Work

Single-cell RNA-seq analysis reveals the role of PGRN in modulating fibroblast heterogeneity and enhancing wound healing

Year: 2026

Type: article

Abstract: Introduction: The wound microenvironment plays a crucial role in the skin wound healing process. Fibroblasts, one of the critical cell types within this microenvironment, exhibit significant heterogeneity that influences healing outcomes. Previous studies have demonstrated that progranulin (PGRN) may reduce fibrogenesis during wound healing. However, the underlying mechanisms remain poorly understood. Thus, this study aims to investigate the specific effects of PGRN on fibroblast heterogeneity of skin wound healing in mice. Methods: To assess the role of PGRN, BALB/c mice were subcutaneously injected with PGRN overexpression or control lentivirus. Excisional skin surgery was performed a week later on the dorsal skin of the mice. Wound healing was monitored up to 21 days post-wounding, and specimens were collected at 7 days post-wounding for single-cell RNA sequencing. Bioinformatics analysis included cell clustering, marker gene identification, GO/KEGG enrichment, pseudotime trajectory, and cell-cell communication. Spatial transcriptomic data were used to analyze the spatial expression of genes of interest. Multi-immunohistochemical staining, western blot, and laser speckle blood flow imaging were performed to validate key molecular findings. Results: Our analysis revealed 33 distinct cell clusters, with a notable increase in the proportions of macrophages, neutrophils, NK cells, and myofibroblasts in the PGRN overexpression group, while fibroblasts and epithelial cells were less represented. Importantly, PGRN overexpression specifically stimulated the proliferation of myofibroblast subcluster 2 and induced unidentified fibroblast subclusters 0 and 5 that were enriched in pathways related to extracellular matrix and angiogenesis. Subcluster 0 fibroblasts were further characterized using the markers Crabp1, Prss35, Fabp5, and Acta2. Their location in the upper layer of the neodermis in the wound was confirmed by spatial transcriptomic data and multi-immunohistochemical staining. Additionally, PGRN treatment significantly enhanced blood flow in newly formed tissue and accelerated wound closure. Conclusion: Our findings indicated that PGRN regulated wound healing by promoting specific fibroblast populations, particularly those associated with angiogenesis and extracellular matrix remodeling. This study provides valuable insights into the complex regulatory mechanisms of PGRN in wound healing and highlights its potential as a therapeutic target for enhancing skin regeneration in clinical settings.

Source: Frontiers in Bioengineering and Biotechnology

DOI: 10.3389/fbioe.2026.1930259