Background: Esophageal squamous cell carcinomas remains a particularly intractable disease due to limited therapeutic options as well as a lack of targeted therapies. The genetic landscape of esophageal squamous cell carcinoma is varied, but a common means of tumorigenesis in squamous cancers are alterations in copy number of putative oncogenic loci that result in upregulated mRNA. Within this subset of loci with copy number alterations, however, it has been difficult to distinguish between driver and passenger mutations using traditional in vitro models. Methods: We functionally screened a data set of amplified “outlier” candidates nominated through a bioinformatics pipeline that scored chromosomal amplifications with linked mRNA overexpression. Screening and validation was performed in a novel three-dimensional organoid model of esophageal squamous cell carcinoma that served as a tabula rasa by virtue of a single p53 null alteration. Results: Functional screening uncovered a novel oncogenic mechanism that serves to drive esophageal squamous cell cancer tumorigenesis both in vitro and in vivo when subcutaneously transplanted into immunodeficient mice. In addition, pharmacologic perturbation of this mechanism resulted in attenuation of the oncogenic phenotype in vitro. Conclusions: We have paired bioinformatics analysis of TCGA data with a p53 null organoid model to functionally validate a novel driver mechanism of esophageal squamous cell carcinoma. Studies are underway to translate these findings and hopefully generate additional therapeutic strategies for esophageal squamous cell carcinoma patients.