45 Our study establishes the utility of two different AAV serotypes (AAV8 and AAV2) for hepatic gene targeting of both adult and neonatal mice in vivo. Interestingly, the biology of these two serotypes differed considerably in terms of gene targeting. Different kinetics for the two serotypes have been described previously with gene addition approaches wherein higher doses of AAV correlated NVP-BKM120 concentration with higher levels of gene expression.36 Here, we observed a similar phenomenon where the highest doses administered produced the greatest gene repair frequencies
in vivo. Targeting was confirmed by immunohistochemistry, RT-PCR, and functional measures of liver correction using serum liver function tests. We also evaluated the frequency of random integration in cells with proper gene repair
using coinjection with a second, nonselectable AAV vector. The average copy number of the irrelevant vector corrected for repopulation efficiency indicated that 0.5%-1% of targeted cells also had a random integration. This number is similar to multiple estimates of random integration of AAV8 from the literature.35 Therefore, it can be concluded that gene repair does not result in a higher random integration frequency. In summary, our experiments demonstrated stable hepatic gene repair in both adult and neonatal mice with AAV-Fah serotypes 2 selleck compound and 8. Serial transplantation was possible without difficulty and serially reconstituted animals had normal hepatic function. Most importantly, this work was the first to show functional
metabolic correction of a disease model using AAV-mediated gene repair and can be envisioned as a therapeutic strategy for disorders with a selective advantage in corrected cells. Although these experiments focused on correcting the metabolic disease HTI, the novel approach described herein can serve as a model for gene repair in any monogenic disease caused by point mutations. We thank Angela Major for histology support and Terry Storm for AAV preparations. “
“Background and Aims: Non-invasive diagnosis of compensated cirrhosis is important. We therefore compared liver stiffness by transient elastography, APRI score, AST/ALT ratio, hyaluronic acid and clinical signs to determine which modality Methamphetamine performed best at identifying compensated cirrhosis. Methods: Patients undergoing evaluation at a single center were recruited and had clinical, serological, endoscopy, radiological imaging, liver stiffness measurement and liver biopsy. Patients were stratified into cirrhotic and non-cirrhotic. Results: In 404 patients (124 cirrhosis), transient elastography was diagnostically superior to the other modalities yielding an AUC 0.9 ± 0.04 compared with hyaluronic acid (AUC 0.81 ± 0.04: P < 0.05), clinical signs (AUC 0.74 ± 0.04: P < 0.05), APRI score (AUC 0.71 ± 0.03: P < 0.05) and AST/ALT ratio (AUC 0.66 ± 0.03: P < 0.05).