Naji, F.; Oterino-Sogo, S.; Beltzung, F.; Garciaruano, D.; Mahfouf, W.; Guegan, J.-P.; Bohec, M.; Groppi, A.; Beylot-Barry, M.; Dousset, L.; Nikolski, M.; Rezvani, H.-R.

Cutaneous squamous cell carcinoma (cSCC) is a common skin cancer associated with substantial morbidity and mortality in advanced stages. Despite its well-described stepwise progression from actinic keratosis to invasive disease, robust molecular markers for stage discrimination and clinical decision-making remain limited. We sought to define the transcriptional continuum underlying cSCC progression, identify stage-associated biomarkers, and assess the broader relevance of these programs across human malignancies. Bulk RNA sequencing (HTG EdgeSeq) and spatial transcriptomics (GeoMx) were performed on biopsies from eight patients, each presenting multiple disease stages (healthy skin, premalignant lesion, tumor core, and invasive front) within the same lesion field, enabling within-patient analysis of progression. Spatial transcriptomic analyses identified more than 2,000 differentially expressed genes whose expression varied across disease stages. These genes were organized into 18 coordinated expression programs reflecting progressive biological rewiring during tumor evolution. Proliferation, extracellular matrix remodeling, inflammation, and stress-response pathways were progressively upregulated, whereas epithelial differentiation and metabolic processes, including lipid and amino acid metabolism, were downregulated. Macrophages exhibited distinct metabolic reprogramming, with increased purine metabolism, glycolysis, and pyruvate metabolism across progression. To evaluate the broader clinical relevance of these progression-associated programs, we developed a reproducible Snakemake pipeline to systematically screen 32 solid and hematologic malignancies from The Cancer Genome Atlas (TCGA). A combined cSCC-progression signature was significantly associated with poor overall survival (P < 0.05) in 10 additional cancer types. Finally, we identified 12 stage-informative biomarkers, whose spatially restricted expression patterns were validated using Visium HD. This study provides a spatially resolved and stage-aware transcriptomic map of cSCC progression, identifies coordinated gene programs underlying disease evolution, and defines progression-associated signatures with prognostic relevance across multiple cancers, highlighting their potential translational value.