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Updated: Sep 5, 2026
Murine Dermal Fibroblast Isolation by FACS
Published on: January 7, 2016
Single-cell transcriptomics and a boundary wound model reveal temporal decoupling and microenvironmental
Bingyue Cao1,2,3, Pan Liu4, Yujia Tan1,2,3
1Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
Background:
Radiotherapy is a key treatment method for cancer, but high-dose treatment can lead to damage to normal tissues and chronic fibrosis. There is a lack of systematic temporal analysis of the dynamic progression from early damage signals to late extracellular matrix (ECM) deposition.
Methods And Materials:
We reanalyzed publicly available scRNA-seq count matrices from GSE211713 to explore microenvironmental remodeling during fibrosis. Then, we established an in vivo local irradiation (12 Gy) mouse model with fully irradiated wounds and boundary-spanning wounds. For tissue repair and molecular phenotyping evaluations, we used tissue staining, IHC, qPCR, and WB.
Results:
scRNA-seq revealed temporal decoupling between ECM-related transcription and myofibroblast function. Our mouse model showed spatial heterogeneity: although the fully irradiated wound exhibited typical inhibition of healing, the wound across the irradiation boundary showed a rapid, matrix-rich response within 14 days. TGF-β/SMAD-related signals, myofibroblast markers, and collagen deposition in the animal model were not strictly synchronous. Additionally, CellChat predicted SPP1-centered communication at the late fibrosis stage.
Conclusions:
Fibrosis after radiotherapy may be influenced not only by local damage but also by the boundary microenvironment. These findings support a shift toward ECM-dominant remodeling and provide a hypothesis-generating basis for targeted intervention.
