A novel human model to deconvolve cell-intrinsic phenotypes of genetically dysregulated pathways in lung squamous cell carcinoma

Tractable, patient relevant models are needed to investigate cancer progression and heterogeneity. Here, we report an alternative and unique in vitro model of lung squamous cell carcinoma (LUSC) using primary human bronchial epithelial cells (hBECs) from three donors. The co-operation of ubiquitous alterations (TP53 and CDKN2A loss) and components of commonly deregulated pathways including squamous differentiation (SOX2), PI3K signalling (PTEN) and the oxidative stress response (KEAP1) was investigated using mutant hBECs harbouring cumulative alterations. Our analyses confirmed that SOX2-overexpression initiates early preinvasive phases, and the co-operation with the oxidative stress response and PI3K pathways to drive more aggressive phenotypes, with expansion of cells expressing LUSC biomarkers and invasiveness. This cooperation was consistent with the classical LUSC subtype. Importantly, we connected pathway dysregulation with expression changes associated with cell-intrinsic processes and immunomodulation. In summary, our approach constitutes a powerful system to model LUSC and unravel genotype-phenotype causations of clinical relevance. ### Competing Interest Statement The authors have declared no competing interest.