Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury
Purpose: Precise molecular and cellular mechanisms of radiation-induced dermatitis are incompletely understood. Histone variant H2A.J is associated with cellular senescence and modulates senescence-associated secretory phenotype (SASP) after DNA-damaging insults, such as ionizing radiation (IR). Usi...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-08-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/23/17/9830 |
| _version_ | 1797495140384768000 |
|---|---|
| author | Yanick Hippchen Gargi Tewary Daniela Jung Zoé Schmal Stephan Meessen Jan Palm Claudia E. Rübe |
| author_facet | Yanick Hippchen Gargi Tewary Daniela Jung Zoé Schmal Stephan Meessen Jan Palm Claudia E. Rübe |
| author_sort | Yanick Hippchen |
| collection | DOAJ |
| description | Purpose: Precise molecular and cellular mechanisms of radiation-induced dermatitis are incompletely understood. Histone variant H2A.J is associated with cellular senescence and modulates senescence-associated secretory phenotype (SASP) after DNA-damaging insults, such as ionizing radiation (IR). Using ex vivo irradiated cultured foreskin, H2A.J was analyzed as a biomarker of radiation-induced senescence, potentially initiating the inflammatory cascade of radiation-induced skin injury. Methods: Human foreskin explants were collected from young donors, irradiated ex vivo with 10 Gy, and cultured in air-liquid interphase for up to 72 h. At different time-points after ex vivo IR exposure, the foreskin epidermis was analyzed for proliferation and senescence markers by immunofluorescence and immunohistochemical staining of sectioned tissue. Secretion of cytokines was measured in supernatants by ELISA. Using our mouse model with fractionated in vivo irradiation, H2A.J expression was analyzed in epidermal stem/progenitor cell populations localized in different regions of murine hair follicles (HF). Results: Non-vascularized foreskin explants preserved their tissue homeostasis up to 72 h (even after IR exposure), but already non-irradiated foreskin epithelium expressed high levels of H2A.J in all epidermal layers and secreted high amounts of cytokines. Unexpectedly, no further increase in H2A.J expression and no obvious upregulation of cytokine secretion was observed in the foreskin epidermis after ex vivo IR. Undifferentiated keratinocytes in murine HF regions, by contrast, revealed low H2A.J expression in non-irradiated skin and significant radiation-induced H2A.J upregulations at different time-points after IR exposure. Based on its staining characteristics, we presume that H2A.J may have previously underestimated the importance of the epigenetic regulation of keratinocyte maturation. Conclusions: Cultured foreskin characterized by highly keratinized epithelium and specific immunological features is not an appropriate model for studying H2A.J-associated tissue reactions during radiation-induced dermatitis. |
| first_indexed | 2024-03-10T01:44:11Z |
| format | Article |
| id | doaj.art-612340066494480fa4c7d0f94b78cc18 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T01:44:11Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-612340066494480fa4c7d0f94b78cc182023-11-23T13:17:45ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012317983010.3390/ijms23179830Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin InjuryYanick Hippchen0Gargi Tewary1Daniela Jung2Zoé Schmal3Stephan Meessen4Jan Palm5Claudia E. Rübe6Department of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyDepartment of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyDepartment of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyDepartment of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyDepartment of Urology, Klinikum Saarbrücken, 66119 Saarbrücken, GermanyDepartment of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyDepartment of Radiation Oncology, Saarland University Medical Center, 66421 Homburg, GermanyPurpose: Precise molecular and cellular mechanisms of radiation-induced dermatitis are incompletely understood. Histone variant H2A.J is associated with cellular senescence and modulates senescence-associated secretory phenotype (SASP) after DNA-damaging insults, such as ionizing radiation (IR). Using ex vivo irradiated cultured foreskin, H2A.J was analyzed as a biomarker of radiation-induced senescence, potentially initiating the inflammatory cascade of radiation-induced skin injury. Methods: Human foreskin explants were collected from young donors, irradiated ex vivo with 10 Gy, and cultured in air-liquid interphase for up to 72 h. At different time-points after ex vivo IR exposure, the foreskin epidermis was analyzed for proliferation and senescence markers by immunofluorescence and immunohistochemical staining of sectioned tissue. Secretion of cytokines was measured in supernatants by ELISA. Using our mouse model with fractionated in vivo irradiation, H2A.J expression was analyzed in epidermal stem/progenitor cell populations localized in different regions of murine hair follicles (HF). Results: Non-vascularized foreskin explants preserved their tissue homeostasis up to 72 h (even after IR exposure), but already non-irradiated foreskin epithelium expressed high levels of H2A.J in all epidermal layers and secreted high amounts of cytokines. Unexpectedly, no further increase in H2A.J expression and no obvious upregulation of cytokine secretion was observed in the foreskin epidermis after ex vivo IR. Undifferentiated keratinocytes in murine HF regions, by contrast, revealed low H2A.J expression in non-irradiated skin and significant radiation-induced H2A.J upregulations at different time-points after IR exposure. Based on its staining characteristics, we presume that H2A.J may have previously underestimated the importance of the epigenetic regulation of keratinocyte maturation. Conclusions: Cultured foreskin characterized by highly keratinized epithelium and specific immunological features is not an appropriate model for studying H2A.J-associated tissue reactions during radiation-induced dermatitis.https://www.mdpi.com/1422-0067/23/17/9830cultured foreskinionizing radiationradiation-induced dermatitishistone variant H2A.Jcellular senescencesenescence-associated secretory phenotype (SASP) |
| spellingShingle | Yanick Hippchen Gargi Tewary Daniela Jung Zoé Schmal Stephan Meessen Jan Palm Claudia E. Rübe Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury International Journal of Molecular Sciences cultured foreskin ionizing radiation radiation-induced dermatitis histone variant H2A.J cellular senescence senescence-associated secretory phenotype (SASP) |
| title | Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury |
| title_full | Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury |
| title_fullStr | Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury |
| title_full_unstemmed | Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury |
| title_short | Cultured Human Foreskin as a Model System for Evaluating Ionizing Radiation-Induced Skin Injury |
| title_sort | cultured human foreskin as a model system for evaluating ionizing radiation induced skin injury |
| topic | cultured foreskin ionizing radiation radiation-induced dermatitis histone variant H2A.J cellular senescence senescence-associated secretory phenotype (SASP) |
| url | https://www.mdpi.com/1422-0067/23/17/9830 |
| work_keys_str_mv | AT yanickhippchen culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT gargitewary culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT danielajung culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT zoeschmal culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT stephanmeessen culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT janpalm culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury AT claudiaerube culturedhumanforeskinasamodelsystemforevaluatingionizingradiationinducedskininjury |