Abstract 280: ALDH2 knockout dysregulates arginine and proline metabolism in SK-BR-3 breast cancer cells

Abstract Aldehyde dehydrogenase 2 (ALDH2) mitigates cellular stress by detoxifying reactive aldehydes produced during alcohol exposure and endogenous metabolic processes. The ALDH2*2 variant, prevalent in East Asian populations, has decreased catalytic activity, which is linked to an increased risk of cancer and other diseases. While the effects of ALDH2 deficiency on cellular stress responses are well documented, the specific metabolic dysregulations arising from this deficiency require further investigation. To explore the impact of ALDH2 deficiency on metabolism and cellular stress in breast cancer, we generated the ALDH2 knockout subline of the SKBR-3 cell line (SKBR-3/ADKO) using CRISPR/Cas9 technology. Our findings indicate that ALDH2 knockout leads to heightened cellular stress, evidenced by elevated levels of 8-OHdG, reactive oxygen species (ROS), and malondialdehyde (MDA). Untargeted metabolomic analysis of SKBR-3 and SKBR-3/ADKO cells using Ultra High Performance Liquid Chromatography/High Resolution Mass Spectrometry assigned a total of 5, 033 metabolites, with 615 metabolites being significantly different (fold change ≥ 2, p-value < 0.05) between the two cell lines. The Mummichog approach has identified six key metabolic pathways altered upon ALDH2 knockout, including: arginine biosynthesis; D-amino acid metabolism; arginine and proline metabolism; alanine, aspartate and glutamate metabolism; pentose and glucuronate interconversions; pantothenate and CoA biosynthesis. The most significantly dysregulated pathway in response to the ALDH2 loss was arginine and proline metabolism, which is an integral pathway to redox homeostasis, ROS production, and cellular stress responses. Our study provides a novel insight into the role of ALDH2 in cancer metabolism, which might have a broader implication for cancer biology. Citation Format: Ming Zhao, Blake R. Rushing, Zhikun Ma, Rachel A. Coble, Amanda B. Parris, Sergey Krupenko, Susan J. Sumner, Xiaohe Yang. ALDH2 knockout dysregulates arginine and proline metabolism in SK-BR-3 breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 280.