Hyperglycemia-induced P300/CBP acetyltransferase drives ZEB2-mediated proinflammatory macrophages and delays wound healing
Soumyajit Roy, Debarun Patra, Palla Ramprasad, Shivam Sharma, Parul Katiyar, Ashvind Bawa, Kanhaiya Singh, Kulbhushan Tikoo, Suman Dasgupta, Chandan K. Sen, Durba Pal
Soumyajit Roy, Debarun Patra, Palla Ramprasad, Shivam Sharma, Parul Katiyar, Ashvind Bawa, Kanhaiya Singh, Kulbhushan Tikoo, Suman Dasgupta, Chandan K. Sen, Durba Pal
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Research Article Dermatology Immunology Inflammation

Hyperglycemia-induced P300/CBP acetyltransferase drives ZEB2-mediated proinflammatory macrophages and delays wound healing

Abstract

Chronic hyperglycemia changes the expression of various transcription factors and mRNA transcripts that impair cellular functionality and delay wound healing. Zinc finger E-box–binding homeobox 2 (ZEB2), a key transcription factor, maintains tissue-specific macrophage identities; however, its role in regulating macrophage polarization during wound healing under hyperglycemic conditions remains unclear. Here, we found that persistent hyperglycemia increases ZEB2 expression in wound macrophages via histone acetylation, contributing to chronic inflammation and delayed wound healing. Exposure to high glucose levels activated P300/CBP, a transcriptional coactivator involved in histone acetylation, which enhanced ZEB2 expression in wound macrophages. The forced expression of ZEB2 shifted macrophage polarity toward a proinflammatory state by upregulating myeloid lineage–directed transcription factors. Conversely, silencing Zeb2 at the wound site reduced hyperglycemia-induced macrophage inflammation. Topical application of C646, an inhibitor of P300, at the wound edges of streptozotocin-induced high-fat diet–fed diabetic mice significantly decreased ZEB2 expression, reduced inflammation, and accelerated wound healing. Therefore, targeted inhibition of P300 represents a promising therapeutic strategy for improving diabetic wound healing by modulating ZEB2-driven inflammation in wound macrophages.

Authors

Soumyajit Roy, Debarun Patra, Palla Ramprasad, Shivam Sharma, Parul Katiyar, Ashvind Bawa, Kanhaiya Singh, Kulbhushan Tikoo, Suman Dasgupta, Chandan K. Sen, Durba Pal

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Figure 1

Elevated glucose levels induce the expression of ZEB2 and activate proinflammatory pathways within macrophage populations.

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Elevated glucose levels induce the expression of ZEB2 and activate proin...
Protein expression was assessed by (A) immunostaining for ZEB2 (red) and CD68 (green) (scale bars: 100 μm [higher magnification] and 500 μm [lower magnification]; n = 3/group) and (B) Western blot analysis of ZEB2, CD68, and p-NF-κB proteins (n = 4/group) in human wound tissue. **P < 0.01 by paired, 2-tailed Student’s t test. Expression of ZEB2 and F4/80 were measured by (C) immunostaining (scale bars: 500 μm, n = 3/group) and (D) Western blot analysis (n = 4/group) in SD-fed and HFD-fed wound tissue from mice on day 7 (d7). *P < 0.05 by paired, 2-tailed Student’s t test. Effect of HG (25 mM D-glucose) induction on ZEB2 and p-NF-κB was examined by (E) Western blot and (F) immunostaining (scale bars: 30 μm). **P < 0.01 by paired, 2-tailed Student’s t test (n = 3). Inflammatory status of HG-treated RAW264.7 cells was assessed by (G) immunofluorescence analysis of ARG1 and iNOS (scale bars: 30 μm, n = 3) and (H and I) flow cytometric analysis of macrophage surface markers (n = 3). (J) Representative images of immunostaining for iNOS, ARG1, and IL-1β in PG-treated, IL-4–treated (20 ng/mL for 24 hours), and IL-4 + HG–treated RAW264.7 cells (scale bars: 30 μm; n = 3). Experiments were repeated 3 times independently. Data are expressed as mean ± standard deviation. ND, nondiabetic; DFU, diabetic foot ulcer; SD, standard diet; HFD, high-fat diet; PG, physiological glucose level; HG, hyperglycemia.