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Volume 41 Issue 6
Jun.  2025
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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2025  >  41(6): 577-586
Chen Yan, Cheng Zhuo, Ma Le, et al. Analysis of the number, type, and functional heterogeneity of senescent cells in the radiation-induced skin wounds in mice[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2025, 41(6): 577-586. Doi: 10.3760/cma.j.cn501225-20240604-00209
Citation: Chen Yan, Cheng Zhuo, Ma Le, et al. Analysis of the number, type, and functional heterogeneity of senescent cells in the radiation-induced skin wounds in mice[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2025, 41(6): 577-586. Doi: 10.3760/cma.j.cn501225-20240604-00209
shu

Analysis of the number, type, and functional heterogeneity of senescent cells in the radiation-induced skin wounds in mice

doi: 10.3760/cma.j.cn501225-20240604-00209

  • Institute of Rocket Force Medicine, State Key Laboratory of Trauma and Chemical Poisoning, Army Medical University, Chongqing 400038, China

Funds:

Key Program of National Natural Science Foundation of China 82030056

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  • Corresponding author: Shi Chunmeng, Email: shicm@tmmu.edu.cn
  • Received Date: 2024-06-04
    Abstract
  •   Objective  To investigate the number, type, and functional heterogeneity of senescent cells in the radiation-induced skin wounds in mice.  Methods  The study was an experimental study. Forty male p16-diphtheria toxin receptor-tdTomato (p16DTR/Tom) transgenic mice aged 6-8 weeks, which could be used to trace senescent cells, were divided into 35 Gy group and 50 Gy group (with 20 mice in each group) according to the random number table method, and 35 or 50 Gy X-ray irradiation was applied to the skin of the right hind limb of the mice to establish 3 or 4 degree of radiation-induced skin injury model, respectively. The positive area percentage of senescent cells in the wound tissue of mice in two groups was detected before irradiation and at 10, 20, and 30 d after irradiation; at 10 d after irradiation, the co-localization of endothelial cells (ECs), mononuclear macrophages (MMs), keratinocytes (KCs), fibroblasts (Fbs) and senescent cells in the wound tissue of mice in 50 Gy group was observed by immunofluorescence method. Nine male p16DTR/Tom transgenic mice aged 6-8 weeks were divided into unirradiated group without any treatment and 35 Gy group and 50 Gy group with the same treatment as above (with 3 mice in each group) according to the random number table method. The wound tissue of mice in 35 Gy group and 50 Gy group at 10 d after irradiation and the normal skin tissue of mice in unirradiated group at the corresponding time point was taken, and the senescence percentages of KCs, Fbs, ECs, and MMs were detected by flow cytometry. Bioinformatics analysis was performed on publicly available single-cell transcriptome sequencing data from normal skin tissue of healthy rats (setting as control group) and mixed wound tissue of rats for 7 and 14 d after irradiation with 30 Gy electron beams (setting as irradiated group), and the two groups of cells were subjected to senescence assessment to screen for senescent cells, the correlation between the expression profiles of senescence-associated secretory phenotypes (SASPs) of various types of senescent cells in irradiated group was analyzed, and the differentially expressed genes (DEGs) with significantly differential expression between the senescent cells in irradiated group and the corresponding normal cells in control group were screened for gene ontology (GO) enrichment analysis.  Results  The positive area percentage of senescent cells in the wound tissue of mice in 50 Gy group was significantly higher than that in 35 Gy group at 20 and 30 d after irradiation (with t values of -5.56 and -5.48, respectively, P < 0.05). ECs, MMs, KCs, and Fbs co-localized with senescent cells in the wound tissue of mice in 50 Gy group at 10 d after irradiation. The senescence percentages of KCs, Fbs, ECs, and MMs in the wound tissue of mice in 50 Gy group at 10 d after irradiation were (21.07±9.49)%, (16.10±3.27)%, (16.90±5.29)%, and (34.13±8.76)%, respectively, which were significantly higher than (3.58±1.13)%, (4.13±0.19)%, (3.86±1.28)%, and (10.14±4.95)% in the normal skin tissue of mice in unirradiated group at the corresponding time point, with P values all < 0.05. Bioinformatics analysis showed that the senescence scores of ECs, Fbs, KCs, macrophages, monocytes, and Schwann cells in the wound tissue of rats in irradiated group were significantly higher than those in the normal skin tissue of rats in control group (with Z values of -8.71, -9.58, -7.19, -8.82, -6.66, and -2.70, respectively, P < 0.05), i.e., 6 types of senescent cells were screened. The SASPs expression profiles of monocytes and macrophages in the wound tissue of rats in irradiated group were significantly correlated (r=0.83, P < 0.05), but there was no statistically significant correlation between the SASPs expression profiles of the remaining types of senescent cells (P > 0.05). GO enrichment analysis showed that, compared with the corresponding normal cells in control group, the significantly up-regulated DEGs of the 6 types of senescent cells in the wound tissue of rats in irradiated group were significantly enriched in the regulation of apoptosis signaling pathway, the significantly up-regulated DEGs of multiple senescent cells were significantly enriched in the myeloid cell differentiation pathway, and the significantly down-regulated DEGs of multiple senescent cells were significantly enriched in the cell division-related pathway, with P values all < 0.05.  Conclusions  The number of senescent cells in the wounds of mice with radiation-induced skin injury is up-regulated, and the accumulation of senescent cells is radiation dose- and time-dependent; multiple types of cells including ECs, Fbs, KCs, and MMs can undergo senescence, and there are obvious differences in the function and SASP expression profiles among various types of senescent cells.

     

    • (1) The dose- and time-dependent accumulation of senescent cells in radiation-induced skin wounds in mice was revealed.
    (2) The functional alterations of senescent cells and the difference in the expression profiles of senescence-associated secretory phenotypes after radiation-induced injury to the skin were elucidated.
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    • References(40)
  • [1]
    Iddins CJ, DiCarlo AL, Ervin MD, et al. Cutaneous and local radiation injuries[J]. J Radiol Prot, 2022, 42(1): 011001. DOI: 10.1088/1361-6498/ac241a.
    [2]
    Yang X, Ren H, Guo X, et al. Radiation-induced skin injury: pathogenesis, treatment, and management[J]. Aging (Albany NY), 2020, 12(22): 23379-23393. DOI: 10.18632/aging.103932.
    [3]
    Davan-Wetton CSA, Pessolano E, Perretti M, et al. Senescence under appraisal: hopes and challenges revisited[J]. Cell Mol Life Sci, 2021, 78(7): 3333-3354. DOI: 10.1007/s00018-020-03746-x.
    [4]
    Adjemian S, Oltean T, Martens S, et al. Ionizing radiation results in a mixture of cellular outcomes including mitotic catastrophe, senescence, methuosis, and iron-dependent cell death[J]. Cell Death Dis, 2020, 11(11): 1003. DOI: 10.1038/s41419-020-03209-y.
    [5]
    Demaria M, Ohtani N, Youssef SA, et al. An essential role for senescent cells in optimal wound healing through secretion of PDGF-AA[J]. Dev Cell, 2014, 31(6): 722-733. DOI: 10.1016/j.devcel.2014.11.012.
    [6]
    Wei X, Li M, Zheng Z, et al. Senescence in chronic wounds and potential targeted therapies[J/OL]. Burns Trauma, 2022, 10: tkab045[2024-06-04]. https://pubmed.ncbi.nlm.nih.gov/35187179/. DOI: 10.1093/burnst/tkab045.
    [7]
    Admasu TD, Kim K, Rae M, et al. Selective ablation of primary and paracrine senescent cells by targeting iron dyshomeostasis[J]. Cell Rep, 2023, 42(2): 112058. DOI: 10.1016/j.celrep.2023.112058.
    [8]
    Huang W, Hickson LJ, Eirin A, et al. Cellular senescence: the good, the bad and the unknown[J]. Nat Rev Nephrol, 2022, 18(10): 611-627. DOI: 10.1038/s41581-022-00601-z.
    [9]
    Chen Y, Ma L, Cheng Z, et al. Senescent fibroblast facilitates re-epithelization and collagen deposition in radiation-induced skin injury through IL-33-mediated macrophage polarization[J]. J Transl Med, 2024, 22(1): 176. DOI: 10.1186/s12967-024-04972-8.
    [10]
    Iwakawa M, Noda S, Ohta T, et al. Different radiation susceptibility among five strains of mice detected by a skin reaction[J]. J Radiat Res, 2003, 44(1): 7-13. DOI: 10.1269/jrr.44.7.
    [11]
    Rodgers KE, Tan A, Kim L, et al. Development of a guinea pig cutaneous radiation injury model using low penetrating X-rays[J]. Int J Radiat Biol, 2016, 92(8): 434-443. DOI: 10.1080/09553002.2016.1186302.
    [12]
    Tu W, Tang S, Yan T, et al. Integrative multi-omic analysis of radiation-induced skin injury reveals the alteration of fatty acid metabolism in early response of ionizing radiation[J]. J Dermatol Sci, 2022, 108(3): 178-186. DOI: 10.1016/j.jdermsci.2023.01.001.
    [13]
    Stuart T, Butler A, Hoffman P, et al. Comprehensive integration of single-cell data[J]. Cell, 2019, 177(7): 1888-1902.e21. DOI: 10.1016/j.cell.2019.05.031.
    [14]
    Wu T, Hu E, Xu S, et al. clusterProfiler 4.0: a universal enrichment tool for interpreting omics data[J]. Innovation (Camb), 2021, 2(3): 100141. DOI: 10.1016/j.xinn.2021.100141.
    [15]
    中国老年医学学会烧创伤分会, 中华医学会组织修复与再生分会, 中国康复医学会再生医学与康复专业委员会, 等. 放射性皮肤损伤的诊断和治疗专家共识(2024版)[J]. 中华烧伤与创面修复杂志, 2024, 40(8): 701-712. DOI: 10.3760/cma.j.cn501225-20240126-00033.
    [16]
    Dai S, Wen Y, Luo P, et al. Therapeutic implications of exosomes in the treatment of radiation injury[J/OL]. Burns Trauma, 2022, 10: tkab043[2024-06-04]. https://pubmed.ncbi.nlm.nih.gov/35071650/. DOI: 10.1093/burnst/tkab043.
    [17]
    Huayllani MT, Ruiz-Garcia H, Boczar D, et al. Adipose-derived stem cells therapy for radiation-induced skin injury[J]. Ann Plast Surg, 2021, 87(6): 639-649. DOI: 10.1097/SAP.0000000000003039.
    [18]
    夏成德, 杨阳. 放射性皮肤溃疡的治疗和预防策略[J]. 中华烧伤与创面修复杂志, 2024, 40(8): 719-724. DOI: 10.3760/cma.j.cn501225-20240415-00134.
    [19]
    DiCarlo AL, Bandremer AC, Hollingsworth BA, et al. Cutaneous radiation injuries: models, assessment and treatments[J]. Radiat Res, 2020, 194(3): 315-344. DOI: 10.1667/RADE-20-00120.1.
    [20]
    Chen W, Wang Y, Zheng J, et al. Characterization of cellular senescence in radiation ulcers and therapeutic effects of mesenchymal stem cell-derived conditioned medium[J/OL]. Burns Trauma, 2023, 11: tkad001[2024-06-04]. https://pubmed.ncbi.nlm.nih.gov/37188110/. DOI: 10.1093/burnst/tkad001.
    [21]
    Wang H, Wang Z, Huang Y, et al. Senolytics (DQ) mitigates radiation ulcers by removing senescent cells[J]. Front Oncol, 2020, 9: 1576. DOI: 10.3389/fonc.2019.01576.
    [22]
    Haston S, Gonzalez-Gualda E, Morsli S, et al. Clearance of senescent macrophages ameliorates tumorigenesis in KRAS-driven lung cancer[J]. Cancer Cell, 2023, 41(7): 1242-1260.e6. DOI: 10.1016/j.ccell.2023.05.004.
    [23]
    Moiseeva V, Cisneros A, Cobos AC, et al. Context-dependent roles of cellular senescence in normal, aged, and disease states[J]. FEBS J, 2023, 290(5): 1161-1185. DOI: 10.1111/febs.16573.
    [24]
    Wang W, Luo J, Sheng W, et al. Proteomic profiling of radiation-induced skin fibrosis in rats: targeting the ubiquitin-proteasome system[J]. Int J Radiat Oncol Biol Phys, 2016, 95(2): 751-760. DOI: 10.1016/j.ijrobp.2016.01.021.
    [25]
    Campbell RA, Docherty MH, Ferenbach DA, et al. The role of ageing and parenchymal senescence on macrophage function and fibrosis[J]. Front Immunol, 2021, 12: 700790. DOI: 10.3389/fimmu.2021.700790.
    [26]
    Nguyen HQ, To NH, Zadigue P, et al. Ionizing radiation-induced cellular senescence promotes tissue fibrosis after radiotherapy. A review[J]. Crit Rev Oncol Hematol, 2018, 129: 13-26. DOI: 10.1016/j.critrevonc.2018.06.012.
    [27]
    Paramos-de-Carvalho D, Jacinto A, Saúde L. The right time for senescence[J]. Elife, 2021, 10: e72449. DOI: 10.7554/eLife.72449.
    [28]
    Kirschner K, Rattanavirotkul N, Quince MF, et al. Functional heterogeneity in senescence[J]. Biochem Soc Trans, 2020, 48(3): 765-773. DOI: 10.1042/BST20190109.
    [29]
    Wilkinson HN, Hardman MJ. Cellular senescence in acute and chronic wound repair[J]. Cold Spring Harb Perspect Biol, 2022, 14(11): a041221. DOI: 10.1101/cshperspect.a041221.
    [30]
    Alessio N, Acar MB, Squillaro T, et al. Progression of irradiated mesenchymal stromal cells from early to late senescence: changes in SASP composition and anti-tumour properties[J]. Cell Prolif, 2023, 56(6): e13401. DOI: 10.1111/cpr.13401.
    [31]
    Cohn RL, Gasek NS, Kuchel GA, et al. The heterogeneity of cellular senescence: insights at the single-cell level[J]. Trends Cell Biol, 2023, 33(1): 9-17. DOI: 10.1016/j.tcb.2022.04.011.
    [32]
    Wu F, Zhang Z, Wang M, et al. Cellular atlas of senescent lineages in radiation-or immunotherapy-induced lung injury by single-cell RNA-sequencing analysis[J]. Int J Radiat Oncol Biol Phys, 2023, 116(5): 1175-1189. DOI: 10.1016/j.ijrobp.2023.02.005.
    [33]
    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[J]. Theranostics, 2024, 14(15): 5809-5825. DOI: 10.7150/thno.100417.
    [34]
    Guerrero-Juarez CF, Dedhia PH, Jin S, et al. Single-cell analysis reveals fibroblast heterogeneity and myeloid-derived adipocyte progenitors in murine skin wounds[J]. Nat Commun, 2019, 10(1): 650. DOI: 10.1038/s41467-018-08247-x.
    [35]
    Paldor M, Levkovitch-Siany O, Eidelshtein D, et al. Single-cell transcriptomics reveals a senescence-associated IL-6/CCR6 axis driving radiodermatitis[J]. EMBO Mol Med, 2022, 14(8): e15653. DOI: 10.15252/emmm.202115653.
    [36]
    Lucas V, Cavadas C, Aveleira CA. Cellular senescence: from mechanisms to current biomarkers and senotherapies[J]. Pharmacol Rev, 2023, 75(4): 675-713. DOI: 10.1124/pharmrev.122.000622.
    [37]
    Muñoz-Espín D, Cañamero M, Maraver A, et al. Programmed cell senescence during mammalian embryonic development[J]. Cell, 2013, 155(5): 1104-1118. DOI: 10.1016/j.cell.2013.10.019.
    [38]
    Zhang J, Yu H, Man MQ, et al. Aging in the dermis: fibroblast senescence and its significance[J]. Aging Cell, 2024, 23(2): e14054. DOI: 10.1111/acel.14054.
    [39]
    Sharpless NE, Sherr CJ. Forging a signature of in vivo senescence[J]. Nat Rev Cancer, 2015, 15(7): 397-408. DOI: 10.1038/nrc3960.
    [40]
    Gorgoulis V, Adams PD, Alimonti A, et al. Cellular senescence: defining a path forward[J]. Cell, 2019, 179(4): 813-827. DOI: 10.1016/j.cell.2019.10.005.
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