Published OnlineFirst February 29, 2016; DOI: 10.1158/1078-0432.CCR-15-2296 Biology of Human Tumors Clinical Cancer Research Clinical and Biologic Significance of MYC Genetic Mutations in De Novo Diffuse Large B-cell Lymphoma Zijun Y. Xu-Monette1, Qipan Deng2, Ganiraju C. Manyam3, Alexander Tzankov4, Ling Li1, Yi Xia1, Xiao-xiao Wang1, Dehui Zou1, Carlo Visco5, Karen Dybkær6, Jun Li3, Li Zhang3, Han Liang3, Santiago Montes-Moreno7, April Chiu8, Attilio Orazi9, Youli Zu10, Govind Bhagat11, Kristy L. Richards12, Eric D. Hsi13, William W.L. Choi14, J. Han van Krieken15, Jooryung Huh16, Maurilio Ponzoni17, Andres J.M. Ferreri17, Ben M. Parsons18, Michael B. Møller19, Sa A. Wang1, Roberto N. Miranda1, Miguel A. Piris7, Jane N. Winter20, L. Jeffrey Medeiros1, Yong Li2, and Ken H. Young1,21 Abstract Purpose: MYC is a critical driver oncogene in many cancers, and synonymous mutations). Most of the nonsynonymous muta- its deregulation in the forms of translocation and overexpression tions correlated with better survival outcomes; in contrast, T58 has been implicated in lymphomagenesis and progression of and F138 mutations (which were associated with MYC rear- diffuse large B-cell lymphoma (DLBCL). The MYC mutational rangements), as well as several mutations occurred at the 30 profile and its roles in DLBCL are unknown. This study aims to untranslated region, correlated with significantly worse sur- determine the spectrum of MYC mutations in a large group of vival outcomes. However, these mutations occurred infre- patients with DLBCL, and to evaluate the clinical significance quently (only in approximately 2% of DLBCL). A germline of MYC mutations in patients with DLBCL treated with rituxi- SNP encoding the Myc-N11S variant (observed in 6.5% of the mab, cyclophosphamide, doxorubicin, vincristine, and predni- study cohort) was associated with significantly better patient sone (R-CHOP) immunochemotherapy. survival, and resulted in reduced tumorigenecity in mouse Experimental Design: We identified MYC mutations in 750 xenografts. patients with DLBCL using Sanger sequencing and evaluated the Conclusions: Various types of MYC gene mutations are present prognostic significance in 602 R-CHOP–treated patients. in DLBCL and show different impact on Myc function and clinical Results: The frequency of MYC mutations was 33.3% at the outcomes. Unlike MYC gene translocations and overexpression, DNA level (mutations in either the coding sequence or the most MYC gene mutations may not have a role in driving untranslated regions) and 16.1% at the protein level (non- lymphomagenesis. Clin Cancer Res; 22(14); 3593–605. Ó2016 AACR. Introduction resulted from aberrant class-switch recombination during ger- MYC is a proto-oncogene encoding the Myc protein, a minal center (GC) reaction and leading to Myc overexpression, transcription factor critical for cell proliferation, metabolism, underlies the pathogenesis of Burkitt lymphoma, and the differentiation, apoptosis, microenvironment remodeling, and poorer prognosis of approximately 10% of diffuse large B-cell immune responses. MYC-IGH chromosomal rearrangement, lymphoma (DLBCL) associated with MYC translocation (1). 1Department of Hematopathology,The University of Texas MD Ander- University Hospital, Odense, Denmark. 20Feinberg School of Medicine, son Cancer Center, Houston, Texas. 2Department of Cancer Biology, Northwestern University, Chicago, Illinois. 21The University of Texas Cleveland Clinic, Cleveland, Ohio. 3Department of Bioinformatics School of Medicine, Graduate School of Biomedical Sciences, Hous- and Computational Biology, The University of Texas MD Anderson ton, Texas. Cancer Center, Houston, Texas. 4University Hospital, Basel, Switzer- Note: Supplementary data for this article are available at Clinical Cancer land. 5San Bortolo Hospital,Vicenza, Italy. 6Aarhus University Hospital, Aalborg, Denmark. 7Hospital Universitario Marques de Valdecilla, Research Online (http://clincancerres.aacrjournals.org/). Santander, Spain. 8Memorial Sloan-Kettering Cancer Center, New Z.Y. Xu-Monette and Q. Deng contributed equally to this article. York, New York. 9Weill Medical College of Cornell University, New 10 11 York, New York. The Methodist Hospital, Houston, Texas. Columbia Corresponding Authors: Ken H. Young, The University of Texas MD Anderson University Medical Center and New York Presbyterian Hospital, New 12 Cancer Center, 1515 Holcombe Boulevard, Houston, Texas 77030. Phone: York, New York. University of North Carolina School of Medicine, 713-745-2598; Fax: 713-792-7273; E-mail: [email protected]; Yong Li, Chapel Hill, North Carolina. 13Cleveland Clinic, Cleveland, Ohio. 14Uni- Cleveland Clinic, Cleveland, OH 44106. Phone: 216-372-6889; E-mail: [email protected] versity of Hong Kong Li Ka Shing Faculty of Medicine, Hong Kong, China. 15Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands. 16Asan Medical Center, Ulsan University College of Med- doi: 10.1158/1078-0432.CCR-15-2296 icine, Seoul, Korea. 17San Raffaele H. Scientific Institute, Milan, Italy. 18Gundersen Lutheran Health System, La Crosse, Wisconsin. 19Odense Ó2016 American Association for Cancer Research. www.aacrjournals.org 3593 Downloaded from clincancerres.aacrjournals.org on October 1, 2021. © 2016 American Association for Cancer Research. Published OnlineFirst February 29, 2016; DOI: 10.1158/1078-0432.CCR-15-2296 Xu-Monette et al. mutations (absent in GC-derived follicular lymphoma, pre-GC Translational Relevance mantle-cell lymphoma, post-GC multiple myeloma, and MYC mutations in diffuse large B-cell lymphoma (DLBCL) VH-mutated chronic lymphocytic leukemia as well as normal are not as well studied as MYC translocations, another form tissues) in the 50UTR and CDS regions of the MYC gene of MYC genetic aberrations. This study fills this knowledge harbored by 12 (32%) of 37 patients with DLBCL; these gap by profiling MYC gene mutations and germline variations mutations were proposed to originate from aberrant somatic in a large group of patients with DLBCL and attempts to under- hypermutation processes during DLBCL lymphomagenesis stand their impact on Myc function and clinical outcomes. (25). Through next-generation sequencing, 6 of 111 DLBCL We found a wide range of single-nucleotide variations of biopsies were found to have MYC mutations (26). However, MYC genes in DLBCL which correlated with different clini- the clinical relevance of MYC mutationsinDLBCLhasnotbeen cal outcomes. Mutations known to have gain-of-functions addressed. implicated in the pathogenesis of Burkitt lymphoma by pre- To fill this knowledge gap, this study aims to profile the vious studies were not frequent in DLBCL, whereas most spectrum and frequency of MYC mutations in a large cohort of MYC mutations were associated with better clinical outcomes. patients with DLBCL, to study the functional consequences and to These results suggested that most MYC mutations in DLBCL evaluate the prognostic significance of these MYC mutations. were probably passenger mutations instead of driver muta- tions during lymphomagenesis. This study showed, for the Patients and Methods first time, the clinical significance of MYC mutations in DLBCL, and supported the oncogenic role of MYC. Patients The study cohort consists of 750 patients with de novo DLBCL between 2000 and 2010 according to the World Health Orga- nization classification criteria as a part of the International DLBCL R-CHOP Consortium Program. Patients with trans- Paradoxically, Myc overexpression is also a potent inducer of formed DLBCL, primary mediastinal, cutaneous, or central apoptosis through the modulation of both p53-dependent and nervous system large B-cell lymphomas, or human immunode- p53-independent pathways, including the activation of TP53, ficiency virus infection were excluded. Cell-of-origin classifica- ARF, CD95/FAS,andBAX, and the inhibition of BCL2, BCLX, tion by either gene expression profiling or immunohistochem- and CFLAR/FLIP (2). Therefore, in tumors, deregulation of MYC ical algorithms has been described previously (1, 28). Survival is often concomitant with other abnormalities (e.g., Bcl-2 over- analysis was performed for 602 patients treated with standard expression) that cooperate with Myc during tumor onset, rituximab, cyclophosphamide, doxorubicin, vincristine, and progression, and chemoresistance (3–5). prednisone (i.e., R-CHOP) chemotherapy whose follow-up data In addition to MYC rearrangement, MYC mutation is another were available, randomly divided into a training set (n ¼ 368) form of genetic abnormality found in Burkitt lymphoma. Mul- and a validation set (n ¼ 234). At last follow-up, 208 of the 602 tiple nonsynonymous mutations in the coding sequence (CDS) of patients had died. The rest (394) patients were censored and had the MYC gene have been found in approximately 40% to 70% of a median follow-up time of 54 months (range, 3–187 months). Burkitt lymphoma leading to a mutated Myc protein with amino This study was conducted in accordance with the Declaration of acid changes (6–9). These Myc mutations cluster in the Myc Helsinki and was approved either as minimal to no risk or as transactivation domain with hotspots in the Myc box I (MBI) exempt from review by the Institutional Review Boards of all motif (44-63aa; Fig. 1A), and have been proposed to have a role in participating centers. lymphomagenesis by enhancing the oncogenicity of Myc (9–12). The clinicopathologic features of the patients with or without Functional studies indicated that Myc T58 mutants had increased mutations