Development of an RNAi therapeutic for alpha-1-antitrypsin liver disease
Christine I. Wooddell, … , Steven B. Kanner, Jeffrey H. Teckman
Christine I. Wooddell, … , Steven B. Kanner, Jeffrey H. Teckman
Published June 18, 2020; First published May 7, 2020
Citation Information: JCI Insight. 2020;5(12):e135348. https://doi.org/10.1172/jci.insight.135348.
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Research Article Hepatology Therapeutics

Development of an RNAi therapeutic for alpha-1-antitrypsin liver disease

Abstract

The autosomal codominant genetic disorder alpha-1 antitrypsin (AAT) deficiency (AATD) causes pulmonary and liver disease. Individuals homozygous for the mutant Z allele accumulate polymers of Z-AAT protein in hepatocytes, where AAT is primarily produced. This accumulation causes endoplasmic reticulum (ER) stress, oxidative stress, damage to mitochondria, and inflammation, leading to fibrosis, cirrhosis, and hepatocellular carcinoma. The magnitude of AAT reduction and duration of response from first-generation intravenously administered RNA interference (RNAi) therapeutic ARC-AAT and then with next-generation subcutaneously administered ARO-AAT were assessed by measuring AAT protein in serum of the PiZ transgenic mouse model and human volunteers. The impact of Z-AAT reduction by RNAi on liver disease phenotypes was evaluated in PiZ mice by measuring polymeric Z-AAT in the liver; expression of genes associated with fibrosis, autophagy, apoptosis, and redox regulation; inflammation; Z-AAT globule parameters; and tumor formation. Ultrastructure of the ER, mitochondria, and autophagosomes in hepatocytes was evaluated by electron microscopy. In mice, sustained RNAi treatment reduced hepatic Z-AAT polymer, restored ER and mitochondrial health, normalized expression of disease-associated genes, reduced inflammation, and prevented tumor formation. RNAi therapy holds promise for the treatment of patients with AATD-associated liver disease. ARO-AAT is currently in phase II/III clinical trials.

Authors

Christine I. Wooddell, Keith Blomenkamp, Ryan M. Peterson, Vladimir M. Subbotin, Christian Schwabe, James Hamilton, Qili Chu, Dawn R. Christianson, Julia O. Hegge, John Kolbe, Holly L. Hamilton, Maria F. Branca-Afrazi, Bruce D. Given, David L. Lewis, Edward Gane, Steven B. Kanner, Jeffrey H. Teckman

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Figure 6

ARO-AAT treatment reduced Z-AAT protein and mRNA in young PiZ mice.

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ARO-AAT treatment reduced Z-AAT protein and mRNA in young PiZ mice. Five...
Five-week-old male PiZ mice given 4 subcutaneous injections (Q2W) of 4 mg/kg ARO-AAT, saline, or RNAi-Control and were euthanized 2 weeks after the final injection (n = 9–16). Untreated baseline mice euthanized at 5 weeks of age (n = 15). (A) Plasma was collected at the indicated times and Z-AAT protein measured, shown as the group mean ± SEM relative to pretreatment. (B) Amounts of Z-AAT mRNA in homogenized liver tissue were quantified relative to the geometric mean of the baseline group. (C and D) The amounts of monomeric and polymeric Z-AAT protein in liver lysates of mice at baseline (5 weeks old) or after receiving 4 Q2W injections and then euthanized (13 weeks old) were measured by semiquantitative Western blotting, shown relative to the baseline group as the mean ± SEM. (E) Representative PAS-D–stained liver sections from mice at baseline or injected Q2W with saline, RNAi-Control, or ARO-AAT. Globule number, globule size, and the area of the liver containing globules were measured by a densitometric method (F, G, and H). Comparisons between groups in panels B–D and F–H were performed using nonparametric Wilcoxon’s rank sum test.