Common AAV gene therapy vectors show nonselective transduction of ex vivo human brain tissue

We have adapted an ex vivo organotypic model to evaluate AAV transduction properties in living slices of human brain tissue. Using single-nucleus RNA sequencing, we found that common AAV vectors show broad transduction of normal cell types, with protein expression most apparent in astrocytes; this work introduces a pipeline for identifying and optimizing AAV gene therapy vectors in human brain samples. Overall design: Sequencing data includes 10X 3' single-nucleus RNA sequencing of patient samples flash frozen: day 0 control tissue, day 14 cultured tissue without vectors, and day 14 tissue to which vectors were applied. Tissues slices underwent Miltenyi Biotec nuclei isolation followed by nuclei FACS sorting, and were processed with the 3' 10X Genomics single-nucleus RNA isolation protocol.