Letters to the Editor 719 1,2 1 1 3 1 C Schaab , FS Oppermann , M Klammer , H Pfeifer , A Tebbe , 4 Klammer M, Dybowksi JN, Hoffmann D, Schaab C. Identification of significant 3,4,5 6 7 8 1 3 T Oellerich , J Krauter , M Levis , AE Perl , H Daub , B Steffen , features by a global mean rank test. BMC Bioinfo 2013 (Submitted). 1 3,4,5 K Godl and H Serve 5 Saiki Y, Yamazaki Y, Yoshida M, Katoh O, Nakamura T. Human EVI9, a homologue 1Evotec (Mu¨nchen) GmbH, Am Klopferspitz 19a, Martinsried, of the mouse myeloid leukemia gene, is expressed in the hematopoietic Germany; progenitors and downregulated during myeloid differentiation of HL60 cells. 2Max Planck Institute of Biochemistry, Am Klopferspitz 18, Martinsried, Genomics 2000; 70: 387–391. Germany; 6 Yin B, Delwel R, Valk PJ, Wallace MR, Loh ML, Shannon KM et al. 3Department of Medicine, Hematology/Oncology, Goethe University, A retroviral mutagenesis screen reveals strong cooperation between Bcl11a overexpression and loss of the Nf1 tumor suppressor gene. Blood 2009; 113: Theodor-Stern-Kai 7, Frankfurt, Germany; 4 1075–1085. German Cancer Consortium (DKTK), Heidelberg, Germany; 7 Dai F, Lin X, Chang C, Feng XH. Nuclear export of Smad2 and Smad3 by RanBP3 5 German Cancer Research Center (DKFZ), Heidelberg, Germany; facilitates termination of TGF-beta signaling. Dev Cell 2009; 16: 345–357. 6 Department of Medicine, Hematology/Oncology, Medizinische 8 Yoon SO, Shin S, Liu Y, Ballif BA, Woo MS, Gygi SP et al. Ran-binding protein 3 Hochschule Hannover, Hannover, Germany; phosphorylation links the Ras and PI3-kinase pathways to nucleocytoplasmic 7Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins transport. Mol Cell 2008; 29: 362–375. University, Baltimore, MD, USA and 9 Choudhary C, Olsen JV, Brandts C, Cox J, Reddy PN, Bohmer FD et al. Mislocalized 8Hematologic Malignancies Program, Abramson Cancer Center, activation of oncogenic RTKs switches downstream signaling outcomes. Mol Cell University of Pennsylvania, Philadelphia, PA, USA 2009; 36: 326–339. 10 Klammer M, Kaminski M, Zedler A, Oppermann F, Blencke S, Marx S et al. E-mail: [email protected] or [email protected] Phosphosignature predicts dasatinib response in non-small cell lung cancer. Mol Cell proteomics 2012; 11: 651–668. 11 Gakovic M, Shu X, Kasioulis I, Carpanini S, Moraga I, Wright AF. The role of RPGR in REFERENCES cilia formation and actin stability. Hum mol genet 2011; 20: 4840–4850. 1 Zarrinkar PP, Gunawardane RN, Cramer MD, Gardner MF, Brigham D, Belli B et al. 12 Meier R, Muller PR, Hirt A, Leibundgut K, Ridolfi-Luthy A, Wagner HP. Differential AC220 is a uniquely potent and selective inhibitor of FLT3 for the treatment of phosphorylation of lamin B2 in normal and leukemic cells. Leuk Res 1997; 21: acute myeloid leukemia (AML). Blood 2009; 114: 2984–2992. 841–847. 2 Cortes JE, Perl AE, Dombret H, Kayser S, Steffen B, Rousselot P et al. 13 Kitteringham NR, Jenkins RE, Lane CS, Elliott VL, Park BK. Multiple reaction Final results of a phase 2 open-label, monotherapy efficacy and safety study of monitoring for quantitative biomarker analysis in proteomics and metabolomics. Quizartinib (AC220) in patientsZ60 years of age with FLT3 ITD positive or J Chromatogr B Analyt Technol Biomed Life Sci 2009; 877: 1229–1239. negative relapsed/refractory. AML Blood (ASH Ann Meeting Abstr) 2012; 120: Abstract 48. 3 Cox J, Mann M. MaxQuant enables high peptide identification rates, individua- This work is licensed under a Creative Commons Attribution 3.0 lized p.p.b.-range mass accuracies and proteome-wide protein quantification. Unported License. To view a copy of this license, visit http:// Nat Biotechnol 2008; 26: 1367–1372. creativecommons.org/licenses/by/3.0/

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High prevalence of oncogenic MYD88 and CD79B mutations in primary testicular diffuse large B-cell lymphoma

Leukemia (2014) 28, 719–720; doi:10.1038/leu.2013.348 signaling were shown to promote NF-kB and JAK-STAT3 signaling in this lymphoma type.8 Intriguingly, recent studies indicate that the prevalence of oncogenic MYD88 mutations varies greatly among the ABC–DLBCL presenting at different anatomical Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous class of sites: whereas MYD88 mutations show a high prevalence in lymphomas, comprising of molecularly distinct subtypes that primary-CNS-lymphomas (PCNSLs) as well as in lymphomas arising differ in gene-expression profile (GEP), genetic aberrations, clinical at some other extra-nodal sites, they are relatively uncommon in presentation and disease outcome.1,2 Within this lymphoma class, primary nodal and gastro-intestinal DLBCL.9–11 In a survey of primary testicular lymphoma (PTL) is a distinctive entity genomic alterations in a large panel of DLBCL, we recently found characterized by unique clinical and molecular features, an activating MYD88 mutation in 10 out of 14 PTLs studied,11 including its exclusive manifestation in the immune-privileged suggesting a high mutation prevalence. Here we extended these microenvironment of the testis and frequent dissemination to the series to obtain robust evidence for a role of deregulated MYD88 contralateral testis and the central nervous system (CNS).3,4 signaling in PTLs. Although the incidence of PTL has significantly increased over The study material comprised a panel of 37 PTL diagnosed as the last decades, there is at present no consensus on a standard DLBCLs according the World Health Organization classification, 14 therapeutic regimen.3,4 A current GEP-based molecular of which have been reported previously.11 All tumors were classification of DLBCL distinguishes two main subtypes: extensively immuno-phenotyped, including antibodies against activated B-cell-like (ABC) lymphoma and germinal-center B-cell- CD20, CD10, MUM1, BCL-2 and BCL-6, and tested for Epstein–Barr like lymphoma.1 PTLs belong to the ABC-DLBCL subtype that is virus (EBV) expression by EBV-encoded RNA in-situ hybridization, characterized by constitutively active nuclear factor (NF)-kB and tested for translocations of BCL-2, BCL-6 and c-MYC by signaling.1,2 NF-kB pathway activation in DLBCL may result from fluorescence in-situ hybridization (Supplementary Table 1).11 oncogenic CARD11 mutations and/or of CD79 mutations causing To detect somatic mutations in MYD88 and CD79B, a panel of chronic active B-cell receptor (BCR) signaling.5–7 In addition, allele-specific PCRs covering all major mutation (hot) spots8 was somatically acquired mutations in MYD88, an adaptor protein that employed. As recently reported, this strategy permits efficient and mediates toll-like receptor (TLR) and interleukin-1 receptor sensitive detection of mutations using DNA extracted from the

Accepted article preview online 20 November 2013; advance online publication, 13 December 2013

& 2014 Macmillan Publishers Limited Leukemia (2014) 694 – 725 Letters to the Editor 720 paraffin-embedded tissue, even in samples with relatively low CONFLICT OF INTEREST 11 tumor load. The detected mutations were verified by Sanger The authors declare no conflict of interest. sequencing. Of the 37 PTLs studied, 25 tumors (68%) were found to harbor a MYD88 mutation (Supplementary Table 1). All these W Kraan1, M van Keimpema1, HM Horlings1, EJM Schilder-Tol1, mutations concerned a leucine to proline exchange at position MECM Oud1, LA Noorduyn2, PM Kluin3, MJ Kersten4, 265 (L265P). Among other MYD88 mutations reported, the L265P M Spaargaren1,5 and ST Pals1,5 mutant has been shown to be biologically the most potent 1Department of Pathology, Academic Medical Center, University of and was unique in its ability to organize a stable signaling 8 Amsterdam, Amsterdam, The Netherlands; complex containing phosphorylated IRAK1. These characteristics 2Pathology Laboratory, Dordrecht, The Netherlands; presumably explain its ‘hotspot’ status in lymphomas, including 3 8,12 Department of Pathology, University Medical Center, Groningen, DLBCL and Waldenstro¨ms macroglobulinemia. A CD79B The Netherlands and mutation was found in 7 of the 37 PTLs (Supplementary 4Department of Hematology, Academic Medical Center, University of Table 1). Interestingly, these tumors all harbored a coexisting Amsterdam, Amsterdam, The Netherlands MYD88 mutation. E-mail: [email protected] Our finding that MYD88 mutations are highly prevalent in PTLs 5These authors share last authorship. confirm and extend previous studies reporting a remarkable site-specific variation in the prevalence of MYD88 mutations. Although MYD88 mutations were relatively infrequent in ABC REFERENCES DLBCLs arising in lymph nodes or gut, tumors arising outside 1 Shaffer 3rd Al, Young RM, Staudt LM. The biology of human lymphoid these ‘professional’ lymphoid tissues frequently contained malignancies revealed by gene expression profiling. Ann Rev Immunol 2012; 30: MYD88 mutations, either with or without a coexisting CD79B 565–610. mutation. Interestingly, they were found to be present in up to 2 Pasqualucci L. The genetic basis of diffuse large B-cell lymphoma. Curr Opin 75% of PCNSLs, which together with testicular lymphomas Hemmatol 2013; 20: 336–344. represent the immune-privileged site-associated diffuse large B 3 Ahmad SS, Idris SF, Follows GA, Williams MV. Primary testicular lymphoma. cell lymphomas (IP-DLBCL). Our current finding of a prevalence Clin Oncol 2012; 24: 358–365. of MYD88 mutation in PTL that is comparable to that reported for 4 Vitolo U, Chiappella A, Ferreri AJ, Martelli M, Baldi I, Balzarotti M et al. First-line 9–11 treatment for primary testicular diffuse large B-cell lymphoma with rituximab- PCNSLs supports the concept that IP-DLBCLs present a CHOP, CNS prophylaxis, and contralateral testis irradiation: final results of an molecularly distinct group of lymphomas with shared international phase II trial. J Clin Oncol 2011; 29: 2766–2772. pathogenetic features. Conceivably, mutational activation of 5 Lenz G, Davis RE, Ngo VN, Lam L, George TC, Wright GW et al. Oncogenic TLR/MYD88 signaling may endow lymphoma-initiating cells with CARD11 mutations in human diffuse large B cell lymphoma. Science 2008; 319: a selective growth advantage at immune-privileged sites. Other 1676–1679. than lymph nodes and mucosa-associated lymphoid tissues, 6 Compagno M, Lim WK, Grunn A, Nandula SV, Brahmachary M, Shen Q et al. IP-sites are barrier-protected and immunologically silent and, Mutations of multiple genes cause deregulation of NF-kappaB in diffuse large consequently, will likely under normal circumstances provide B-cell lymphoma. Nature 2009; 459: 717–721. only limited stimulation by TLR ligands. Coexistent CD79B (or 7 Davis RE, Ngo VN, Lenz G, Tolar P, Young RM, Romesser PB et al. Chronic active B-cell-receptor signalling in diffuse large B-cell lymphoma. Nature 2010; 463: other BCR pathway) mutations, causing chronic active BCR 88–92. signaling, may further promote the selective outgrowth of the 8 Ngo VN, Young RM, Schmitz R, Jhavar S, Xiao W, Lim KH et al. Oncogenically active tumor cells within the stimulus-low IP microenvironment. In view MYD88 mutations in human lymphoma. Nature 2011; 470: 115–119. of the crucial role of adhesion and chemokine receptors in tissue- 9 Gonzalez-Aguilar A, Idbaih A, Boisselier B, Habbita N, Rossetto M, Laurenge A et al. specific lymphoma dissemination,13,14 dysregulated ‘homing’ of Recurrent mutations of MYD88 and TBL1XR1 in primary central nervous system lymphoma cells carrying oncogenic MYD88 and/or CD79 lymphomas. Clin Cancer Res 2012; 18: 5203–5211. mutations could present an alternative mechanism underlying 10 Montesinos-Rongen M, Godlewska E, Brunn A, Wiestler OD, Siebert R, Deckert M. the observed site-specific differences in prevalence of these Activating L265P mutations of the MYD88 gene are common in primary central mutations in DLBCL.13 nervous system lymphoma. Acta Neuropathol 2011; 122: 791–792. 11 Kraan W, Horlings HM, van Keimpema M, Schilder-Tol EJM, Oud MECM, In conclusion, our results suggest that MYD88 mutations, and Scheepstra C et al. High prevalence of oncogenic MYD88 and CD79B mutations in to a lesser extent CD79B mutations, are important drivers of diffuse large B-cell lymphomas presenting at immuneprivileged sites. Blood lymphomagenesis in PTL. Presumably, PTL patients may benefit Cancer J 2013; 3: e139. from therapies targeting MYD88 signaling components, includ- 12 Treon SP, Xu L, Yang G, Zhou Y, Liu X, Cao Y et al. MYD88 L265P somatic ing IRAK kinase inhibitors, either alone or in combination with mutation in Waldenstro¨ m’s macroglobulinemia. N Engl J Med 2012; 367: drugs blocking key mediators of BCR signaling such as Bruton’s 826–833. tyrosine kinase.14,15 It will be of interest to explore if patients 13 Pals ST, de Gorter DJ, Spaargaren M. Lymphoma dissemination: the other face with DLBCLs arising in lymph nodes and MALT and lacking these of lymphocyte homing. Blood 2007; 110: 3102–3111. mutations, nevertheless also show evidence of active MYD88 14 de Rooij MF, Kuil A, Geest CR, Eldering E, Chang BY, Buggy JJ et al. The clinically and/or BCR signaling. Such activation might be triggered by active BTK inhibitor PCI-32765 targets B-cell receptor- and chemokine-controlled adhesion and migration in chronic lymphocytic leukemia. Blood 2012; 119: environmental ligands and be associated with non-oncogene 2590–2594. addiction to these pathways. Thus, like PTL patients, these 15 Lim KH, Romero DL, Chaudhary D, Robinson SD, Staudt LM. IRAK4 kinase as a patients may also benefit from therapy targeting MYD88 and/or novel therapeutic target in the abc subtype of diffuse large B cell lymphoma. ASH BCR signaling. Annual Meeting Abstracts 2012; 120: 62.

Supplementary Information accompanies this paper on the Leukemia website (http://www.nature.com/leu)

Leukemia (2014) 694 – 725 & 2014 Macmillan Publishers Limited