Chimeric AAV Capsids for High Efficiency Nucleic Acid Transfer

Researchers in Prof. Mark Kay's laboratory have developed recombinant adeno-associated viral (AAV) capsid proteins that transduce human primary hepatocytes at high efficiency in vitro and in vivo. This vector (AAV-LK03) was engineered through a unique protocol of DNA shuffling and selection using a humanized murine model. This method facilitated the final selection of a vector that is capable of highly efficient functional transduction, leading to a high level of transgene expression. The AAV-LK03 vector is a highly promising clinical candidate for gene therapy applications and could also be used for research that requires genetic manipulation of xenotransplanted cells.

AAV-LK03 is selective for human hepatocytes in vivo. The newly engineered AAV chimera (AAV-LK03) selectively transduced human hepatocytes (in humanized FRG animals – hFRG) but not in non-humanized controls (Ctr). Conversely, the AAV3 vector did not transduce either type of mouse and the AAV8 vector was not specific for human hepatocytes (as shown by transduction of the Ctr mouse). Human albumin (hAlb) levels and estimated percentages of repopulation with human hepatocytes are given for each animal.

Stage of Research
The inventors have engineered the chimeric AAV vector and demonstrated its efficacy in vitro and in vivo (43% mean transduction in human hepatocytes).