Mouse digit fibroblast AAV gene delivery platform regulates regenerative outcome

The mouse digit tip robustly regenerates post-amputation, a process facilitated by the formation of a blastema. Fibroblasts constitute most of this structure, with specific subtypes contributing significantly more to the blastema. However, it remains unclear whether these fibroblast subpopulations and their associated genes are unique to regeneration or part of a broader wound-healing response. In this study, we performed single-cell RNA sequencing of non-regenerating digits to computationally identify fibroblast subpopulations and genes associated with regeneration and fibrosis. Furthermore, we developed a robust adeno-associated virus (AAV)-based gene delivery system to functionally assess the impact of newly identified candidate pro-fibrotic and pro-regenerative genes on wound healing outcomes. Overall design: Cohorts of adult wildtype FVB/NJ mice underwent P2 mid-digital (non-regenerative) digit amputation; 12, 17, and 28 days post-amputation, fibrosing digit tissues were collected and used for single cell RNA sequencing and analysis. An unamputated, homeostatic mid-digital sample was also collected.