Novel repurposing of sulfasalazine for the treatment of adrenocortical carcinomas, probably through the SLC7A11/xCT-hsa-miR-92a-3p-OIP5-AS1 network pathway

Chitra Subramanian; Kelli McNamara; Seth W Croslow; Yanqi Tan; Daniel Hess; Katja Kiseljak-Vassiliades; Margaret E Wierman; Jonathan V Sweedler; Mark S Cohen

doi:10.1016/j.surg.2024.07.075

Novel repurposing of sulfasalazine for the treatment of adrenocortical carcinomas, probably through the SLC7A11/xCT-hsa-miR-92a-3p-OIP5-AS1 network pathway

Surgery. 2024 Oct 17:S0039-6060(24)00702-5. doi: 10.1016/j.surg.2024.07.075. Online ahead of print.

Authors

Chitra Subramanian¹, Kelli McNamara¹, Seth W Croslow², Yanqi Tan², Daniel Hess¹, Katja Kiseljak-Vassiliades³, Margaret E Wierman³, Jonathan V Sweedler², Mark S Cohen⁴

Affiliations

¹ Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, University of Illinois Urbana-Champaign, Champaign, IL.
² Department of Chemistry, University of Illinois Urbana-Champaign, Champaign, IL.
³ Division of Endocrinology, Metabolism and Diabetes, University of Colorado Anschutz School of Medicine, Aurora, CO.
⁴ Department of Biomedical and Translational Sciences, Carle Illinois College of Medicine, University of Illinois Urbana-Champaign, Champaign, IL; Department of Surgery, Carle Illinois College of Medicine, University of Illinois Urbana-Champaign, Champaign, IL. Electronic address: [email protected].

PMID: 39424480
DOI: 10.1016/j.surg.2024.07.075

Abstract

Background: Recent multigenomic analysis of adrenocortical carcinomas (ACCs) identified SLC7A11/xCT as a novel biomarker. The Food and Drug Administration-approved anti-inflammatory drug, sulfasalazine (SAS), induces ferroptosis by blocking SLC7A11 expression. We hypothesize that SAS could be repurposed to target ACC cells.

Methods: Expression of SLC7A11 and its association with ACC survival was analyzed using Gene Expression Profiling Interactive Analysis (GEPIA). The validated ACC cell lines NCI-H295R, ACC1, and ACC2 were grown in 2D culture. In vitro studies included the CellTiter-Glo assay to calculate viability, Western blot (WB) analysis for apoptosis and other target protein changes, reverse transcriptase polymerase chain reaction for steroidogenic enzyme changes, C11BODIPY for lipid peroxidation, and mass spectrometry for changes in lipids.

Results: The Cancer Genome Atlas Program database analysis in GEPIA showed that SLC7A11 and linked long noncoding RNA OAP5-AS1 are highly expressed in ACC tumors versus normal adrenals (n = 77 vs 128; P < .05). This was associated with poor overall and disease-free survival with hazard ratios of 4.3 and 5.2 for SLC7A11 and 4.8 and 2.7 for OAP5-AS1, respectively. ACC cell line half-inhibitory maximum concentration values after 72-hour SAS treatment ranged from 412 nM (ACC1) to 799 nM (ACC2), and all showed cleavage of poly (ADP-ribose) polymerase, upregulation of p-Akt and p-ERK, and downregulation of GPX4 and SLC7A11 (P < .05) by WB analysis. Sphere formation, migration, and invasion assay showed inhibition, and lipid peroxidation using C11BODIPY, increase in intracellular iron, induction of oxidative stress, and significant upregulation of oxidized polyunsaturated fatty acid phospholipids (P < .05 each) by mass spectrometry suggests induction of ferroptosis.

Conclusion: SAS downregulates tSLC7A11 in ACCs, targets the Akt/ERK pathway and lipid metabolism, and induces cell death in vitro, warranting additional translational studies to define its therapeutic potential in ACC.