Functionally distinct cancer-associated fibroblast subpopulations establish a tumor-promoting environment in squamous cell carcinoma

Cutaneous squamous cell carcinoma (cSCC) is a serious public health problem due to its high incidence and metastatic potential. It may progress from actinic keratosis (AK), a precancerous lesion, or the in situ carcinoma Bowen's disease (BD). During this progression, malignant keratinocytes activate dermal fibroblasts into tumor-promoting cancer-associated fibroblasts (CAFs), whose origin and emergence remain largely unknown. Here, we generate and analyze >115,000 single-cell transcriptomes from healthy skin, BD and cSCC of male donors. Our results reveal immunoregulatory and matrix-remodeling CAF subtypes that might derive from pro-inflammatory and mesenchymal fibroblasts, respectively. These CAF subtypes are largely absent in AK and interact with different cell types to establish a pro-tumorigenic microenvironment. These findings are cSCC-specific and could not be recapitulated in basal cell carcinomas. Our study provides important insights into the functionalities of dermal CAFs that will be highly beneficial for the specific targeting of the cSCC microenvironment. scRNA-seq of three human healthy UVR-exposed skin samples, three human Bowen's disease samples and five human cSCC samples using the Chromium Single Cell Reagent kit (v3.1 chemistry) from 10X Genomics.