Selective tropism of the recombinant adeno‐associated virus 9 serotype for rat cardiac tissue

Abstract

Cardiac gene transfer may serve as a novel therapeutic approach for heart disease. Numerous serotypes of recombinant adeno-associated virus (rAAV) have been identified with variable tropisms to cardiac tissue. Both in vitro and in vivo experiments were undertaken to compare cardiac tropisms of rAAV-2, 5, 7, 8 and 9. For the in vitro studies, 10(7) vector genome (vg) of rAAV-2, 5, 7, 8 or 9 were used to transduce both rat neonatal cardiac myocytes (RNCM) and fibroblasts (RNCF). For the in vivo studies, 4 x 10(10) vg of rAAV-2, 5, 7, 8 or 9, and 4 x 10(11) vg of rAAV8 or 9 were administered in 5-day-old rats via a relatively non-invasive intracardiac injection. One and two months post-administration, green fluorescent protein (GFP) expression in tissues was visualized and GFP mRNA was quantified by the real-time polymerase chain reaction. At 3 days post-viral transduction, rAAV9 and rAAV2 produced the highest transducing efficiency in RNCM. Only rAAV2 elicited any transduction in the RNCF. The results obtained in vivo indicated that the order for transduction efficiency in the heart was: rAAV9 > rAAV8 > rAAV7 > rAAV2 = rAAV5. The transduction efficiency order in the liver was: rAAV2 > rAAV5 > rAAV7 > rAAV8 > rAAV9. Injection of a higher dose (4 x 10(11) vg) of rAAV9 provided more widespread and highly cardiac-selective GFP expression in the heart than rAAV8. Zero to minimal expression of GFP was found in the lung and kidney for both doses of all rAAV serotypes utilized. Collectively, the results obtained in the present study suggest that rAAV9 provides the most selective and stable transduction efficiency in cardiac tissue, and this expression was primarily exhibited in cardiac myocytes.