T505A variant of ETV5 promotes proliferation of precursor B cells in a mouse model of acute lymphoblastic leukemia

Abstract

Precursor B cell acute lymphoblastic leukemia (pre-B-ALL) arises as a result of precursor B cells acquiring driver mutations that lead to arrested differentiation and increased proliferation. Identification of driver mutations and understanding their biological function is critical to understanding pre-B-ALL development and advancing disease treatment. Using a mouse model of pre-B-ALL driven by deletion of genes encoding the related E26-transformation-specific (ETS) transcription factors PU.1 and Spi-B, we performed whole exome sequencing to identify secondary driver mutations. We identified recurrent variants in E26 transformation-specific transcription variant 5 (ETV5) resulting in R392P, V444I, and T505A amino acid changes. We found that the R392P and V444I variants altered the ability of ETV5 to bind to DNA using electrophoretic mobility shift assay. R392P and V444I variants did not activate a Dual-Specificity-Phosphatase 6 (DUSP6) reporter. In contrast, T505A ETV5 could interact with DNA and activate the DUSP6 promoter. To determine biological function, we forced expression of wild type, R392P, V444I, or T505A ETV5 in an interleukin-7-dependent pre-B cell line. Proliferation and apoptosis assays showed that T505A ETV5 conferred a proliferative advantage to pre-B cells. RNA sequencing showed that expression of ETV5 variants significantly altered gene expression in cultured cells. Through gene set enrichment analysis, T505A was suggested to downregulate the p53 pathway and the anti-proliferative protein, B cell translocation gene 2 (encoded by Btg2). In summary, these data suggest that ETV5 mutations play a role in pre-B-ALL by affecting proliferation and cell survival.