Antisense oligonucleotide treatment ameliorates alpha-1 antitrypsin–related liver disease in mice
Shuling Guo, … , Michael L. McCaleb, Brett P. Monia
Shuling Guo, … , Michael L. McCaleb, Brett P. Monia
Published December 20, 2013
Citation Information: J Clin Invest. 2014;124(1):251-261. https://doi.org/10.1172/JCI67968.
View: Text | PDF
Research Article Article has an altmetric score of 23

Antisense oligonucleotide treatment ameliorates alpha-1 antitrypsin–related liver disease in mice

Abstract

Alpha-1 antitrypsin deficiency (AATD) is a rare genetic disease that results from mutations in the alpha-1 antitrypsin (AAT) gene. The mutant AAT protein aggregates and accumulates in the liver leading to AATD liver disease, which is only treatable by liver transplant. The PiZ transgenic mouse strain expresses a human AAT (hAAT) transgene that contains the AATD-associated Glu342Lys mutation. PiZ mice exhibit many AATD symptoms, including AAT protein aggregates, increased hepatocyte death, and liver fibrosis. In the present study, we systemically treated PiZ mice with an antisense oligonucleotide targeted against hAAT (AAT-ASO) and found reductions in circulating levels of AAT and both soluble and aggregated AAT protein in the liver. Furthermore, AAT-ASO administration in these animals stopped liver disease progression after short-term treatment, reversed liver disease after long-term treatment, and prevented liver disease in young animals. Additionally, antisense oligonucleotide treatment markedly decreased liver fibrosis in this mouse model. Administration of AAT-ASO in nonhuman primates led to an approximately 80% reduction in levels of circulating normal AAT, demonstrating potential for this approach in higher species. Antisense oligonucleotides thus represent a promising therapy for AATD liver disease.

Authors

Shuling Guo, Sheri L. Booten, Mariam Aghajan, Gene Hung, Chenguang Zhao, Keith Blomenkamp, Danielle Gattis, Andrew Watt, Susan M. Freier, Jeffery H. Teckman, Michael L. McCaleb, Brett P. Monia

×

Figure 6

Robust AAT reduction in cynomolgus hepatocytes and in vivo after AAT-ASO treatment.

Options: View larger image (or click on image) Download as PowerPoint
Robust AAT reduction in cynomolgus hepatocytes and in vivo after AAT-ASO...
(A) Dose-dependent reduction of AAT mRNA in primary hepatocytes isolated from cynomolgus monkeys. Cells were electroporated in growth medium in the presence of AAT-ASO at the indicated concentrations and plated. Twenty-four hours after transfection, total cellular RNA was isolated and the amount of AAT mRNA present was quantitated using a qRT-PCR assay (TaqMan). Results represent mean ± SD (n = 3) compared to untransfected control. (B) AAT mRNA levels in livers. (C) Plasma AAT protein levels during the treatment period. Cynomolgus monkeys were treated with AAT-ASO for 12 weeks at 25 mg/kg 3 times in the first week and twice a week in weeks 2 to 12. Liver total RNA was purified, and qRT-PCR was carried out to determine the levels of AAT mRNA in the liver. The plasma protein levels were measured by the immunoturbidimetry method using a clinical analyzer. Results represent the mean ± SD (n = 4). *P < 0.05, **P < 0.01 by 2-way ANOVA with Bonferroni’s post-hoc tests for A and C and Student’s t test for B.