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[cat] La leucèmia limfoblàstica aguda (LLA) és l’expansió clonal de cèl·lules malignes originades a partir de progenitors hematopoètics i és el càncer pediàtric més freqüent. La supervivència dels pacients pediàtrics afectes de LLA ha augmentat de forma important en les últimes dècades, actualment supera el 90% en països de renda alta. La intensitat terapèutica dels règims quimioteràpics actuals ha arribat al límit de la tolerància. D’aquesta manera, per a millorar el pronòstic és més important disminuir-ne la toxicitat, més que incrementar-ne la intensitat. L’asparaginasa (ASP) és un quimioteràpic essencial en el tractament de la LLA però s’associa a toxicitats freqüents i a vegades greus, com hipersensibilitat i pancreatitis, que poden tenir un impacte en la supervivència. En la pràctica clínica s’utilitzen 3 formulacions d’ASP, dues derivades d’E. coli (una nativa i una pegilada: E. coli-ASP nativa, PEG-ASP) i una d’Erwinia chrysanthemi (Erwinia-ASP), amb un patró similar de toxicitat excepte una potencial menor immunogenicitat de PEG-ASP respecte E. coli-ASP nativa. La hipersensibilitat és la toxicitat més freqüent i pot manifestar-se com al·lèrgia real (clínica d’al·lèrgia amb inactivació del fàrmac), pseudoal·lèrgia (clínica d’al·lèrgia sense inactivació del fàrmac) i inactivació silent (IS) (inactivació del fàrmac sense cap simptomatologia associada). S’han relacionat certs polimorfismes genètics amb la pancreatitis i la hipersensibilitat associades a ASP. Per a avaluar l’efectivitat de l’ASP, la tècnica gold stantard és la determinació d’activitat d’asparaginasa en sèrum (AAS). Un nivell ≥ 100 UI/L defineix una AAS en rang terapèutic. Partint d’aquest context, en aquest treball de tesi vam plantejar les següents hipòtesis: l’anàlisi de polimorfismes genètics relacionats amb toxicitat a ASP podria identificar els pacients amb major risc de pancreatitis o al·lèrgia; la incidència d'al·lèrgia en pacients pediàtrics que reben un esquema seqüencial d’ASP (E. coli nativa-ASP seguida de PEG-ASP) podria ser major a la dels que reben únicament PEG-ASP dins del mateix protocol i, finalment; que amb la dosi més baixa de PEG-ASP (1.000 UI/m2) es podrien aconseguir nivells d’AAS suficients per a una correcta efectivitat clínica. Per a donar resposta a aquestes hipòtesis, vam enfocar els següents objectius: estudiar la correlació entre els polimorfismes en el gen de l’asparagin-sintetasa (ASNS) relacionats amb toxicitat a ASP en una sèrie de pacients pediàtrics afectes de LLA i comparar la incidència d’al·lèrgia clínica en pacients pediàtrics amb LLA que han rebut diferents esquemes d'administració d’ASP dins el mateix protocol terapèutic. En la nostra població d’estudi, els polimorfismes rs3757676 i rs3832526, descrits prèviament com a haplotip de risc i protector del gen ASNS enfront al·lèrgia i pancreatitis, no van tenir una relació clara amb l’aparició d’aquestes toxicitats. El risc de presentar al·lèrgia a ASP es va associar tant amb el fet de rebre un protocol d’alt risc de recaiguda com el fet de pertànyer a l’ètnia llatinoamericana. El risc de presentar pancreatitis secundària a ASP es va associar amb el fet de tenir una edat ≥ 10 anys en el moment del diagnòstic. En el nostre protocol de tractament LAL/SEHOP-PETHEMA 2013, l’ús de PEG-ASP en primera línia es va associar amb una menor incidència d'al·lèrgia clínica que l'observada en l’ús seqüencial d’E. coli-ASP nativa seguida de PEG-ASP. PEG-ASP a una dosi de 1.000 UI/m2 intramuscular va ser eficaç per assolir una AAS suficient en més del 90% dels pacients. La incidència d'inactivació silent en els pacients tractats amb PEG-ASP en primera línia va ser molt baixa. Vam detectar una associació significativa entre els nivells d’AAS i l’increment dels nivells d’amoni sèric, la disminució dels nivells d’albúmina, i l’increment dels nivells de triglicèrids. No es van observar diferències significatives en la incidència de pancreatitis en funció de la formulació d’ASP utilitzada., [eng] Acute lymphoblastic leukaemia (ALL) is the clonal expansion of malignant cells originating from haematopoietic progenitors and is the most common paediatric cancer. Survival of paediatric ALL patients has increased significantly in recent decades, now exceeding 90% in high-income countries. The therapeutic intensity of current chemotherapy regimens has reached the limit of tolerance. Thus, to improve prognosis, it is more important to decrease toxicity than to increase intensity. Asparaginase (ASP) is an essential chemotherapy drug in the treatment of ALL but is associated with frequent and sometimes severe toxicities, such as hypersensitivity and pancreatitis, which can impact survival. In clinical practice, 3 ASP formulations are used, two derived from E. coli (one native and one pegylated: native E. coli-ASP, PEG-ASP) and one from Erwinia chrysanthemi (Erwinia-ASP), with a similar pattern of toxicity except for a potential lower immunogenicity of PEG-ASP compared to native E. coli-ASP. Hypersensitivity is the most common toxicity and can manifest as allergy (clinical allergy with drug inactivation), allergic-like reaction (clinical allergy without drug inactivation) and silent inactivation (SI) (drug inactivation without any associated symptoms). Certain genetic polymorphisms have been linked to ASP-associated pancreatitis and hypersensitivity. To assess the effectiveness of ASP, the gold standard technique is the determination of serum asparaginase activity (SAA). A level ≥ 100 IU/L defines a SAA in therapeutic range. Against this background, we hypothesise that: the analysis of genetic polymorphisms related to ASP toxicity could identify patients at higher risk of pancreatitis or allergy; the incidence of allergy in paediatric patients receiving a sequential ASP scheme (native E. coli-ASP followed by PEG-ASP) could be higher than those receiving only PEG-ASP within the same clinical protocol and, finally, that the lowest dose of PEG-ASP (1,000 IU/m2) could achieve sufficient SAA levels for clinical effectiveness. To answer these hypotheses, we focused on the following objectives: to study the correlation between polymorphisms in the asparagin synthetase gene (ASNS) related to ASP toxicity in a series of paediatric patients with ALL and to compare the incidence of clinical allergy in paediatric patients with ALL who have received different ASP administration schemes in the same therapeutic protocol. In our study population, the rs3757676 and rs3832526 polymorphisms, previously described as risk and protective haplotypes of the ASNS gene for allergy and pancreatitis, were not clearly associated with the occurrence of these toxicities. The risk of ASP allergy was associated with both receiving a high-risk relapse protocol and being of Latin American ethnicity. The risk of pancreatitis was associated with age ≥ 10 years at diagnosis. In our LAL/SEHOP-PETHEMA 2013 treatment protocol, first-line use of PEG-ASP was associated with lower incidence of clinical allergy than that observed with sequential use of native E. coli-ASP followed by PEG-ASP. PEG-ASP at a dose of 1,000 IU/m2 intramuscularly was effective in achieving sufficient SAA in more than 90% of patients. The incidence of silent inactivation in patients treated with PEG-ASP first-line was very low. We detected a significant association between SAA levels and increased serum ammonium levels, decreased albumin levels, and increased triglyceride levels. No significant differences in the incidence of pancreatitis were observed depending on the ASP formulation used.

[eng] Next-generation sequencing (NGS) has revolutionized biomedical sciences, especially in the area of cancer. It has nourished genomic research with extensive collections of sequenced genomes that are investigated to untangle the molecular bases of disease, as well as to identify potential targets for the design of new treatments. To exploit all this information, several initiatives have emerged worldwide, among which the Pan-Cancer project of the ICGC (International Cancer Genome Consortium) stands out. This project has jointly analyzed thousands of tumor genomes of different cancer types in order to elucidate the molecular bases of the origin and progression of cancer. To accomplish this task, new emerging technologies, including virtualization systems such as virtual machines or software containers, were used and had to be adapted to various computing centers. The portability of this system to the supercomputing infrastructure of the BSC (Barcelona Supercomputing Center) has been carried out during the first phase of the thesis. In parallel, other projects promote the application of genomics discoveries into the clinics. This is the case of MedPerCan, a national initiative to design a pilot project for the implementation of personalized medicine in oncology in Catalonia. In this context, we have centered our efforts on the methodological side, focusing on the detection and characterization of somatic variants in tumors. This step is a challenging action, due to the heterogeneity of the different methods, and an essential part, as it lays at the basis of all downstream analyses. On top of the methodological section of the thesis, we got into the biological interpretation of the results to study the evolution of chronic lymphocytic leukemia (CLL) in a close collaboration with the group of Dr. Elías Campo from the Hospital Clínic/IDIBAPS. In the first study, we have focused on the Richter transformation (RT), a transformation of CLL into a high-grade lymphoma that leads to a very poor prognosis and with unmet clinical needs. We found that RT has greater genomic, epigenomic and transcriptomic complexity than CLL. Its genome may reflect the imprint of therapies that the patients received prior to RT, indicating the presence of cells exposed to these mutagenic treatments which later expand giving rise to the clinical manifestation of the disease. Multiple NGS- based techniques, including whole-genome sequencing and single-cell DNA and RNA sequencing, among others, confirmed the pre-existence of cells with the RT characteristics years before their manifestation, up to the time of CLL diagnosis. The transcriptomic profile of RT is remarkably different from that of CLL. Of particular importance is the overexpression of the OXPHOS pathway, which could be used as a therapeutic vulnerability. Finally, in a second study, the analysis of a case of CLL in a young adult, based on whole genome and single-cell sequencing at different times of the disease, revealed that the founder clone of CLL did not present any somatic driver mutations and was characterized by germline variants in ATM, suggesting its role in the origin of the disease, and highlighting the possible contribution of germline variants or other non-genetic mechanisms in the initiation of CLL.

The authors thank the Hematopathology Collection registered at the Biobank of Hospital Clínic—Institut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS) as well as Sílvia Martín for the technical support. This study was supported by the “la Caixa” Foundation (CLLEvolution-LCF/PR/HR17/52150017, Health Research 2017 Program HR17-00221, to EC), the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (810287, BCLLatlas, to EC, and HH), CERCA Program/Generalitat de Catalunya, Generalitat de Catalunya Suport Grups de Recerca AGAUR 2017-SGR-1142 (to EC), CIBERONC (CB16/12/00225 to EC), Ministerio de Ciencia e Innovación PID2020-117185RB-I00 (to XSP), FEDER: European Regional Development Fund “Una manera de hacer Europa”, and Fundación Asociación Española Contra el Cáncer FUNCAR-PRYGN211258SUÁR (to XSP). The authors thankfully acknowledge the computer resources at MareNostrum4 and the technical support provided by Barcelona Supercomputing Center (RES activity BCV-2018-3-0001). FN acknowledge research support from the American Association for Cancer Research (2021 AACR-Amgen Fellowship in Clinical/Translational Cancer Research, Grant Number 21-40-11-NADE), the European Hematology Association (EHA Junior Research Grant 2021, Grant Number RG-202012-00245), and the Lady Tata Memorial Trust (International Award for Research in Leukemia 2021–2022, Grant Number LADY_TATA_21_3223). EC is an Academia Researcher of the “Institució Catalana de Recerca i Estudis Avançats” (ICREA) of the Generalitat de Catalunya. This work was partially developed at the Centre Esther Koplowitz (CEK, Barcelona, Spain). Peer Reviewed "Article signat per 11 autors/es: Romina Royo, Laura Magnano, Julio Delgado, Sara Ruiz-Gil, Josep Ll. Gelpí, Holger Heyn, Malcom A. Taylor, Tatjana Stankovic, Xose S. Puente, Ferran Nadeu & Elías Campo"

Anti-CD19 chimeric antigen receptor (CAR) T-cell therapy is a newer and effective therapeutic option approved for patients with relapsed/refractory acute lymphoblastic leukemia and diffuse large B-cell lymphoma. Acute kidney injury is a complication of CAR T-cell therapy that can result in kidney failure. In most cases, it is thought to be related to hemodynamic changes due to cytokine release syndrome. Kidney biopsy in this clinical scenario is usually not performed. We report on a kidney transplant recipient in his 40s who developed a posttransplant lymphoproliferative disorder of B-cell origin refractory to conventional treatments and received anti-CD19 CAR T-cell therapy as compassionate treatment. Beginning on day 12 after CAR T-cell infusion, in the absence of clinical symptoms, a progressive decline in estimated glomerular filtration rate of the kidney graft occurred. A subsequent allograft biopsy showed mild tubulointerstitial lymphocyte infiltrates, falling into a Banff borderline-changes category and resembling an acute immunoallergic tubulointerstitial nephritis. Neither CAR T cells nor lymphomatous B cells were detected within the graft cellular infiltrates, suggesting an indirect mechanism of kidney injury. Although kidney graft function partially recovered after steroid therapy, the posttransplant lymphoproliferative disorder progressed and the patient died 7 months later.

This work was partially supported by the European Union's Horizon 2020 research and innovation program (grant No 856620); grants from the Instituto de Salud Carlos III (Madrid, Spain; PI17/02256 and PI20/01845); Consejeria de Economia, Conocimiento, Empresas y Universidad (Granada, Spain; A-CTS-448-UGR18); Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (Sevilla, Spain; PY20/01282); Generalitat de Catalunya (17SGR437); Gilead Sciences Fellowship (GLD17/00282); the "Xarxa de Bancs de tumors" sponsored by Pla Director d'Oncologia de Catalunya (XBTC); the Associazione Italiana per la Ricerca sul Cancro and Fondazione Cariplo (TRIDEO 16923 and AIRC IG21436); the Spanish Association Against Cancer (AECC) Scientific Foundation grant GCTRA18022MORE; and the Consortium for Biomedical Research in Epidemiology and Public Health (CIBERESP), action Genrisk. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript., In conclusion, this study confirmed the association of 31 GWASidentified SNPs with CLL risk and shed some light on the function of some of these biomarkers in the modulation of TReg, B, and T cell differentiation and proliferation, blood concentrations of B cell-related proteins, cell survival, and the expression of immuneand non-immune-related loci. Though outside the scope of the current study, it is important to mention that additional functional studies using blood samples from CLL patients are still required to validate our findings and to decipher the exact biological mechanisms behind the observed associations. A potential limitation of this work was the relatively small population size of the CRuCIAL cohort that hampered the validation of the SNPs showing modest associations., European Union's Horizon 2020 research and innovation program 856620, Instituto de Salud Carlos III PI17/02256 PI20/01845, Consejeria de Economia, Conocimiento, Empresas y Universidad (Granada, Spain) A-CTS-448-UGR18 Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (Sevilla, Spain) PY20/01282, Generalitat de Catalunya, General Electric 17SGR437, Gilead Sciences GLD17/00282, "Xarxa de Bancs de tumors" - Pla Director d'Oncologia de Catalunya (XBTC), Fondazione AIRC per la ricerca sul cancro Fondazione Cariplo TRIDEO 16923 AIRC IG21436, Spanish Association Against Cancer (AECC) Scientific Foundation grant GCTRA18022MORE, Consortium for Biomedical Research in Epidemiology and Public Health (CIBERESP), action Genrisk

The authors would like to thank the patients who participated in the study and their families, friends, caregivers and referring physicians. The authors are very grateful to Drs. Miguel Ángel Perales (Memorial Sloan Kettering Cancer Center, New York), Shannon Maude (Children's Hospital of Philadelphia), Juan José Lasarte (Centro de Investigación Médica Aplicada, Navarra) and Michael Schmitt (Universitäts Klinikum Heidelberg), members of the CART19-BE-01 trial DSMB. The authors are also indebted to the study staff and health care providers at Hospital Clínic, specifically to Anna Boronat, Raquel Martín-Ibáñez, Unai Perpiñá, Josep M. Canals, Vanina Rodríguez, Guillermo Suñé, Esteve Trias and Jordi Yagüe who participated in the development of the CART19 construct; Sara Varea, Judit Pich, Gabriela Recalde, Leticia Pereira and Joan Albert Arnáiz, from the Clinical Trials Unit; Gonzalo Calvo, Andrea Scalise, Joaquín Sáez-Peñataro, Ferran Torres and Sandra Castaño, from the Department of Pharmacology; and Josep Maria Campistol, Antoni Castells, Aurea Mira and Marc Roda as the main representatives from the institution. This study was funded by CatSalut, Projecte ARI and grants co-financed by the Instituto de Salud Carlos III-Subdirección General de Evaluación y Fomento de la Investigación Sanitaria-and Fondo Europeo de Desarrollo Regional (FEDER) PICI14/122, PI13/676, PIE13/33, PI18/775; and Fundació La Caixa CP042702/LCF/PR/GN18/50310007. VO-M and JD were recipients of research grants from FEHH and Generalitat de Catalunya (PERIS IPFE SLT006/17/301), respectively. CART19 cells are emerging as an alternative therapy for patients with chronic lymphocytic leukemia (CLL). Here we report the outcome of nine consecutive patients with CLL treated with ARI-0001 CART19 cells, six of them with Richter's transformation (RT). One patient with RT never received therapy. The cytokine release syndrome rate was 87.5% (12.5% grade ≥3). Neurotoxicity was not observed in any patient. All patients experienced absolute B-cell aplasia, and seven (87.5%) responded to therapy. With a median follow-up of 5.6 months, two patients with RT experienced a CD19-negative relapse. In conclusion, ARI-0001 cell therapy was feasible, safe, and effective in patients with high-risk CLL or RT.

The integration of multi-omics data can greatly facilitate the advancement of research in Life Sciences by highlighting new interactions. However, there is currently no widespread procedure for meaningful multi-omics data integration. Here, we present a robust framework, called InterTADs, for integrating multi-omics data derived from the same sample, and considering the chromatin configuration of the genome, i.e. the topologically associating domains (TADs). Following the integration process, statistical analysis highlights the differences between the groups of interest (normal versus cancer cells) relating to (i) independent and (ii) integrated events through TADs. Finally, enrichment analysis using KEGG database, Gene Ontology and transcription factor binding sites and visualization approaches are available. We applied InterTADs to multi-omics datasets from 135 patients with chronic lymphocytic leukemia (CLL) and found that the integration through TADs resulted in a dramatic reduction of heterogeneity compared to individual events. Significant differences for individual events and on TADs level were identified between patients differing in the somatic hypermutation status of the clonotypic immunoglobulin genes, the core biological stratifier in CLL, attesting to the biomedical relevance of InterTADs. In conclusion, our approach suggests a new perspective towards analyzing multi-omics data, by offering reasonable execution time, biological benchmarking and potentially contributing to pattern discovery through TADs. ‘Hellenic Network for Precision Medicine’ in the framework of the Hellenic Republic – Siemens Settlement Agreement; European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH – CREATE – INNOVATE [GenOptics, project code: T2E1DK-00407]

Several gene expression profiles with a strong correlation with patient outcomes have been previously described in chronic lymphocytic leukemia (CLL), although their applicability as biomarkers in clinical practice has been particularly limited. Here we describe the training and validation of a gene expression signature for predicting early progression in patients with CLL based on the analysis of 200 genes related to microenvironment signaling on the NanoString platform. In the training cohort (n = 154), the CLL15 assay containing a 15-gene signature was associated with the time to first treatment (TtFT) (hazard ratio [HR], 2.83; 95% CI, 2.17-3.68; P < .001). The prognostic value of the CLL15 score (HR, 1.71; 95% CI, 1.15-2.52; P = .007) was further confirmed in an external independent validation cohort (n = 112). Notably, the CLL15 score improved the prognostic capacity over IGHV mutational status and the International Prognostic Score for asymptomatic early-stage (IPS-E) CLL. In multivariate analysis, the CLL15 score (HR, 1.83; 95% CI, 1.32-2.56; P < .001) and the IPS-E CLL (HR, 2.23; 95% CI, 1.59-3.12; P < .001) were independently associated with TtFT. The newly developed and validated CLL15 assay successfully translated previous gene signatures such as the microenvironment signaling into a new gene expression–based assay with prognostic implications in CLL., This work was supported by research funding from the Asociacion Espa ´ nola Contra el C ˜ ancer grant [5U01CA157581- 05, ECRIN-M3 - A29370] and in part by the Instituto de Salud Carlos III, Fondo de Investigaciones Sanitarias [PI17/00950, M.C., PI17/00943, F.B, PI18/01392, P.A.], and the Spanish Ministry of Economy and Competitiveness [CIBERONC-CB16/12/00233], the Education Council or Health Council of the Junta de Castilla y Leon [GRS 2036/A/19], and Gilead Sciences [GLD15/00348]. ´ This work was supported by research funding from the Asociacion´ Espanola Contra el C ˜ ancer grant [5U01CA157581-05, ECRIN-M3 ´ - A29370] and in part by the Instituto de Salud Carlos III, Fondo de Investigaciones Sanitarias [PI17/00950, M.C., PI17/00943, F.B, PI18/01392, P.A.], and the Spanish Ministry of Economy and Competitiveness [CIBERONC-CB16/12/00233], the Education Council or Health Council of the Junta de Castilla y Leon [GRS ´ 2036/A/19], Gilead Sciences [GLD15/00348] and Gilead Fellowships [GLD16/00144, GLD18/00047, F.B.], and Fundacio la ´Marato de TV3 [201905-30-31 F.B]. All Spanish funding was ´ cosponsored by the European Union FEDER program “Una manera de hacer Europa”. M.C. holds a contract from Ministerio de Ciencia, Innovacion y Universidades [RYC-2012-2018].

The combination of a calcineurin inhibitor (CNI) such as tacrolimus (TAC) or cyclosporine (CYSP) with methotrexate (MTX) or with mycophenolate mofetil (MMF) has been commonly used for graft-versus-host disease (GVHD) prophylaxis after reduced-intensity conditioning (RIC) allogeneic hematopoietic cell transplantation (alloHCT), but there are limited data comparing efficacy of the 2 regimens. We evaluated 1564 adult patients who underwent RIC alloHCT for acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML), and myelodysplastic syndrome (MDS) from 2000 to 2013 using HLA-identical sibling (matched related donor [MRD]) or unrelated donor (URD) peripheral blood graft and received CYSP or TAC with MTX or MMF for GVHD prophylaxis. Primary outcomes of the study were acute and chronic GVHD and overall survival (OS). The study divided the patient population into 4 cohorts based on regimen: MMF-TAC, MMF-CYSP, MTX-TAC, and MTX-CYSP. In the URD group, MMF-CYSP was associated with increased risk of grade II to IV acute GVHD (relative risk [RR], 1.78; P < .001) and grade III to IV acute GVHD (RR, 1.93; P = .006) compared with MTX-TAC. In the URD group, use of MMF-TAC (versus MTX-TAC) lead to higher nonrelapse mortality. (hazard ratio, 1.48; P = .008). In either group, no there was no difference in chronic GVHD, disease-free survival, and OS among the GVHD prophylaxis regimens. For RIC alloHCT using MRD, there are no differences in outcomes based on GVHD prophylaxis. However, with URD RIC alloHCT, MMF-CYSP was inferior to MTX-based regimens for acute GVHD prevention, but all the regimens were equivalent in terms of chronic GVHD and OS. Prospective studies, targeting URD recipients are needed to confirm these results. ispartof: BIOLOGY OF BLOOD AND MARROW TRANSPLANTATION vol:25 issue:1 pages:73-85 ispartof: location:United States status: published

Ibrutinib demonstrated robust efficacy, regardless of high-risk features, in previously untreated or relapsed/refractory chronic lymphocytic leukemia (CLL). The IBRORS-CLL study supports the effectiveness and the manageable safety profile of single-agent ibrutinib, which was not adversely affected by high-risk characteristics in real-world CLL patients in Spain. We also found a high molecular testing rate of del(17p)/TP53 mutation and IGHV mutation status. Background: Ibrutinib demonstrated remarkable efficacy and favorable tolerability in patients with untreated or relapsed/refractory (R/R) chronic lymphocytic leukemia (CLL), including those with high-risk genetic alterations. The IBRORS-CLL study assessed the characteristics, clinical management and outcome of CLL patients receiving ibrutinib in routine clinical practice in Spain.Patients: Observational, retrospective, multicenter study in CLL patients who started single-agent ibrutinib as first-line treatment or at first or second relapse between January 2016 and January 2019. Results: A total of 269 patients were included (median age: 70.9 years; cardiovascular comorbidity: 55.4%, including hypertension [47.6%] and atrial fibrillation [AF] [7.1%]). Overall, 96.7% and 69% of patients underwent molecular testing for del(17p)/TP53 mutation and IGHV mutation status. High-risk genetic features included unmutated IGHV (79%) and del(17p)/TP53 mutation (first-line: 66.3%; second-line: 23.1%). Overall, 84 (31.2%) patients received ibrutinib as first-line treatment, and it was used as second- and third-line therapy in 121 (45.0%) and 64 (23.8%) patients. The median progression-free survival and overall survival were not reached irrespective of del(17p)/TP53, or unmutated IGHV. Common grade >= 3 adverse events were infections (122%) and bleeding (3%). Grade >= 3 AF occurred in 1.5% of patients. Conclusion: This real-world study shows that single-agent ibrutinib is an effective therapy for CLL, regardless of age and high-risk molecular features, consistent with clinical trials. Additionally, single-agent ibrutinib was well tolerated, with a low rate of cardiovascular events. This study also emphasized a high molecular testing rate of del(17p)/TP53 mutation and IGHV mutation status in clinical practice according to guideline recommendations. (C) 2021 The Author(s). Published by Elsevier Inc.

Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults in Western countries. Its etiology is largely unknown but increasing incidence rates observed worldwide suggest that lifestyle and environmental factors such as diet might play a role in the development of CLL. Hence, we hypothesized that the consumption of ultra-processed food and drinks (UPF) might be associated with CLL. Data from a Spanish population-based case-control study (MCC-Spain study) including 230 CLL cases (recruited within three years of diagnosis) and 1634 population-based controls were used. The usual diet during the previous year was collected through a validated food frequency questionnaire and food and drink consumption was categorized using the NOVA classification scheme. Logistic regression models adjusted for potential confounders were used. Overall, no association was reported between the consumption of UPF and CLL cases (OR per each 10% increase of the relative contribution of UPF to total dietary intake = 1.09 (95% CI: 0.94; 1.25)), independently of the Rai stage at diagnosis. However, when analyses were restricted to cases diagnosed within, Spanish Ministry of Economy and Competitiveness Carlos III Institute of Health, FEDER funds/European Regional Development Fund (ERDF) (grants PI17/01280, PI11/01810, PI14/01219, PI11/02213, PI09/1662, PI15/00966, PI18/00909, RCESP C03/09, RTICESP C03/10, RTIC RD06/0020/0095, RD12/0036/0056, Rio Hortega CM13/00232, Juan de la Cierva de Incorporacion IJCI-2016-29502 and SV-09-CLINIC-1), Centro de Investigación Biomédica en Red: Epidemiología y Salud Pública (CIBERESP, Spain), Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR), CERCA Programme/Generalitat de Catalunya for institutional support(2017SGR1085), European Regional Development FundERDF/FEDER, Spanish Ministerio de Economía y Competitividad (MINECO), ISCIII-Sub-Directorate General for Evaluation and the European Regional Development Fund (FEDER-”Una manera de Hacer Europa”) and CIBERONC., Spanish Ministry of Science and Innovation through the “Centro de Excelencia Severo Ochoa 2019–2023” Program (CEX2018-000806-S), Generalitat de Catalunya - CERCA Program

Chronic Lymphocytic Leukemia (CLL) is the most prevalent leukemia in Western countries and is notable for its variable clinical course. This variability is partly reflected by the mutational status of IGHV genes. Many CLL samples have been studied in recent years by next-generation sequencing. These studies have identified recurrent somatic mutations in NOTCH1, SF3B1, ATM, TP53, BIRC3 and others genes that play roles in cell cycle, DNA repair, RNA metabolism and splicing. In this study, we have taken a deep-targeted massive sequencing approach to analyze the impact of mutations in the most frequently mutated genes in patients with CLL enrolled in the REM (rituximab en mantenimiento) clinical trial. The mutational status of our patients with CLL, except for the TP53 gene, does not seem to affect the good results obtained with maintenance therapy with rituximab after front-line FCR treatment.

Summary The manageable toxicity profile of obinutuzumab (GA101; G) alone or with chemotherapy in first‐line (1L; fit and non‐fit) and relapsed/refractory (R/R) patients with chronic lymphocytic leukaemia (CLL) was established in the primary analysis of the Phase IIIb GREEN trial (Clinicaltrials.gov: NCT01905943). The final analysis (cut‐off, 31 January 2019) is reported here. Patients received G (1000 mg) alone (G‐mono; fit and non‐fit patients) or with chemotherapy [fludarabine and cyclophosphamide (FC; fit patients); chlorambucil (non‐fit patients); bendamustine (any patient)]. Study endpoints were safety (primary) and efficacy (secondary). Subgroup analyses were performed on prognostic biomarkers in 1L CLL. Overall, 630 patients received 1L and 341 received R/R CLL treatment. At the final analysis, no new safety signals were observed [Grade ≥ 3 adverse events (AEs): 1L 82·7%, R/R 84·5%; serious AEs: 1L 58·1%, R/R 62·5%]. Neutropenia (1L 50·5%, R/R 53·4%) and thrombocytopenia (1L 14·6%, R/R 19·1%) were the most common Grade 3–5 AEs. G‐mono‐, G‐bendamustine and G‐FC‐treated patients with unmutated immunoglobulin heavy chain trended towards shorter progression‐free survival. Achievement of minimal residual disease negativity was greatest in 1L patients treated with G‐FC. In this final analysis of the GREEN trial, the safety profile of G was consistent with current risk management strategies. Biomarker analyses supported efficacy in the specific subgroups., publishedVersion

Targeting Notch signaling has emerged as a promising therapeutic strategy for chronic lymphocytic leukemia (CLL), particularly in NOTCH1-mutated patients. We provide first evidence that the Notch ligand DLL4 is a potent stimulator of Notch signaling in NOTCH1-mutated CLL cells while increases cell proliferation. Importantly, DLL4 is expressed in histiocytes from the lymph node, both in NOTCH1-mutated and -unmutated cases. We also show that the DLL4-induced activation of the Notch signaling pathway can be efficiently blocked with the specific anti-Notch1 antibody OMP-52M51. Accordingly, OMP-52M51 also reverses Notch-induced MYC, CCND1, and NPM1 gene expression as well as cell proliferation in NOTCH1-mutated CLL cells. In addition, DLL4 stimulation triggers the expression of protumor target genes, such as CXCR4, NRARP, and VEGFA, together with an increase in cell migration and angiogenesis. All these events can be antagonized by OMP-52M51. Collectively, our results emphasize the role of DLL4 stimulation in NOTCH1-mutated CLL and confirm the specific therapeutic targeting of Notch1 as a promising approach for this group of poor prognosis CLL patients., Ministerio de Ciencia e Innovación (SAF2015-31242-R and RTI2018-094584-B-I00 to DC and SAF2016-75442-R to MLT), Departament de Salut (SLT002-16-00350) to DC, the ISCII-Sub-Directorate General for Evaluation (PM15/00007), which are part of Plan Nacional de I+D+I and are co-financed by and the European Regional Development Fund (FEDER-“Una manera de hacer Europa”), Generalitat de Catalunya (2014SGR967 and 2017SGR1009) and CIBERONC (CB16/12/00334, CB16/12/00225 and CB16/12/00236

[eng] Resistance to standard therapy and relapses are observed in several cases of both myeloid and lymphoid cell malignancies. Such phenomenon poses unquestionable limits to the utility of current therapeutic approaches, and remains a critical endeavor for the clinical management of patients with advanced cancers. To counteract resistance and to prevent relapse, the deployment of a wide portfolio of agents was put into action, providing at times only marginal benefits with large trade off in toxicity and tolerability. With the advent of immunotherapy huge improvements in patients’ management and overall survival was achieved, particularly in the field of CAR T-cells. Engineering T-cells to redirect them against a tumor antigen have granted impressive responses in the treatment of B-cell malignancies. On the other hand, the clinical implementation of CARTs for other blood cancers is lagging behind. Major reasons lay within the biology, expression and localization of the antigen to be targeted, as it can play vital roles in cell regulation and development in healthy tissues. The present work have provided extensive contribution to the field of CAR T-cells in two different manners: i) by elucidating with robust in vitro and in vivo comparative experiments whether CD123 selective targeting qualifies as myeloablative or not ii) by validating CD1a as a feasible therapeutic antigen for T-ALL and by creating a novel anti CD1a CAR, which showed robust antileukemic activity while preserving safety towards healthy T-cells and T-cells precursors. The development of innovative therapeutic tools that permit to tackle refractory-relapsed cancer with otherwise no further therapeutic options, opens the doors for important advances that will enable society to win the battle against blood cancers.

On behalf of the PETHEMA Group, Spanish Society of Hematology., [Background]: Pediatric‐based or ‐inspired trials have improved the prognosis of adolescents and young adults (AYA) with Philadelphia chromosome‐negative (Ph‐neg) acute lymphoblastic leukemia (ALL)., [Methods]: This study reports the results of treatment of the ALLRE08 trial, a full pediatric trial for AYA aged 15‐30 years with standard‐risk (SR) ALL., [Results]: From 2008 to 2018, 89 patients (38 adolescents [15‐18 years] and 51 young adults [YA, 19‐30 years], median age: 20 [15‐29] years) were enrolled in the ALLRE08 trial. The complete response (CR) was 95%. Twenty‐two patients were transferred to a high‐risk (HR) protocol because of poor marrow response on day 14 (n = 20) or high‐level of end‐induction minimal residual response (MRD ≥ 0.25%, n = 2). Cumulative incidence of relapse (CIR) at 5 years was 35% (95%CI: 23%‐47%), with significant differences between adolescents and YA: 13% (4%‐28%) vs 52% (34%‐67%), P = .012. No treatment‐related mortality was observed in 66/66 patients following the ALLRE08 trial vs 3/23 patients moved to a HR trial. The estimated 5‐year overall survival (OS) was 74% (95%CI: 63%‐85%), with significantly higher rates for adolescents vs YA: 87% (95%CI: 74%‐100%) vs 63% (46%‐80%), P = .021. Although CIR or OS were lower in patients who were transferred to a HR trial, the differences were not statistically significant (CIR: 34% [21%‐47%] vs 37% [14%‐61%]; OS: 78% [66%‐90%] vs 61% [31%;91%])., [Conclusion]: A full pediatric trial is feasible and effective for AYA with Ph‐neg, SR‐ALL, with better results for adolescents than for YA. Outcome of patients with poor early response rescued with a HR trial was not significantly inferior., We thanks to the following institutions from Spain their support for this paper: ICO‐Hospital Germans Trias i Pujol. Josep Carreras Research Institute. Universitat Autònoma de Barcelona. Hospital Universitari i Politècnic La Fe, Valencia. Hospital Clínico Valencia. Hospital Central de Asturias, Oviedo. Hospital Universitario Virgen del Rocío, Sevilla. Hospital General Universitario de Alicante. Hospital Universitari Vall d'Hebron, Universitat Autònoma de Barcelona. Hospital General Universitario de Castellón. ICO‐Hospital Josep Trueta, Girona. Hospital Dr Peset, Valencia. ICO‐Hospital Duran i Reynals, L'Hospitalet de Llobregat. Hospital Son Espases, Palma de Mallorca. Hospital Clínico Universitario, Cancer Research Center (IBMCC‐CSIC/USAL), Cytometry Service, Salamanca. Hospital San Pedro de Alcántara, Cáceres. Hospital Morales Meseguer, Murcia. Hospital Mútua de Terrassa, Terrassa. Hospital Doce de Octubre, Madrid. Hospital Ramón y Cajal, Madrid. Hospital de Sant Pau, Barcelona. Hospital Arnau de Vilanova, Lleida. Hospital Lucus Augusti, Lugo. ICO‐Hospital Joan XXIII, Tarragona.

BCR-ABL1-like B-cell precursor acute lymphoblastic leukemia (BCP-ALL) remains poorly characterized in adults. We sought to establish the frequency and outcome of adolescent and adult BCR-ABL1-like ALL using a novel RNA-Seq signature in a series of patients with BCP-ALL. To this end, we developed and tested an RNA-Seq custom panel of 42 genes related to a BCR-ABL1-like signature in a cohort of 100 patients with BCP-ALL and treated with risk-adapted ALL trials. Mutations related to BCR-ABL1-like ALL were studied in a panel of 33 genes by next-generation sequencing (NGS). Also, CRLF2 overexpression and IKZF1/CDKN2A/B deletions were analyzed. Twenty out of 79 patients (12–84 years) were classified as BCR-ABL1-like (25%) based on heatmap clustering, with significant overexpression of ENAM, IGJ, and CRLF2 (P ≤ 0.001). The BCR-ABL1-like subgroup accounted for 29% of 15–60-year-old patients, with the following molecular characteristics: CRLF2 overexpression (75% of cases), IKZF1 deletions (64%), CDKN2A/B deletions (57%), and JAK2 mutations (57%). Among patients with postinduction negative minimal residual disease, those with the BCR-ABL1-like ALL signature had a higher rate of relapse and lower complete response duration than non-BCR-ABL1-like patients (P = 0.007). Thus, we have identified a new molecular signature of BCR-ABL1-like ALL that correlates with adverse prognosis in adult patients with ALL.

[eng] Chronic lymphocytic leukemia (CLL) is the most common form of adult leukemia in Western countries. Although the disease might follow an indolent course, it rapidly progresses in a fraction of cases, become resistant to treatment, and eventually transform to a more aggressive B-cell lymphoma, known as Richter syndrome. The mechanisms underlying these distinct clinical courses are not fully understood. In this Thesis, we aimed to elucidate the genomic determinants of CLL progression, to provide tools to characterize these tumors from next-generation sequencing data, and to extract key biological findings that could improve the management of the patients. In the first chapter (Studies 1 and 2), we characterized a noncoding mutation effecting the small nuclear RNA U1, a component of the spliceosome involved in the 5’ splice site recognition via base paring. Mutations in this gene altered the splicing and expression of multiple genes, were found in CLL tumors lacking clinically relevant genomic alterations, and were independently associated with patients’ outcome. In the next chapter (Studies 3, 4, and 5), we aimed to deeper into the subclonal architecture of CLL. We identified mutations present in small subpopulations associated with disease progression, recognized common evolutionary trajectories, and showed that the integration of the whole tumor architecture into prognostic models could improve the stratification of the patients. In the third chapter (Study 6), we analyzed the whole genome of CLL patients undergoing Richter syndrome and observed that this transformation was accompanied by an increased mutational and genomic complexity. We identified a unifying mutational process that could orchestrate this genomic chaos. In the fourth chapter (Studies 7 and 8), we developed a bioinformatic algorithm aimed to reconstruct the immunoglobulin gene rearrangements in lymphoid neoplasms from whole-genome sequencing, which might facilitate the use of this methodology in the future clinical practice. By applying this algorithm, we studied a recurrent mutation in the IGLV3-21 gene associated with an aggressive disease with a strong influence on the current and future risk stratification of CLL patients. Altogether, this Thesis has contributed to understand the genomic determinants of CLL progression through the analysis of its dynamic and heterogeneous genetic makeup., [cat] La leucèmia limfocítica crònica (LLC) és la forma més freqüent de leucèmia en adults als països occidentals. Malgrat que la malaltia pot seguir un curs clínic indolent, aquesta pot progressar ràpidament en una fracció dels casos, tornant-se resistent al tractament i, fins i tot, transformar a un limfoma de cèl·lules B més agressiu, fenomen conegut com síndrome de Richter. Els mecanismes que condicionen aquests diferents cursos clínics no són del tot coneguts. Els objectius d’aquesta tesi doctoral van ser elucidar els determinants genòmics de la progressió de la LLC, proporcionar eines per caracteritzar aquests tumors a partir de dades de seqüenciació de nova generació i extreure aspectes biològics clau d’aquests tumors que permetin millorar el maneig dels pacients. Al primer capítol (estudis 1 i 2), vam caracteritzar una mutació no codificant que afectava l’ARN nuclear petit U1, un component de l’espliceosoma implicat en el reconeixement del lloc 5’ d’empalmament (o splicing) per complementarietat de seqüència. Les mutacions d’aquest gen van alterar l’splicing i l’expressió de múltiples gens, es van trobar en tumors de LLC mancats d’alteracions genòmiques clínicament rellevants i es van associar de forma independentment amb el pronòstic dels pacients. En el següent capítol (estudis 3, 4 i 5), vam intentar aprofundir en l’arquitectura subclonal de la LLC. Es van identificar mutacions presents en petites subpoblacions associades a la progressió de la malaltia, es van reconèixer trajectòries evolutives comunes i es va demostrar que la integració de l'arquitectura tumoral en models pronòstics podria millorar l'estratificació dels pacients. En el tercer capítol (estudi 6), vam analitzar el genoma complet de pacients amb LLC que desenvolupaven la síndrome de Richter i vam observar que aquesta transformació anava acompanyava d’un augment de la complexitat mutacional i genòmica. Vam identificar un procés mutacional unificador que podria orquestrar aquest caos genòmic. Al quart capítol (estudis 7 i 8), vam desenvolupar un algoritme bioinformàtic dirigit a reconstruir els reordenaments genètics de les immunoglobulines en les neoplàsies limfoides a partir de la seqüenciació del genoma complet, cosa que podria facilitar l’ús d’aquesta metodologia en la pràctica clínica. Mitjançant l’aplicació d’aquest algoritme, vam estudiar una mutació recurrent en el gen IGLV3-21 associada a una malaltia agressiva amb una forta influència en la present i futura estratificació dels pacients amb LLC. En conjunt, aquesta tesi ha contribuït a entendre els determinants genòmics de la progressió de la LLC mitjançant l’anàlisi de la seva dinàmica i heterogènia composició genètica.

Although the evidence is not consistent, epidemiologic studies have suggested that taller adult height may be associated with an increased risk of some non-Hodgkin lymphoma (NHL) subtypes. Height is largely determined by genetic factors, but how these genetic factors may contribute to NHL risk is unknown. We investigated the relationship between genetic determinants of height and NHL risk using data from eight genome-wide association studies (GWAS) comprising 10,629 NHL cases, including 3,857 diffuse large B-cell lymphoma (DLBCL), 2,847 follicular lymphoma (FL), 3,100 chronic lymphocytic leukemia (CLL), and 825 marginal zone lymphoma (MZL) cases, and 9,505 controls of European ancestry. We evaluated genetically predicted height by constructing polygenic risk scores using 833 height-associated SNPs. We used logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) for association between genetically determined height and the risk of four NHL subtypes in each GWAS and then used fixed-effect meta-analysis to combine subtype results across studies. We found suggestive evidence between taller genetically determined height and increased CLL risk (OR = 1.08, 95% CI = 1.00–1.17, p = 0.049), which was slightly stronger among women (OR = 1.15, 95% CI: 1.01–1.31, p = 0.036). No significant associations were observed with DLBCL, FL, or MZL. Our findings suggest that there may be some shared genetic factors between CLL and height, but other endogenous or environmental factors may underlie reported epidemiologic height associations with other subtypes.

We aimed to study the association between occupational exposure to pesticides and chronic lymphocytic leukemia (CLL) in Spain. Occupational exposure to pesticides (four insecticides, four herbicides and two fungicides) was evaluated using a job-exposure matrix for the Spanish population (MatEmESp) among 302 CLL cases and 1567 population controls in five regions of Spain, 2010–2013. Cumulative exposure scores (CES) were obtained by summing across the exposed jobs the product of prevalence, intensity and duration of exposure to each active substance. Principal components analysis (PCA) and logistic regression models adjusted for age, sex, region, education and occupational exposure to solvents were used. Around 20% of controls and 29% of cases were exposed to one or more pesticides. Compared to non-exposed, subjects in the highest tertile (3rd tertile) of CES of insecticides, herbicides, fungicides were more likely to have CLL [OR (95% CI), P-trend; 2.10 (1.38; 3.19), 0.002; 1.77 (1.12; 2.80), 0.12; and 1.67 (1.06; 2.64), 0.10, respectively). Following PCA, the first component (PC1, explaining 70% of the variation) equally led by seven active substances (the insecticide pyrethrin, all herbicides, all fungicides) was associated with a 26% higher odds of having CLL for 1-standard deviation increase in PC1 (95% CI: 1.14 to 1.40). These results confirm previous associations between CLL and exposure to pesticides and provide additional evidence by application groups and active substance. However, more research is needed to disentangle independent effects of individual active substances., Spanish Ministry of Economy and Competitiveness-Carlos III Institute of Health, European Union (EU) PI08/1770 PI08/0533 PI08/1359 PS09/00773 PI11/01403 PI11/02213 PI14/1219 PI15/00966 PI17/01280 RTIC RD06/0020/0095 RD12/0036/0036 SV-09-CLINIC-1, Instituto de Salud Carlos III IJCI-2016-29502, Instituto de Salud Carlos III API 10/09, Obra Social CAJASTUR (SV-CAJASTUR-1), La Caixa Foundation 2010ACUP 00310, Secretariat for Universities and Research of the Ministry of Business and Knowledge of the Government of Catalonia 2017SGR1085, Spanish Ministerio de Economia y Competitividad (MINECO) through the Instituto de Salud Carlos III (ISCIII) PMP15/00007, CIBERONC del ISCIII

Chronic lymphocytic leukemia (CLL) is a B lymphoid malignancy highly dependent on the microenvironment. Despite new targeted therapies such as ibrutinib and venetoclax, disease progression and relapse remain an issue. CLL cell interactions with the supportive tissue microenvironment play a critical role in disease pathogenesis. We used a platform for drug discovery based on systems biology and artificial intelligence, to identify drugs targeting key proteins described to have a role in the microenvironment. The selected compounds were screened in CLL cell lines in the presence of stromal cells to mimic the microenvironment and validated the best candidates in primary CLL cells. Our results showed that the commercial drug simvastatin was the most effective and selective out of the tested compounds. Simvastatin decreased CLL cell survival and proliferation as well as cell adhesion. Importantly, this drug enhanced the antitumor effect of venetoclax and ibrutinib. We proposed that systems biology approaches combined with pharmacological screening could help to find new drugs for CLL treatment and to predict new combinations with current therapies. Our results highlight the possibility of repurposing widely used drugs such as statins to target the microenvironment and to improve the efficacy of ibrutinib or venetoclax in CLL cells. The study was supported by research funding from the Spanish Ministry of Economy and Competitiveness through the Plan Estatal de Investigación Científica y Técnica y de Innovación (MINECO) RTI2018-094584-B-I00 [to DC] and was cofunded by the European Regional Development Fund [ERDF] and the CERCA program from Generalitat de Catalunya, Centro de Investigación Biomédica en Cáncer [CIBERONC] [CB16/12/00334 and CB16/12/00225] and Generalitat de Catalunya Suport Grups de Recerca [2017 SGR 1009]. This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 306240. NG is a recipient of a predoctoral fellowship from AGAUR, L.R. is a recipient of a postdoctoral fellowship from AGAUR (PERIS), ILO is a recipient of the PhD4MD program and HPA is a recipient of a predoctiral fellowship from Spanish Ministry of Economy and Competitiveness (FPU)

Insulin‐like growth factor levels and chronic lymphocytic leukaemia Spanish Ministry of Economy and Competitiveness–CarlosIII Institute of Health cofunded by FEDER funds/European Regional Develpment Fund (ERDF)–a way to build Europe (PI17/01280, PI11/01810, PI14/01219, PI11/02213, PI15/00966, RCESP C03/09, RTICESP C03/10, RTIC RD06/0020/0095, RD12/0036/0056, Rio Hortega CM13/00232, SV-09-CLINIC-1, CIBERESP and SAF2016-79347-R) and the Agència de Gestió d’Ajuts Universitaris i de Recerca AGAUR(2017SGR1085, 2014SGR756). The ICGC CLL-Genome Project was funded by Spanish Ministerio de Economía y Competitividad (MINECO) through the Instituto de Salud CarlosIII (ISCIII), PMP15/00007 and CIBERONC

This study was supported by the Ministerio de Economía y Competitividad, Grant n. SAF2015-67633-R ,and PI16/00420 which are part of Plan Nacional de I+D+I and are co-financed by the European Regional Development Fund (FEDER-“Una manera de hacer Europa”) and the CERCA program from Generalitat Catalunya. European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement n. 306240; Generalitat de Catalunya Suport Grups de Recerca AGAUR 2017-SGR1009, and Departament de Salut (SLT002-16-00350), Instituto de Salud Carlos III (ISCIII) International Cancer Genome Consortium for Chronic Lymphocytic Leukemia (ICGC-CLL Genome Project), and project PM15/00007, which is part of Plan Nacional de I+D+I and are co-financed by FEDER. NG is a recipient of a predoctoral fellowship from Agaur and EC is an Academia Researcher of the "Institució Catalana de Recerca i Estudis Avançats" (ICREA) of the Generalitat de Catalunya., Giménez, N., Martínez-Trillos, A., Montraveta, A., Lopez-Guerra, M., Rosich, L., Nadeu, F., Valero, J.G., Aymerich, M., Magnano, L., Rozman, M., Matutes, E., Delgado, J., Baumann, T., Gine, E., González, M., Alcoceba, M., Terol, M.J., Navarro, B., Colado, E., Payer, A.R., Puente, X.S., López-Otín, C., Lopez-Guillermo, A., Campo, E., Colomer, D., Villamor, N.

Chronic lymphocytic leukemia is associated with a highly heterogeneous disease course in terms of clinical outcomes and responses to chemoimmunotherapy. This heterogeneity is partly due to genetic aberrations identified in chronic lymphocytic leukemia cells such as mutations of TP53 and/or deletions in chromosome 17p [del(17p)], resulting in loss of one TP53 allele. These aberrations are associated with markedly decreased survival and predict impaired response to chemoimmunotherapy thus being among the strongest predictive markers guiding treatment decisions in chronic lymphocytic leukemia. Clinical trials demonstrate the importance of accurately testing for TP53 aberrations [both del(17p) and TP53 mutations] before each line of treatment to allow for appropriate treatment decisions that can optimize patients’ outcomes. The current report reviews the diagnostic methods to detect TP53 disruption better, the role of TP53 aberrations in treatment decisions and current therapies available for patients with chronic lymphocytic leukemia carrying these abnormalities. The standardization in sequencing technologies for accurate identification of TP53 mutations and the importance of continued evaluation of TP53 aberrations throughout initial and subsequent lines of therapy remain unmet clinical needs as new therapeutic alternatives become available.

Introduction: Chronic inflammation plays a critical role in lymphomagenesis and several dietary factors seem to be involved its regulation. The aim of the current study was to assess the association between the inflammatory potential of the diet and the risk of lymphoma and its subtypes in the European Investigation into Cancer and Nutrition (EPIC) study. Methods: The analysis included 476,160 subjects with an average follow-up of 13.9 years, during which 3,136 lymphomas (135 Hodgkin lymphoma (HL), 2606 non-Hodgkin lymphoma (NHL) and 395 NOS) were identified. The dietary inflammatory potential was assessed by means of an inflammatory score of the diet (ISD), calculated using 28 dietary components and their corresponding inflammatory weights. The association between the ISD and lymphoma risk was estimated by hazard ratios (HR) and 95% confidence intervals (CI) calculated by multivariable Cox regression models adjusted for potential confounders. Results: The ISD was not associated with overall lymphoma risk. Among lymphoma subtypes, a positive association between the ISD and mature B-cell NHL (HR for a 1-SD increase: 1.07 (95% CI 1.01; 1.14), p trend = 0.03) was observed. No statistically significant association was found among other subtypes. However, albeit with smaller number of cases, a suggestive association was observed for HL (HR for a 1-SD increase = 1.22 (95% CI 0.94; 1.57), p trend 0.13). Conclusions: Our findings suggested that a high ISD score, reflecting a pro-inflammatory diet, was modestly positively associated with the risk of B-cell lymphoma subtypes. Further large prospective studies on low-grade inflammation induced by diet are warranted to confirm these findings Grant sponsor: Spanish Ministry of Economy and Competitiveness-Carlos III Institute of Health cofunded by FEDER funds/European Regional Development Fund (ERDF)—a way to build Europe, Grant numbers: [PI13/00061 (to Granada), PI13/01162 (to EPIC-Murcia, Regional Governments of Andalucıa, Asturias, Basque Country, Murcia and Navarra), PI17/01280 and PI14/01219 (to Barcelona), Centro de Investigación Biomédica en Red: Epidemiología y Salud Pública (CIBERESP, Spain)]; Grant sponsor: Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR), CERCA Programme / Generalitat de Catalunya for institutional support; Grant number (2017SGR1085)

Chronic inflammation plays a role in the development of chronic lymphocytic leukaemia (CLL), and diet might modulate chronic inflammation. This study aims to evaluate the association between the dietary inflammatory index (DII&reg, ) and CLL. A total of 366 CLL cases and 1643 controls of the Spanish multicase-control (MCC) Spain study were included. The inflammatory potential of the diet was assessed using the energy-adjusted dietary inflammatory index (E-DII) based on 30 items from a validated semi-quantitative food frequency questionnaire. Odds ratios (OR) and 95% confidence intervals (CI) were estimated using logistic regression models controlling for potential confounders. Overall, a modest, non-statistically significant, positive association was observed between CLL and E-DII scores (OR for a one-unit increase in E-DII: 1.05 (CI 95%: 0.99, 1.12), p-value = 0.09 and by tertiles: ORT2vsT1: 1.20 (CI 95%: 0.90, 1.59), OR T3vsT1: 1.21 (CI 95%: 0.90, 1.62), p trend = 0.21). These results were independent from disease severity (p-het: 0.70), time from diagnosis (p-het: 0.67) and CLL treatment received (p-het: 0.56). No interactions were detected. In conclusion, the consumption of a diet with high pro-inflammatory components was not significantly associated with CLL. Changes towards a more pro-inflammatory dietary pattern in younger generations not included here warrant future research.

Chronic lymphocytic leukemia patients with differential somatic hypermutation status of the immunoglobulin heavy variable genes, namely mutated or unmutated, display fundamental clinicobiological differences. Considering this, we assessed prognosis separately within mutated and unmutated chronic lymphocytic leukemia in 3015 patients, hypothesizing that the relative significance of relevant indicators may differ between these two categories. Within Binet-A mutated chronic lymphocytic leukemia patients, besides TP53 abnormalities, trisomy 12 and stereotyped subset #2 membership were equivalently associated with the shortest time-to-first-treatment and a treatment probability at 5- and 10-years after diagnosis of 40% and 55%, respectively; the remaining cases exhibited 5-year and 10-year treatment probability of 12% and 25%, respectively. Within Binet-A unmutated chronic lymphocytic leukemia patients, besides TP53 abnormalities, del(11q) and/or SF3B1 mutations were associated with the shortest time-to-first-trearment (5- and 10-year treatment probability: 78% and 98%, respectively); in the remaining cases, males had a significantly worse prognosis than females. In conclusion, the relative weight of indicators that can accurately risk stratify early-stage chronic lymphocytic leukemia patients differs depending on the somatic hypermutation status of the immunoglobulin heavy variable genes of each patient. This finding highlights the fact that compartmentalized approaches based on immunogenetic features are necessary to refine and tailor prognostication in chronic lymphocytic leukemia.

Genome-wide association studies have provided evidence for inherited genetic predisposition to chronic lymphocytic leukemia (CLL). To gain insight into the mechanisms underlying CLL risk we analyze chromatin accessibility, active regulatory elements marked by H3K27ac, and DNA methylation at 42 risk loci in up to 486 primary CLLs. We identify that risk loci are significantly enriched for active chromatin in CLL with evidence of being CLL-specific or differentially regulated in normal B-cell development. We then use in situ promoter capture Hi-C, in conjunction with gene expression data to reveal likely target genes of the risk loci. Candidate target genes are enriched for pathways related to B-cell development such as MYC and BCL2 signalling. At 14 loci the analysis highlights 63 variants as the probable functional basis of CLL risk. By integrating genetic and epigenetic information our analysis reveals novel insights into the relationship between inherited predisposition and the regulatory chromatin landscape of CLL., The definition of regulatory landscape at chronic lymphocytic leukaemia (CLL) risk loci is limited. Here, the authors perform an epigenomic characterisation of 42 known risk loci in CLL and normal B cells at different developmental stages and show active chromatin and target genes in the risk loci.

// Burcu Yigit 1 , Peter J. Halibozek 1 , Shih-Shih Chen 2 , Michael S. O’Keeffe 1 , Jon Arnason 3 , David Avigan 3 , Valter Gattei 4 , Atul Bhan 5 , Osman Cen 6 , Richard Longnecker 6 , Nicholas Chiorazzi 2 , Ninghai Wang 1 , Pablo Engel 7 , Cox Terhorst 1 1 Division of Immunology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA 2 Karches Center for Chronic Lymphocytic Leukemia Research, The Feinstein Institute for Medical Research, Manhasset, NY, USA 3 Division of Hematology/Oncology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA 4 Clinical and Experimental Onco-Hematology Unit, Centro di Riferimento Oncologico, I.R.C.C.S., Aviano, Italy 5 Department of Pathology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA 6 Department of Microbiology and Immunology, Northwestern University Feinberg School of Medicine, Chicago, IL, USA 7 Immunology Unit, Department of Cell Biology, Immunology and Neurosciences, Medical School, University of Barcelona, Barcelona, Spain Correspondence to: Burcu Yigit, email: byigit@bidmc.harvard.edu Cox Terhorst, email: cterhors@bidmc.harvard.edu Keywords: CLL, TCL1-192, SLAMF6, BCR, ibrutinib Received: January 29, 2016 Accepted: March 07, 2016 Published: March 25, 2016 ABSTRACT The signaling lymphocyte activation molecule family [SLAMF] of cell surface receptors partakes in both the development of several immunocyte lineages and innate and adaptive immune responses in humans and mice. For instance, the homophilic molecule SLAMF6 (CD352) is in part involved in natural killer T cell development, but also modulates T follicular helper cell and germinal B cell interactions. Here we report that upon transplantation of a well-defined aggressive murine B220 + CD5 + Chronic Lymphocytic Leukemia (CLL) cell clone, TCL1-192, into SCID mice one injection of a monoclonal antibody directed against SLAMF6 (αSlamf6) abrogates tumor progression in the spleen, bone marrow and blood. Similarly, progression of a murine B cell lymphoma, LMP2A/λMyc, was also eliminated by αSlamf6. But, surprisingly, αSLAMF6 neither eliminated TCL1-192 nor LMP2A/λMyc cells, which resided in the peritoneal cavity or omentum. This appeared to be dependent upon the tumor environment, which affected the frequency of sub-populations of the TCL1-192 clone or the inability of peritoneal macrophages to induce Antibody Dependent Cellular Cytotoxicity (ADCC). However, co-administering αSlamf6 with the Bruton tyrosine kinase (Btk) inhibitor, ibrutinib, synergized to efficiently eliminate the tumor cells in the spleen, bone marrow, liver and the peritoneal cavity. Because an anti-human SLAMF6 mAb efficiently killed human CLL cells in vitro and in vivo , we propose that a combination of αSlamf6 with ibrutinib should be considered as a novel therapeutic approach for CLL and other B cell tumors.

In chronic lymphocytic leukemia (CLL), the non-hematopoietic stromal microenvironment plays a critical role in promoting tumor cell recruitment, activation, survival, and expansion. However, the nature of the stromal cells and molecular pathways involved remain largely unknown. Here, we demonstrate that leukemic B lymphocytes induce the activation of retinoid acid synthesis and signaling in the microenvironment. Inhibition of RA-signaling in stromal cells causes deregulation of genes associated with adhesion, tissue organization and chemokine secretion including the B-cell chemokine CXCL13. Notably, reducing retinoic acid precursors from the diet or inhibiting RA-signaling through retinoid-antagonist therapy prolong survival by preventing dissemination of leukemia cells into lymphoid tissues. Furthermore, mouse and human leukemia cells could be distinguished from normal B-cells by their increased expression of Rarγ2 and RXRα, respectively. These findings establish a role for retinoids in murine CLL pathogenesis, and provide new therapeutic strategies to target the microenvironment and to control disease progression., The stromal microenvironment plays a key role in the expansion of chronic lymphocytic leukemia. Here, the authors use the Eµ-TCL1 mouse model to show that leukemic B-cells induce the activation of retinoic acid synthesis in stromal cells of the lymphoid microenvironment, and that impacting on retinoic acid signalling via diet or chemical inhibition prolonged survival by preventing leukemia dissemination and accumulation in lymphoid tissues.

Predoctoral contract to MS (CIBERESP), Spanish Ministry of Economy and Competitiveness Juan de la Cierva de Incorporación grant IJCI-2014-20900. Spanish Ministry of Economy and Competitiveness - Carlos III Institute of Health cofunded by FEDER funds/European Regional Develpment Fund (ERDF) - a way to build Europe [(grants PI17/01280, PI11/01810, PI14/01219, PI11/02213, PI09/1662, PI15/00966, RCESP C03/09, RTICESP C03/10, RTIC RD06/0020/0095, RD12/0036/0056, Rio Hortega CM13/00232, SV-09-CLINIC-1 and Centro de Investigación Biomédica en Red: Epidemiología y Salud Pública (CIBERESP))] and the Agència de Gestió d'Ajuts Universitaris i de Recerca AGAUR (2017SGR1085, 2014SGR756). The ICGC CLL-Genome Project was funded by Spanish Ministerio de Economía y Competitividad (MINECO) through the Instituto de Salud Carlos III (ISCIII), PMP15/00007 and Centro de Investigación Biomédica en Red: Oncología (CIBERONC)., Solans, M., Castelló, A., Benavente, Y., Marcos-Gragera, R., Amiano, P., Gracia-Lavedan, E., Costas, L., Robles, C., Gonzalez-Barca, E., Banda, E.L., Alonso, E., Aymerich, M., Campo, E., Dierssen-Sotos, T., Fernández-Tardón, G., Olmedo-Requena, R., Gimeno, E., Castaño-Vinyals, G., Aragonés, N., Kogevinas, M., Sanjose, S., Pollán, M., Casabonne, D.

Clinical responses to bendamustine in chronic lymphocytic leukemia (CLL) are highly heterogeneous and no specific markers to predict sensitivity to this drug have been reported. In order to identify biomarkers of response, we analyzed the in vitro activity of bendamustine and the gene expression profile in primary CLL cells. We observed that mRNA expression of CD69 (CD69) and ITGAM (CD11b) constitute the most powerful predictor of response to bendamustine. When we interrogated the predictive value of the corresponding cell surface proteins, the expression of the activation marker CD69 was the most reliable predictor of sensitivity to bendamustine. Importantly, a multivariate analysis revealed that the predictive value of CD69 expression was independent from other clinico-biological CLL features. We also showed that when CLL cells were co-cultured with distinct subtypes of stromal cells, an upregulation of CD69 was accompanied by a reduced sensitivity to bendamustine. In agreement with this, tumor cells derived from lymphoid tumor niches harbored higher CD69 expression and were less sensitive to bendamustine than their peripheral blood counterparts. Furthermore, pretreatment of CD69 high CLL cases with the B-cell receptor (BCR) pathway inhibitors ibrutinib and idelalisib decreased CD69 levels and enhanced bendamustine cytotoxic effect. Collectively, our findings indicate that CD69 could be a predictor of bendamustine response in CLL patients and the combination of clinically-tested BCR signaling inhibitors with bendamustine may represent a promising strategy for bendamustine low responsive CLL cases.

Several chronic lymphocytic leukaemia (CLL) susceptibility loci have been reported; however, much of the heritable risk remains unidentified. Here we perform a meta-analysis of six genome-wide association studies, imputed using a merged reference panel of 1,000 Genomes and UK10K data, totalling 6,200 cases and 17,598 controls after replication. We identify nine risk loci at 1p36.11 (rs34676223, P=5.04 × 10−13), 1q42.13 (rs41271473, P=1.06 × 10−10), 4q24 (rs71597109, P=1.37 × 10−10), 4q35.1 (rs57214277, P=3.69 × 10−8), 6p21.31 (rs3800461, P=1.97 × 10−8), 11q23.2 (rs61904987, P=2.64 × 10−11), 18q21.1 (rs1036935, P=3.27 × 10−8), 19p13.3 (rs7254272, P=4.67 × 10−8) and 22q13.33 (rs140522, P=2.70 × 10−9). These new and established risk loci map to areas of active chromatin and show an over-representation of transcription factor binding for the key determinants of B-cell development and immune response.

Fluorizoline is a new synthetic molecule that induces apoptosis by selectively targeting prohibitins. In the study herein, the pro-apoptotic effect of fluorizoline was assessed in 34 primary samples from patients with chronic lymphocytic leukemia. Fluorizoline induced apoptosis in chronic lymphocytic leukemia cells at concentrations in the low micromolar range. All primary samples were sensitive to fluorizoline irrespective of patients' clinical or genetic features, whereas normal T lymphocytes were less sensitive. Fluorizoline increased the protein levels of the pro-apoptotic B-cell lymphoma 2 family member NOXA in chronic lymphocytic leukemia cells. Furthermore, fluorizoline synergized with ibrutinib, 5-aminoimidazole-4-carboxamide riboside or venetoclax to induce apoptosis. These results suggest that targeting prohibitins could be a new therapeutic strategy for chronic lymphocytic leukemia.

Background Non-coding RNAs have been drawing increasing attention in recent years as functional data suggest that they play important roles in key cellular processes. N-BLR is a primate-specific long non-coding RNA that modulates the epithelial-to-mesenchymal transition, facilitates cell migration, and increases colorectal cancer invasion. Results We performed multivariate analyses of data from two independent cohorts of colorectal cancer patients and show that the abundance of N-BLR is associated with tumor stage, invasion potential, and overall patient survival. Through in vitro and in vivo experiments we found that N-BLR facilitates migration primarily via crosstalk with E-cadherin and ZEB1. We showed that this crosstalk is mediated by a pyknon, a short ~20 nucleotide-long DNA motif contained in the N-BLR transcript and is targeted by members of the miR-200 family. In light of these findings, we used a microarray to investigate the expression patterns of other pyknon-containing genomic loci. We found multiple such loci that are differentially transcribed between healthy and diseased tissues in colorectal cancer and chronic lymphocytic leukemia. Moreover, we identified several new loci whose expression correlates with the colorectal cancer patients’ overall survival. Conclusions The primate-specific N-BLR is a novel molecular contributor to the complex mechanisms that underlie metastasis in colorectal cancer and a potential novel biomarker for this disease. The presence of a functional pyknon within N-BLR and the related finding that many more pyknon-containing genomic loci in the human genome exhibit tissue-specific and disease-specific expression suggests the possibility of an alternative class of biomarkers and therapeutic targets that are primate-specific. Electronic supplementary material The online version of this article (doi:10.1186/s13059-017-1224-0) contains supplementary material, which is available to authorized users.

[eng] Follicular Lymphoma (FL) and Chronic Lymphocytic Leukemia (CLL) share several features in common: 1) they are indolent B-cell neoplasms, 2) patients usually relapse after treatment, 3) both pathologies as yet remain incurable. The initial driving event in both malignancies is the early acquisition of genetic alterations; however, the proliferative drive for malignant cells is largely dependent on external signals from the tumor microenvironment, which favor the survival of malignant cells. For this reason, the main aim of this thesis was to explore new therapies targeting the interactions between the tumor cell and its microenvironment in FL and CLL. For the work in FL, we chose to use the Follicular Dendritic Cell (FDC) line HK, isolated from human tonsils. The results obtained indicate that the microenvironment in FL, and more specifically FDCs, can profoundly modify the FL transcriptome by activating different pathways among that: angiogenesis, migration, adhesion and serum-like responses. Then, it was demonstrated that the PI3K pathway is a common signaling mediator of the identified pathways and may constitute a good target for therapy in this setting. The results obtained demonstrated that PI3K inhibition by buparlisib significantly reduced FDC- induced expression and secretion of proangiogenic factors in FL cells and also reduced vasculature formation in vivo. Moreover, it was demonstrated that buparlisib hampers FDC-derived serum-like responses and survival signaling. It was also uncovered that FDCs increase the motility properties of FL cells and consequently lymphoma dissemination, by two complementary mechanisms: first, by increasing the expression of adhesion molecules, which favor the adhesion of FL cells to the extracellular matrix (ECM), and second, by secreting chemotactic molecules such as CXCL12. Buparlisib also shows inhibitory activity in this scenario; on the one hand, it reduces FL adhesion to ECM, and reduces CXCL12 secretion and induced migration and signaling. Moreover, buparlisib shows in vivo activity in subcutaneous and systemic mouse models. In summary, this work demonstrates that FDCs contribute to FL lymphomagenesis trough the modulation of a plethora of events leading to lymphoma growth and dissemination, and how the inhibition of PI3K/AKT axis could interfere with this crosstalk and constitute a valuable therapeutic tool. For the work in CLL we chose to work with daratumumab, an anti-CD38 monoclonal antibody CD38 expression is dynamic and regulated by the tumor microenvironment, and is considered a negative prognostic marker for CLL. The results obtained with daratumumab show that it induces lysis of CLL cells mainly via Antibody-Dependent Cellular Cytotoxicity (ADCC) and Antibody-Dependent Cellular Phagocytosis (ADCP) in the presence of external effectors both in vitro and in vivo. However, daratumumab is not able to engage significant Complement-Dependent Cytotoxicity (CDC) on CLL cells. Daratumumab activity may extend beyond its effect on the tumor cells, as we have demonstrated its potential to counteract microenvironment-derived signaling in protective cancer niches, such as lymph nodes and bone marrow. In this context, daratumumab interferes with in vitro cell migration and in vivo homing of CLL cells to spleen in NSG mice. Moreover, daratumumab interferes with transendothelial migration and organ invasion of malignant cells by significantly reducing CD49d/CD29-mediated adhesion to VCAM-1, indicating that daratumumab treatment may impede CLL tissue infiltration that leads to progressive disease. Daratumumab also demonstrated impressive in vivo activity in a CLL systemic mouse model, where it prevented tumor progression and significantly improved overall survival. Taken together, daratumumab immunotherapy opens a new horizon as a novel therapeutic approach for CD38+ CLL by not only inducing the classical antibody-mediated cytotoxicity but also by harnessing microenvironment-derived survival signaling and blocking CLL dissemination to secondary lymphoid organs., [spa] El linfoma folicular (FL) y la leucemia linfática crónica (CLL) son dos neoplasias indolentes de células B que siguen siendo incurables. En los dos tipos tumorales, la progresión de la enfermedad depende de las señales de supervivencia recibidas del microambiente tumoral. El objetivo principal de esta tesis ha sido explorar nuevas terapias terapéuticas dirigidas a las interacciones entre las células tumorales y su microambiente en FL y CLL. Para el trabajo en FL, se optó por utilizar la línea celular HK, que son un tipo de células dendríticas foliculares (FDCs) aisladas de amígdalas humanas. Los resultados obtenidos indican que el microambiente en FL, y más concretamente las FDCs, puede modificar profundamente el transcriptoma del FL mediante la activación de diferentes vías, entre ellas: la angiogénesis, la migración, la adhesión y la respuesta a suero de las células. Seguidamente, se demostró que la vía de PI3K es un denominador común de las diferentes vías identificadas, constituyendo una buena diana terapéutica para interferir en la interacción del FL con su microambiente. Los resultados obtenidos demostraron que la inhibición de PI3K por buparlisib (pan-inhibidor de PI3K) reduce significativamente la activación de estas vías, después del contacto con el microambiente, en células de FL tanto in vitro como in vivo. Para el trabajo en CLL se optó por trabajar con daratumumab, el primer anticuerpo monoclonal humano anti-CD38. La expresión de CD38 es dinámica en la CLL y está regulada por el microambiente tumoral, considerándose un marcador de mal pronóstico. Los resultados obtenidos con daratumumab muestran que induce lisis celular de las CLL principalmente mediante ADCC y ADCP en presencia de efectores externos, tanto in vitro como in vivo. La actividad de daratumumab va más allá de su efecto directo sobre las células tumorales, demostrando su potencial para contrarrestar la interacción de las células de CLL con el microambiente y las señales protectoras derivadas de éste, interferiendo en procesos de migración, adhesión e invasión celular, indicando que el tratamiento con daratumumab puede impedir la infiltración de las células de CLL en los tejidos, impidiendo así la diseminación de la enfermedad.

[spa] La Leucemia Linfática Crónica (LLC) es la neoplasia linfoide más frecuente con una incidencia creciente a partir de los 40 años. Se trata de una enfermedad heterogénea, indolente en la mayoría de casos pero con un menor grupo de pacientes que presentan una evolución clínica desfavorable y necesidad precoz de tratamiento. Hay diversas variables descritas con valor pronóstico en la LLC aunque muy pocas de ellas validadas en análisis prospectivos. El presente estudio consistió en el seguimiento prospectivo de 307 pacientes diagnosticados de LLC en el Hospital Duran i Reynals entre los años de 2003 y 2010. Al momento del diagnóstico se recogieron los datos demográficos, clínicos, de laboratorio así como los principales factores pronósticos utilizados en la práctica clínica. Se hizo seguimiento de los pacientes de cara a progresión de la LLC, necesidad de tratamiento o exitus. Se estudió la incidencia de las principales variables pronosticas en estos pacientes y posteriormente se realizaron dos grandes análisis enfocados en las poblaciones con estadios iniciales de la enfermedad: (1) De los 135 casos diagnosticados entre 2003 y 2006, se analizaron los factores pronósticos más relevantes para supervivencia libre de progresión (SLP) en los pacientes con estadios 0 y 1 de Rai (n=107); (2) De los 307 casos diagnosticados entre 2003 y 2010 se estudiaron las mismas variables pronosticas para SLP y supervivencia libre de tratamiento (SLT) en los 231 pacientes con linfocitos clonales con fenotipo de LLC en sangre periférica sin ningún otro signo o síntoma de la enfermedad (pacientes con estadío 0 de Rai y pacientes con linfocitosis B monoclonal, LBM). El principal objetivo del estudio fue identificar factores pronósticos presentes en el momento del diagnóstico de las formas iniciales de LLC, analizando variables ampliamente utilizadas en la práctica clínica diaria y todo ello en un estudio prospectivo con un largo seguimiento., [eng] Chronic lymphocytic leukemia (CLL) is the most common leukemia in western countries and has a heterogeneous clinical course. The majority of patients are diagnosed with early stage CLL and it is difficult to predict which of them will present a more unfavorable clinical behavior with early disease progression and need for treatment. In the past 25 years, many studies tried to identify prognostic variables in CLL although most of them are retrospective, include patients at various stages of the disease and/or analyze variables that are not available in daily clinical practice in most centers. We describe a prospective study conducted during 10 years in the Institut Català d'Oncologia, Hospital Duran i Reynals that includes 307 patients diagnosed with lymphoproliferative syndromes with LLC phenotype. Clinical data and laboratory findings were collected at diagnosis and all prognostic variables analyzed are available in clinical practice. The aim of the study was to identify prognostic factors regarding progression free survival and treatment free survival in early stages CLL patients.

Summary Chronic lymphocytic leukemia (CLL) is an adult B cell malignancy. Genome-wide association studies show that variation at 15q15.1 influences CLL risk. We deciphered the causal variant at 15q15.1 and the mechanism by which it influences tumorigenesis. We imputed all possible genotypes across the locus and then mapped highly associated SNPs to areas of chromatin accessibility, evolutionary conservation, and transcription factor binding. SNP rs539846 C>A, the most highly associated variant (p = 1.42 × 10−13, odds ratio = 1.35), localizes to a super-enhancer defined by extensive histone H3 lysine 27 acetylation in intron 3 of B cell lymphoma 2 (BCL2)-modifying factor (BMF). The rs539846-A risk allele alters a conserved RELA-binding motif, disrupts RELA binding, and is associated with decreased BMF expression in CLL. These findings are consistent with rs539846 influencing CLL susceptibility through differential RELA binding, with direct modulation of BMF expression impacting on anti-apoptotic BCL2, a hallmark of oncogenic dependency in CLL., Graphical Abstract, Highlights • SNP rs539846 underlies 15q15.1 association with chronic lymphocytic leukemia • rs539846 resides in a B cell super-enhancer, disrupting a conserved RELA-binding site • The rs539846 risk allele (A) reduces enhancer activity and RELA binding in CLL • rs539846-A confers lower BMF expression, Kandaswamy et al. find that SNP rs539846 underlies the 15q15.1 chronic lymphocytic leukemia risk locus. Follow-up data demonstrate that rs539846 resides within a transcriptional enhancer and alters RELA binding at a conserved site. The rs539846-A risk allele results in reduced RELA-mediated enhancer activity and lower expression of BCL-2-modifying factor.

Chronic lymphocytic leukemia (CLL) is a common lymphoid malignancy with strong heritability. To further understand the genetic susceptibility for CLL and identify common loci associated with risk, we conducted a meta-analysis of four genome-wide association studies (GWAS) composed of 3,100 cases and 7,667 controls with follow-up replication in 1,958 cases and 5,530 controls. Here we report three new loci at 3p24.1 (rs9880772, EOMES, P=2.55 × 10−11), 6p25.2 (rs73718779, SERPINB6, P=1.97 × 10−8) and 3q28 (rs9815073, LPP, P=3.62 × 10−8), as well as a new independent SNP at the known 2q13 locus (rs9308731, BCL2L11, P=1.00 × 10−11) in the combined analysis. We find suggestive evidence (P, Chronic lymphocytic leukemia is a highly inheritable cancer. Here the authors conduct a metaanalysis of four genome-wide association studies and identify three novel loci located near EOMES, SERPINB6 and LPP associated with risk of this disease.

Background: Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults in Europe and in North America. It is caused by the neoplastic transformation of a population of B-lymphocytes coexpressing CD5, CD19 and CD23. Although the etiology is not well known there is evidence of the existence of a genetical base. CLL is not only caused by the accumulation of B-lymphocytes due to a failure in the apoptosis. It is also caused by the active proliferation of the B-cells in response to the signs sent by the environment. Its clinical course is variable. One third of CLL patients are affected by an indolent form that does not require treatment. Another third of patients with leukemia will require iterative therapies and a small fraction patients develop Richter syndrome that reduces its overall survival to 5 to 8 months. This variability is due to the heterogeneity of the molecules that participate in the pathogenesis. One of these molecules is CD38. Some studies have demonstrated that the presence of this marker confers a worse prognostic, specially the presence of one polymorphism of CD38, rs6449182. Fluradabine, clyclophosphamide and rituximab (FCR) is considered the gold standard therapy for young patients under 65 years with CLL because achieves high percentage of minimal residual disease (MRD). But it has not been studied if in CLL patients with this polymorphism, FCR continues offering these good results. Objective: To analyze the relationship between the polymorphisms of CD38 and treatment outcomes in CLL patients treated with FCR. Methods: The design is a multicenter prospective cohort study. 338 patients will be recruited during 4 years. We will analyze the presence of the polymorphism and evaluate the minimal residual disease (MRD) after the treatment with FCR

on behalf of ERIC, the European Research Initiative on CLL; International audience; We report on markedly different frequencies of genetic lesions within subsets of chronic lymphocytic leukemia patients carrying mutated or unmutated stereotyped B-cell receptor immunoglobu-lins in the largest cohort (n=565) studied for this purpose. By combining data on recurrent gene mutations (BIRC3, MYD88, NOTCH1, SF3B1 and TP53) and cytogenetic aberrations, we reveal a subset-biased acquisition of gene mutations. More specifically, the frequency of NOTCH1 mutations was found to be enriched in subsets expressing unmutated immunoglobu-lin genes, i.e. #1, #6, #8 and #59 (22-34%), often in association with trisomy 12, and was significantly different (P

Allogeneic hematopoietic cell transplantation has become a viable option for younger patients with poor-risk chronic lymphocytic leukemia. The results obtained with either conventional or reduced-intensity conditioning regimens have been recently evaluated and compared with alternative nontransplant strategies. This manuscript deals with practical aspects of the procedure, including patient and donor selection, conditioning regimen, GVHD prophylaxis, disease monitoring, infectious and noninfectious complications, and timing of the procedure. Finally, we speculate on how we could improve the results obtained with the procedure and new advances currently in clinical trials.

Plasmacytic differentiation may occur in almost all small B cell lymphomas (SBLs), although it varies from being uniformly present (as in lymphoplasmacytic lymphoma (LPL)) to very uncommon (as in mantle cell lymphomas (MCLs)). The discovery of MYD88 L265P mutations in the vast majority of LPLs has had a major impact on the study of these lymphomas. Review of the cases contributed to the 2014 European Association for Haematopathology/Society for Hematopathology slide workshop illustrated how mutational testing has helped refine the diagnostic criteria for LPL, emphasizing the importance of identifying a clonal monotonous lymphoplasmacytic population and highlighting how LPL can still be diagnosed with extensive nodal architectural effacement, very subtle plasmacytic differentiation, follicular colonization, or uncommon phenotypes such as CD5 or CD10 expression. MYD88 L265P mutations were found in 11/11 LPL cases versus only 2 of 28 other SBLs included in its differential diagnosis. Mutational testing also helped to exclude other cases that would have been considered LPL in the past. The workshop also highlighted how plasmacytic differentiation can occur in chronic lymphocytic leukemia/small lymphocytic lymphoma, follicular lymphoma, SOX11 negative MCL, and particularly in marginal zone lymphomas, all of which can cause diagnostic confusion with LPL. The cases also highlighted the difficulty in distinguishing lymphomas with marked plasmacytic differentiation from plasma cell neoplasms. Some SBLs with plasmacytic differentiation can be associated with amyloid, other immunoglobulin deposition, or crystal-storing histiocytosis, which may obscure the underlying neoplasm. Finally, although generally indolent, LPL may transform, with the workshop cases suggesting a role for TP53 abnormalities.

[cat] Un dels factors pronòstics més importants en la leucèmia limfàtica crònica (LLC) són les categories de risc citogenètic, basades en la detecció de del(13q), trisomia 12, del(11q) i del(17p) per hibridació in situ fluorescent (FISH). Aquests grups són alhora heterogenis, i s'han descrit altres alteracions recurrents de valor pronòstic incert no incloses als panells de FISH utilitzats. L'objectiu global de la tesi doctoral és realitzar la caracterització citogenètica dels pacients amb LLC i correlacionar les dades obtingudes amb el pronòstic d'aquests. Els primers treballs estudien la variabilitat entre els pacients amb LLC i deleció de 13q. El treball 1 compara les característiques dels pacients amb del(13q) monoal•èlica (13qx1) i bial•èlica (13qx2), o amb coexistència d'ambdós clons (13qM) d'una cohort de 627 pacients amb LLC i del(13q) aïllada per FISH. Es va demostrar que, malgrat no presentar diferències significatives al curs clínic, la mediana de nuclis alterats per FISH era superior als pacients amb 13qx2 i 13qM respecte al grup 13qx1, i que les pèrdues bial•èliques podien sorgir com a evolució clonal de les monoal•èliques. D'altra banda, el percentatge de del(13q) per FISH tenia un impacte significatiu al pronòstic. Es va concloure que el pronòstic associat a la del(13q) aïllada es pot estratificar segons el percentatge d'alteració per FISH, i que la pèrdua del segon al•lel de 13q no és suficient per aportar un pitjor pronòstic. El treball 2 s'ha centrat en l'impacte de la detecció d'alteracions de 13q per citogenètica convencional (CC). Es van comparar les característiques dels pacients amb alteracions de 13q14 al cariotip, 25 pacients amb translocacions [t(13q)] i 62 amb delecions intersticials [i-del(13q)], amb un grup de 295 pacients amb del(13q) únicament detectada per FISH [F-del(13q)]. Es van estudiar les alteracions incloses al panell de FISH rutina en LLC i les delecions de RB1. En total es van identificar 25 t(13q) diferents; totes mostraven deleció de D13S319, però només un 32% també presentava pèrdua de RB1. Les medianes de nuclis amb del(13q) dels grup t(13q) i i-del(13q), eren significativament superiors que la del F-del(13q). A més, els pacients amb t(13q) mostraven una incidència major de del(13q) bial•èliques i del(17p) concomitant. La taxa de pèrdua de RB1 era significativament superior al grup i-del(13q). Respecte a l’evolució clínica, el temps fins requerir tractament dels t(13q) i i-del(13q) era significativament més curt que als F-del(13q). En conclusió, l’anàlisi per CC als pacients amb del(13q) permet identificar un subgrup de pacients amb un curs clínic més agressiu. La segona part de la tesi, recollida al treball 3, pretén definir l’aplicabilitat a la pràctica clínica dels microarrays genòmics en la rutina diagnòstica de la LLC. Es van analitzar 70 pacients per microarrays, i es van comparar els resultats amb els obtinguts amb les tècniques utilitzades a la rutina diagnòstica, CC i FISH. La tècnica amb una major taxa de detecció d’alteracions van ser els microarrays. No obstant, els microarrays oferien una menor sensibilitat i no van detectar del(11q) o del(17) identificades per FISH en quatre pacients. A més, la significança clínica de les altres anomalies identificades microarrays no va poder ser demostrada. Es va concloure que l’anàlisi per microarrays és una tècnica vàlida per a l’estudi citogenètic en LLC, però que la completa substitució de la CC i el FISH a la pràctica clínica podia afectar negativament al maneig d’alguns pacients. Globalment, els resultats d’aquesta tesi demostren que l’aplicació combinada de diferents tècniques d’anàlisi citogenètic en LLC permet la identificació d’un ampli ventall d’anomalies, així com una millor caracterització de les alteracions incloses al panell de FISH de la LLC, que poden ser útils per a una millor estratificació pronòstica dels pacients amb LLC., [eng] One of the most important prognostic factors in chronic lymphocytic leukemia (CLL) are cytogenetic risk categories based on the detection of del(13q), trisomy 12, del(11q) and del(17p) by fluorescent in situ hybridization (FISH). However, they are heterogeneous and other alterations have been described. The aim was to perform a cytogenetic characterization of CLL patients and correlate the data with the prognosis. Two of the papers included in the thesis were focused on del(13q). The first analyzes 627 patients with isolated del(13q) by FISH, and compares those who carried monoallelic, biallelic or mosaicism of both 13q deletions. While no significant differences in the outcome was observed, the percentage of del(13q) by FISH had a significant impact on prognosis. In conclusion, del(13q) prognosis can be stratified according to the percentage by FISH, and the loss of the second 13q allele is not enough to trigger a worse outcome. In the second, we compared 25 patients with translocations [t(13q)] and 62 with interstitial deletions [i-del (13q)], with 295 patients with del(13q) only detected by FISH [F-del (13q)]. All the t(13q) showed deletion of the locus D135319, but only 32% also lost RB1. Significant differences in cytogenetic features were observed among groups, this could contribute to the significant shorter time to first treatment observed in t(13q) and i-del(13q) patients. In conclusion, a subgroup of 13q-patients with a poorer outcome can be identified by conventional cytoge n et ics (CC). The third paper compares CC, FISH and microarrays results from 70 patients. Microarray analysis obtained the highest abnormalities detection rate, but showed lower sensitivity and failed to detect some abnormalities identified by FISH. Moreover, clinical significance of other anomalies detected by microarrays could not be demonstrated. In conclusion, microarrays are valid for cytogenetic characterization in CLL. However, the complete replacement of the CC and FISH by microarrays in routine laboratories could negatively affect the management of some patients. Overall, the results show that the combined application of cytogenetic techniques in LLC allow the identification of a wide range of abnormalities, and a better characterization of alterations included in the CLL FISH panel, that may be useful for better prognostic stratification of CLL patients.

[spa] La leucemia linfática crónica (LLC) es la leucemia más frecuente en la edad adulta en los países occidentales, y se caracteriza por la acumulación y proliferación de linfocitos clonales B maduros CD5+ en la sangre, médula ósea, y ganglios linfáticos. La mediana de supervivencia de los pacientes es de alrededor de 10 años aunque su curso clínico es heterogéneo. Durante varias décadas el tratamiento de esta enfermedad se había basado en el empleo de agentes alquilantes como el clorambucilo, en los análogos de la purinas, o en la asociación de ambos, como la combinación de fludarabina y ciclofosfamida (FC). Posteriormente las combinaciones de rituximab, un anticuerpo monoclonal anti-CD20, con quimioterapia, en general FC, lo que ha pasado a denominarse tratamiento inmunoquimioterápico, revolucionaron el tratamiento de esta enfermedad al alcanzar intervalos de supervivencia libre de progresión (SLP) superiores a cinco años, lo que se tradujo en una mayor supervivencia de los enfermos. Sin embargo, a pesar de estos buenos resultados, los pacientes siguen recayendo de su enfermedad y la LLC continúa siendo incurable más allá del trasplante de progenitores hematopoyéticos. Con el fin de mejorar estos resultados se han diseñado nuevas combinaciones de quimioterapia como la asociación de FC con mitoxantrone (FCM), una anthracenedione con capacidad de inhibir la topoisomerasa II. Por otro lado, las estrategias de mantenimiento o las terapias de consolidación dirigidas a controlar la enfermedad mínima residual (EMR) persistente tras el tratamiento inicial, podrían derivar en una mejora en la evolución de los pacientes con LLC. Con estos antecedentes, diseñamos un ensayo clínico multicéntrico fase II en pacientes jóvenes (= 70 años) con LLC previamente no tratados que consistió en un tratamiento inicial con rituximab 500 mg/m2 (día 1) (375mg/m2 en el primer ciclo), fludarabina 25mg/m2 (días 1-3), ciclofosfamida 200mg/m2 (días 1-3), y mitoxantrone 6mg/m2 (día 1), (R-FCM), cada cuatro semanas, hasta un total de seis ciclos. Posteriormente, aquellos pacientes que obtuvieron al menos una respuesta parcial (RP) tras el tratamiento inicial fueron elegibles para la fase de mantenimiento, que consistió en 375mg/m2 de rituximab cada tres meses durante dos años. Además, se analizó la EMR mediante citometría de flujo de forma simultánea en sangre periférica y médula ósea. El objetivo principal del estudio fue el determinar la eficacia de R-FCM medida a través de la tasa de respuestas, incluyendo la tasa de respuestas con EMR negativa. Como objetivos secundarios se planteó el determinar la duración de la respuesta y la SLP, el análisis de las variables clínico-biológicas que influyen en la respuesta, y el perfil de toxicidad del tratamiento. Se incluyeron 81 pacientes en el estudio, con una mediana de edad de 60 años (rango, 40 a 70 años). La tasa de respuesta global obtenida tras el tratamiento inicial con R-FCM fue del 93%, con una tasa de respuesta completa (RC) con EMR negativa, tasa de RC con EMR positiva y de RP del 46%, 36% y 11%, respectivamente. Los factores que se correlacionaron con una menor tasa de RC fueron el estadio clínico avanzado al inicio del tratamiento, la deleción de 17p, y un valor elevado de ß2-microglobulina. En general, el tratamiento con R-FCM fue bien tolerado y aunque se observó algún grado de neutropenia en un 41% de los ciclos administrados, ésta fue grave únicamente en una 13% de los ciclos. En cuanto a la toxicidad infecciosa, se observó un evento infeccioso mayor en un 8% de los ciclos administrados. Sesenta y siete pacientes recibieron el tratamiento de mantenimiento con rituximab. Al finalizar el mantenimiento, un 40.6% de los pacientes presentaban una RC con EMR negativa, un 40.6% una RC con EMR positiva, un 4.8% un RP, y un 14% se consideraron como fallo de tratamiento. Seis de los 29 pacientes (21%) que se hallaban en RC con EMR positiva o en RP tras R-FCM mejoraron su respuesta mediante el tratamiento de mantenimiento. La SLP y la supervivencia global estimada a los cuatro años fueron del 74.8% y 93.7%, respectivamente. El nivel de EMR obtenido tras el tratamiento con R-FCM fue la variable con mayor poder predictivo de la SLP. Se observó neutropenia grave en el 8.5% de los ciclos de mantenimiento y 16 pacientes tuvieron un episodio infeccioso grado 3–4. En conclusión, la combinación de R-FCM es un tratamiento de primera línea altamente eficaz en pacientes jóvenes con LLC. Esta combinación obtuvo una alta tasa de RC, consiguiendo en gran parte de ellas la negativización de la EMR. El tratamiento de mantenimiento con rituximab tras R-FCM consiguió una SLP prolongada y mejoró la calidad de las respuestas en un porcentaje de pacientes, particularmente en aquellos con persistencia de enfermedad detectable tras R-FCM., [eng] Chronic lymphocytic leukemia (CLL) is a frequent malignancy composed of CD5+ B-lymphocytes, is predominant in older people, and has a variable clinical course. The median survival of patients with CLL is approximately 10 years, but the individual prognosis is extremely variable. The addition of monoclonal antibodies to chemotherapy has significantly improved treatment of CLL. Based on excellent results with the chemotherapy-only regimen fludarabine, cyclophosphamide, and mitoxantrone (FCM), we built a new chemoimmunotherapy combination—rituximab plus FCM (R-FCM). The effectiveness of R-FCM followed by rituximab maintenance therapy as first-line treatment for younger patients with CLL (age = 70) has been investigated in a phase 2 clinical trial that included an initial treatment with rituximab 500 mg/m2 on day 1 (375 mg/m2 the first cycle), fludarabine 25 mg/m2 on days 1 to 3, cyclophosphamide 200 mg/m2 on days 1 to 3, and mitoxantrone 6 mg/m2 on day 1 (R-FCM), for 6 cycles. Patients achieving response received maintenance with rituximab 375 mg/m2 every 3 months for 2 years. Eighty-one patients (median age, 60 years; range, 40 to 70 years) were enrolled in the study. The overall response, minimal residual disease (MRD)–negative complete response (CR), MRD positive CR, and partial response (PR) rates were 93%, 46%, 36%, and 11%, respectively. Severe neutropenia developed in 13% of patients. Major and minor infections were reported in 8% and 5% of cycles, respectively. Advanced clinical stage, del(17p), or increased serum ß2-microglobulin levels correlated with a lower CR rate. Sixty-seven patients having achieved CR or PR with R-FCM were given maintenance therapy. At the end of maintenance, 40.6% of patients were in CR MRD-negative, 40.6% were in CR MRD-positive, 4.8% remained in PR, and 14% were considered failures. Six of 29 patients (21%) who were in CR MRD-positive or in PR after R-FCM improved their response upon rituximab maintenance. The 4-year progression-free survival (PFS) and overall survival rates were 74.8% and 93.7%, respectively. MRD status after R-FCM induction was the strongest predictor of PFS. R-FCM is highly effective in previously untreated CLL, with an 82% CR rate and a high proportion of MRD-negative CRs (46%). Treatment toxicity is acceptable. Maintenance with rituximab after R-FCM improved the quality of the response, particularly in patients MRD-positive after initial treatment, and obtained a prolonged PFS.

Polyomaviruses are suspected to cause cancer in humans although to date Merkel cell polyomavirus (MCPyV) is the only proven human carcinogen. Potential associations of up to nine polyomaviruses with 468 lymphoproliferative disorders, 1135 bladder cancer and 359 chronic lymphocytic leukemia (CLL) subjects using three case-control studies in Spain has been explored. Viral exposure was measured as seroreactivity against these viruses by virus-like-particles enzyme linked immunosorbent assay and fluorescent bead-based multiplex serology technology. In explored lymphomas, only diffuse large b-cell lymphoma (DLBCL) showed a significant six-times higher MCPyV seroprevalence (OR=6.10; 95%CI=1.88-19.75) than controls. Bladder cancer cases showed significant higher seroreactivities against BKPyV (OR= 1.37; 95%CI=1.04-1.80) and MCPyV (OR=1.48; 95%CI=1.16-1.88). In CLL, lower seroprevalences for the nine polyomaviruses tested were obtained (OR range=0.21-0.70). The serological patterns obtained in DLBCL and bladder cancer cannot exclude reactive patterns due to disease. Confirmation by means of prospective and molecular studies is required to adequately interpret the associations obtained. Our results in CLL are inconclusive for carcinogenesis due to strong lower seroreactivities. It is unlikely the polyomaviruses explored could play a role in CLL., Es sospita que els Polyomaviruses causen càncer en humans però només el poliomavirus de cèl•lules de Merkel (MCPyV) es carcinògen humà confirmat. Fins a nou poliomavirus en 468 trastorns limfoproliferatius, 1135 càncers de bufeta i 359 leucèmies limfocítica crònica (LLC) s’han explorat mitjançant tres estudis cas-control a Espanya. L’exposició viral es va mesurar mitjançant enzyme linked immunosorbent assay amb virus-like-particles i tecnologia múltiplex de serologia fluorescent basada en beads. Dels limfomes explorats, nomes el limfoma difús de cèl•lules b grans (LDCBG) va mostrar una MCPyV seroprevalença significativament 6-cops mes alta (OR=6.10; 95%CI=1.88-19.75) versus controls. Els casos de càncer de bufeta van mostrar elevades seroreactivitats envers BKPyV (OR= 1.37; 95%CI=1.04-1.80) i MCPyV (OR=1.48; 95%CI=1.16-1.88). En LLC, es van observar seroprevalences més baixes per als 9 poliomavirus mesurats (OR rang=0.21-0.70). Els patrons serològics obtinguts en LDCBG y càncer de bufeta no permeten descartar que siguin deguts a la malaltia. Es necessita confirmació mitjançant estudis prospectius i moleculars per interpretar correctament les associacions obtingudes. Els resultats en LLC son inconclusius respecte carcinogènesi donades les baixes seroreactivitats observades. Es poc probable que els poliomavirus estudiats juguin algun paper en LLC.

This work was supported by research funding from the Spanish Ministry of Science and Innovation(MICINN) through the Instituto de Salud Carlos III(ISCIII)(ICGC-CLL Genome Project)and Red Temática de Investigación Cooperativa en Cáncer(RTICC)grants RD06/0020/0039, RD06/0020/0051 and RD12/0036/0023., Santacruz, R., Villamor, N., Aymerich, M., Martínez-Trillos, A., López, C., Navarro, A., Rozman, M., Beà, S., Royo, C., Cazorla, M., Colomer, D., Giné, E., Pinyol, M., Puente, X.S., López-Otín, C., Campo, E., López-Guillermo, A., Delgado, J.

We investigated the clinico-biological features, outcomes, and prognosis of 949 patients with chronic lymphocytic leukemia according to age. No biological differences (cytogenetics by fluorescent in situ hybridization, IGHV, ZAP-70, CD38, NOTCH1, SF3B1) were found across age groups. Elderly patients (>70 years; n=367) presented more frequently with advanced disease (Binet C/Rai III-IV: 10/12% versus 5/5%; P4; hazard ratio 2.2, P

La via de supervivència PI3K/AKT/mTOR es troba constitutivament activada en molts tumors, fet que ha afavorit el desenvolupament de nombrosos inhibidors al considerar-se una possible opció terapèutica. Tant el MCL com la CLL són dues neoplàsies limfoides per les quals encara no hi ha cap teràpia completament efectiva i en les que la via de PI3K/AKT/mTOR està sobreactivada. En la CLL s’han descrit resultats esperançadors amb l’inhibidor específic de la isoforma p110δ de PI3K, idelalisib. Ara bé, s’han descrit possibles redundàncies entre les diferents isoformes de PI3K i el rol de la isoforma p110α en la progressió tumoral, fets que han suggerit la importància d’inhibir totes les isoformes de PI3K. En aquest context, el primer objectiu d’aquesta tesi es centra en estudiar l’efecte antitumoral de NVP-BKM120, un inhibidor de PI3K, en la CLL. El nostre estudi estableix que NVP-BKM120 inhibeix efectivament la senyalització de PI3K i que interfereix en l’efecte protector del microambient amb la subseqüent inducció de l’apoptosi en les cèl•lules de CLL. Els nostres resultats proporcionen una visió del mecanisme d’acció de NVP-BKM120 en la CLL, suggerint que la inducció de BIM és un factor clau en la citotoxicitat de NVP-BKM120. A més, NVP-BKM120 inhibeix la migració i la polimerització d’actina de les cèl•lules de CLL, el que pot ser particularment important per la mobilització de les cèl•lules tumorals i la sensibilització a la quimioteràpia. Finalment, considerem que NVP-BKM120 podria ser una estratègia terapèutica atractiva tant com a monoteràpia com en combinació en la CLL. D’altra banda, s’ha aprovat la utilització de temsirolimus, un inhibidor de mTORC1, en el tractament del MCL. La estratègia d’inhibir la via de PI3K/AKT/mTOR a un nivell inferior de la cascada de senyalització ha demostrat una bona tolerabilitat i certa activitat antitumoral com a monoteràpia, especialment en el MCL. L’efectivitat d’aquests agents, però, ha quedat reduïda en part per la presència de bucles retroactius que paradoxalment, promouen la supervivència de les cèl•lules tumorals. Per aquest motiu, aquests compostos són bons candidats en combinació amb altres fàrmacs que permetin superar aquests mecanismes de resistència. El segon objectiu d’aquesta tesi es basa en analitzar els mecanismes relacionats amb la sensibilitat i la resistència a everolimus, un inhibidor de mTORC1, en el MCL. Els nostres resultats mostren per primera vegada que l’ús d’un inhibidor de l’autofàgia pot superar la resistència a la combinació d’everolimus i un inhibidor d’AKT isoselectiu en línies cel•lulars i mostres primàries de MCL. El rol de prosupervivència proposat per l’autofàgia en cèl•lules resistents a la inhibició d’AKT i mTOR posa en rellevància diferents possibilitats potencials pel desenvolupament clínic d’aquesta estratègia combinada. Amb el desenvolupament d’inhibidors de segona generació, com els inhibidors catalítics duals PI3K/mTOR, es pretén suprimir la reactivació compensatòria de la via i solucionar els possibles mecanismes de resistència. Per aquest motiu, el tercer objectiu d’aquesta tesi pretén determinar la millor estratègia terapèutica en el MCL mitjançant l’estudi comparatiu dels inhibidors everolimus (inhibidor de mTORC1), NVP-BKM120 (inhibidor de PI3K) i NVP-BEZ235 (inhibidor dual PI3K/mTOR). Les nostres dades donen suport a l’ús de l’inhibidor dual de PI3K/mTOR NVP-BEZ235 com una prometedora aproximació per interferir en els processos relacionats amb el microambient en el MCL. El MCL és un dels limfomes més difícils de tractar, la qual cosa és, en part, deguda a les recaigudes freqüents i a la progressiva resistència al tractament, suggerint que el microambient podria mantenir cèl•lules tumorals residuals resistents a la quimioteràpia. Els nostres resultats plantegen l’ús de NVP-BEZ235 com a estratègia terapèutica per interferir en les interaccions entre les cèl•lules malignes i el microambient, gràcies a la seva contribució en processos essencials com l’angiogènesi, la migració, la invasió i la senyalització per citocines. Finalment, proposem que inhibir la via PI3K/AKT/mTOR per múltiples nivells pot proporcionar una activitat antitumoral més eficaç que les actuals estratègies utilitzant inhibidors de mTOR en el MCL., The phosphatidylinositol 3-kinase (PI3K)/Akt/mTOR survival pathway is constitutively activated in chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL). CLL and MCL are B-lymphoid neoplasms with poor response to conventional chemotherapy, thereby making the PI3K/Akt/mTOR inhibition an attractive therapeutic strategy. Antitumor activity has been observed with mTOR inhibitors as monotherapy. However, several signaling feedback loops might attenuate the effectiveness of these compounds. We found that the activity of the mTOR kinase inhibitor everolimus in MCL cells is closely linked to Akt phosphorylation status, and that the prevention of Akt rephosphorylation upon everolimus treatment by means of a selective Akt inhibitor greatly enhances everolimus activity in MCL cells. Furthermore, our data show that an accumulation of autophagic vacuoles may limit the efficacy of dual Akt/mTOR targeting in resistant cells, and that secondary inhibition of autophagy completes the therapeutic potential of this strategy. These results suggest that autophagy induction protects MCL cells from Akt/mTOR targeting and counteracting autophagy may represent an attractive strategy for sensitizing MCL cells to everolimus-based therapy. Selective PI3K inhibition or dual PI3K/mTOR catalytic inhibition are different therapeutic approaches developed to achieve effective pathway blockage. We investigated the activity of the NVP-BKM120, an orally available pan class I phosphatidylinositol-3-kinase inhibitor in CLL. Our findings establish that NVP-BKM120 effectively inhibits the PI3K signaling pathway and disturbs the protective effect of the tumor microenvironment with the subsequent apoptosis induction through the Akt/FoxO3a/Bim axis. Targeting the PI3K/Akt/mTOR pathway at multiple levels with the dual PI3K/mTOR inhibitor NVP‐BEZ235 is likely to be a more effective strategy for the treatment of MCL than single inhibition of these kinases. Our findings suggest the use of NVP‐BEZ235 as a therapeutic strategy to interfere with malignant cell/microenvironment interactions, thanks to its involvement in essential processes such as angiogenesis, migration, invasiveness and cytokine signaling. We propose that targeting the PI3K/Akt/mTOR pathway at multiple levels may therefore provide a more effective antitumor activity than the current strategies using mTOR inhibitors in MCL.

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the West and is an incurable malignancy. No firmly established evidence exists for environmental risk factors in the etiology of CLL. However, CLL is estimated to have one of the highest familial risks for a hematologic malignancy; this along with other evidence strongly supports an inherited genetic component. In the past 5 years, genome-wide association studies (GWAS) have provided the foundation for new avenues in the investigation of pathogenesis of this disease with 22 susceptibility loci currently identified. We review here the advances made in identifying these loci, the potential to translate these findings into clinical practice, and future directions needed to advance our understanding of the genetic susceptibility of CLL. (C) 2013 Elsevier Inc. All rights reserved.