post-injection in mice. Microdystrophin are visible around 139 kDa. (F) Representative microdystrophin IHF pictures of the TA, EDL, and DIA at 6 weeks post-injection in mice. Images include a scale bar. (G) Quantification of dystrophin positive myofibers in TA, EDL, and DIA at 6 weeks post-injection in D2.mdx mice. The percentage of dystrophin+ fibers is represented of number of dystrophin positive fibers over fibers positive for laminin.
Article Snippet: The DBA2 (B6; 129S4-DBA2tm1Cpr/J, strain no. 000671) (D2) controls mice were purchased directly from Charles River Laboratories) while the DBA2/mdx (D2.B10-Dmdmdx/J, strain no. 013141) (D2.mdx) mice were bred and obtained from the Center d’Exploration et de Recherche Fonctionnelle Expérimentale (CERFE) in Evry, France.
Techniques: In Vivo, Injection, Control, Gene Expression, Expressing, Western Blot