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Investigation of the keratinocyte transcriptome altered in high-glucose environment: An in-vitro model system for precision medicine

  • Yang-Yi Chen
    Affiliations
    Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
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  • Shu-Mei Huang
    Affiliations
    Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
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  • Yu-Wen Cheng
    Affiliations
    Department of Neurosurgery, Kaohsiung Veterans General Hospital, Kaohsiung, Taiwan
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  • Meng-Chi Yen
    Affiliations
    Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Department of Emergency Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan
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  • Ya-Ling Hsu
    Affiliations
    Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
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  • Cheng-Che E. Lan
    Correspondence
    Correspondence to: Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, No.100 Shih-Chuan 1 st Rd, Kaohsiung, Taiwan.
    Affiliations
    Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Department of Dermatology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
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      Highlights

      • High-glucose environment altered the keratinocyte transcriptome responses to wounding.
      • In high-glucose cultivated keratinocytes, TNF, CYP24A1, NR4A3 and GGT1 were key overexpressed genes after wounding.
      • In high-glucose environment, wounding suppressed keratinocyte differentiation, while promoted myeloid leukocyte activation.
      • Keratinocytes from different individuals demonstrated diverse cellular responses to wounding in high-glucose environment.
      • The model is a valuable tool to explore the heterogeneity in the pathomechanism of diabetic wounds among individuals.

      Abstract

      Background

      Impaired wound healing is a serious diabetes complication compromising patients’ quality of life. However, the pathogenesis of diabetic wounds (DWs) remains incompletely understood. Human epidermal keratinocyte (HEK) is the sentinel cell that initiates healing processes after the epidermal integrity has been disrupted.

      Objective

      This study aimed to investigate the functional roles of HEKs in wound healing and to identify candidate genes, signaling pathways and molecular signatures contributing to the DWs.

      Methods

      HEKs were cultured in normal or high-glucose environment, followed by scratch, to mimic the microenvironment of normal wounds and DWs. Subsequently, we performed RNA sequencing and systematically analyzed the expression profiles by bioinformatics approaches.

      Results

      High-glucose environment altered the keratinocyte transcriptome responses to wounding. In experimental model of DWs, we found that TNF, CYP24A1, NR4A3 and GGT1 were key overexpressed genes in keratinocytes and were implicated in multiple cellular responses. Further analysis showed that wounding in high-glucose environment activated G-protein-coupled receptor (GPCR) signaling, cAMP response element-binding protein (CREB) signaling, and adrenomedullin signaling in keratinocytes, while dysregulated skin development and immune responses as compared to their counterpart in normal glucose settings.

      Conclusion

      This simplified in-vitro model serves as a valuable tool to gain insights into the molecular basis of DWs and to facilitate establishment of personalized therapies in clinical practice

      Keywords

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