Fig. 3

E571K and E571G XPO1 mutations accelerate disease onset in the Eµ-TCL1 mouse model. a Schematic representing the Eμ-XPO1xTCL1 double transgenic mouse model, overexpressing either WT-, E571K-, or E571G-XPO1 under immunoglobulin heavy chain (Eμ-) promoter/enhancer elements in c57bl/6 mice. b Eμ-XPO1xTCL1 and Eμ-TCL1 mice were followed monthly by flow cytometry analysis for development of a CLL-like disease circulating in the peripheral blood. Mice from each Eμ-XPO1^{WT/E571K/E571G}xTCL1 colonies spontaneous developed an aggressive CD19 + /CD5 + and CD19 + /B22dim CLL-like disease similar to that observed in the Eμ-TCL1 model. c Eμ-XPO1xTCL1 and Eμ-TCL1 mice were followed monthly by flow cytometry analysis for development of a CLL-like disease circulating in the peripheral blood, and the time to disease onset (> 20% CD19 + /CD5 + and CD19 + /B22dim population in peripheral blood) was recorded for each mouse. Mice from Eμ-XPO1^E571KxTCL1 (n = 188) and Eμ-XPO1^E571GxTCL1 (n = 174) colonies displayed an accelerated time to disease onset compared to mice in the Eμ-TCL1 (n = 247) model (p = 0.093, p = 0.001, respectively). No change in rate of disease onset was observed between Eμ-XPO1^WTxTCL1 (n = 151) and Eμ-TCL1 mice (p = 0.713). Regardless of disease status, mice were continuously monitored for emergence of symptoms qualified as reaching early removal criteria (ERC), recording the survival time for each mouse. No significant difference in overall survival was noted between Eμ-XPO1xTCL1 (WT, n = 151, E571K, n = 189; E571G, n = 173) and Eμ-TCL1 mice (n = 246). A slight survival advantage was observed in Eμ-XPO1^WTxTCL1 mice compared to Eμ-TCL1 mice, although no level of significance was reached (p = 0.331). Statistical significance of Kaplan–Meier plots were determined via log-rank (Mantel-Cox) test. *p < 0.05. **p < 0.01. ***p < 0.001. d Comparative histopathology from Eμ-XPO1xTCL1 and Eμ-TCL1 transgenic mice. All mice examined developed lymphoid neoplasia, similar in character to the Eµ-TCL1 transgenic mouse model. Neoplastic lymphocytes were widely disseminated, often affecting the spleen (× 40), lymph nodes (× 40), bone marrow (× 60), and liver (× 20). e Comparison of IGHV gene usage between splenic B cells from Eµ-XPO1xTCL1 (XPO1-WT, XPO1-E571K, and XPO1-E571G) and Eμ-TCL1 mice (n = 3 per group). All mice demonstrated remarkably low diversity in BCR usage, suggesting a distinct clonal expansion comprises the tumor burden in these animals. Usage of BCR reads were binned by heavy chain V gene names, with each V gene normalized by the total number of heavy-chain reads. The top 10 most abundant genes are shown in triplicate mice with shared V genes maintaining the same color across all triplicates and samples. Genes not in the top 10 in terms of usage were grouped as “others,” and, if present, are shown as the bottom most segment in a dark forest-green color (see label). Gene names of the top 10 V genes in order of abundance are presented in Additional file 4: Table S1