(WHS-L2.06) A longitudinal single-cell transcriptomic atlas of diabetic porcine wounds reveals distinct fibroblast subtypes and trajectories in response to tissue mechanical manipulation
Thursday, May 16, 2024
10:30 AM – 11:30 AM East Coast USA Time
Background Diabetic foot ulceration (DFU) is a complex complication of diabetes, and its pathogenesis remains elusive. Drawing on the success of a strain-programmed patch in expediting wound closure, we conducted a groundbreaking single-cell analysis of diabetic Yucatan minipig skin injuries to unravel the intricate mechanisms of diabetic wound healing at distinct time points. Methods In a longitudinal study, we examined various time points (3, 7, 14, and 28 days post-wounding) in Alloxan-induced diabetic swine treated with either Tegaderm, a non-strain patch formulation, or our improved strain-programmable patch formulation. Three wounds per group per timepoint underwent mechanical and enzymatic digestion, generating highly viable single-cell suspensions. Subsequently, we performed detailed single-cell analyses to unveil the molecular landscape of diabetic porcine wound healing. Results Sequencing 280,000 cells enabled the identification of expected cell types, including keratinocytes, fibroblasts, endothelial cells, smooth muscle cells, myeloid, and lymphoid cells. We identified several separate subsets of fibroblasts, which are transcriptionally and functionally distinct and change in abundance along the time and treatment axes, with some changes reaching statistical significance. Through differential expression and pathway analyses, we deeply characterize the fibroblast subtypes uncover their functional significance, and reveal, through cell-cell communication analysis, extensive inflammatory pathway activation on Day 7, consistent with an enhanced inflammatory wound healing process. Finally, through trajectory analysis, we propose a model of fibroblast differentiation for each subtype. In addition, leveraging our diabetic foot ulcer dataset and comparing it against mouse wounding studies, we highlight the porcine model's superior resemblance to human skin wounds. Conclusion Our study unveils distinct fibroblast subsets in diabetic porcine wounds undergoing transcriptional and functional changes over time and treatments. Porcine wound cells closely resemble human wounded tissue in both cellular proportions and activated pathways. This comprehensive understanding of diabetic wound healing mechanisms opens avenues for targeted interventions and emphasizes the translational relevance of the porcine model in advancing diabetic wound research.