{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T18:41:13Z","timestamp":1783104073658,"version":"3.54.6"},"reference-count":26,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T00:00:00Z","timestamp":1770681600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background\/Objectives: Females are generally more resistant to ischemia-related ferroptosis than males, due to differences in iron metabolism, antioxidant pathways, and sex hormone-mediated regulation of ferroptosis suppressors. This has not been systematically studied in a human donor liver model. To investigate the effect of sex on ferroptosis and oxidative stress pathways in non-utilized donor livers (NDLs), we assessed patterns of gene expression in NDLs under ex vivo normothermic machine perfusion (NMP). Methods: We utilized the PROTECT dual-circuit ex vivo NMP system to assess three male and two female NDLs undergoing 6 h NMP. Perfusate and tissue samples were collected at baseline and 6 h of NMP. Malondialdehyde (MDA) levels were quantified as biochemical markers of iron overload and lipid peroxidation, respectively. Ferroptosis-related gene expression was assessed using molecular assays. Comparisons between male and female NDLs were used to determine the influence of sex on ferroptosis and oxidative injury during NMP. Results: NMP was successfully performed on NDLs (n = 5) from three male (56.3 \u00b1 5.7 years) and two female donors (46.5 \u00b1 0.7 years, p = 0.15). The fold-change in the oxidative stress marker MDA was comparable between female (1.2 \u00b1 0.6) and male (1.0 \u00b1 0.4) NDLs after 6 h NMP (p = 0.76). All livers showed upregulation of ferroptosis-related genes (Hypoxia-inducible factor 1 alpha, Iron Responsive Binding Elements 2, Ribosomal Protein L8, Ferritin Heavy Chain 1, Acyl-CoA synthetase family member 2, ATP synthase membrane subunit c locus 3, Heme-oxygenase 1, NAD(P)H Quinone Dehydrogenase 1, Tetratricopeptide Repeat Domain 35, Nuclear Factor Erythroid 2 Related Factor 2). ACSF2 expression was significantly higher in female NDLs compared with males undergoing 6 h NMP (3.6 \u00b1 3.0 vs. 1.0 \u00b1 0.7-fold change, p = 0.04). There were no sex-based significant differences observed in the expression of other ferroptosis-related genes (HIF-1\u03b1, IREB2, RPL8, FTH-1, ATP5G3, HO-1, NQO1, TTC35, and NRF2) between male and female NDLs. No gene reached statistical significance after false-discovery-rate (FDR) correction. Conclusions: Normothermic machine perfusion of NDLs was feasible, and no sex-related differences were observed in MDA levels or most ferroptosis-related gene expression after 6 h. Although ACSF2 showed higher expression in female livers, this was not significant after multiple testing correction, highlighting the need for larger studies to explore sex-dependent ferroptosis signaling during liver preservation.<\/jats:p>","DOI":"10.3390\/genes17020220","type":"journal-article","created":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T09:16:08Z","timestamp":1770801368000},"page":"220","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Gender-Specific Gene Regulation of Ferroptosis in Non-Utilized Liver Donors"],"prefix":"10.3390","volume":"17","author":[{"given":"Hala","family":"Nazzal","sequence":"first","affiliation":[{"name":"College of Arts and Sciences, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Halil Kaan","family":"Caliskan","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-7387-2212","authenticated-orcid":false,"given":"Meghan","family":"Unes","sequence":"additional","affiliation":[{"name":"School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6750-3240","authenticated-orcid":false,"given":"Chandrashekhara","family":"Manithody","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shun","family":"Onishi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pranjali","family":"Rajalakshmi","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3816-8047","authenticated-orcid":false,"given":"Yasar","family":"Caliskan","sequence":"additional","affiliation":[{"name":"Division of Nephrology, SSM Health Saint Louis University Hospital, 1201 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mustafa","family":"Nazzal","sequence":"additional","affiliation":[{"name":"Department of Surgery, SSM Health Saint Louis University Hospital, 1201 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ajay","family":"Jain","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, School of Medicine, Saint Louis University, 1402 S. Grand Blvd, St. Louis, MO 63104, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"115920","DOI":"10.1016\/j.celrep.2025.115920","article-title":"DTL Dose-Dependent Control of Sex-Dimorphic Ferroptosis in Liver Ischemia Reperfusion Injury","volume":"44","author":"Huang","year":"2025","journal-title":"Cell Rep."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Granata, S., Votrico, V., Spadaccino, F., Catalano, V., Netti, G.S., Ranieri, E., Stallone, G., and Zaza, G. (2022). Oxidative Stress and Ischemia\/Reperfusion Injury in Kidney Transplantation: Focus on Ferroptosis, Mitophagy and New Antioxidants. 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