Figure 4.

Cerebrospinal Fluid-Directed rAAV9-rsATP7A Gene Therapy Plus Subcutaneous Copper Histidinate Increases Brain Copper Levels in mo-br Mutant Mice

(A) Brain copper levels, measured by ICP-MS, are significantly higher in 12-day-old mutant mice treated with rAAV9-rsATP7A (1.6 × 10¹⁰ vg) plus sc copper histidinate (AAV9 + Cu) compared to untreated mutants (UT). A two-tailed, paired Student’s t test was used to calculate p. (B) X-ray fluorescence microscopy (XFM) of representative 12-day-old mo-br mouse brain sections from untreated (top) and high-dose rAAV9-rsATP7A plus sc copper histidinate-treated (AAV9 + Cu, bottom) mice. Left, H&E-stained tissue sections. Boxed areas in white (left) correspond to the X-ray fluorescence microscopy elemental maps for Cu (middle) and phosphorus (P, right), respectively, the latter obtained as control. The elemental map for Cu shows diffusely increased signals within the choroid plexus of the lateral ventricle (box indicates region of focus) and subventricular zone of the rAAV9-rsATP7A plus sc copper histidinate-treated brain, whereas the phosphorus signals (right) are comparable. Red temperature scale represents copper concentration (0–53 μg/g) and phosphorus concentration 0–10 mg/g or 0–12 mg/g, as noted. Error bars, SEM.