ROMO1 Promotes the Repair of Radiation-induced Skin Injury by Regulating Mitochondrial Function and Keratinocyte Proliferation
Abstract
Objective
To investigate the molecular mechanisms of early mitochondrial stress in keratinocytes induced by ionizing radiation (IR), focusing on the key role of ROMO1 in radiation-induced skin injury (RISI).
Methods
HaCaT keratinocytes were used to assess cellular status, mitochondrial dysfunction and oxidative stress at various time points after different irradiation doses. Mitochondrial proteomics at 24 hours post-20 Gy irradiation identified ROMO1 as the protein with the greatest reduction in expression. ROMO1-overexpressing HaCaT cells and skin-specific Romo1 knockout (Romo1-sKO) mice were established to investigate its functional mechanisms. ROMO1 expression and localization were analyzed by Western blot and immunofluorescence; flow cytometry measured ROS, mtROS, and mitochondrial membrane potential in ROMO1-overexpressing cells, while CCK-8, LDH release, and colony formation assays evaluated viability and proliferation. The role of Romo1 in the progression and repair of radiation-induced skin injury was further examined in Romo1-sKO mice.
Results
At 24 hours post-20 Gy irradiation, HaCaT cells exhibited inhibited proliferation (P < 0.05) and mitochondrial dysfunction (P < 0.0001). Mitochondrial proteomics identified 43 differentially expressed proteins (26 upregulated, 17 downregulated), with enrichment analysis indicating their involvement in oxidative phosphorylation, mitochondrial assembly and stress response. Among these, reactive oxygen species modulator 1 (ROMO1) was identified as the most significantly downregulated key protein in the early radiation response (P < 0.01, FDR = 0.03). In vitro, ROMO1 overexpression increased mtROS levels and mitochondrial membrane potential (P < 0.0001) but suppressed cell viability and promoted reproductive death (P < 0.05) after radiation. In vivo, Romo1-sKO mice showed accelerated wound healing, with a reduction in skin injury score by approximately 1 point, along with enhanced tissue regeneration at 25 days post-irradiation compared to control mice.
Conclusion
The mitochondrial protein ROMO1 is a key regulator of the early keratinocyte stress response to radiation. It impedes the repair of radiation-induced skin injury by suppressing cell proliferation via modulation of mitochondrial function. Targeted inhibition of ROMO1 may represent a novel therapeutic strategy to promote healing and prevent chronic progression of radiation-induced skin injury.
Keywords: ROMO1, Radiation-induced skin injury, Mitochondrial stress response, Keratinocyte proliferation
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LEE J, JANG W H, SHIM S, et al. Characterization of early-stage cutaneous radiation injury by using optical coherence tomography angiography. Biomed Opt Express, 2020, 11(5): 2652-2664. doi: 10.6084/M9.FIGSHARE.C.4891266.V1.
LÓPEZ E, GUERRERO R, NÚÑEZ M I, et al. Early and late skin reactions to radiotherapy for breast cancer and their correlation with radiation-induced DNA damage in lymphocytes. Breast Cancer Res, 2005, 7(5): R690-R698. doi: 10.1186/bcr1277.
YANG X, REN H, GUO X, et al. Radiation-induced skin injury: pathogenesis, treatment, and management. Aging (Albany NY), 2020, 12(22): 23379-23393. doi: 10.18632/aging.103932.
CUI J, WANG T J, ZHANG Y X, et al. Molecular biological mechanisms of radiotherapy-induced skin injury occurrence and treatment. Biomed Pharmacother, 2024, 180: 117470. doi: 10.1016/j.biopha.2024.117470.
TUIENG R J, CARTMELL S H, KIRWAN C C, et al. The effects of ionising and non-ionising electromagnetic radiation on extracellular matrix proteins. Cells, 2021, 10(11): 3041. doi: 10.3390/cells10113041
WANG P, FAN L, TIAN M. Progress on the mechanism of radiation-induced skin injury. Chin J Radiol Health, 2022, 31(4): 524-529. doi: 10. 13491/j.issn.1004-714X.2022.04.027.
YU H, ZHONG T, XU Y, et al. Molecular profiling of skin cells identifies distinct cellular signatures in radiation-induced skin injury across various stages in the murine dataset. Exp Hematol Oncol, 2025, 14(1): 18. doi: 10. 1186/s40164-025-00596-w.
YAN T, YANG P, BAI H, et al. Single-cell RNA-Seq analysis of molecular changes during radiation-induced skin injury: the involvement of Nur77. Theranostics, 2024, 14(15): 5809-5825. doi: 10.7150/thno.100417.
. TIAN L C, YUAN Y Y, DANG X H. Mitochondria: the target of ionizing radiation damage. Progress in Biochemistry and Biophysics, 2025, 52(4 ): 836-844. doi: 10.16476/j.pibb.2024.0394.
ZHANG C, GAO X, LI M, et al. The role of mitochondrial quality surveillance in skin aging: focus on mitochondrial dynamics, biogenesis and mitophagy. Ageing Res Rev, 2023, 87: 101917. doi: 10.1016/j.arr.2023. 101917.
YORDANOV A, TSONEVA E. ROMO1: a distinct mitochondrial protein with dual roles in dynamics and function. Antioxidants (Basel), 2025, 14(5): 540. doi: 10.3390/antiox14050540.
CHUNG Y M, KIM J S, YOO Y D. A novel protein, Romo1, induces ROS production in the mitochondria. Biochem Biophys Res Commun, 2006, 347(3): 649-655. doi: 10.1016/j.bbrc.2006.06.140.Epub2006Jul5.
CHUNG J S, LEE S B, PARK S H, et al. Mitochondrial reactive oxygen species originating from Romo1 exert an important role in normal cell cycle progression by regulating p27(Kip1) expression. Free Radic Res, 2009, 43(8): 729-737. doi: 10.1080/10715760903038432.
TANG S K, CHEN J H, GENG F H, et al. The radiosensitization effect of STING agonist on cutaneous melanoma cells. Chin J Radiol Med Prot, 2023, 43(1): 1-7. doi: 10.3760/cma.j.cn112271-20220826-00350.
SHENG N, ZHANG Z, ZHENG H, et al. Scutellarin rescued mitochondrial damage through ameliorating mitochondrial glucose oxidation via the Pdk-Pdc Axis. Adv Sci (Weinh), 2023, 10(32): e2303584. doi: 10.1002/advs.202303584.
ZHANG Y H, LIU H F, ZHANG S Y, et al. Proteomic analysis of radiation-induced esophagitis in rats, Chin J Radiol Med Prot, 2024, 44(9): 725-733. doi: 10.3760/cma.j.cn112271-20240103-00003.
GENG F, CHEN J, SONG B, et al. Chaperone- and PTM-mediated activation of IRF1 tames radiation-induced cell death and the inflammatory response. Cell Mol Immunol, 2024, 21(8): 856-872. doi: 10. 1038/s41423-024-01185-3.
GENG F, ZHONG L, YANG T, et al. A frog skin-derived peptide targeting SCD1 exerts radioprotective effects against skin injury by inhibiting STING-mediated inflammation. Adv Sci (Weinh), 2024, 11(25): e2306253. doi: 10.1002/advs.202306253.
LEE G Y, YOU D G, LEE H R, et al. Romo1 is a mitochondrial nonselective cation channel with viroporin-like characteristics. J Cell Biol, 2018, 217(6): 2059-2071. doi: 10.1083/jcb.201709001.
JOMOVA K, RAPTOVA R, ALOMAR S Y, et al. Reactive oxygen species, toxicity, oxidative stress, and antioxidants: chronic diseases and aging. Arch Toxicol, 2023, 97(10): 2499-2574. doi: 10.1007/s00204-023-03562-9.
WANG K, TEPPER J E. Radiation therapy-associated toxicity: Etiology, management, and prevention. CA Cancer J Clin, 2021, 71(5): 437-454. doi: 10.3322/caac.21689.
SORIANO J L, CALPENA A C, SOUTO E B, et al. Therapy for prevention and treatment of skin ionizing radiation damage: a review. Int J Radiat Biol, 2019, 95(5): 537-553. doi: 10.1080/09553002.2019.1562254.
AZZAM E I, JAY-GERIN J P, PAIN D. Ionizing radiation-induced metabolic oxidative stress and prolonged cell injury. Cancer Lett, 2012, 327(1/2): 48-60. doi: 10.1016/j.canlet.2011.12.012.
RÜBE C E, FREYTER B M, TEWARY G, et al. Radiation dermatitis: radiation-induced effects on the structural and immunological barrier function of the epidermis. Int J Mol Sci, 2024, 25(6): 3320. doi: 10.3390/ijms25063320.
SEO M D, KANG T J, LEE C H, et al. HaCaT keratinocytes and primary epidermal keratinocytes have different transcriptional profiles of cornified envelope-associated genes to T helper cell cytokines. Biomol Ther (Seoul), 2012, 20(2): 171-176. doi: 10.4062/biomolther.2012.20.2.171.
XU F, HUANG H, PENG K, et al. ROMO1 overexpression protects the mitochondrial cysteinome from oxidations in aging. Nat Commun, 2025, 16(1): 5133. doi: 10.1038/s41467-025-60503-z.
CHUNG J S, PARK S, PARK S H, et al. Overexpression of Romo1 promotes production of reactive oxygen species and invasiveness of hepatic tumor cells. Gastroenterology, 2012, 143(4): 1084-1094.e7. doi: 10. 1053/j.gastro.2012.06.038.
TSONEVA E, DIMITROVA P D, METODIEV M, et al. The effects of ROMO1 on cervical cancer progression. Pathol Res Pract, 2023, 248: 154561. doi: 10.1016/j.prp.2023.154561.
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