Blimp1 Is a Tumor Suppressor Gene in Diffuse Large B Cell Lymphoma.

Abstract 446

The PRDM1/ BLIMP1 gene encodes a zinc finger transcriptional repressor that is expressed in a subset of germinal center (GC) B cells and in all plasma cells, and is required for terminal B cell differentiation. The BLIMP1 locus is biallelically inactivated by structural alterations in approximately one third of activated B cell-like diffuse large B cell lymphoma (ABC-DLBCL) (Pasqualucci et al, J Exp Med 2006). Moreover, the expression of the Blimp1 protein is absent in up to 80% of ABC-DLBCL due to alternative genetic and epigenetic mechanisms. These findings suggest that BLIMP1 may function as a tumor suppressor gene whose loss may contribute to the pathogenesis of this lymphoma type by blocking terminal B cell differentiation. To investigate the role of BLIMP1 inactivation in lymphomagenesis in vivo, we tested whether conditional deletion of the Blimp1 gene in mouse B cells can promote the growth of lymphomas recapitulating the features of ABC-DLBCL. Toward this end, a mouse model carrying a loxP-flanked exon 5 of the Blimp1 gene that can be deleted by Cre-mediated recombination (Ohinata et al, Nature 2005) was crossed with a CD19-Cre deletor strain, expressing the Cre recombinase in all B cells. The resulting mice were monitored for tumor development and survival. Consistent with previous observations in a similar model (Shapiro-Shelef et al, Immunity 2003), Blimp1 conditional knockout (Blimp1^CD19KO) mice showed a severe impairment in the generation of CD138⁺ plasma cells and had decreased serum immunoglobulin levels of all isotypes, together with a two-fold increase in the number of PNA^hiCD95⁺ GC B cells. Over time, significantly reduced survival was observed in the Blimp1^CD19KO cohort, with only 27% of the animals being alive at 15 months of age (LogRank p value<0.0001). Macroscopic and flow cytometric analysis of the lymphoid compartments revealed the presence of splenomegaly in 32/38 (84%) Blimp1^CD19KO, as compared to 1/25 (4%) age-matched wildtype (WT) littermates, and a significant increase in IgM⁺IgD^-CD21⁺CD23^lo splenic B cells, indicative of marginal zone B cell expansion. In addition, 79% (n=30/38) of Blimp1^CD19KO mice showed markedly hyperplastic bronchus-associated lymphoid tissue (BALT). Notably, between 10 and 16 months of age 34% (13/38) of these animals developed clonal lymphoproliferative disorders with a mature B cell phenotype (B220+Pax5+) and histologic features of DLBCL (n=6) or less aggressive lymphoid proliferations (LPD: n=6; marginal zone lymphoma: n=1), in contrast with 1/27 heterozygous and 0/25 WT animals. Sequencing analysis of the rearranged immunoglobulin variable region genes in lymphoma biopsies revealed the presence of somatic mutations in 6/8 samples investigated, demonstrating their origin from a GC-experienced B cell. Moreover, immunohistochemical staining for Bcl6 and Irf4 documented a late-GC “activated” B cell phenotype (Bcl6-Irf4+) in all tumors tested (n=4), consistent with the expansion of cells that had been committed to plasma cell differentiation. These data demonstrate that Blimp1 is a bona-fide tumor suppressor gene whose B-cell specific inactivation in vivo promotes the development of lymphomas sharing features of the human ABC-DLBCL.