Extended Data Fig. 10: AAV-NoSTOP(UGA) restores TPP1 activity in Cln2-R207X(TGA) mice and in patient-derived fibroblasts.
From: An engineered UGA suppressor tRNA gene for disease-agnostic AAV delivery
(a) Workflow of in vivo assessment of the therapeutic efficacy of MyoAAV.2xCuO-hNFS-R8(VG4) in Cln2KI/KI mice. (b) Bar graph showing the TPP1 enzymatic activities in the indicated tissues collected from untreated wild-type mice (WT), untreated Tpp1KI/KI mice, or Tpp1KI/KI mice treated with MyoAAV.2xCuO-hNFS-R8(VG4) and sacrificed at 4 weeks post-treatment. Data are normalized to the WT levels as 100%. (c) Bar graph showing the ratios of mouse Tpp1 mRNA to Tbp mRNA (mTpp1/mTbp) in the indicated tissues collected from untreated wild-type mice (WT), untreated Tpp1KI/KI mice, or treated Tpp1KI/KI mice. In b and c, each dot represents an individual animal (n = 5 or 6). Data are presented as mean and standard deviation. Statistical analysis was performed with two-sided unpaired t-test in b, or one-way ANOVA followed by Dunnett’s multiple comparisons test in c. (d) Schematics showing the workflow of delivering the CuO-hNFS-R8 construct to patient-derived fibroblasts via lentiviral vector transduction. (e) Bar graph showing the TPP1 activity in untreated normal fibroblasts, or the fibroblasts from a CLN2 patient (CLN2R127X/R208X) treated with a control lentiviral vector expressing EGFP (Lenti.EGFP), Lenti.CuO-hNFS-R8, or G418. Each dot represents a biological repeat (n = 3). Data are presented as mean and standard deviation. Statistical analysis was performed with one-way ANOVA followed by Dunnett’s multiple comparisons test. Schematic in a and d created in BioRender. Zou, H. (2025) https://BioRender.com/dqjni51.
