Journal: Molecular Therapy
Article Title: Reversion of metabolic dysfunction-associated steatohepatitis by skeletal muscle-directed FGF21 gene therapy
doi: 10.1016/j.ymthe.2024.10.023
Figure Lengend Snippet: AAV1-FGF21 biodistribution and biological activity in dogs Healthy Beagle dogs (dog-1 and dog-2) were treated with 7 × 10 12 vg/kg of AAV1-canine optimized FGF21 and followed up for 4 months. (A) Vector genome copy number (left panel, blue) and canine optimize FGF21 ( coFGF21 ) expression (right panel, orange) were analyzed in tissue punches from multiple regions of the skeletal muscle and the liver obtained during necropsy of dogs. (B) Schematic representation indicating the mean coFGF21 expression in hindlimb skeletal muscles. Image courtesy of IMAIOS️ (Micheau A, Hoa D, e-Anatomy, www.imaios.com , https://doi.org/10.37019/e-anatomy ). (C) Levels of biologically active FGF21 in fasted conditions measured using a cell-based reporter gene assay (iLite). (D) Hepatic expression levels of KLB ( n = 2–3/group). (E) Serum triglyceride levels pre-AAV and 4 months post-AAV ( n = 1–3). (F and G) Hepatic expression levels of ACADM (F) and ACADL (G) ( n = 2–3/group). (H) Serum adiponectin levels pre-AAV and 4 months post-AAV ( n = 1–3). (I–L) Quantification of ADIPOQ (I), UCP1 (J), ELOVL3 (K), and PPARGC1A (L) expression in perirenal (prWAT) and gluteal WAT (gWAT) ( n = 2–3/group). (M) Representative images of the hematoxylin and eosin staining of prWAT sections. Insets show multilocular adipocytes in prWAT of AAV-FGF21-treated dogs. Scale bars, 100 μm. Inset scale bars, 25 μm. Data are presented as mean ± SD. Data were analyzed using a Mann-Whitney two-tailed test. coFGF21, canine optimized FGF21; ND, non-detected; vg/dg, vector genomes/diploid genome; AU, arbitrary units; FC, fold change; G, gluteus; T, tensor; Q, quadriceps; V, vastus; LLL, left lateral lobe; LML, left medial lobe; RML, right medial lobe.
Article Snippet: Canine adiponectin in serum was measured using the Human adiponectin ELISA High sensitivity kit (RD191023100, BioVendor Group), which has been previously validated for reliable measurement of canine adiponectin., Serum FGF21 levels in dogs remained undetectable when measured with four different canine FGF21 ELISAs (E08F0016, Blue Gene; MBS028669, MyBiosource; ABIN1053687, Antibodies-online; KT-100218, Kamiya Biomedical Company).
Techniques: Activity Assay, Plasmid Preparation, Expressing, Muscles, Reporter Gene Assay, Staining, MANN-WHITNEY, Two Tailed Test