Single cell analysis of naïve or AAV transduced NSG-PiZ mouse livers

To determine whether different hepatotropic AAV capsids can mediate transduction of human hepatocytes in liver-humanized chimeric mice we performed an experiment in NSG-PiZ mice. NSG-PiZ mice were simultaneously administered barcoded AAV vectors with capsids derived from AAV8, AAV9, AAVrh10, AAVDJ/8 and AAV.LK03 at a total dose of 1E12 vg per mouse. At 14 days post delivery naïve and AAV treated mice were sacrificed and livers perfused to generate single cell suspensions for scRNA-seq. We transplanted mice with human hepatocytes from 2 donors (4 mice per donor) and half of the mice from each donor were adminsitered AAV. Overall design: Four NSG-PiZ mice were transplanted with human hepatocytes from Donor A and 4 NSG-PiZ mice were transplanted with human hepatocytes from Donor B. Two Donor A and 2 Donor B mice were kept as untransduced contols. The remaining 2 Donor A and 2 Donor B mice were transduced with a mixture of AAV8 (2E11 vgs/mouse), AAV9 (2E11 vgs/mouse), AAVrh10 (2E11 vgs/mouse), AAVDJ/8 (2E11 vgs/mouse) and AAV.LK03 (2E11 vgs/mouse) for a total dose of 1E12 vgs by retroorbital IV injection. The AAVs carried a barcoded GFP gene specific to each serotype. At 14 days post transduction, hepatic cells were collected and pooled from the 4 untransduced mice and separately from the 4 AAV transduced mice and immediately processed for 10X single cell RNA seq.