不同AAV突变衣壳在食蟹猴脑的转导效果分析OA

Objective To systematically evaluate the neuronal transduction efficiency of adeno-associated virus(AAV)capsid variants,including AAV-PHP.B and AAV-PHP.eB,in the brain of cynomolgus monkeys(Macaca fascicularis),and to analyze the effects of different injection doses and routes on whole-brain infection.Methods Recombinant AAVs carrying the hSyn-EYFP expression cassette packaged into AAV9,AAV-PHP.B,and AAV-PHP.eB capsids were administered to cynomolgus monkeys through different routes(intracerebroventricular injection(ICV),intracisternal magna injection(ICM),and intravenous injection(IV))and at different doses.After 6 weeks of transgene expression,the animals were perfused,and brain tissues were collected for cryosectioning and immunofluorescence staining.The ratio of EYFP and NeuN co-labeled cells to the total NeuN-positive cells in regions including the prefrontal cortex,caudate/putamen nucleus,hippocampus,and thalamus was quantified to assess neuronal transduction efficiency.Results Following ICV injection,low-dose AAV-PHP.B(1×1013 vg/kg)demonstrated significantly higher neuronal transduction efficiency than AAV9 in the prefrontal cortex(8.4％vs.1.2％,P<0.000 1)and caudate/putamen nucleus(6.3％vs.0.5％,P<0.000 1).Increasing the AAV-PHP.B dose to 5×1013 vg/kg further enhanced its efficiency in the caudate/putamen nucleus(P<0.000 1).Combined ICV and ICM injection synergistically enhanced the whole-brain transduction of AAV-PHP.B,particularly in the prefrontal cortex(14.7％)and thalamus(2.6％).When delivered using the combined route,AAV-PHP.eB showed significantly superior transduction efficiency in deep brain structures such as the hippocampus and thalamus compared with AAV-PHP.B(P<0.01).In contrast,intravenous injection of AAV-PHP.eB failed to achieve efficient transduction(<1％)in any brain region examined.Conclusions The AAV-PHP.B and AAV-PHP.eB capsids exhibited superior transduction capabilities in the cynomolgus monkey brain compared with AAV9.Combined ICV and ICM injection represents a highly effective approach for achieving widespread neuronal transduction throughout the brain.This study provides crucial experimental evidence for selecting appropriate AAV delivery tools and optimizing administration protocols for non-human primate studies and future clinical translation.