Transcriptomic Landscape and Immune Microenvironment Around Wound Bed Define Regenerative versus Non-regenerative Outcomes in Mouse Digit Amputation

作者：Archana Prabahar, Connie S. Chamberlain, Ray Vanderby, William L. Murphy, William D’Angelo, Kulkarni Mangesh, Bryan N. Brown, Barsanjit Mazumder, Stephen F. Badylak, Peng Jiang · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2024 · DOI：10.1101/2024.12.04.626910 · 研究领域：Tissue Engineering and Regenerative Medicine、Wound Healing and Treatments、Mesenchymal stem cell research

Abstract In the mouse distal terminal phalanx (P3), it remains unclear why amputation at less than 33% of the digit results in regeneration, while amputation exceeding 67% leads to non-regeneration. Unraveling the molecular mechanisms underlying this disparity could provide crucial insights for regenerative medicine. In this study, we aim to investigate the tissues within the wound bed to understand the tissue microenvironment associated with regenerative versus non-regenerative outcomes. We employed a P3-specific amputation model in mice, integrated with time-series RNA-seq and a macrophage assay challenged with pro- and anti-inflammatory cytokines, to explore these mechanisms. Our findings revealed that non-regenerative digits exhibit a greater intense early transcriptional response in the wound bed compared to regenerative ones. Furthermore, early macrophage phenotypes differ distinctly between regenerative and non-regenerative outcomes. Regenerative digits also display unique co-expression modules related to Bone Morphogenetic Protein 2 (BMP2). The differentially expressed genes (DEGs) between regenerative and non-regenerative digits are enriched in targets of several transcription factors, such as HOXA11 and HOXD11 from the HOX gene family, showing a time-dependent pattern of enrichment. These transcription factors, known for their roles in bone regeneration, skeletal patterning, osteoblast activity, fracture healing, angiogenesis, and key signaling pathways, may act as ...