Single-cell RNA-seq of mouse androgenetic alopecia models treated with CXCL12-neutralizing antibody

Androgenetic alopecia (AGA) is a common form of hair loss characterized by follicular miniaturization, dermal fibrosis, and low-grade immune activation, yet the precise mediators linking androgen signaling to these tissue-level changes remain poorly defined. Here, we use single-cell RNA sequencing (scRNA-seq) to map the skin microenvironment in a testosterone-induced mouse model of AGA and assess the therapeutic impact of a CXCL12-neutralizing antibody. We find that androgen receptor-positive dermal fibroblasts are the principal source of CXCL12, which acts in an autocrine manner via ACKR3 to drive TGF-ß–mediated extracellular matrix deposition. In parallel, CXCL12 signals through CXCR4 to reprogram a Sox2+ Twist1+ dermal papilla cell population toward a fibrotic, low-inductive state and expand a pro-fibrotic Trem2+ macrophage subset. Blockade of CXCL12 reverses these transcriptional and cellular alterations across stromal, follicular, and immune compartments, restoring a regenerative skin niche and supporting hair regrowth. These findings position CXCL12 as a central mediator of fibroimmune remodeling in AGA and establish its neutralization as a promising therapeutic strategy. Overall design: Skin samples were collected from three groups of mice: control (Con), androgenetic alopecia (AGA) model induced by testosterone propionate (TP), and TP-induced AGA model treated with a CXCL12-neutralizing antibody (TP+Ab). At the end of the treatment period, dorsal skin tissues were harvested (n = 15 per group) and enzymatically dissociated to generate single-cell suspensions for single-cell RNA sequencing (scRNA-seq).