Fig. 5: Derivation, empirical training, and validation of in silico gene therapy efficacy model (GETEM) for Pompe disease correction in a developing infant.

a Schematic showing enzymatic cross-correction of somatic tissues by injected rhGAA (enzyme replacement therapy, ERT). For a given developing liver of a patient, progenitor cells proliferate and can asymmetrically divide into mature cells. Injected rhGAA is absorbed by the liver as well as striated muscle tissue (both heart and skeletal muscle). b Percentage of normal cardiac tissue within a heart of a Pompe diseased infant after one year of biweekly ERT at 20 mg/kg. Percentages of normal skeletal muscle tissue and hepatocytes are in Supplementary Fig. 9. c Schematic for in silico gene editing model for previously published precise correction of a single base pair in mouse disease models, d Flowchart for training and validating the GETEM model using previously published studies of in vivo somatic cell genome editing of a mouse liver by intravenous injection. e (top) Absolute change of edited cells over time predicted by the GETEM for the studies utilized in (d). For studies that employed selection in the liver, arrows indicate the time at which the selective pressure favoring edited cells was applied (for Fah^−/− models, this indicates the removal of NTBC supplementation in the diet; and, for Otc editing, this indicates the induction of a high protein diet.). f Validation of the model using additional published editing strategies. Arrows on days 21 and 24 indicate removal of selection pressure on the treated mice as per published experimental protocol. Arrow on day 2 in the mRNA LNP indicates redosing of the LNP. (n represents biological replicates as previously reported in the literature, mean ± s.d). g Percentage of total sequencing reads from primary fibroblasts of Pompe diseased patients that were treated with S1mplexes targeting W746X D645N or R660H GAA mutations. Results indicate gene correction and imprecise editing for three different mutations. h Percentage of edited alleles that are precisely edited in (g).