The advent of massive parallel sequencing highlighted the genetic complexity of cancer, and unveiled the putative contribution of several unsuspected genes to the process of malignant transformation. These advances also indicated a need for the systematic examination of newly-found mutant genes as to determine whether they truly contribute to cancer pathogenesis, or simply represent innocuous variants irrelevant to the observed phenotype. Diffuse large B cell lymphoma (DLBCL), the most common lymphoid malignancy in adults, is curable in only ~60% of cases. This clinical challenge is associated with the disease?s complex genetics, which include disruption of epigenetic modifiers, constitutive activation of the NF-?B pathway, and deregulation of B cell relevant transcription factors. The latter, can derive from chromosomal translocation, genomic amplification/deletions and somatic mutations. Recently, we developed an IGH-targeted capture/sequencing strategy and discovered novel aberrant gene fusions in DLBCL including the juxtaposing of IRF8 (interferon regulatory factor 8) to the IGH locus, t(14;16)(q32;q24), a prototypical model of transcription factor deregulation in B cell lymphomas. Independently, IRF8 was found to be somatically mutated in ~10% of DLBCL biopsies. Interestingly, close to 50% of the IRF8-mutant DLBCLs also have mutations in KMT2D, a histone methyltransferase that when inactivated enhances the lymphomagenic potential of other genetic lesions. IRF8, a member of the interferon family of transcription factors, is expressed in the germinal center (GC) where it directly influences the expression of several key regulators of the GC reaction, including BCL6, AICDA and PRDM1. The central objective of this proposal is to test the hypothesis that IRF8 is a bona fide oncogene in DLBCL, which can be deregulated by chromosomal translocation and somatic mutations. To advance this concept, we found that ectopic expression of IRF8 in DLBCL cell lines promotes a lymphomagenic profile characterized by induction of BCL6 and AICDA, suppression of PRDM1 and resistance to apoptosis. In addition, we generated preliminary data to show that the missense IRF8 mutants found in DLBCL are all gain-of-function. Lastly, we created a mouse with B cell restricted transgenic expression of Irf8. Building on our preliminary data, and on the availability of this novel animal model, we propose to address the following specific aims: 1) Determine the mechanism for Irf8 lymphomagenesis in a mouse model that mimics the IGH/IRF8 fusion found in human DLBCL; 2) Characterize in vivo the pro-lymphoma cooperation between the oncogenic Irf8 and the tumor suppressor Kmt2d; 3) Define the functional consequences of the somatic IRF8 mutations found in DLBCL. When this project is completed, we will show that IRF8 functions as an oncogene that cooperates with KMT2D loss for the development of DLBCL. We also expect to mechanistically link IRF8?s lymphomagenesis to the deregulation of BCL6, AICDA and PRDM1.

Public Health Relevance

Lymphoma, a tumor of the blood cells, is the fifth most common cancer type in the western world. Importantly, lymphoma is one of the few cancers for which the risk was significantly increased among Vietnam veterans. In addition, more recent data (1996-2007) from Italian army peacekeeping troops in the Balkans, also preliminarily suggested an increase in lymphoma incidence. Although the mechanism associated with this elevated risk of lymphoma development have not been fully defined, chemical exposure in the battlefield as well as contact with infectious agents are likely possibilities. Thus, it should be expected that with the current extended deployment of US military forces in foreign lands, a new wave of increased lymphoma diagnosis in military personnel may ensue. To specially attend to the needs of this population, we critically need research that: a) leads to a better understanding of lymphoma biology, b) identifies novel biomarkers for diagnosis and early detection, and c) facilitates the development of more effective and less toxic therapeutics.

Agency
National Institute of Health (NIH)
Institute
Veterans Affairs (VA)
Type
Non-HHS Research Projects (I01)
Project #
5I01BX001882-07
Application #
9685098
Study Section
Hematology (HEMA)
Project Start
2013-04-01
Project End
2021-03-31
Budget Start
2019-04-01
Budget End
2020-03-31
Support Year
7
Fiscal Year
2019
Total Cost
Indirect Cost
Name
South Texas Veterans Health Care System
Department
Type
DUNS #
078493228
City
San Antonio
State
TX
Country
United States
Zip Code
78229
Butler, Matthew J; Aguiar, Ricardo C T (2017) Biology Informs Treatment Choices in Diffuse Large B Cell Lymphoma. Trends Cancer 3:871-882
Elkashef, Sara M; Lin, An-Ping; Myers, Jamie et al. (2017) IDH Mutation, Competitive Inhibition of FTO, and RNA Methylation. Cancer Cell 31:619-620
Yuan, Hongfeng; He, Meilan; Cheng, Fan et al. (2017) Tenovin-6 inhibits proliferation and survival of diffuse large B-cell lymphoma cells by blocking autophagy. Oncotarget 8:14912-14924
Jeong, D; Kim, J; Nam, J et al. (2015) MicroRNA-124 links p53 to the NF-?B pathway in B-cell lymphomas. Leukemia 29:1868-74
Bouamar, Hakim; Jiang, Daifeng; Wang, Long et al. (2015) MicroRNA 155 control of p53 activity is context dependent and mediated by Aicda and Socs1. Mol Cell Biol 35:1329-40
Ortega, M; Bhatnagar, H; Lin, A-P et al. (2015) A microRNA-mediated regulatory loop modulates NOTCH and MYC oncogenic signals in B- and T-cell malignancies. Leukemia 29:968-76
Jiang, Daifeng; Aguiar, Ricardo C T (2014) MicroRNA-155 controls RB phosphorylation in normal and malignant B lymphocytes via the noncanonical TGF-?1/SMAD5 signaling module. Blood 123:86-93
Kim, Sang-Woo; Ramasamy, Kumaraguruparan; Bouamar, Hakim et al. (2012) MicroRNAs miR-125a and miR-125b constitutively activate the NF-?B pathway by targeting the tumor necrosis factor alpha-induced protein 3 (TNFAIP3, A20). Proc Natl Acad Sci U S A 109:7865-70