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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: No.100 Shih-Chuan 1 st Rd, Kaohsiung, Taiwan; Department of Dermatology, Kaohsiung Medical University Hospital, Kaohsiung Medical University.
    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 down-regulated keratinocyte differentiation and skin development, while up-regulated myeloid cell differentiation and myeloid leukocyte activation.
      • Keratinocyte samples from different individuals demonstrated diverse cellular responses to wounding in high-glucose environment.
      • The in-vitro model serves as a valuable tool to explore the heterogeneity in the pathomechanism of diabetic wounds among individuals and to facilitate the establishment of personalized therapies.

      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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      References

      1. IDF DIABETES ATLAS. International Diabetes Federation. 10th ed. p.32-34. ISBN 978-2-930229-98-0.2021 p. 32-34.
        • Bowling F.L.
        • Rashid S.T.
        • Boulton A.J.M.
        Preventing and treating foot complications associated with diabetes mellitus.
        Nature Reviews Endocrinology. 2015; 11: 606-616
        • Lima A.L.
        • Illing T.
        • Schliemann S.
        • Elsner P.
        Cutaneous Manifestations of Diabetes Mellitus: A Review.
        Am J Clin Dermatol. 2017; 18: 541-553
        • Yamamoto T.
        • Orikasa R.
        Multiple porokeratosis developed in association with the worsening of diabetes mellitus.
        Dermatologica Sinica. 2021; 39: 109-110
        • Lin E.S.
        • Chang W.A.
        • Chen Y.Y.
        • Wu L.Y.
        • Chen Y.J.
        • Kuo P.L.
        Deduction of Novel Genes Potentially Involved in Keratinocytes of Type 2 Diabetes Using Next-Generation Sequencing and Bioinformatics Approaches.
        Journal of clinical medicine. 2019; 8: 1
        • Berbudi A.
        • Rahmadika N.
        • Tjahjadi A.I.
        • Ruslami R.
        Type 2 Diabetes and its Impact on the Immune System.
        Curr Diabetes Rev. 2020; 16: 442-449
        • Hu S.C.
        • Lan C.E.
        High-glucose environment disturbs the physiologic functions of keratinocytes: Focusing on diabetic wound healing.
        Journal of dermatological science. 2016; 84: 121-127
        • Baltzis D.
        • Eleftheriadou I.
        • Veves A.
        Pathogenesis and Treatment of Impaired Wound Healing in Diabetes Mellitus: New Insights.
        Advances in Therapy. 2014; 31: 817-836
        • Lan C.C.
        • Wu C.S.
        • Huang S.M.
        • Wu I.H.
        • Chen G.S.
        High-glucose environment enhanced oxidative stress and increased interleukin-8 secretion from keratinocytes: new insights into impaired diabetic wound healing.
        Diabetes. 2013; 62: 2530-2538
        • Wetzler C.
        • Kämpfer H.
        • Stallmeyer B.
        • Pfeilschifter J.
        • Frank S.
        Large and Sustained Induction of Chemokines during Impaired Wound Healing in the Genetically Diabetic Mouse: Prolonged Persistence of Neutrophils and Macrophages during the Late Phase of Repair.
        Journal of Investigative Dermatology. 2000; 115: 245-253
        • Raziyeva K.
        • Kim Y.
        • Zharkinbekov Z.
        • Kassymbek K.
        • Jimi S.
        • Saparov A.
        Immunology of Acute and Chronic Wound Healing.
        Biomolecules. 2021; 11: 5
        • Geng K.
        • Ma X.
        • Jiang Z.
        • Huang W.
        • Gao C.
        • Pu Y.
        • et al.
        Innate Immunity in Diabetic Wound Healing: Focus on the Mastermind Hidden in Chronic Inflammatory.
        Frontiers in Pharmacology. 2021; 12: 509
        • Zhang E.
        • Miramini S.
        • Patel M.
        • Richardson M.
        • Ebeling P.
        • Zhang L.
        Role of TNF-α in early-stage fracture healing under normal and diabetic conditions.
        Computer Methods and Programs in Biomedicine. 2022; 213106536
        • Lan C.C.
        • Liu I.H.
        • Fang A.H.
        • Wen C.H.
        • Wu C.S.
        Hyperglycaemic conditions decrease cultured keratinocyte mobility: implications for impaired wound healing in patients with diabetes.
        The British journal of dermatology. 2008; 159: 1103-1115
        • Huang S.M.
        • Wu C.S.
        • Chiu M.H.
        • Wu C.H.
        • Chang Y.T.
        • Chen G.S.
        • et al.
        High glucose environment induces M1 macrophage polarization that impairs keratinocyte migration via TNF-α: An important mechanism to delay the diabetic wound healing.
        Journal of dermatological science. 2019; 96: 159-167
        • Perrault D.P.
        • Bramos A.
        • Xu X.
        • Shi S.
        • Wong A.K.
        Local Administration of Interleukin-1 Receptor Antagonist Improves Diabetic Wound Healing.
        Ann Plast Surg. 2018; 80 (S317-s21)
        • Jimi S.
        • Jaguparov A.
        • Nurkesh A.
        • Sultankulov B.
        • Saparov A.
        Sequential Delivery of Cryogel Released Growth Factors and Cytokines Accelerates Wound Healing and Improves Tissue Regeneration.
        Frontiers in Bioengineering and Biotechnology. 2020; : 8
        • Lan C.C.
        • Huang S.M.
        • Wu C.S.
        • Wu C.H.
        • Chen G.S.
        High-glucose environment increased thrombospondin-1 expression in keratinocytes via DNA hypomethylation.
        Translational research: the journal of laboratory and clinical medicine. 2016; 169 (e1-3): 91-101
        • Huang S.M.
        • Wu C.S.
        • Chao D.
        • Wu C.H.
        • Li C.C.
        • Chen G.S.
        • et al.
        High-glucose-cultivated peripheral blood mononuclear cells impaired keratinocyte function via reduced IL-22 expression: implications on impaired diabetic wound healing.
        Experimental dermatology. 2015; 24: 639-641
        • Huang S.M.
        • Wu C.S.
        • Chiu M.H.
        • Yang H.J.
        • Chen G.S.
        • Lan C.E.
        High-glucose environment induced intracellular O-GlcNAc glycosylation and reduced galectin-7 expression in keratinocytes: Implications on impaired diabetic wound healing.
        Journal of dermatological science. 2017; 87: 168-175
        • Thomas K.
        • Kiwit M.
        • Kerner W.
        Glucose concentration in human subcutaneous adipose tissue: comparison between forearm and abdomen.
        Exp Clin Endocrinol Diabetes. 1998; 106: 465-469
        • Wiegand C.
        • Hipler U.-C.
        • Elsner P.
        • Tittelbach J.
        Keratinocyte and Fibroblast Wound Healing In Vitro Is Repressed by Non-Optimal Conditions but the Reparative Potential Can Be Improved by Water-Filtered Infrared A.
        Biomedicines. 2021; 9: 1802
        • Pertea M.
        • Kim D.
        • Pertea G.M.
        • Leek J.T.
        • Salzberg S.L.
        Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown.
        Nature protocols. 2016; 11: 1650-1667
        • Ashburner M.
        • Ball C.A.
        • Blake J.A.
        • Botstein D.
        • Butler H.
        • Cherry J.M.
        • et al.
        Gene Ontology: tool for the unification of biology.
        Nature Genetics. 2000; 25: 25-29
        • Huang da W.
        • Sherman B.T.
        • Lempicki R.A.
        Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources.
        Nature protocols. 2009; 4: 44-57
        • Yu G.
        • Wang L.G.
        • Han Y.
        • He Q.Y.
        clusterProfiler: an R package for comparing biological themes among gene clusters.
        Omics: a journal of integrative biology. 2012; 16: 284-287
        • Krämer A.
        • Green J.
        • Pollard Jr., J.
        • Tugendreich S.
        Causal analysis approaches in Ingenuity Pathway Analysis.
        Bioinformatics (Oxford, England). 2014; 30: 523-530
        • León C.
        • García-García F.
        • Llames S.
        • García-Pérez E.
        • Carretero M.
        • Arriba M.D.C.
        • et al.
        Transcriptomic Analysis of a Diabetic Skin-Humanized Mouse Model Dissects Molecular Pathways Underlying the Delayed Wound Healing Response.
        Genes (Basel). 2020; 12
        • Subramanian A.
        • Tamayo P.
        • Mootha V.K.
        • Mukherjee S.
        • Ebert B.L.
        • Gillette M.A.
        • et al.
        Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles.
        Proceedings of the National Academy of Sciences of the United States of America. 2005; 102: 15545-15550
        • Falcone M.
        • Meier J.J.
        • Marini M.G.
        • Caccialanza R.
        • Aguado J.M.
        • Del Prato S.
        • et al.
        Diabetes and acute bacterial skin and skin structure infections.
        Diabetes Research and Clinical Practice. 2021; 174108732
        • Sawaya A.P.
        • Pastar I.
        • Stojadinovic O.
        • Lazovic S.
        • Davis S.C.
        • Gil J.
        • et al.
        Topical mevastatin promotes wound healing by inhibiting the transcription factor c-Myc via the glucocorticoid receptor and the long non-coding RNA Gas5.
        J Biol Chem. 2018; 293: 1439-1449
        • Lindley L.E.
        • Stojadinovic O.
        • Pastar I.
        • Tomic-Canic M.
        Biology and Biomarkers for Wound Healing.
        Plast Reconstr Surg. 2016; 138: 18s-28s