1. Gene therapy rescues GALC activity in CNS tissues of Krabbe‐affected NHP
Gene therapy rescues GALC activity in CNS tissues of Krabbe‐affected NHP ASpecific GALC enzymatic activity in single brain slices of WT untreated (UT; white bars, physiological levels), WT LV.hGALC‐treated (JV02), and Krabbe‐affected LV.hGALC‐treated (JV02) animals (filled gray bars, injected hemisphere; striped gray bars, contralateral hemisphere). Two‐way ANOVA followed by Bonferroni's multiple comparison tests; *P < 0.05, **P < 0.01 versus matched slices of WT UT.B, CSummary of GALC activity in brain (B) and spinal cord (C; n = 3 blocks/animal) samples from JT02 and JV02, as well as from WT and Krabbe UT controls.DChromatographic profile of GALC in selected brain slices and spinal cord (SC) tissue of untreated (UT) WT and LV.hGALC‐treated Krabbe NHP (JT02). Extracts were run through a Sephadex S‐300 gel filtration column. Fractions (0.2 ml) were assayed for enzyme activity using MUGAL substrate in the presence (○) or absence of 11 μM AgNO3 (●).EGALC activity in the CSF of JT02 (pre‐ and post‐treatment) and JV02 (post‐treatment). Dots represent technical replicates.FBeginning at 2 months (m) of age, animals were evaluated monthly for motor performance using a modified Bayley's scale for infant development. For the study animals (JV02 and JT02), scores are reported as pre‐surgery and post‐surgery. Affected animals score significantly lower than normal animals at any age considered. JT02's motor scores increase over time post‐LV.GALC injection, being close to the mean observed in normal juveniles at 5 months. Historical data for normal and affected animals are presented as means plus one standard deviation. Historical data are analyzed by two‐way ANOVA and Bonferroni's posttests, P < 0.05 at 6 months, P < 0.001 at all other ages in affected versus normal animals; n = 5–20 animals/group, except for normal animals at 6 months (n = 1). See also Appendix Table S9.Data information: Data in (A‐C) are represented as floating bars (min to max, line at mean; n = 2–6 blocks/slice). Number of blocks analyzed/animal: 16 (WT UT), 24 (JT02), and 22 (JV02). GALC activity in Krabbe UT samples is < 0.001 nmol/h*mg; n = 3 blocks. One‐way ANOVA and Bonferroni's multiple comparison test (B) or Dunnet's multiple comparison test (C); *P < 0.05, **P < 0.001 versus WT UT.
Vasco Meneghini et al. EMBO Mol Med. 2016;8:
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