Figure 2.

In vitro testing of the pAAV9.hSynI.GBA plasmid and assessment of neuroinflammatory response to the AAV9.hSYNI.hGBA vector in wild-type animals. (A) Schematic of the AAV9.hSYNI.hGBA expression cassette, where hGBA represents the human GBA gene. (B and C) Western blot analysis and protein quantification following transfection with the pAAV9.hSynI.GBA plasmid showed increased expression of GCase protein compared to controls. (D) Enzymatic assay on cell lysate and supernatant demonstrated that GCase was enzymatically functional, with increased activity in transfected samples. (E) Assessment of neuroinflammation following systemic administration of AAV9.hSYNI.hGBA to neonatal wild-type mice. Immunohistochemical staining for the marker GFAP did not show increase in astrogliosis in brains from injected mice (WT + AAV) in different regions compared to controls (WT—AAV). (F) Staining quantification confirmed normal GFAP levels in cortex (S1BF), hippocampus (CA1/CA2) and thalamus (VPM/VPL) in injected mice (WT + AAV).