Fig. 8: A LICA1 showed cross-species enhanced transduction in the skeletal muscle.
From: An engineered AAV targeting integrin alpha V beta 6 presents improved myotropism across species
Scheme of comparative study between multiple AAV capsids in multiple species. Transgene expression cassette is ITR_tMCK_hCAPN3-inactive_BC_SV40pA_ITR. Barcodes (BCs) allow quantitative measurements of transgene expression mRNA level. 2/3 barcodes were used for each capsid to minimize the sequence bias. AAV production of each capsid variant was done separately, before pooled together at the equimolar amount before AAV transduction in vitro (C, human myotube, n = 2 biological replicates, dose: 2E10/2E11 vg per 12wp well, duration: 48 h), or in vivo injection in C57Bl6 mice (B, n = 3 mice, dose: 5E12 vg/kg per capsid variant) and Macaca fascicularis NHP (D, n = 3 NHPs, dose: 3.2E12 vg/kg per capsid variant). Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en). B The mRNA enrichment (BC_mRNA/Rplp0/ BC_AAV) of different capsid variants in the liver and three skeletal muscles in C57Bl6 mice measured by RT-qPCR. Each dot correponds to the individual barcode. The color of each barcode corresponds to the individual mouse in Fig. S6C. The statistics were performed on the average on different barcodes (using for the same capsid variant in each mouse) by using two-way ANOVA ( ~ mice * capsids) followed by FDR correction. C The mRNA enrichment, measured by NGS, of different capsid variants in in vitro human myotubes at two AAV concentration. The log2FC compared to AAV9 of all variants is presented. Each dot correponds to the individual barcode. The colors correpsond to different biological replicates. D The mRNA enrichment (BC_mRNA/Rplp0/BC_AAV) of different capsid variants in the liver and skeletal muscles in NHP measured by RT-qPCR. Each dot correponds to the individual barcode. The data was presented for individual primates (P1-3). The statistics were to compare the mRNA level of LICA1 with natural capsids, AAV8/AAV9, performed on the fold change of LICA1 to AAV8/AAV9 (averages of barcodes using for the same capsid variant in each primate were used to calculate the fold-change) by using two-tailed one-sample t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001; ns not significant. Data in (B) and (D) are presented as mean ± SEM. Source data are provided as a Source Data file.
