Author(s): Ferreira, Inês C. ; Torrejón, Estefania ; Abecasis, Bernardo ; Alexandre, Bruno M. ; Gomes, Ricardo A. ; Verslype, Chris ; van Pelt, Jos ; Barbas, Ana ; Simão, Daniel ; Bandeiras, Tiago M. ; Bortoluzzi, Alessio ; Rebelo, Sofia P.
Date: 2024
Persistent ID: http://hdl.handle.net/10362/172685
Origin: Repositório Institucional da UNL
Subject(s): ALDH2; CETSA-MS; HCC; Sorafenib; Target validation; Biotechnology; Analytical Chemistry; Biochemistry; Molecular Medicine
Funding Information: We thank UniMS–Mass Spectrometry Unit from ITQB/iBET (Oeiras, Portugal) for MS data acquisition, and particular to Patrícia Alves, Ana Guerreiro and Catarina Correia for discussions and technical support on mass spectrometry. We thank Pedro Lamosa for discussions and technical support on NMR. We also thank Joerg Bomke, Pedro Sousa, Carolina Cassona, Inês Isidro and Inês Carrondo for fruitful discussions. Funding Information: This work was supported by the Fundação para a Ciência e Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT/MCTES, Portugal) through national funds to iNOVA4Health (UIDB/04462/2020 and UIDP/04462/2020) and the Associate Laboratory LS4FUTURE (LA/P/0087/2020). The NMR data were acquired at CERMAX, ITQB-NOVA, Oeiras, Portugal with equipment funded by FCT, project AAC 01/SAICT/2016. Publisher Copyright: © 2024
Sorafenib is a multikinase inhibitor indicated for first-line treatment of unresectable hepatocellular carcinoma. Despite its widespread use in the clinic, the existing knowledge of sorafenib mode-of-action remains incomplete. To build upon the current understanding, we used the Cellular Thermal Shift Assay (CETSA) coupled to Mass Spectrometry (CETSA-MS) to monitor compound binding to its target proteins in the cellular context on a proteome-wide scale. Among the potential sorafenib targets, we identified aldehyde dehydrogenase 2 (ALDH2), an enzyme that plays a major role in alcohol metabolism. We validated the interaction of sorafenib with ALDH2 by orthogonal methods using pure recombinant protein, proving that this interaction is not mediated by other cellular components. Moreover, we showed that sorafenib inhibits ALDH2 activity, supporting a functional role for this interaction. Finally, we were able to demonstrate that both ALDH2 protein expression and activity were reduced in sorafenib-resistant cells compared to the parental cell line. Overall, our study allowed the identification of ALDH2 as a novel sorafenib target and sheds light on its potential role in both hepatocellular carcinoma and sorafenib resistance condition.
