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

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

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Figure 1

Characterization of an ASO that effectively reduces AAT levels in cells.

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Characterization of an ASO that effectively reduces AAT levels in cells....
(A) Dose-dependent reduction of AAT mRNA in HepG2 cells. (B) Dose-dependent reduction of hAAT mRNA in primary hepatocytes isolated from PiZ mice. 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 (UTC). (C) Dose-dependent reduction of hAAT mRNA levels in PiZ mice after AAT-ASO treatment. (D) Dose-dependent reduction of circulating hAAT protein in PiZ mice after AAT-ASO treatment. Six-week-old PiZ mice were treated for 4 weeks with the indicated doses of AAT-ASO via subcutaneous injection. hAAT mRNA levels in livers were quantified by qRT-PCR (TaqMan), and plasma AAT levels were determined by an immunoturbidimetric method. Results represent mean ± SD (n = 4). *P < 0.05, **P < 0.01 by 2-way ANOVA with Bonferroni’s post-hoc tests for A, B, and D; 1-way ANOVA with Tukey’s comparisons for C.