Fluphenazine Reduces Proteotoxicity in C. elegans and Mammalian Models of Alpha-1-Antitrypsin Deficiency

• Published in: PloS one Vol. 9; no. 1; p. e87260
• Main Authors: Li, Jie, Pak, Stephen C., O’Reilly, Linda P., Benson, Joshua A., Wang, Yan, Hidvegi, Tunda, Hale, Pamela, Dippold, Christine, Ewing, Michael, Silverman, Gary A., Perlmutter, David H.
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 31.01.2014; Public Library of Science (PLoS)
• Subjects: a1-antitrypsin; Aberration; Accumulation; alpha 1-Antitrypsin - genetics; alpha 1-Antitrypsin - metabolism; alpha 1-Antitrypsin Deficiency - genetics; alpha 1-Antitrypsin Deficiency - metabolism; alpha 1-Antitrypsin Deficiency - prevention & control; Analysis; Animal models; Animals; Animals, Genetically Modified; Antipsychotic Agents - pharmacology; Autophagy; Autophagy - drug effects; Autophagy - genetics; Biology; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Cell culture; Cell cycle; Cell death; Cells (Biology); CHO Cells; Cricetinae; Cricetulus; Deficiency diseases; Disease Models, Animal; Drug discovery; Drug screening; Drugs; Endoplasmic reticulum; Fibrosis; Fluphenazine; Fluphenazine - pharmacology; Gene expression; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; HeLa Cells; Hep G2 Cells; Hepatocellular carcinoma; Hepatocytes; Hepatology; Hospitals; Humans; Immunoblotting; Influenza; Intracellular; Kinases; Liver; Liver - drug effects; Liver - metabolism; Liver - pathology; Liver diseases; Load distribution; Mammals; Medicine; Mice; Mice, Transgenic; Microscopy, Fluorescence; Mood; Mutation; Pediatrics; Phagocytosis; Phagosomes; Phagosomes - drug effects; Phagosomes - metabolism; Pharmacology; Phenothiazine; Phenothiazines; Physicians; Physiology; Proteins; Rodents; Screening; Survival Analysis; United States > US; Pittsburgh Pennsylvania; Pennsylvania
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0087260

The classical form of α1-antitrypsin deficiency (ATD) is associated with hepatic fibrosis and hepatocellular carcinoma. It is caused by the proteotoxic effect of a mutant secretory protein that aberrantly accumulates in the endoplasmic reticulum of liver cells. Recently we developed a model of this deficiency in C. elegans and adapted it for high-content drug screening using an automated, image-based array scanning. Screening of the Library of Pharmacologically Active Compounds identified fluphenazine (Flu) among several other compounds as a drug which reduced intracellular accumulation of mutant α1-antitrypsin Z (ATZ). Because it is representative of the phenothiazine drug class that appears to have autophagy enhancer properties in addition to mood stabilizing activity, and can be relatively easily re-purposed, we further investigated its effects on mutant ATZ. The results indicate that Flu reverses the phenotypic effects of ATZ accumulation in the C. elegans model of ATD at doses which increase the number of autophagosomes in vivo. Furthermore, in nanomolar concentrations, Flu enhances the rate of intracellular degradation of ATZ and reduces the cellular ATZ load in mammalian cell line models. In the PiZ mouse model Flu reduces the accumulation of ATZ in the liver and mediates a decrease in hepatic fibrosis. These results show that Flu can reduce the proteotoxicity of ATZ accumulation in vivo and, because it has been used safely in humans, this drug can be moved rapidly into trials for liver disease due to ATD. The results also provide further validation for drug discovery using C. elegans models that can be adapted to high-content drug screening platforms and used together with mammalian cell line and animal models.