Octamer Binding Protein-2 (Oct-2): A non-oncogene addiction in germinal center derived lymphomas and a promising therapeutic target

STELLA MARIS RANUNCOLO; VU NGO; WENMING XIAO; GEORGE WRIGHT; LOUIS M. STAUDT

Washington DC

Congreso; 101st Annual Meeting of the American Association for Cancer Research (AACR); 2010

To identify genes required for the proliferation and survival of Diffuse Large B Cell Lymphomas (DLBCL) we conducted an ?Achilles Heel? RNA interference screen in cell lines model of ABC (Activated B Cell-like) and GCB (Germinal Center B-cell like) DLBCL subtypes. One of the most toxic shRNAs in this screen targeted Oct2, encoding a POU domain transcriptional activator, primarily lymphoid restricted. It was identified due its ability to bind the DNA octamer motif (ATGCAAAT) within immunoglobulin (Ig) genes promoters. The B cell specific coactivator OCA-B interacts with Oct-2 enhancing its transactivation potential. Although Oct2 and OCA-B are dipensable for Ig transcription, they are essential for germinal center (GC) development. To understand the apoptotic cell death of DLBCL cells following shRNA Oct2 induction we investigated the genetic pathways controlled by Oct2. We profiled gene expression changes in DLBCL cells upon Oct2 knocked down and merged this data with genome-wide assessment of Oct2 and OCA-B binding sites, coupling chromatin immunoprecipitation (ChiP) with high-throughput sequencing (ChIPSeq). More than 60% of the Oct2 target genes also showed OCA-B biding. The Oct2/OCA-B overlapping set of targets was enriched for genes selectively expressed in GC B cells. We found that Oct2/OCA-B lie upstream many transcription factors known to play critical roles in GC development such as BCL6, MTA3, PU.1, IRF8 and SpiB. Oct2 regulates these genes in DLBCL cells and in centroblasts. Among Oct-2 downstream effectors, BCL6 could rescue DLBCL cells from the Oct2 shRNA lethal effect. The Oct2/OCA-B binding of BCL6 promoter was confirmed by single locus ChIP in primary GC B-cells and derived lymphomas. Gel shifts experiments showed that Oct2 binds a non canonical octamer site at the BCL6 transcription start site. Our findings uncovered a novel aspect of the Oct2 biology believed to regulate the activity of octamer containing promoters. Interestingly, Oct2 controls the expression of GC specific genes that do not harbor a canonical octamer motif. By array CGH, amplification of Oct2 and OCA-B was found in less than 1% of non Hodgkin lymphoma (NHL) patient samples. Nonetheless, lymphoma cells become dependent on the Oct2 controlled network to survive, being an example of non oncogene addiction. This turns Oct2 into an attractive therapeutic target for NHL treatment. Oct2 and OCA-B lie upstream of BCL6, widely considered a master regulator of the GC response, suggesting that Oct2 directed therapy should kill the same DLBCLs as BCL6 Peptide Inhibitor. Furthermore, all GC and Post-GC B cells tested require Oct2 for survival, indicating that Oct2 directed therapy might have a broder activity spectrum than the BCL6 directed therapy. The Oct2/OCA-B binding interface would be amenable to attack with potential manageable toxicity since this interaction is exclusively required in GC B cells. Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3898. ©2010 American Association for Cancer Research