RT Journal Article SR Electronic T1 Aldehyde Oxidase Contributes to All-Trans-Retinoic Acid Biosynthesis in Human Liver JF Drug Metabolism and Disposition JO Drug Metab Dispos FD American Society for Pharmacology and Experimental Therapeutics SP 202 OP 211 DO 10.1124/dmd.120.000296 VO 49 IS 3 A1 Guo Zhong A1 Chris J. Seaman A1 Erickson M. Paragas A1 Huaqing Xi A1 Karla-Luise Herpoldt A1 Neil P. King A1 Jeffrey P. Jones A1 Nina Isoherranen YR 2021 UL http://dmd.aspetjournals.org/content/49/3/202.abstract AB All-trans-retinoic acid (atRA) is a critical endogenous signaling molecule. atRA is predominantly synthesized from retinaldehyde by aldehyde dehydrogenase 1A1 (ALDH1A1), but aldehyde oxidase (AOX) may also contribute to atRA biosynthesis. The goal of this study was to test the hypothesis that AOX contributes significantly to atRA formation in human liver. Human recombinant AOX formed atRA from retinaldehyde (Km ∼1.5 ± 0.4 µM; kcat ∼3.6 ± 2.0 minute−1). In human liver S9 fractions (HLS9), atRA formation was observed in the absence of NAD+, suggesting AOX contribution to atRA formation. In the presence of NAD+, Eadie-Hofstee plots of atRA formation in HLS9 indicated that two enzymes contributed to atRA formation. The two enzymes were identified as AOX and ALDH1A1 based on inhibition of atRA formation by AOX inhibitor hydralazine (20%–50% inhibition) and ALDH1A1 inhibitor WIN18,446 (50%–80%inhibition). The expression of AOX in HLS9 was 9.4–24 pmol mg−1 S9 protein, whereas ALDH1A1 expression was 156–285 pmol mg−1 S9 protein measured by liquid chromatography–tandem mass spectrometry (LC-MS/MS) quantification of signature peptides. The formation velocity of atRA in the presence of NAD+ correlated significantly with the expression of ALDH1A1 and AOX protein. Taken together, the data show that both AOX and ALDH1A1 contribute to atRA biosynthesis in the human liver, with ALDH1A1 being the high-affinity, low-capacity enzyme and AOX being the low-affinity, high-capacity enzyme. The results suggest that in the case of ALDH1A dysfunction or excess vitamin A, AOX may play an important role in regulating hepatic vitamin A homeostasis and that inhibition of AOX may alter atRA biosynthesis and signaling.SIGNIFICANCE STATEMENT This study provides direct evidence to show that human AOX converts retinaldehyde to atRA and contributes to hepatic atRA biosynthesis. The finding that AOX may be responsible for 20%–50% of overall hepatic atRA formation suggests that alterations in AOX activity via drug-drug interactions, genetic polymorphisms, or disease states may impact hepatic atRA concentrations and signaling and alter vitamin A homeostasis.