Received: 31 January 2020 Accepted: 8 February 2020 DOI: 10.1002/hon.2725 INVITED REVIEW Genetics and epigenetics of leukemia and lymphoma: from knowledge to applications, meeting report of the Josep Carreras Leukaemia Research Institute Maribel Parra1 | Maria Joao Baptista2 | Eulàlia Genescà3 | Pere Llinàs-Arias4 | 4,5,6,7 Manel Esteller 1 Lymphocyte Development and Disease Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Spain 2 Lymphoid neoplasms Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Spain 3 Acute lymphoblastic leukemia (ALL) Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Spain 4 Cancer Epigenetics Group, Josep Carreras Leukaemia Research Institute (IJC), Badalona, Spain 5 Centro de Investigacion Biomedica en Red Cancer (CIBERONC), Madrid, Spain 6 Institucio Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain 7 Physiological Sciences Department, School of Medicine and Health Sciences, University of Barcelona (UB), Barcelona, Spain Correspondence Manel Esteller, Cancer Epigenetics Group, Abstract Josep Carreras Leukaemia Research Institute The meeting, which brought together leading scientists and clinicians in the field of (IJC), IJC Building, Campus ICO-Germans Trias i Pujol, Badalona, Spain. leukemia and lymphoma, was held at the new headquarters of the Josep Carreras Email: mesteller@carrerasresearch.org; Leukaemia Research Institute (IJC) in Badalona, Catalonia, Spain, September 19-20, mparra@carrerasresearch.org 2019. Its purpose was to highlight the latest advances in our understanding of the Funding information molecular mechanisms driving blood cancers, and to discuss how this knowledge can The Company of Biologists, Grant/Award Number: EA145 be translated into an improved management of the disease. Special emphasis was placed on the role of genetic and epigenetic heterogeneity, and the exploitation of Peer Review The peer review history for this article is epigenetic regulation for developing biomarkers and novel treatment approaches. available at https://publons.com/publon/10. 1002/hon.2725. KEYWORDS DNA methylation, epi genetics, genetics, leukemia, lymphoma, meeting report 1 | I N T RO DU CT I O N of Leukemia and Lymphoma: From Knowledge to Applications.” The program featured 20 keynote presentations, 9 short talks, and One in eight cases of cancer in the world arises from blood cells, the 30 posters, which represented the status quo of current research in lymphatic system, or bone marrow. Malignant hemopathies, such as genetics and epigenetics of malignant hemopathies. Manel Esteller, leukemia and lymphomas, fall into more than a 100 subtypes with Director of the Institute, welcomed the participants. The meeting very different survival rates. What differentiates one type from was co-organized by Anna Bigas (Institut Hospital del Mar [IMIM], another? These questions were debated during the Inaugural Sympo- Barcelona, Spain), and Marcus Buschbeck, Pablo Menéndez, and sium at the IJC headquarters at Badalona: “Genetics and Epigenetics Francesc Solé (Josep Carreras Leukaemia Research Institute [IJC], Maria Joao Baptista and Eulàlia Genescà contributed equally to the report. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2020 The Authors. Hematological Oncology published by John Wiley & Sons, Ltd 432 wileyonlinelibrary.com/journal/hon Hematological Oncology. 2020;38:432–438. PARRA ET AL. 433 Badalona, Spain). The highlights of the meeting are illustrated in demonstrates how methylation at a specific single CpG dinucleotide Figure 1. in the ZAP-70 50 regulatory sequence is a highly predictive and repro- ducible biomarker of poor prognosis in CLL.10 At the moment, the researchers in Plass's laboratory are trying to distinguish CLL-specific 2 | RECENT ADVANCES IN LEUKEMIA epigenetic alterations from developmental-epi alterations, a task that requires different normal B cells at the various stages of maturation. Pablo Menéndez (Leader of the Stem Cell, IJC-Campus Clinic, Spain) Anna Bigas's research (Coordinator of the Stem Cell and Cancer set the session on acute leukemias in motion. His research has con- Group, IMIM, Barcelona, Spain) aims to decipher the molecular mech- centrated on the specific subset of MLL-AFA4 infant leukemia. Infant anisms regulating stem cells in the hematopoietic system, addressing BCP-ALL (B Cell Precursor Acute Lymphoblastic Leukemia) patients in particular the role of Notch1 in these processes. She explained her harboring the fusion transcript MLL-AF4 (t[4;11]) have a particularly group's most recent findings regarding the mechanism by which poor prognosis and he summarized his laboratory's efforts to identify β-catenin contributes to leukemic initiating cell regulation in Notch- the cell of origin (COO).1-3 Taking a broad, multilayer omics approach, induced T acute lymphoblastic leukemia (T-ALL) murine models.11 his findings have revealed the genetic and epigenetic evolution of a They found that Notch 1 depends on β-catenin for the leukemogenic TCF3-67 ZNF384/PTPN11-driven clone in twins with BCP-ALL, which activity associated with Myc upregulation. supports the hypothesis that a pre-VDJ primitive fetal hematopoietic Meinrad Busslinger (Research Institute of Molecular Pathology, progenitor or stem cell is the COO of TCF3-ZNF384 and PTPN11 Vienna, Austria) spoke about the role of the transcription factor PAX5 4 mutations. Menéndez also drew attention to the great effort expe- in BCP-ALL. He explained that, by generating transgenic mice mimick- 5-8 nded on mimicking this leukemia in vitro. ing the PAX5-ETV6 translocation that occurs in BCP-ALL, the PAX5 Christoph Plass (Head of the Division of Cancer Epigenomics, fusion protein arrests lymphopoiesis at the pro-B-to-pre-B cell transi- German Cancer Research Center, Heidelberg, Germany) spoke about tion and did not interfere with the expression of most of the regulated the epigenetics of leukemia, giving an overview on chronic lympho- PAX5 target genes. Regulated PAX5-ETV6 target genes identified in 9 cytic leukemia (CLL). He also reviewed his previous work, which these BCP-ALLs encode proteins involved in pre-B cell receptor F I G U R E 1 Illustration of the scientific highlights in the genetics and epigenetics of leukemia and lymphoma presented at the Josep Carreras reseach meeting. Meeting sessions were divided in groups according to the hemopathy and basic research advances. ABC, activated B cell; AID, activation-induced deaminase; BCP, B Cell Precursor; GCB, Germinal center B cell; MDS, Myelodysplastic syndrome; MGUS, monoclonal gammopathy of uncertain significance; MMS, smoldering multiple myeloma; PTCL, Peripheral T cell lymphoma; UNG, Uracil-N-glycosylase 434 PARRA ET AL. signaling and migration/adhesion, which could contribute to the pro- Dr Miguel Angel Piris (Fundación Jimenez Díaz, Madrid, Spain) liferation, survival, and tissue infiltration of leukemic B cells.12 began his talk with an overview of peripheral T-cell lymphomas Guillermo García-Manero (MD Anderson Cancer Center, Hous- (PTCLs). He spoke about the constraints on the diagnosis of this sub- ton, TX, USA) reviewed the treatment of myelodysplastic syndromes group of lymphomas and the fruits of a Spanish study including (MDS) and acute myeloblastic leukemia (AML), two very closely 200 cases (PTCL-SPANISH-T-REAL STUDY). The distribution by his- related hematological malignances. He explained that since many tological subtype was identical to that in other series: 34% 13 driver alterations in MDS, such as epigenetic modifications, are angioimmunoblastic T-cell lymphoma (AITL), 14% PTCL with T follicu- common to primary AML, the use of hypomethylating agents has been lar helper (TFH) phenotype, 12% PTCL NOS, 7% intestinal T-cell lym- helpful in prolonging the survival of these patients.14 He detailed clini- phoma, 13% extranodal NK/T, 8% ALK+ anaplastic large cell cal schedules for the different risk-classified MDS subtypes and AML lymphoma (ALCL), and 11% ALK− ALCL. A signature of 13 genes was and drew attention to the importance of using immunotherapy in defined, expressed not only by neoplastic T cells but also by stromal combination with hypomethylating agents.15 He concluded by cells. The signature indicates that deregulated expression of TFH, T- broaching the question of whether patients with low-risk MDS need cytotoxic, and Treg markers may play a role in AITL/PTCL, and that to be treated.16 non-neoplastic B cells, fibroblastic reticular cells, and follicular den- Adolfo Ferrnando (Institute for Cancer Genetics, Columbia Uni- dritic cells are probably involved in AITL/PTCL survival.29,30 versity, New York, NY, USA) has pioneered the application of omics Dr Almudena Ramiro (Spanish National Center for Cardiovascular techniques in pediatric T-ALL. He highlighted the importance of acti- Research, Madrid, Spain) talked about activation-induced deaminase vating mutations in N/KRAS genes that drive sensitivity to vincristine (AID)-induced mutagenesis in normal and lymphoma B cells. She and resistance to methotrexate, which are two chemotherapeutic reviewed the central role of AID in the GC, and the somatic hyper- agents currently in use.17 He also explained how gain-of-function mutation (SHM) and class switch recombination (CSR) processes. In B mutations in the cytosolic 50 nucleotidase II gene (NT5C2) are associ- cells, AID deaminates cytosines in immunoglobulin genes, generating ated with 6-MP resistance.18 Since NT5C2 mutations are found in deoxyuracil. The resulting U:G mismatch is processed by base excision 18,19 20% of relapsed T-ALLs, he concluded that chemotherapy repair or mismatch repair and can lead to point mutations (SHM) or imposes a major evolutionary bottleneck and selects for resistance- double-strand breaks followed by a recombination reaction (CSR). AID associated mutations at relapse. alone is not sufficient to promote lymphomagenesis.31 Indeed, UNG Michel Sadelain (Center for Cell Engineering, Memorial Sloan Ket- regulates the specificity of AID-induced mutations by originating tering Cancer Center, New York, NY, USA) researches engineered error-free and error-prone repair, depending on the sequence con- human cell therapy, particularly that involving chimeric antigen recep- text.32 UNG inhibition gives rise to an increased mutational load and tor (CAR) technology, a cell-based therapy that is changing the way accounts for AID-induced lymphomagenesis through SHM (Figure 1). we treat cancer patients, especially those with leukemia. He explained AID can also induce off-target mutations or chromosome transloca- the most recent advances made in his laboratory to improve CAR-T tions. Almudena's group has identified nine novel genes, including cell therapy by: (a) increasing CAR-T cell efficacy and overcoming pos- MEF2B, LYN, TNFAIP3, GNA13, and IRF8, that accumulate AID- sible mechanisms of CAR-T resistance20-22; (b) preventing CAR-T cell induced mutations and that are mutated in DLBCL.33 cytotoxic release23; and (c) promoting access to patients, decreasing Dr Manel Esteller (Director of the IJC, Badalona, Catalonia, Spain) production costs, and exploring alternative manufacturing and cell reviewed the current state of cancer epigenetics, particularly DNA sources.24,25 methylation in hematological malignancies, some of which have been discovered by his group. Mutations in DNMT3A, that catalyzes 5-methylcytosine methylation occur in a range of hematological malig- 2.1 | Research advances in lymphoma nancies, such as AML and MDS.34 DNA methyltransferase inhibitors, such as azacytidine and decitabine, were the first FDA-approved epi- Dr. Louis Staudt (Center for Cancer Research, National Cancer Institute, drugs and are used in first-line treatment of MDS. Some post- NIH, Bethesda, MD, USA) reviewed the contribution of genomic stud- translational modifications of histones are responsible for nucleosome ies to the improved diagnosis and treatment of lymphomas. Microarray compactation and chromatin conformation. In this context, Dr Esteller studies showed that diffuse large B cell lymphomas (DLBCL) have two presented a new inhibitor of Histone deacetylase 6 that is effective in distinct gene expression profiles that mimic the COO of germinal cen- lymphomas like DLBCL and mantle cell lymphoma35 (Figure 1). Dr ter B cell (GCB) or of activated B cell (ABC). These subtypes have very Esteller further commented on the relevance of epigenetic biomarkers different outcomes with conventional rituximab plus chemotherapy as predictors of response to immunotherapy. For example, the EPI- schemes.26 Recently, DLBCL genetic subtypes have been described MMUNE signature, a DNA methylation signature that includes the T- that provide a better understanding of DLBCL pathogenesis.27 Some cell transcription factor FOXP1, identified the patients with non- genetic DLBCL subtypes share features of other lymphomas, for small-cell lung cancer who were most likely to respond to anti-PD-1 instance the MCD subtype with primary extranodal lymphomas, and inhibitors.36 RNA modifications are also a hot scientific topic. the EZB subtype with germinal center (GC)-derived lymphomas such as NUDT16 removes methylated caps from RNAs, thereby preparing follicular lymphoma (FL) and Burkitt lymphoma (BL).28 them for degradation. Dr Esteller showed that NUDT16 promoter PARRA ET AL. 435 CpG island methylation in T-ALL altered gene transcripts that stabilize disease, in which GC B lymphocytes are the cells of origin of the dis- the C-MYC oncogene, a key gene in T-ALL leukemogenesis.37 To con- ease, driving full transformation of malignant plasma cells at advanced clude, Dr Esteller talked about the transition from one mature cell ages in mice. They found that NF-kB signaling is essential for MM type to another. He showed that pre-B cell-to-macrophage trans- development while Myc deregulation as well as other genetic abnor- differentiation is associated with DNA hypomethylation events in pro- malities markedly accelerate the onset of the disease. The immune moters and distal regulatory regions.38 profiling approach revealed that the immune cell composition in the Dr Ari Melnick (Weill Cornell Medicine, New York, NY, USA) talked bone marrow microenvironment of mice developing MM is similar to about the importance of BCL6 in the GC phenotype and the concur- that of MM patients. These newly generated GEM might be promising rent epigenomic events. Lymphoma cells derive from different stages tools to guide next immunotherapy trials. of the GC reaction and often show mutations and/or aberrant expres- sion of transcription factors and epigenetic modifiers. The following chromatin modifier genes are recurrently mutated in GCB-DLBCL and 4 | A D V A N C E S I N BA S I C R E S E A R C H FL: KMT2D (30-80% of cases), CREBBP (30-40%), EZH2 (30%), TET2 (10% and founder mutation in DLBCL), EP300 (10%).39-41 CREBBP and The basic research talks started with a presentation by Matthias HDAC3 oppose each other via BCL6 during the GC reaction and con- Merkenschlager (MRC London Institute of Medical Sciences, Imperial trol the transition from the dark to light zone of GC to its exit.42 College, London, UK) that focused on the role of three-dimensional CREBBP and EP300 mutations disrupt enhancer switches, causing the (3D) chromatin structure in gene regulation. Cohesin and the architec- sustained repression of enhancers targets by BCL6/HDAC3 com- tural protein CTCF cooperate in genome folding. Merkenschlager 43,44 plexes. (Figure 1). TET2 mutations lead to protein loss, 5mC is not explained how the loss of cohesin downregulates inducible genes, converted in 5hmC, and promoters are kept repressed.45,46 Loss of since it primarily affects the frequency of bursts, which are intense TET2 results in GC arrest and renders cells dependent on HDAC3, like periods of activity between others of inactivity. This frequency is CREBBP mutations. Notably, TET2 and CREBBP mutations are mutu- determined by chromatin looping, enhancer activity, and transcription 45 ally exclusive in DLBCL. EZH2 mediates the transient repression of factor concentrations. Merkenschlager also described how macro- gene promoters in a BCL6-dependent manner.47 EZH2 mutation phage activation leads to the repression and activation of genes and results in enzymatic gain of function, and in vivo EZH2 mutant models enhancers. Mouse macrophages deficient in cohesin have a lower show GC hyperplasia and lymphomagenesis.48 level of expression of inflammatory genes and primary human AML with cohesin mutations have a similar pattern of gene expression.53 Jane Skok (NYU School of Medicine, New York, NY, USA) spoke 3 | RESEARCH ADVANCES IN MULTIPLE about how genes and chromosomes are organized in discreet terri- MYELOMA tories in the interphase nucleus, with little intermingling between them. The genome is folded into regulatory units, known as topologically Maria-Victoria Mateos (University Hospital of Salamanca, Salamanca, associated domains (TADs). There are two main TAD compartments, A Spain) presented a roadmap for patients with multiple myeloma (MM). She and B, that are associated with active and inactive chromatin, respec- explained the differential diagnostic criteria of MM, monoclonal tively. CTCF plays a crucial role organizing chromatin into TADs by gammopathy of uncertain significance (MGUS), and smoldering MM promoting the formation of the loops and boundaries important for (SMM), pointing out that the overall risk of progression from SMM to MM gene regulation. Skok talked about CTCFL (CTCF-like protein) that was 10% per year in the first 5 years.49 Treatment failure/relapses in MM appears to be altered in cancer. Using an inducible complementation can be explained by the escape of particular tumor cells and the accumula- system, they analyzed the impact of expressing CTCFL in the presence tion of oncogenic events. In fact, minimal residual disease status proves to or absence of endogenous CTCF. Skok showed that unique and over- be more important than achieving complete response (CR) for an MM lapping CTCF and CTCFL binding sites differ in their DNA motifs, dis- prognosis.50 The most active treatments must be administered to standard tribution in promoters, intronic/intergenic regions, and local chromatin risk patients as soon as possible.51 High-risk patients, such as those pre- folding, reflecting disparities in each factor's function. CTCFL does not senting del(17p), barely achieve CR with standard treatment approaches share CTCF's insulating properties, nevertheless CTCFL can disrupt (Figure 1) and are good candidates for experimental therapies.52 CTCF-mediated looping, linked to gene expression changes.  Jose Angel Martínez-Climent (Center for Applied Medical Thomas Graf (Center for Genomic Regulation, Barcelona, Spain) Research [CIMA], University of Navarra, Pamplona, Spain) describe introduced a highly efficient cellular transdifferentiation system devel- the progress that has been made in MM immunotherapy. With cur- oped in his laboratory. This approach consists of B cell precursors rent therapies, the median overall survival of MM patients is of up to (pre-B cells) that express an inducible form of the myeloid transcrip- 10 years, but the disease remains incurable. Martínez-Climent's group tion factor C/EBPα causing the direct transdifferentiation into have generated a large collection of genetically heterogeneous mice macrophage-like cells.54,55 Transient expression of C/EBPa in pre-B (GEM) carrying different combinations of eight of the frequent genetic cells followed by the induction of the Yamanaka factors Oct4, Sox2, changes observed in MM patients (Figure 1). At young age, GEM mice Klf4, and Myc, enhances 100-fold the efficiency of the reprogramming with different mutations develop premalignant (MGUS-like) stage into induced pluripotent stem cells (iPSCs).56,57 More recently, using 436 PARRA ET AL. single-cell RNA sequencing to study the transdifferentiation of murine Symposium has represented a remarkable opportunity to discuss with pre-B cells into macrophages and their reprogramming into iPSCs, his worldwide basic, clinical and translational researchers, their recent laboratory has noted a variation in cell fate conversion, which they advances, and novel discoveries in lymphocyte biology, leukemia, and could trace to two subtypes of pre-B cells in the starting population. lymphoma. Overall, the findings presented could pave the way to Large pre-BII cells are highly reprogrammable and express high levels improve the stratification of the diseases, to identify novel biomarkers of Myc, while small pre-BII cells are more susceptible to trans- and to develop new therapeutic targets. differentiation and express low Myc levels. High levels of Myc are a strong predictor of the reprogrammability of a variety of somatic cell ACKNOWLEDG MENTS types into iPSCs58 (Figure 1). The Inaugural Symposium of the Josep Carreras Leukaemia Research Veronika Sexl (Institute of Pharmacology and Toxicology, Vienna, Institute (IJC) entitled “Genetics and Epigenetics of Leukemia and Austria) talked about CDK6, explaining that it is highly expressed in B- Lymphoma: From Knowledge to Applications” was supported by The ALL and ALCL. Her laboratory has demonstrated that CDK6 is a tran- Company of Biologists, Ferrer, CELGENE, Promega, METTLER scriptional regulator with kinase-dependent and kinase-independent TOLEDO, Incyte, DELTACLON, bioNova, Novartis, STEMCELL Tech- roles.59-65 Genome-wide approaches have enabled them to show that nologies, Thermo Fisher Scientific, and DD BIOLAB. CDK6 regulates transcription by binding to chromatin co-localizing with SP1 and NFYA, among others.64 She went on to explain how CONFLIC T OF INT ER E ST CDK6 regulates transcriptional responses that interfere with M.E. is a consultant of Ferrer International and Quimatryx. JAK2V617F-driven disease.65 Sexl also presented her group's findings from a drug screen using wild-type and CDK6-deficient cells that OR CID uncovered that CDK6-deficient cells lack an intact p53 response. Maria Joao Baptista https://orcid.org/0000-0003-3471-2535 Reiner Siebert (Institute of Human Genetics, Ulm University, and Manel Esteller https://orcid.org/0000-0003-4490-6093 Ulm University Medical Center, Ulm, Germany) spoke about the multi- layer sequence-based omics analysis in GC-derived B cell lymphomas. RE FE RE NCE S In particular, using data obtained by several omics approaches in 1. Bueno C, Roldan M, Anguita E, et al. Bone marrow mesenchymal stem cells from patients with aplastic anemia maintain functional and 255 GC B cell lymphomas, he not only showed how DLBCL but also immune properties and do not contribute to the pathogenesis of the all analyses non-Burkitt lymphomas could be divided into different disease. Haematologica. 2014;99(7):1168-1175. genomic subtypes. The second part of his talk dealt on MYC-positive 2. Bueno C, Catalina P, Melen GJ, et al. Etoposide induces MLL lymphomas. He talked about IGH breakpoints in IG-MYC transloca- rearrangements and other chromosomal abnormalities in human tions in the regions of the IGA (22%), IGG (19%), IGM (40%), and VDJ embryonic stem cells. Carcinogenesis. 2009;30(9):1628-1637. 3. Bueno C, Calero-Nieto FJ, Wang X, et al. Enhanced hemato- segments (19%). Few pathways or complexes other than MYC are endothelial specification during human embryonic differentiation deregulated in BLs, but this deregulation seems to be essential and through developmental cooperation between AF4-MLL and MLL-AF4 can occur in the DNA, epigenome, and transcriptome level.66 Finally, fusions. Haematologica. 2019;104(6):1189-1201. Siebert presented results about 8207 differentially methylated regions 4. Bueno C, Tejedor JR, Bashford-Rogers R, et al. Natural history and cell of origin of TC F3-ZN F384 and PTPN11 mutations in monozygotic associated with differential expression between BL and FL.67 twins with concordant BCP-ALL. Blood. 2019;134(11):900-905. Xabier Agirre (CIMA, University of Navarre, Pamplona, Spain) 5. Sanjuan-Pla A, Romero-Moya D, Prieto C, Bueno C, Bigas A, began his talk with a overview of DNA methylation and histone modi- Menendez P. Intra-bone marrow transplantation confers fications. To identify new small molecules that target epigenetic regu- superior multilineage engraftment of murine aorta-gonad Mesoneph- ros cells over intravenous transplantation. Stem Cells Dev. 2016;25(3): lators, they performed a screen in AML and ALL after knocking down 259-265. 134 genes. Agirre reported that G9a and DNMT1 had been identified 6. Prieto C, Stam RW, Agraz-Doblas A, et al. Activated KRAS cooperates as potential therapeutic targets.68 They have designed dual small mol- with MLL-AF4 to promote Extramedullary engraftment and migration ecules that act against G9a and DNMT activity. The lead compound, of cord blood CD34+ HSPC but is insufficient to initiate leukemia. Cancer Res. 2016;76(8):2478-2489. CM-272, inhibits cell proliferation and promotes apoptosis in AML, 7. Prieto C, Marschalek R, Kuhn A, Bursen A, Bueno C, Menendez P. ALL, and DLBCL, inducing interferon-stimulated genes and immuno- The AF4-MLL fusion transiently augments multilineage hematopoietic genic cell death. CM-272 also prolongs survival in xenogeneic models engraftment but is not sufficient to initiate leukemia in cord blood (Figure 1).69 Furthermore, G9a and DNMT1 inhibition enhances the CD34(+) cells. Oncotarget. 2017;8(47):81936-81941. 8. Bueno C, Menendez P. Human acute leukemia induced pluripotent response to PD-L1 blockade, leading to tumor regression and the stem cells: a unique model for investigating disease development and metastasis in a new cancer mouse model.70 pathogenesis. Stem Cell Investig. 2017;4:55. 9. Oakes CC, Seifert M, Assenov Y, et al. DNA methylation dynamics during B cell maturation underlie a continuum of disease phenotypes 5 | CONCLUDING REMARKS in chronic lymphocytic leukemia. Nat Genet. 2016;48(3):253-264. 10. Claus R, Lucas DM, Stilgenbauer S, et al. Quantitative DNA methyla- tion analysis identifies a single CpG dinucleotide important for ZAP- The Inaugural Symposium of the IJC has gathered pioneering scientific 70 expression and predictive of prognosis in chronic lymphocytic leu- experts of the fields of immunology and hematological diseases. The kemia. J Clin Oncol. 2012;30(20):2483-2491. PARRA ET AL. 437 11. Gekas C, D'Altri T, Aligue R, Gonzalez J, Espinosa L, Bigas A. Beta- 32. Pérez-Durán P, Belver L, de Yébenes VG, et al. UNG shapes the spec- catenin is required for T-cell leukemia initiation and MYC transcrip- ificity of AID-induced somatic hypermutation. J Exp Med. 2012;209 tion downstream of Notch1. Leukemia. 2016;30(10):2002-2010. (7):1379-1389. 12. Smeenk L, Fischer M, Jurado S, et al. Molecular role of the  33. Alvarez-Prado  Pérez-Durán P, Pérez-García A, et al. A broad atlas AF, PAX5-ETV6 oncoprotein in promoting B-cell acute lymphoblastic leu- of somatic hypermutation allows prediction of activation-induced kemia. EMBO J. 2017;36(6):718-735. deaminase targets. J Exp Med. 2018;215(3):761-771. 13. Ogawa S. Genetics of MDS. Blood. 2019;133(10):1049-1059. 34. Ley TJ, Ding L, Walter MJ, et al. DNMT3A mutations in acute myeloid 14. Chamseddine AN, Jabbour E, Kantarjian HM, Bohannan ZS, Garcia- leukemia. N Engl J Med. 2010;363:2424-2433. Manero G. Unraveling myelodysplastic syndromes: current knowl- 35. Pérez-Salvia M, Aldaba E, Vara Y, et al. In vitro and in vivo activity of edge and future directions. Curr Oncol Rep. 2016;18(1):4. a new small-molecule inhibitor of HDAC6 in mantle cell lymphoma. 15. Daver N, Boddu P, Garcia-Manero G, et al. Hypomethylating agents Haematologica. 2018;103(11):e537-e540. in combination with immune checkpoint inhibitors in acute myeloid 36. Duruisseaux M, Martínez-Cardús A, Calleja-Cervantes ME, et al. Epi- leukemia and myelodysplastic syndromes. Leukemia. 2018;32(5): genetic prediction of response to anti-PD-1 treatment in non-small- 1094-1105. cell lung cancer: a multicentre, retrospective analysis. Lancet Respir 16. Jabbour E, Short NJ, Montalban-Bravo G, et al. Randomized phase Med. 2018;6(10):771-781. 2 study of low-dose decitabine vs low-dose azacitidine in lower-risk 37. Anadón C, van Tetering G, Ferreira HJ, et al. Epigenetic loss of MDS and MDS/MPN. Blood. 2017;130(13):1514-1522. the RNA decapping enzyme NUDT16 mediates C-MYC activation in 17. Oshima K, Khiabanian H, da Silva-Almeida AC, et al. Mutational land- T-cell acute lymphoblastic leukemia. Leukemia. 2017;31(7):1622- scape, clonal evolution patterns, and role of RAS mutations in 1625. relapsed acute lymphoblastic leukemia. Proc Natl Acad Sci USA. 2016; 38. Bueno-Costa A, Piñeyro D, Soler M, et al. B-cell leukemia Trans- 113(40):11306-11311. differentiation to macrophage involves reconfiguration of DNA meth- 18. Tzoneva G, Perez-Garcia A, Carpenter Z, et al. Activating mutations in ylation for long-range regulation. Leukemia. 2019. https://doi.org/10. the NT5C2 nucleotidase gene drive chemotherapy resistance in 1038/s41375-019-0643-1. relapsed ALL. Nat Med. 2013;19(3):368-371. 39. Pasqualucci L, Trifonov V, Fabbri G, et al. Analysis of the coding 19. Meyer JA, Wang J, Hogan LE, et al. Relapse-specific mutations in genome of diffuse large B-cell lymphoma. Nat Genet. 2011;43(9): NT5C2 in childhood acute lymphoblastic leukemia. Nat Genet. 2013; 830-837. 45(3):290-294. 40. Morin RD, Mendez-Lago M, Mungall AJ, et al. Frequent mutation of 20. Zhao Z, Condomines M, van der Stegen SJC, et al. Structural design histone-modifying genes in non-Hodgkin lymphoma. Nature. 2011; of engineered costimulation determines tumor rejection kinetics and 476(7360):298-303. persistence of CAR T cells. Cancer Cell. 2015;28(4):415-428. 41. Cerchietti LC, Hatzi K, Caldas-Lopes E, et al. BCL6 repression of 21. Hamieh M, Dobrin A, Cabriolu A, et al. CAR T cell trogocytosis and EP300 in human diffuse large B cell lymphoma cells provides a basis cooperative killing regulate tumour antigen escape. Nature. 2019;568 for rational combinatorial therapy. J Clin Invest. 2010;120(12):4569- (7750):112-116. 4582. 22. Feucht J, Sun J, Eyquem J, et al. Calibration of CAR activation poten- 42. Hatzi K, Jiang Y, Huang C, et al. A hybrid mechanism of action tial directs alternative T cell fates and therapeutic potency. Nat Med. for BCL6 in B cells defined by formation of functionally distinct 2019;25(1):82-88. complexes at enhancers and promoters. Cell Rep. 2013;4(3): 23. Giavridis T, van der Stegen SJC, Eyquem J, Hamieh M, Piersigilli A, 578-588. Sadelain M. CAR T cell-induced cytokine release syndrome is medi- 43. Zhang J, Vlasevska S, Wells VA, et al. The CREBBP acetyltransferase ated by macrophages and abated by IL-1 blockade. Nat Med. 2018;24 is a haploinsufficient tumor suppressor in B-cell lymphoma. Cancer (6):731-738. Discov. 2017;7(3):322-337. 24. Themeli M, Riviere I, Sadelain M. New cell sources for T cell engineer- 44. Jiang Y, Ortega-Molina A, Geng H, et al. CREBBP inactivation pro- ing and adoptive immunotherapy. Cell Stem Cell. 2015;16(4):357-366. motes the development of HDAC3-dependent lymphomas. Cancer 25. Riviere I, Sadelain M. Chimeric antigen receptors: a cell and gene Discov. 2017;7(1):38-53. therapy perspective. Mol Ther. 2017;25(5):1117-1124. 45. Dominguez PM, Ghamlouch H, Rosikiewicz W, et al. TET2 deficiency 26. Lenz G, Wright G, Dave SS, et al. Stromal gene signatures in large-B- causes germinal center hyperplasia, impairs plasma cell differentia- cell lymphomas. Lymphoma/leukemia molecular profiling project. N tion, and promotes B-cell lymphomagenesis. Cancer Discov. 2018;8 Engl J Med. 2008;359(22):2313-2323. (12):1632-1653. 27. Schmitz R, Wright GW, Huang DW, et al. Genetics and pathogenesis 46. Rampal R, Alkalin A, Madzo J, et al. DNA hydroxymethylation profil- of diffuse large B-cell lymphoma. N Engl J Med. 2018;378(15):1396- ing reveals that WT1 mutations result in loss of TET2 function in 1407. acute myeloid leukemia. Cell Rep. 2014;9(5):1841-1855. 28. Young RM, Phelan JD, Wilson WH, Staudt LM. Pathogenic B-cell 47. Beguelin W, Teater M, Gearhart MD, et al. EZH2 and BCL6 cooperate receptor signaling in lymphoid malignancies: new insights to improve to assemble CBX8-BCOR complex to repress bivalent promoters, treatment. Immunol Rev. 2019;291(1):190-213. mediate germinal center formation and lymphomagenesis. Cancer Cell. 29. Manso R, González-Rincón J, Rodríguez-Justo M, et al. Overlap at the 2016;30(2):197-213. molecular and immunohistochemical levels between angioimmu- 48. Beguelin W, Popovic R, Teater M, et al. EZH2 is required for germinal noblastic T-cell lymphoma and a subgroup of peripheral T-cell lym- center formation and somatic EZH2 mutations promote lymphoid phomas without specific morphological features. Oncotarget. 2018;9 transformation. Cancer Cell. 2013;23(5):677-692. (22):16124-16133. 49. Kyle RA, Remstein ED, Therneau TM, et al. Clinical course and prog- 30. Manso R, Sánchez-Beato M, González-Rincón J, et al. Mutations in nosis of smoldering (asymptomatic) multiple myeloma. N Engl J Med. the JAK/STAT pathway genes and activation of the pathway, a rele- 2007;356(25):2582-2590. vant finding in nodal peripheral T-cell lymphoma. Br J Haematol. 50. Lahuerta JJ, Paiva B, Vidriales MB, et al. Depth of response in multi- 2018;183(3):497-501. ple myeloma: a pooled analysis of three PETHEMA/GEM clinical tri- 31. Ramiro AR, Jankovic M, Callen E, et al. Role of genomic instability and als. GEM (Grupo Español de Mieloma)/PETHEMA (Programa Para el p53 in AID-induced c-myc-Igh translocations. Nature. 2006;440 Estudio de la Terapéutica en Hemopatías Malignas) cooperative study (7080):105-109. group. J Clin Oncol. 2017;35(25):2900-2910. 438 PARRA ET AL. 51. Moreau P, San Miguel J, Sonneveld P, et al. Multiple myeloma: ESMO 63. Uras IZ, Scheicher RM, Kollmann K, et al. Cdk6 contributes to cyto- clinical practice guidelines for diagnosis, treatment and follow-up. skeletal stability in erythroid cells. Haematologica. 2017;102(6):995- ESMO guidelines committee. Ann Oncol. 2017;28(suppl_4):iv52-iv61. 1005. 52. Sonneveld P, Avet-Loiseau H, Lonial S, et al. Treatment of multiple 64. Bellutti F, Tigan AS, Nebenfuehr S, et al. CDK6 antagonizes myeloma with high-risk cytogenetics: a consensus of the international p53-induced responses during tumorigenesis. Cancer Discov. 2018;8 myeloma working group. Blood. 2016;127(24):2955-2962. (7):884-897. 53. Cuartero S, Weiss FD, Dharmalingam G, et al. Control of inducible 65. Uras IZ, Maurer B, Nivarthi H, et al. CDK6 coordinates JAK2 V617F gene expression links cohesin to hematopoietic progenitor self- mutant MPN via NF-κB and apoptotic networks. Blood. 2019;133 renewal and differentiation. Nat Immunol. 2018;19(9):932-941. (15):1677-1690. 54. Xie H, Ye M, Feng R, Graf T. Stepwise reprogramming of B cells into 66. López C, Kleinheinz K, Aukema SM, et al. Genomic and transcriptomic macrophages. Cell. 2004;117(5):663-676. changes complement each other in the pathogenesis of sporadic 55. Bussmann LH, Schubert A, Vu Manh TP, et al. A robust and highly Burkitt lymphoma. Nat Commun. 2019;10(1):1459. efficient immune cell reprogramming system. Cell Stem Cell. 2009;5 67. Kretzmer H, Bernhart SH, Wang W, et al. DNA methylome analysis in (5):554-566. Burkitt and follicular lymphomas identifies differentially methylated 56. Di Stefano B, Sardina JL, van Oevelen C, et al. C/EBPα poises B cells regions linked to somatic mutation and transcriptional control. Nat for rapid reprogramming into induced pluripotent stem cells. Nature. Genet. 2015;47(11):1316-1325. 2014;506(7487):235-239. 68. Estève PO, Chin HG, Smallwood A, et al. Direct interaction between 57. Di Stefano B, Collombet S, Jakobsen JS, et al. C/EBPα creates elite DNMT1 and G9a coordinates DNA and histone methylation during cells for iPSC reprogramming by upregulating Klf4 and increasing the replication. Genes Dev. 2006;20(22):3089-3103. levels of Lsd1 and Brd4. Nat Cell Biol. 2016;18(4):371-381. 69. San José-Enériz E, Agirre X, Rabal O, et al. Discovery of first-in-class 58. Francesconi M, Di Stefano B, Berenguer C, et al. Single cell RNA-seq reversible dual small molecule inhibitors against G9a and DNMTs in identifies the origins of heterogeneity in efficient cell trans- hematological malignancies. Nat Commun. 2017;8:15424. differentiation and reprogramming. Elife. 2019;8:e41627. 70. Segovia C, San José-Enériz E, Munera-Maravilla E, et al. Inhibition of 59. Kollmann K, Heller G, Schneckenleithner C, et al. A kinase- a G9a/DNMT network triggers immune-mediated bladder cancer independent function of CDK6 links the cell cycle to tumor angiogen- regression. Nat Med. 2019;25(7):1073-1081. esis. Cancer Cell. 2013;24(2):167-181. 60. Scheicher R, Hoelbl-Kovacic A, Bellutti F, et al. CDK6 as a key regula- tor of hematopoietic and leukemic stem cell activation. Blood. 2015; 125(1):90-101. How to cite this article: Parra M, Baptista MJ, Genescà E, 61. Tigan AS, Bellutti F, Kollmann K, Tebb G, Sexl V. CDK6-a review of Llinàs-Arias P, Esteller M. Genetics and epigenetics of the past and a glimpse into the future: from cell-cycle control to tran- leukemia and lymphoma: from knowledge to applications, scriptional regulation. Oncogene. 2016;35(24):3083-3091. meeting report of the Josep Carreras Leukaemia Research 62. Uras IZ, Walter GJ, Scheicher R, et al. Palbociclib treatment of Institute. Hematological Oncology. 2020;38:432–438. https:// FLT3-ITD+ AML cells uncovers a kinase-dependent transcriptional regulation of FLT3 and PIM1 by CDK6. Blood. 2016;127(23):2890- doi.org/10.1002/hon.2725 2902.