O20 Spatial transcriptomic profiling reveals interleukin1β+ macrophages as major drivers of senescence in chronic wounds
Lauren Colborn, Holly Wilkinson, Leah Cooksey, Matthew Hardman, Alexander Johns, Nina RochaAbstract
Introduction and aims
Chronic wounds affect over 2.2 million patients in the UK, yet effective therapies remain lacking owing to limited understanding of the cellular mechanisms underlying impaired healing. Our group was the first to mechanistically demonstrate that cellular senescence contributes to delayed wound repair in vivo, and that targeting CXCR2 dampens senescence and promotes tissue regeneration. Thus, in the present study, we integrated spatial transcriptomics and immune profiling to define the senescent wound microenvironment in both human tissue and murine models to uncover cellular and molecular drivers of senescence in human chronic wounds and inform therapeutic targets.
Methods
Spatial transcriptomics (Xenium, 10X Genomics) and protein multiplex imaging (MACSima, Miltenyi Biotec) were conducted on human and murine chronic wound tissue. Immunohistochemistry of selected makers further validated transcriptomic findings. Complimentary in vivo studies were conducted in diabetic mice, with wound healing outcomes and immune cell senescence assessed via histology and flow cytometry.
Results
Transcriptomic analysis revealed significant upregulation of interleukin (IL)1β and CXCL5 in high-senescent wounds, implicating inflammatory chemokine signalling in sustaining the senescent niche. Both IL1β and CXCL5 show strong colocalization and enrichment within a myeloid cluster spatially adjacent to the leading edge of the wound. CellChat analysis confirmed strong predicted ligand–receptor interactions between this myeloid cluster and basal keratinocytes, indicating macrophage priming and paracrine crosstalk driving cellular dysfunction and chronic inflammation. In vivo blockade of IL1β reduced immune cell senescence and significantly improved wound healing, reinforcing its central role in modulating wound inflammation and repair.
Conclusions
Our data validate senescence as a hallmark of chronic wound dysfunction, where immunosenescence drives sustained inflammation and impaired healing. IL1β-driven immune–epithelial crosstalk emerged as a key mechanism in this process, identifying IL1β as a promising therapeutic target. These findings provide a foundation for precision therapies aimed at targeting senescence to improve wound healing outcomes in patients.