Effect of Adeno-Associated Virus Serotype and Genomic Structure on Liver Transduction and Biodistribution in Mice of Both Genders

Abstract

Recombinant adeno-associated viral (AAV) vectors have unique properties, which make them suitable vectors for gene transfer. Here we assess the liver transduction efficiency and biodistribution of AAV-pseudotyped capsids (serotypes) 1, 5, 6, and 8, combined with single-stranded and double-stranded genomic AAV2 structures carrying the luciferase reporter gene after systemic administration. The analysis was performed in vivo and ex vivo, in male and female mice. Gender-related differences in AAV-mediated transduction and biodistribution were shown for the four serotypes. Our data confirm the superiority of AAV8 over the rest of the serotypes, as well as a significant advantage of double-stranded genomes in terms of liver transduction efficiency, particularly in females. Regarding biodistribution, AAV5 displayed a narrower tropism than the other serotypes tested, transducing, almost exclusively, the liver. Interestingly, AAV1 and AAV8, in particular those having single-stranded genomes, showed high transduction efficiency of female gonads. However, no inadvertent germ line transmission of AAV genomes was observed after breeding single-stranded AAV8-injected female mice with untreated males. In conclusion, double-stranded AAV8 vectors led to the highest levels of liver transduction in mice, as demonstrated by luciferase expression. Nevertheless, the transduction of other organs with AAV8 vectors could favor the use of less efficient serotypes, such as AAV5, which display a narrow tropism.

Adeno-associated virus serotype 9-mediated pulmonary transgene expression: effect of mouse strain, animal gender and lung inflammation

“…Besides mouse strain, animal gender has also been shown to influence rAAV-mediated gene delivery, transgene expression and biodistribution. 27,28 In a study addressing liver-directed delivery of vector particles, Davidoff et al 27 reported 5-to 13-fold higher transgene expression in male compared with female mice, irrespective of the promoter, cDNA and mouse strain. Moreover, molecular analysis revealed that the rAAV genome was stably retained in male liver at levels that were sevenfold higher than those observed in females.…”

“…These differences were observed with both rAAV2-and rAAV5-based vectors. In a relatively recent study, Paneda et al 28 reported gender-related differences in rAAV-mediated liver transduction and biodistribution for the serotypes 1, 5, 6 and 8. To the best of our knowledge, there are no prior studies comparing the effect of animal gender on AAV2/9-mediated pulmonary gene transfer.…”

“…26 Selective tropism of AAV2/9 for alveolar epithelium 8,14 makes SPB À/À mice a highly suitable animal model to asses the efficiency of AAV2/9 vector in an inflamed lung. Since animal gender has also been shown to influence AAV-mediated gene delivery, transgene expression and biodistribution, 27,28 experiments in the present study were performed in both male and female mice.…”

AAV vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers

“…The vector of choice was dsAAV1-PBGDLuc, which expresses luciferase from a ubiquitous porphobilinogen deaminase promoter (PBGD) and contains a double-stranded genome (Figure 1a). 20 We considered that transduction of AAV vectors could be different depending on the route of administration used for viral entry. Therefore, we introduced 3.4Â10 10 viral genomes of dsAAV1-PBGD-Luc through the hepatic artery (n¼6), the portal vein (n¼6) or by direct injection into the liver (n¼6).…”

“…The clear benefit is that transduction from dsAAV is faster and 10-to 20-fold higher than transduction from standard single-stranded AAV (ssAAV). [18][19][20] Transgene expression mediated by AAVs can persist for several years in vivo. 21,22 AAV genomes persist in non-dividing cells as episomal DNAs, but a very low level of integration could also occur.…”