Bridging Inflammation and Proliferation: scRNA-seq Analysis of Chemotactic and Growth Factor Signaling in Mouse Skin Wound Repair

Lingzhang Meng^1*Hongmian Li¹Jian Song¹Wenxian Lin¹Xiuli Mao²Xiamin Zhang²Mingyue Yang²Kezhao Wu³Liu Lu⁴Feitang Liang⁴Feng Long⁵Yueyong Li⁶Qiang Tang⁴

• ¹The People's Hospital of GuangXi Zhuang Autonomous Region, Nanning, China
• ²Youjiang Medical University for Nationalities, Baise, China
• ³The University of Sydney, Sydney, Australia
• ⁴Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, China
• ⁵The People's Hospital of Beihai, Beihai, China
• ⁶Changsha Central Hospital, Changsha, China

The transition from inflammation to proliferation is a critical but poorly understood phase in wound healing. To elucidate the cellular and molecular dynamics of this pivotal stage, we performed single-cell RNA sequencing (scRNA-seq) on mouse skin biopsies 4 days after injury. By employing our newly developed R packages, OptiRes for optimized clustering and TidyGenePlot for annotation, we identified 21 distinct cell types. Our analysis revealed a dynamic shift in cellular composition, characterized by an influx of neutrophils, classical monocytes, and M1 macrophages. This recruitment of phagocytes was driven by enhanced chemotactic signaling through CCR5, CCR1, and ACKR1, a finding we validated in vivo. Furthermore, CellChat analysis identified four distinct intercellular communication clusters, highlighting the early activation of VEGF and EGF signaling pathways, which are essential for angiogenesis and re-epithelialization. To confirm the functional significance of our findings, we demonstrated that inhibition of CCR pathways reduced phagocyte infiltration. Together, these findings provide a high-resolution map of the cellular and molecular landscape during the transition from inflammation to proliferation, offering novel insights into the mechanisms that orchestrate tissue repair and identifying potential intervention manner to enhance wound healing.