Key Genetic Determinants Driving Esophageal Squamous Cell Carcinoma Initiation and Immune Evasion

Abstract

AbstractDespite recent progress in identifying aberrant genetic and epigenetic alterations in esophageal squamous cell carcinoma (ESCC), the mechanism of ESCC initiation remain unknown. Using genetically engineered esophageal organoids (EOs), we identified the key genetic determinants that drive ESCC tumorigenesis. A single-cell transcriptomic analysis uncovered thatTrp53, Cdkn2a, andNotch1(PCN) triple knockout (KO) induces neoplastic features of ESCC by generating distinct cell lineage trajectories with multiple root cells and high cell plasticity. AlthoughTrp53andNotch1(PN) double KO was sufficient to induce esophageal neoplasia and cellular heterogeneity, additional inactivation ofCdkn2awas indispensable for immune landscape remodeling forin vivotumorigenesis.PCNKO generated immunosuppressive niche enriched with exhausted T cells and M2 macrophages via the CCL2-CCR2 axis in an autochthonous ESCC mouse model. Moreover, genetic or pharmacological blockade of the CCL2-CCR2 axis suppressed ESCC tumorigenesis. Comparative single-cell transcriptomic analyses classified ESCC patient tumors into three subgroups and identified a specific subset recapitulating PCN-type ESCC signatures, including the high expression of CCL2 and CD274/PD-L1. Our study unveils that loss ofTP53, CDKN2A, andNOTCH1induces esophageal neoplasia and immune evasion for ESCC initiation and proposes the CCL2 blockade as a viable approach to target a subset of ESCC.