Multiscale and cross-species evolution of adeno-associated viruses for kidney gene transfer. Adeno-associated virus

Gene delivery to the kidney has proven difficult, limiting the translation of genetic medicines to over 10% of the global population suffering from chronic kidney disease. Here, we evolve adeno-associated viral (AAV) capsid libraries by cross-species cycling in different kidney models. Multiple new variants, notably, AAV.k13 and AAV.k20 were enriched following sequential intravenous (IV) cycling through mouse and pig kidneys, ex vivo cycling in human organoid cultures, and ex vivo machine perfusion in isolated non-human primate (NHP) kidneys. Following IV administration, these vectors display robust and widespread transduction in murine kidneys, with selective tropism for proximal tubules. Markedly higher transgene expression compared to parental AAV9 vectors was also observed in proximal tubule epithelial cells within human organoid cultures and in transplanted pig kidneys. Moreover, following ureteral delivery, AAV.k20 was capable of efficiently transducing pig and NHP kidneys, underscoring potential for clinical translation. AAV.k13 and AAV.k20 variants are promising vectors for therapeutic gene transfer applications in kidney diseases and transplantation.