56 results on '"Larry Mansouri"'
Search Results
2. Correction : Different prognostic impact of recurrent gene mutations in chronic lymphocytic leukemia depending on IGHV gene somatic hypermutation status: a study by ERIC in HARMONY
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Larry Mansouri, Birna Thorvaldsdottir, Lesley-Ann Sutton, Georgios Karakatsoulis, Manja Meggendorfer, Helen Parker, Ferran Nadeu, Christian Brieghel, Stamatia Laidou, Riccardo Moia, Davide Rossi, Mark Catherwood, Jana Kotaskova, Julio Delgado, Ana E. Rodríguez-Vicente, Rocío Benito, Gian Matteo Rigolin, Silvia Bonfiglio, Lydia Scarfo, Mattias Mattsson, Zadie Davis, Ajay Gogia, Lata Rani, Panagiotis Baliakas, Hassan Foroughi-Asl, Cecilia Jylhä, Aron Skaftason, Inmaculada Rapado, Fatima Miras, Joaquín Martinez-Lopez, Javier de la Serna, Jesús María Hernández Rivas, Patrick Thornton, María José Larráyoz, María José Calasanz, Viktória Fésüs, Zoltán Mátrai, Csaba Bödör, Karin E. Smedby, Blanca Espinet, Anna Puiggros, Ritu Gupta, Lars Bullinger, Francesc Bosch, Bárbara Tazón-Vega, Fanny Baran-Marszak, David Oscier, Florence Nguyen-Khac, Thorsten Zenz, Maria Jose Terol, Antonio Cuneo, María Hernández-Sánchez, Sarka Pospisilova, Ken Mills, Gianluca Gaidano, Carsten U. Niemann, Elias Campo, Jonathan C. Strefford, Paolo Ghia, Kostas Stamatopoulos, and Richard Rosenquist
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Cancer Research ,Oncology ,Genetics research ,Hematology ,Cancer genetics - Published
- 2023
3. RPS15 mutations rewire RNA translation in chronic lymphocytic leukemia
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Stavroula Ntoufa, Nikos Papakonstantinou, Georgios Tsiolas, Šárka Pospíšilová, Richard Rosenquist, Fotis Psomopoulos, Kostas Stamatopoulos, Marina Gerousi, Stamatia Laidou, Karla Plevová, Theodoros Moysiadis, Niki Stavrogianni, Antonios M. Makris, Larry Mansouri, and Achilles Anagnostopoulos
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Ribosomal Proteins ,0301 basic medicine ,Translational efficiency ,Chronic lymphocytic leukemia ,Biology ,03 medical and health sciences ,0302 clinical medicine ,immune system diseases ,Ribosomal protein ,hemic and lymphatic diseases ,medicine ,Humans ,Eukaryotic Small Ribosomal Subunit ,Ribosome profiling ,Gene ,Lymphoid Neoplasia ,RNA ,Translation (biology) ,Hematology ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,030104 developmental biology ,030220 oncology & carcinogenesis ,Mutation ,Cancer research - Abstract
Recent studies of chronic lymphocytic leukemia (CLL) have reported recurrent mutations in the RPS15 gene, which encodes the ribosomal protein S15 (RPS15), a component of the 40S ribosomal subunit. Despite some evidence about the role of mutant RPS15 (mostly obtained from the analysis of cell lines), the precise impact of RPS15 mutations on the translational program in primary CLL cells remains largely unexplored. Here, using RNA sequencing and ribosome profiling, a technique that involves measuring translational efficiency, we sought to obtain global insight into changes in translation induced by RPS15 mutations in CLL cells. To this end, we evaluated primary CLL cells from patients with wild-type or mutant RPS15 as well as MEC1 CLL cells transfected with mutant or wild-type RPS15. Our data indicate that RPS15 mutations rewire the translation program of primary CLL cells by reducing their translational efficiency, an effect not seen in MEC1 cells. In detail, RPS15 mutant primary CLL cells displayed altered translation efficiency of other ribosomal proteins and regulatory elements that affect key cell processes, such as the translational machinery and immune signaling, as well as genes known to be implicated in CLL, hence highlighting a relevant role for RPS15 in the natural history of CLL.
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- 2021
4. Higher-order connections between stereotyped subsets
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Andrea Patriarca, Marco Montillo, Niki Stavroyianni, Claudia Haferlach, Lesley-Ann Sutton, Livio Trentin, Franco Fais, Raphael Sandaltzopoulos, Arnon P. Kater, Anton W. Langerak, Marine Armand, Davide Rossi, Diane F. Jelinek, Davide Bagnara, Lydia Scarfò, Andreas Agathangelidis, Krzysztof Giannopoulos, Eugen Tausch, Andrey Sudarikov, Silvio Veronese, Salem H. Alshemmari, B V Biderman, Zadie Davis, Chrysoula Belessi, Lisa Bonello, Achilles Anagnostopoulos, Gerlinde Mitterbauer-Hohendanner, David Oscier, Sofia Kossida, Lone Bredo Pedersen, Paolo Ghia, Csaba Bödör, Christian Brieghel, Andrea Visentin, Véronique Giudicelli, Matthias Ritgen, Panagiotis Panagiotidis, Panagiotis Baliakas, Stephan Stilgenbauer, Ellen J van Gastel, Renee C. Tschumper, Christiane Pott, Frederic Davi, Katerina Gemenetzi, Valentina Guido, Elias Campo, Gianluca Gaidano, Irina Panovska, Sabine Jeromin, Karla Plevová, Kostas Stamatopoulos, Kamila Brázdilová, Maria Karypidou, Alba Navarro, Christof W. Schneider, Theodoros Moysiadis, Larry Mansouri, Darko Antic, Cristina Tresoldi, Constance Baer, Šárka Pospíšilová, Maria Roumelioti, Katrina Vanura, Xiao-Jie Yan, Hana Skuhrová Francová, Richard Rosenquist, Blanca Espinet, Paola Francia di Celle, Monica Facco, Paul Costeas, Michael Hallek, Carsten Utoft Niemann, Teodora Karan-Djurasevic, Manja Meggendorfer, Kirsten Fischer, Aleksandar Dimovski, Letizia Foroni, Marie-Paule Lefranc, Mark Catherwood, Anne de Septenville, Anastasia Chatzidimitriou, Sarah Lawless, Nicholas Chiorazzi, Agathangelidis, Andrea, Chatzidimitriou, Anastasia, Gemenetzi, Katerina, Giudicelli, Veronique, Karypidou, Maria, Plevova, Karla, Davis, Zadie A, Yan, Xiao-Jie, Jeromin, Sabine, Schneider, Christof, Pedersen, Lone Bredo, Tschumper, Renee, Sutton, Lesley A, Baliakas, Panagioti, Scarfò, Lydia, van Gastel, Ellen J, Armand, Marine, Tausch, Eugen, Biderman, Bella, Baer, Constance, Bagnara, Davide, Navarro, Alba, de Septenville, Anne, Guido, Valentina, Mitterbauer-Hohendanner, Gerlinde, Dimovski, Aleksandar, Brieghel, Christian, Lawless, Sarah, Meggendorfer, Manja, Stranska, Kamila, Ritgen, Matthia, Facco, Monica, Tresoldi, Cristina, Visentin, Andrea, Patriarca, Andrea, Catherwood, Mark, Bonello, Lisa, Sudarikov, Andrey, Vanura, Katrina, Roumelioti, Maria, Skuhrova Francova, Hana, Moysiadis, Theodoro, Veronese, Silvio M, Giannopoulos, Krzysztof, Mansouri, Larry, Karan-Djurasevic, Teodora, Sandaltzopoulos, Raphael, Bödör, Csaba, Fais, Franco, Kater, Arnon P, Panovska-Stavridis, Irina, Rossi, Davide, Alshemmari, Salem, Panagiotidis, Panagioti, Costeas, Paul A, Espinet, Blanca, Antic, Darko, Foroni, Letizia, Montillo, Marco, Trentin, Livio, Stavroyianni, Niki, Gaidano, Gianluca, Francia di Celle, Paola, Niemann, Carsten Utoft, Campo, Elía, Anagnostopoulos, Achille, Pott, Christiane, Fischer, Kirsten, Hallek, Michael, Oscier, David Graham, Stilgenbauer, Stephan, Haferlach, Claudia, Jelinek, Diane F, Chiorazzi, Nichola, Pospisilova, Sarka, Lefranc, Marie-Paule, Kossida, Sofia, Langerak, Anton W, Belessi, Chrysoula, Davi, Frederic, Rosenquist, Richard, Ghia, Paolo, Stamatopoulos, Kostas, Experimental Immunology, Clinical Haematology, AII - Cancer immunology, CCA - Cancer biology and immunology, and Immunology
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Chronic lymphocytic leukemia ,Immunology ,B-cell receptor ,Immunoglobulin Variable Region ,Disease ,Computational biology ,Biology ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Gene Frequency ,hemic and lymphatic diseases ,Immunoglobulin ,medicine ,Humans ,Chronic ,030304 developmental biology ,Gene Rearrangement ,0303 health sciences ,Leukemia ,Lymphoid Neoplasia ,Repertoire ,B-Cell ,breakpoint cluster region ,Cell Biology ,Hematology ,Gene rearrangement ,Somatic Hypermutation ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Lymphocytic ,Stereotypy (non-human) ,Immunoglobulin Heavy Chains ,Somatic Hypermutation, Immunoglobulin ,030220 oncology & carcinogenesis ,IGHV@ - Abstract
Chronic lymphocytic leukemia (CLL) is characterized by the existence of subsets of patients with (quasi)identical, stereotyped B-cell receptor (BcR) immunoglobulins. Patients in certain major stereotyped subsets often display remarkably consistent clinicobiological profiles, suggesting that the study of BcR immunoglobulin stereotypy in CLL has important implications for understanding disease pathophysiology and refining clinical decision-making. Nevertheless, several issues remain open, especially pertaining to the actual frequency of BcR immunoglobulin stereotypy and major subsets, as well as the existence of higher-order connections between individual subsets. To address these issues, we investigated clonotypic IGHV-IGHD-IGHJ gene rearrangements in a series of 29 856 patients with CLL, by far the largest series worldwide. We report that the stereotyped fraction of CLL peaks at 41% of the entire cohort and that all 19 previously identified major subsets retained their relative size and ranking, while 10 new ones emerged; overall, major stereotyped subsets had a cumulative frequency of 13.5%. Higher-level relationships were evident between subsets, particularly for major stereotyped subsets with unmutated IGHV genes (U-CLL), for which close relations with other subsets, termed “satellites,” were identified. Satellite subsets accounted for 3% of the entire cohort. These results confirm our previous notion that major subsets can be robustly identified and are consistent in relative size, hence representing distinct disease variants amenable to compartmentalized research with the potential of overcoming the pronounced heterogeneity of CLL. Furthermore, the existence of satellite subsets reveals a novel aspect of repertoire restriction with implications for refined molecular classification of CLL. Key Points: • In a series of 29 856 CLL patients, the incidence of BcR stereotypy peaked at 41%. • Higher-order relations exist between stereotyped subsets, particularly for those from U-CLL, for which satellite subsets were identified.
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- 2021
5. The Number of Signaling Pathways Altered by Driver Mutations in Chronic Lymphocytic Leukemia Impacts Disease Outcome
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Ane Y. Schmidt, Richard Rosenquist, Lone Bredo Pedersen, Carsten Utoft Niemann, Finn Cilius Nielsen, Mattias Mattsson, Julio Delgado, Larry Mansouri, Caspar da Cunha-Bang, Elias Campo, Line Offenbach Jacobsen, Tycho Baumann, Michael Asger Andersen, Savvas Kinalis, Ferran Nadeu, Mette K. Andersen, Christian Brieghel, and Christina Westmose Yde
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0301 basic medicine ,Oncology ,Cancer Research ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Gene mutation ,03 medical and health sciences ,0302 clinical medicine ,hemic and lymphatic diseases ,Internal medicine ,Databases, Genetic ,Humans ,Medicine ,Prospective Studies ,Prospective cohort study ,Gene ,Aged ,Hematology ,business.industry ,Computational Biology ,High-Throughput Nucleotide Sequencing ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Rate ,030104 developmental biology ,030220 oncology & carcinogenesis ,Mutation ,Cohort ,Biomarker (medicine) ,business ,IGHV@ ,Genes, Neoplasm ,Signal Transduction - Abstract
Purpose: Investigation of signaling pathways altered by recurrent gene mutations and their clinical impact in a consecutive cohort of patients with newly diagnosed chronic lymphocytic leukemia (CLL). The heterogeneous clinical course and genetic complexity of CLL warrant improved molecular prognostication. However, the prognostic value of recurrent mutations at the time of diagnosis remains unclear. Experimental Design: We sequenced samples from 314 consecutive, newly diagnosed patients with CLL to investigate the clinical impact of 56 recurrently mutated genes assessed by next-generation sequencing. Results: Mutations were identified in 70% of patients with enrichment among IGHV unmutated cases. With 6.5 years of follow-up, 15 mutated genes investigated at the time of diagnosis demonstrated significant impact on time to first treatment (TTFT). Carrying driver mutations was associated with shorter TTFT and poor overall survival. For outcome from CLL diagnosis, the number of signaling pathways altered by driver mutations stratified patients better than the number of driver mutations. Moreover, we demonstrated gradual impact on TTFT with increasing number of altered pathways independent of CLL-IPI risk. Thus, a 25-gene, pathway-based biomarker assessing recurrent mutations refines prognostication in CLL, in particular for CLL-IPI low- and intermediate-risk patients. External validation emphasized that a broad gene panel including low burden mutations was key for the biomarker based on altered pathways. Conclusions: We propose to include the number of pathways altered by driver mutations as a biomarker together with CLL-IPI in prospective studies of CLL from time of diagnosis for incorporation into clinical care and personalized follow-up and treatment.
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- 2020
6. PD-L1 and IDO1 are potential targets for treatment in patients with primary diffuse large B-cell lymphoma of the CNS
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Christer Sundström, Andrei Alexsson, Peter Hollander, Gunilla Enblad, Cecilia Lindskog, Claes Ladenvall, Rose-Marie Amini, Larry Mansouri, Maysaa Abdulla, Mattias Berglund, and Lucia Cavelier
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PD-L1 ,Epstein-Barr Virus Infections ,Herpesvirus 4, Human ,PD-L2 ,In situ hybridization ,B7-H1 Antigen ,030218 nuclear medicine & medical imaging ,03 medical and health sciences ,0302 clinical medicine ,Lymphocytes, Tumor-Infiltrating ,IDO1 ,Gene expression ,PD-1 ,Tumor Microenvironment ,Medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,PCNSL ,EBER ,Cancer och onkologi ,Tissue microarray ,biology ,business.industry ,Clinical Laboratory Medicine ,RNA ,Hematology ,General Medicine ,medicine.disease ,Lymphoma ,Klinisk laboratoriemedicin ,Oncology ,030220 oncology & carcinogenesis ,Cancer and Oncology ,biology.protein ,Cancer research ,RNA extraction ,Lymphoma, Large B-Cell, Diffuse ,business ,Diffuse large B-cell lymphoma - Abstract
Background Programmed cell death 1 (PD-1) and its ligands PD-L1 and PD-L2, as well as Indoleamine 2,3-deoxygenase (IDO1) can be expressed both by tumor and microenvironmental cells and are crucial for tumor immune escape. We aimed to evaluate the role of PD-1, its ligands and IDO1 in a cohort of patients with primary diffuse large B-cell lymphoma of the CNS (PCNSL). Material and methods Tissue microarrays (TMAs) were constructed in 45 PCNSL cases. RNA extraction from whole tissue sections and RNA sequencing were successfully performed in 33 cases. Immunohistochemical stainings for PD-1, PD-L1/paired box protein 5 (PAX-5), PD-L2/PAX-5 and IDO1, and Epstein-Barr virus encoding RNA (EBER) in situ hybridization were analyzed. Results High proportions of PD-L1 and PD-L2 positive tumor cells were observed in 11% and 9% of cases, respectively. High proportions of PD-L1 and PD-L2 positive leukocytes were observed in 55% and 51% of cases, respectively. RNA sequencing revealed that gene expression of IDO1 was high in patients with high proportion of PD-L1 positive leukocytes (p = .01). Protein expression of IDO1 in leukocytes was detected in 14/45 cases, in 79% of these cases a high proportion of PD-L1 positive leukocytes was observed. Gene expression of IDO1 was high in EBER-positive cases (p = .0009) and protein expression of IDO1 was detected in five of six EBER-positive cases. Conclusion Our study shows a significant association between gene and protein expression of IDO1 and protein expression of PD-L1 in the tumor microenvironment of PCNSL, possibly of importance for prediction of response to immunotherapies. Title in Thesis: PD-L1 and IDO1 are important immunosuppressive molecules in primary diffuse large B-cell lymphoma of the CNS
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- 2021
7. Risk-adapted bendamustine + rituximab is a tolerable treatment alternative for elderly patients with chronic lymphocytic leukaemia: a regional real-world report on 141 consecutive Swedish patients
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Maria Winqvist, Lotta Hansson, Anders Österborg, Sandra Eketorp Sylvan, Richard Rosenquist, Jeanette Lundin, Anna Asklid, Agnes Mattsson, Larry Mansouri, Joel Wiggh, and Hemming Johansson
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Bendamustine ,Male ,medicine.medical_specialty ,Neutropenia ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Bendamustine Hydrochloride ,Humans ,Dosing ,bendamustine ,Stage (cooking) ,Aged ,Retrospective Studies ,CD20 ,Aged, 80 and over ,Sweden ,Chlorambucil ,biology ,business.industry ,Age Factors ,Haematological Malignancy ‐ Clinical ,Hematology ,Middle Aged ,real‐world ,medicine.disease ,Prognosis ,Adaptation, Physiological ,Leukemia, Lymphocytic, Chronic, B-Cell ,Treatment Outcome ,030220 oncology & carcinogenesis ,outcome ,biology.protein ,Rituximab ,Female ,business ,IGHV@ ,CLL ,Research Paper ,030215 immunology ,medicine.drug - Abstract
Summary Bendamustine + rituximab (BR) is the current first‐line standard‐of‐care for chronic lymphocytic leukaemia (CLL) in fit patients aged 66–70 years, whereas chlorambucil + CD20 antibody is recommended in older patients with co‐morbidities. This retrospective real‐world study investigated whether risk‐adapted BR was safe and effective in elderly patients. All 141 CLL patients in the Stockholm region (diagnosed from 2007 to 2016, identified from regional registries) who had received BR as first (n = 84) or later line (n = 57) were analysed. Median age was 72 years, 49% had Binet stage C, 40% had Cumulative Illness Rating Scale (CIRS) score ≥ 6, 20% Eastern Cooperative Oncology Group (ECOG) score 2. None had del(17p). Only 15% of patients aged ≥80 years received full‐dose bendamustine and 65% of them postponed rituximab until cycle 2. Corresponding numbers in patients 73–79 years were 21% and 36% and in
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- 2020
8. Cell-of-origin determined by both gene expression profiling and immunohistochemistry is the strongest predictor of survival in patients with diffuse large B-cell lymphoma
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Gunilla Enblad, Martin Erlanson, Richard Rosenquist, Maja Fors, Fazila Asmar, Sofie Degerman, Kirsten Grønbæk, Larry Mansouri, Per-Ola Andersson, Susanne Bram Ednersson, Tatjana Pandzic, Peter Hollander, Helga Munch Petersen, Rose-Marie Amini, Lucia Cavelier, Maysaa Abdulla, and Magnus Hultdin
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Adult ,medicine.medical_specialty ,Adolescent ,Cell of origin ,Denmark ,Biology ,Lymphocyte Activation ,Young Adult ,immune system diseases ,hemic and lymphatic diseases ,Internal medicine ,medicine ,Humans ,Hematologi ,Survival analysis ,Research Articles ,Aged ,Retrospective Studies ,Aged, 80 and over ,Sweden ,B-Lymphocytes ,Hematology ,Clinical Laboratory Medicine ,Gene Expression Profiling ,Germinal center ,Middle Aged ,medicine.disease ,Germinal Center ,Prognosis ,Immunohistochemistry ,Survival Analysis ,Lymphoma ,Gene expression profiling ,Klinisk laboratoriemedicin ,Cancer research ,Lymphoma, Large B-Cell, Diffuse ,Diffuse large B-cell lymphoma ,Algorithms ,Research Article - Abstract
The tumor cells in diffuse large B‐cell lymphomas (DLBCL) are considered to originate from germinal center derived B‐cells (GCB) or activated B‐cells (ABC). Gene expression profiling (GEP) is preferably used to determine the cell of origin (COO). However, GEP is not widely applied in clinical practice and consequently, several algorithms based on immunohistochemistry (IHC) have been developed. Our aim was to evaluate the concordance of COO assignment between the Lymph2Cx GEP assay and the IHC‐based Hans algorithm, to decide which model is the best survival predictor. Both GEP and IHC were performed in 359 homogenously treated Swedish and Danish DLBCL patients, in a retrospective multicenter cohort. The overall concordance between GEP and IHC algorithm was 72%; GEP classified 85% of cases assigned as GCB by IHC, as GCB, while 58% classified as non‐GCB by IHC, were categorized as ABC by GEP. There were significant survival differences (overall survival and progression‐free survival) if cases were classified by GEP, whereas if cases were categorized by IHC only progression‐free survival differed significantly. Importantly, patients assigned as non‐GCB/ABC both by IHC and GEP had the worst prognosis, which was also significant in multivariate analyses. Double expression of MYC and BCL2 was more common in ABC cases and was associated with a dismal outcome. In conclusion, to determine COO both by IHC and GEP is the strongest outcome predictor to identify DLBCL patients with the worst outcome.
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- 2020
9. Genomic arrays identify high-risk chronic lymphocytic leukemia with genomic complexity: a multi-center study
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Clemens Mellink, Šárka Pospíšilová, Paolo Ghia, Constantine S. Tam, Mar Bellido, Marie Jarošová, Richard Rosenquist, Eva van den Berg, Jacqueline Schoumans, Claudia Haferlach, Lotta Hansson, Zadie Davis, Blanca Espinet, Anna Puiggros, David Oscier, Eric Eldering, Marian Stevens-Kroef, Jonathan C. Strefford, Panagiotis Baliakas, Karla Plevová, Ana E. Rodríguez-Vicente, Alexander C. Leeksma, Kostas Stamatopoulos, Rebeqa Gunnarsson, Pino J Poddighe, Anne Marie van der Kevie-Kersemaekers, Arnon P. Kater, Meaghan Wall, Florence Nguyen-Khac, Theodoros Moysiadis, Anders Österborg, Anh Nhi Tran, Larry Mansouri, Ilaria Scarpelli, Hidde Posthuma, Gisela Barbany, Loic Ysebaert, Helen Parker, Gilead Sciences, Kay Kendall Leukaemia Fund, Cancer Research UK, Wessex Medical Research, Swedish Research Council, Knut and Alice Wallenberg Foundation, Karolinska Institute, Graduate School, AII - Cancer immunology, CCA - Cancer biology and immunology, Human Genetics, Experimental Immunology, Clinical Haematology, Amsterdam Reproduction & Development (AR&D), Leeksma, A. C., Baliakas, P., Moysiadis, T., Puiggros, A., Plevova, K., van der Kevie-Kersemaekers, A. -M., Posthuma, H., Rodriguez-Vicente, A. E., Tran, A. N., Barbany, G., Mansouri, L., Gunnarsson, R., Parker, H., van den Berg, E., Bellido, M., Davis, Z., Wall, M., Scarpelli, I., Osterborg, A., Hansson, L., Jarosova, M., Ghia, P., Poddighe, P., Espinet, B., Pospisilova, S., Tam, C., Ysebaert, L., Nguyen-Khac, F., Oscier, D., Haferlach, C., Schoumans, J., Stevens-Kroef, M., Eldering, E., Stamatopoulos, K., Rosenquist, R., Strefford, J. C., Mellink, C., Kater, A. P., CCA - Cancer Treatment and quality of life, and Human genetics
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0301 basic medicine ,Oncology ,medicine.medical_specialty ,GENES ,Genomic complexity ,Chronic lymphocytic leukemia ,Cancer development and immune defence Radboud Institute for Molecular Life Sciences [Radboudumc 2] ,FEATURES ,ABERRATIONS ,Article ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,hemic and lymphatic diseases ,medicine ,Humans ,Chronic Lymphocytic Leukemia ,Hematologi ,Cytogenetics and Molecular Genetics ,Lymphoproliferative Disorders ,Chromosome Aberrations ,Hematology ,business.industry ,Hazard ratio ,Cytogenetics ,Cancer ,KARYOTYPE ,Genomics ,CHEMOTHERAPY ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,CYTOGENETICS ,3. Good health ,Leukemia ,030104 developmental biology ,030220 oncology & carcinogenesis ,Multiple comparisons problem ,SURVIVAL ,Medical genetics ,business ,CLL ,HYBRIDIZATION ,RESISTANCE - Abstract
Complex karyotype (CK) identified by chromosome-banding analysis (CBA) has shown prognostic value in chronic lymphocytic leukemia (CLL). Genomic arrays offer high-resolution genome-wide detection of copy-number alterations (CNAs) and could therefore be well equipped to detect the presence of a CK. Current knowledge on genomic arrays in CLL is based on outcomes of single center studies, in which different cutoffs for CNA calling were used. To further determine the clinical utility of genomic arrays for CNA assessment in CLL diagnostics, we retrospectively analyzed 2293 arrays from 13 diagnostic laboratories according to established standards. CNAs were found outside regions captured by CLL FISH probes in 34% of patients, and several of them including gains of 8q, deletions of 9p and 18p (p, This study was partly funded by an unrestricted contribution from Janssen Pharmaceuticals and from GILEAD Sciences SA. A.C.L. is supported by the Peters van der Laan foundation. J.C.S. was funded by Bloodwise (11052, 12036), the Kay Kendall Leukaemia Fund (873), Cancer Research UK (C34999/A18087, ECMC C24563/A15581), Wessex Medical Research and the Bournemouth Leukaemia Fund. K.P., M.J., and S.P. are supported by the project MHCR DRO no. 65269705, the research infrastructures NCMG LM2015091, and EATRIS-CZ LM2015064, and the project CEITEC2020 LQ1601, funded by MEYS CR. R.R. is supported by Swedish Cancer Society, the Swedish Research Council, the Knut and Alice Wallenberg Foundation, Karolinska Institutet, Karolinska University Hospital, and Radiumhemmets Forskningsfonder, Stockholm.
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- 2020
10. Specific T Cell Receptor Gene Repertoire Profiles in Subgroups of CLL Patients with Distinct Genomic Aberrations
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Richard Rosenquist, Panagiotis Baliakas, Electra Sofou, Fotis Psomopoulos, Kostas Stamatopoulos, Achilles Anagnostopoulos, Elisavet Vlachonikola, Paolo Ghia, Pamela Ranghetti, Nikos Pechlivanis, Sabine Jeromin, Niki Stavroyianni, Claudia Haferlach, Cecilia Österholm-Corbascio, Anastasia Chatzidimitriou, Lydia Scarfò, Georgios Karakatsoulis, Glykeria Gkoliou, and Larry Mansouri
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Genetics ,T-Cell Receptor Gene ,Repertoire ,Immunology ,Cell Biology ,Hematology ,Biology ,Biochemistry - Abstract
Chronic lymphocytic leukemia (CLL) B cells engage in multifaceted bi-directional interactions with bystander cells, including T cells. Immunogenetic studies in CLL revealed clonal expansions of T cells and shared T cell clonotypes between different patients, strongly implying clonal selection by antigens. Although the exact nature of these antigens remains largely elusive, evidence exists that the clonotypic B cell receptor immunoglobulin (BcR IG) may serve as a source of antigenic epitopes for T cells. That said, recurrent genomic aberrations associated with distinct abnormal expression profiles could represent an alternative, non mutually exclusive, source of potent immunogenic onco-antigens that might shape the T cell repertoire in CLL. On these grounds, here we interrogated the T cell receptor (TR) gene repertoire of CLL patients with different genomic aberration profiles with the aim to identify unique signatures that would allude to distinct antigen selection pressures. The study group included 46 patients with CLL, sampled before treatment initiation, who were categorized in 5 subgroups defined by a unique genomic aberration, as follows: +12, n=18; del(11q), n=10; del(13q), n=7; del(17p)/TP53mut, n=6; NOTCH1mut, n=5. Confounding effects of multiple aberrations have been minimized, as we previously established through comprehensive characterization (including FISH, SNP arrays and gene panels) that the analyzed patients carried only one of the above aberrations. Starting material was RNA extracted from blood mononuclear cells. TRBV-TRBD-TRBJ gene rearrangements were RT-PCR amplified and subjected to paired-end next generation sequencing (NGS). Raw NGS reads (n=13,213,563| median: 294,757/sample) were processed through a purpose-built bioinformatics pipeline. Only productive rearrangements (n=9,249,546 | median=199,184/sample) were taken into consideration for the computation of clonotypes i.e. TRB rearrangements with identical TRBV gene usage and amino acid complementarity-determining region 3 (CDR3) sequence. Overall, 513,984 distinct clonotypes (median=10,304 clonotypes/sample) were assessed. The main measure of clonality employed in this study was the median cumulative frequency of the 10 most expanded T cell clonotypes/sample (MCF-10). For comparisons of the clonality profiles, a group of 17 aged-matched healthy individuals were used as controls. All patients displayed oligoclonal T cell expansions with the following MCF-10 values: del(11q): 21.6%, +12: 25%, del(13q): 20.6%, NOTCH1mut: 9.1%, del(17p)/TP53mut: 12.9%; the difference between the del(11q) and +12 groups versus the NOTCH1mut group was statistically significant (p Disclosures Scarfo: Janssen: Honoraria, Other: Travel grants; Astra Zeneca: Honoraria; Abbvie: Honoraria. Anagnostopoulos: Abbvie: Other: clinical trials; Sanofi: Other: clinical trials ; Ocopeptides: Other: clinical trials ; GSK: Other: clinical trials; Incyte: Other: clinical trials ; Takeda: Other: clinical trials ; Amgen: Other: clinical trials ; Janssen: Other: clinical trials; novartis: Other: clinical trials; Celgene: Other: clinical trials; Roche: Other: clinical trials; Astellas: Other: clinical trials . Ghia: AbbVie: Consultancy, Honoraria, Research Funding; Acerta/AstraZeneca: Consultancy, Honoraria, Research Funding; AstraZeneca: Consultancy, Honoraria, Research Funding; ArQule/MSD: Consultancy, Honoraria; BeiGene: Consultancy, Honoraria; Celgene/Juno/BMS: Consultancy, Honoraria; Gilead: Consultancy, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Roche: Consultancy, Honoraria; Sunesis: Research Funding. Haferlach: MLL Munich Leukemia Laboratory: Other: Part ownership. Rosenquist: Roche: Honoraria; Janssen: Honoraria; Illumina: Honoraria; AstraZeneca: Honoraria; Abbvie: Honoraria. Stamatopoulos: Gilead: Honoraria, Research Funding; Abbvie: Honoraria, Research Funding; AstraZeneca: Honoraria, Research Funding; Janssen: Honoraria, Research Funding. Baliakas: Janssen: Honoraria; Gilead: Honoraria, Research Funding; Abbvie: Honoraria. Chatzidimitriou: Abbvie: Honoraria, Research Funding; Janssen: Honoraria, Research Funding.
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- 2021
11. Proteogenomic Subtyping of Chronic Lymphocytic Leukemia Identifies a Novel Poor Outcome Subgroup with a Distinct Drug Response Profile
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Junyan Lu, Alexander J. Helmboldt, Wolfgang Huber, Matthias Stahl, Simon Anders, Thorsten Zenz, Larry Mansouri, Rozbeh Jafari, Carsten Mueller-Tidow, Ioannis Siavelis, Mark Kriegsmann, Eugen Tausch, Xiaofang Cao, Georgios Mermelekas, Mattias Vesterlund, Angela Lenze, Richard Rosenquist, Carolin Kolb, Sarah Richter, Sophie A. Herbst, Peter Dreger, Tobias Roider, Mareike Knoll, Nora Liebers, Marc Zapatka, Felix Czernilofsky, Marco Herling, Sascha Dietrich, Eva Schitter, Katharina Kriegsmann, Sebastian Scheinost, Cecilia Österholm-Corbascio, Husen M. Umer, Murat Iskar, Stephan Stilgenbauer, Matthias Ganzinger, Janne Lehtiö, and Peter-Martin Bruch
- Subjects
Oncology ,medicine.medical_specialty ,business.industry ,Chronic lymphocytic leukemia ,Immunology ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Outcome (game theory) ,Subtyping ,Internal medicine ,Drug response ,Medicine ,business ,health care economics and organizations - Abstract
Background: Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the western world and shows a very heterogeneous clinical course. While the genetic landscape of CLL has been well characterized during recent years it can only partially explain the underlying biology of this heterogeneity. Proteogenomics could offer a valuable tool to fill this gap and improve the understanding of CLL biology. Methods: Here, we performed a large proteogenomic analysis (n=263) of three clinically annotated CLL cohorts: For the discovery cohort (Germany_1: n=68) we performed in-depth HiRIEF LC-MS based proteomics (more than 9000 proteins quantified) alongside genome-, transcriptome and ex-vivo drug response-profiling with 43 clinically established drugs. The proteome of two additional validation cohorts (Germany_2: n=44, Sweden_1: n=89), were characterized by data-independent acquisition (DIA) mass spectrometry. Results: To connect the CLL genotype with the molecular phenotype, we investigated associations between recurrent genetic alterations of CLL, mRNA expression and protein abundance. We found that trisomy 12, IGHV status and SF3B1 mutations had the greatest impact on protein abundances. CLL specific recurrent chromosomal deletions and gains (trisomy 12, del17p, del13q, del11q, gain8q24) consistently impacted on gene expression and protein abundance through gene dosage effects. We explored functional consequences of these gene dosage effects and found that the additional copy of chromosome 12 increased the abundance of central B-cell receptor (BCR) protein complexes through cis- and trans-effects, which could explain the increased response of trisomy 12 patient samples to BCR inhibition. Somatic mutations of TP53, ATM and XPO1 were associated with less, but specific and biologically-relevant protein abundance changes. p53 for instance, was the most upregulated protein in TP53 mutated samples, owing to the known stabilisation of mutant p53. This effect was not detectable on transcript level. ATM and XPO1 protein abundances were significantly lower in ATM and XPO1 mutated cases, indicating loss-of-function phenotypes of these mutations. To understand global similarities and differences between CLL patients on the proteomic level, we performed unsupervised clustering and identified clinically meaningful subgroups. Unsupervised clustering of the proteomics data identified six subgroups with contrasting clinical behaviour. TP53 mutations, IGHV status, trisomy 12 and their interactions explained five subgroups. These results show that quantitative mass spectrometry-based proteomics distinguished clinically relevant subgroups of CLL. Most importantly, we identified a previously unappreciated subgroup of CLL, comprising 20% of all cases, which could be uncovered by proteomic profiling and showed no association with frequent genetic or transcriptional alterations. This new CLL subgroup was characterized by accelerated disease progression, SF3B1 mutation-independent splicing alterations, metabolomic reprogramming and increased vulnerability to inhibitors of metabolic enzymes and the proteasome. Surprisingly, major BCR signaling proteins were downregulated in this subgroup, suggesting less dependence on BCR activity. In accordance with this observation, an unsupervised analysis revealed that low levels of many BCR signaling proteins (e.g. PLCG2 and PIK3CD) were associated with short time to next treatment. The existence of this subgroup could be confirmed in the validation cohorts. Finally, we performed an unsupervised multi-omics factor analysis (MOFA) across all omics data sets in parallel. This unsupervised analysis confirmed the existence of the above identified CLL subgroups and an important role of SF3B1 mutation-independent splicing alterations in CLL. Conclusion: Our integrative multi-omics analysis provides the first comprehensive overview of the interplay between genetic variants, the transcriptome, and the proteome, along with functional consequences for drug response and clinical outcome in CLL. Importantly, we identified a new subgroup with accelerated disease progression, a distinct proteomic signature and a clinically exploitable drug sensitivity profile. Figure Disclosures Mueller-Tidow: BiolineRx: Research Funding; Daiichi Sankyo: Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees, Research Funding; BMBF: Research Funding; Wilhelm-Sander-Stiftung: Research Funding; Jose-Carreras-Siftung: Research Funding; Bayer AG: Research Funding; Deutsche Krebshilfe: Research Funding; Deutsche Forschungsgemeinschaft: Research Funding; Janssen-Cilag Gmbh: Membership on an entity's Board of Directors or advisory committees. Dreger:Neovii: Research Funding; Roche: Consultancy, Speakers Bureau; Riemser: Consultancy, Research Funding, Speakers Bureau; Novartis: Consultancy, Speakers Bureau; Janssen: Consultancy; Gilead: Consultancy, Speakers Bureau; AstraZeneca: Consultancy; AbbVie: Consultancy, Speakers Bureau. Stilgenbauer:Pharmacyclics: Consultancy, Honoraria, Other, Research Funding; Novartis: Consultancy, Honoraria, Other, Research Funding; Mundipharma: Consultancy, Honoraria, Other, Research Funding; Janssen-Cilag: Consultancy, Honoraria, Other: travel support, Research Funding; GlaxoSmithKline: Consultancy, Honoraria, Other: travel support, Research Funding; Gilead: Consultancy, Honoraria, Other: travel support, Research Funding; Genzyme: Consultancy, Honoraria, Other: travel support, Research Funding; Genentech: Consultancy, Honoraria, Other: travel support, Research Funding; F. Hoffmann-LaRoche: Consultancy, Honoraria, Other: travel support, Research Funding; Celgene: Consultancy, Honoraria, Other: travel support, Research Funding; Boehringer-Ingelheim: Consultancy, Honoraria, Other: travel support, Research Funding; Amgen: Consultancy, Honoraria, Other: travel support, Research Funding; AbbVie: Consultancy, Honoraria, Other: travel support, Research Funding. Tausch:Roche: Consultancy, Honoraria, Research Funding; AbbVie: Consultancy, Honoraria, Research Funding; Janssen-Cilag: Consultancy, Honoraria, Research Funding. Dietrich:Roche: Membership on an entity's Board of Directors or advisory committees; Janssen: Membership on an entity's Board of Directors or advisory committees, Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; KITE: Membership on an entity's Board of Directors or advisory committees.
- Published
- 2020
12. Frequent NFKBIE deletions are associated with poor outcome in primary mediastinal B-cell lymphoma
- Author
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Niki Stavroyianni, Helga D. Munch-Petersen, Kenichi Yoshida, Markus Schneider, Andreas Rosenwald, German Ott, Lucile Couronné, Michael Hummel, Marita Ziepert, Daniel Noerenberg, Emma Young, Richard Rosenquist, Seishi Ogawa, Thorsten Zenz, Jonathan C. Strefford, Larry Mansouri, Rose-Marie Amini, Hans G. Drexler, Viktor Ljungström, Martin-Leo Hansmann, Mareike Frick, Dido Lenze, Martin Erlanson, Ralf Küppers, Alfredo Rivas-Delgado, Armando López-Guillermo, Elisabeth Ralfkiaer, Christopher Maximilian Arends, Magnus Hultdin, Tatjana Pandzic, Bernd Dörken, Claudia D. Baldus, Maria K. Angelopoulou, Theodora Papadaki, Kirsten Grønbæk, Kostas Stamatopoulos, Anna Tasidou, Maysaa Abdulla, Nils Waldhueter, Frederik Damm, Theodoros P. Vassilakopoulos, Elias Campo, Damien Roos-Weil, Jude Fitzgibbon, Elena Mylonas, Blanca Gonzalez, Penelope Korkolopoulou, George Kanellis, Aron Skaftason, Gunilla Enblad, Olivier A. Bernard, Fazila Asmar, Jessica Okosun, Clemens A. Schmitt, and Christian Bastard
- Subjects
Adult ,Male ,0301 basic medicine ,medicine.medical_specialty ,Lymphoma, B-Cell ,Adolescent ,Chronic lymphocytic leukemia ,Immunology ,Medizin ,Follicular lymphoma ,Biology ,Mediastinal Neoplasms ,Biochemistry ,Disease-Free Survival ,03 medical and health sciences ,NFKBIE Gene ,0302 clinical medicine ,Proto-Oncogene Proteins ,hemic and lymphatic diseases ,Internal medicine ,Biomarkers, Tumor ,medicine ,Humans ,Aged ,Hematology ,Cell Biology ,Middle Aged ,medicine.disease ,NFKBIE ,3. Good health ,Lymphoma ,Survival Rate ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer research ,Female ,I-kappa B Proteins ,Mantle cell lymphoma ,Primary mediastinal B-cell lymphoma ,Gene Deletion - Abstract
We recently reported a truncating deletion in the NFKBIE gene, which encodes IκBε, a negative feedback regulator of NF-κB, in clinically aggressive chronic lymphocytic leukemia (CLL). Because preliminary data indicate enrichment of NFKBIE aberrations in other lymphoid malignancies, we screened a large patient cohort (n = 1460) diagnosed with different lymphoid neoplasms. While NFKBIE deletions were infrequent in follicular lymphoma, splenic marginal zone lymphoma, and T-cell acute lymphoblastic leukemia (
- Published
- 2016
13. Tailored approaches grounded on immunogenetic features for refined prognostication in chronic lymphocytic leukemia
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Zadie Davis, Richard Rosenquist, Chrysoula Belessi, Panagiotis Baliakas, Niki Stavroyianni, Theodoros Moysiadis, Elias Campo, Julio Delgado, Šárka Pospíšilová, Kostas Stamatopoulos, Marta Larrayoz, Davide Rossi, Jonathan C. Strefford, Larry Mansouri, Achilles Anagnostopoulos, Mattias Mattsson, Karin E. Smedby, Diego Cortese, Anastasia Hadzidimitriou, Lesley-Ann Sutton, Neus Villamor, David Oscier, Alba Navarro, Jana Kotašková, Evangelia Stalika, Gianluca Gaidano, Karla Plevová, Mark Catherwood, Gunnar Juliusson, Paolo Ghia, Sabine Jeromin, Oonagh Sheehy, Lydia Scarfò, Andreas Agathangelidis, Emma Young, Helen Parker, Eva Minga, Aliki Xochelli, Claudia Haferlach, Baliakas, Panagioti, Moysiadis, Theodoro, Hadzidimitriou, Anastasia, Xochelli, Aliki, Jeromin, Sabine, Agathangelidis, Andrea, Mattsson, Mattia, Sutton, Lesley-Ann, Minga, Eva, Scarfò, Lydia, Rossi, Davide, Davis, Zadie, Villamor, Neu, Parker, Helen, Kotaskova, Jana, Stalika, Evangelia, Plevova, Karla, Mansouri, Larry, Cortese, Diego, Navarro, Alba, Delgado, Julio, Larrayoz, Marta, Young, Emma, Anagnostopoulos, Achille, Smedby, Karin E., Juliusson, Gunnar, Sheehy, Oonagh, Catherwood, Mark, Strefford, Jonathan C., Stavroyianni, Niki, Belessi, Chrysoula, Pospisilova, Sarka, Oscier, David, Gaidano, Gianluca, Campo, Elia, Haferlach, Claudia, Ghia, Paolo, Rosenquist, Richard, Stamatopoulos, Kostas, and Universitat de Barcelona
- Subjects
Male ,Oncology ,medicine.medical_specialty ,Pronòstic mèdic ,Chronic lymphocytic leukemia ,Somatic hypermutation ,Relative weight ,Kaplan-Meier Estimate ,Immunogenetics ,Article ,Time-to-Treatment ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,hemic and lymphatic diseases ,Biomarkers, Tumor ,medicine ,Humans ,Chronic Lymphocytic Leukemia ,Leucèmia limfocítica crònica ,Hematologi ,Aged ,Neoplasm Staging ,030304 developmental biology ,Aged, 80 and over ,Chromosome Aberrations ,0303 health sciences ,Hematology ,biology ,business.industry ,medicine.disease ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,3. Good health ,Leukemia ,030220 oncology & carcinogenesis ,Mutation ,biology.protein ,Female ,Disease Susceptibility ,Antibody ,business ,Trisomy - Abstract
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.
- Published
- 2019
14. Inhibition of SYK or BTK augments venetoclax sensitivity in SHP1-negative/BCL-2-positive diffuse large B-cell lymphoma
- Author
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Claudio Martines, Malgorzata Bobrowicz, Sven Turkalj, Beata Pyrzynska, Joanna Stachura, Binu K. Sasi, Engin Bojnik, Francesco Bertoni, Dimitar G. Efremov, Magdalena Winiarska, Richard Rosenquist, Elena Xerxa, Hilal Kalkan, Larry Mansouri, Valdemar Priebe, Fabiola Porro, Rosa Fazio, and Abdessamad Zerrouqi
- Subjects
0301 basic medicine ,Cancer Research ,B-cell receptor ,Syk ,Antineoplastic Agents ,Apoptosis ,Mice, SCID ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Phosphatidylinositol 3-Kinases ,0302 clinical medicine ,immune system diseases ,Mice, Inbred NOD ,hemic and lymphatic diseases ,Cell Line, Tumor ,medicine ,Agammaglobulinaemia Tyrosine Kinase ,Bruton's tyrosine kinase ,Animals ,Humans ,Syk Kinase ,MCL1 ,PI3K/AKT/mTOR pathway ,Cell Proliferation ,Sulfonamides ,biology ,Venetoclax ,Protein Tyrosine Phosphatase, Non-Receptor Type 6 ,breakpoint cluster region ,Hematology ,medicine.disease ,Bridged Bicyclo Compounds, Heterocyclic ,Xenograft Model Antitumor Assays ,3. Good health ,030104 developmental biology ,Oncology ,chemistry ,Proto-Oncogene Proteins c-bcl-2 ,030220 oncology & carcinogenesis ,Cancer research ,biology.protein ,Female ,Lymphoma, Large B-Cell, Diffuse ,Diffuse large B-cell lymphoma ,Signal Transduction - Abstract
The BCL-2 inhibitor venetoclax has only limited activity in DLBCL despite frequent BCL-2 overexpression. Since constitutive activation of the B cell receptor (BCR) pathway has been reported in both ABC and GCB DLBCL, we investigated whether targeting SYK or BTK will increase sensitivity of DLBCL cells to venetoclax. We report that pharmacological inhibition of SYK or BTK synergistically enhances venetoclax sensitivity in BCL-2-positive DLBCL cell lines with an activated BCR pathway in vitro and in a xenograft model in vivo, despite the only modest direct cytotoxic effect. We further show that these sensitizing effects are associated with inhibition of the downstream PI3K/AKT pathway and changes in the expression of MCL-1, BIM, and HRK. In addition, we show that BCR-dependent GCB DLBCL cells are characterized by deficiency of the phosphatase SHP1, a key negative regulator of the BCR pathway. Re-expression of SHP1 in GCB DBLCL cells reduces SYK, BLNK, and GSK3 phosphorylation and induces corresponding changes in MCL1, BIM, and HRK expression. Together, these findings suggest that SHP1 deficiency is responsible for the constitutive activation of the BCR pathway in GCB DLBCL and identify SHP1 and BCL-2 as potential predictive markers for response to treatment with a venetoclax/BCR inhibitor combination.
- Published
- 2018
15. No improvement in long-term survival over time for chronic lymphocytic leukemia patients in stereotyped subsets #1 and #2 treated with chemo(immuno)therapy
- Author
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Tatiana Tzenou, Richard Rosenquist, Marie-Paule Lefranc, Karla Plevová, Charles C. Chu, Yorick Sandberg, Gunnar Juliusson, Véronique Giudicelli, Livio Trentin, Mark Catherwood, Frederic Davi, Šárka Pospíšilová, Xiao-Jie Yan, Silvio Veronese, Lesley-Ann Sutton, Carsten Utoft Niemann, Nikos Darzentas, Kostas Stamatopoulos, Achilles Anagnostopoulos, Diane F. Jelinek, David Oscier, Mattias Mattsson, Nicholas Chiorazzi, Karin E. Smedby, Panagiotis Panagiotidis, Chrysoula Belessi, Florence Nguyen-Khac, Marco Montillo, Eva Minga, Paolo Ghia, Anton W. Langerak, Lydia Scarfò, Andreas Agathangelidis, Tait D. Shanafelt, Panagiotis Baliakas, Niki Stavroyianni, Larry Mansouri, Anastasia Hadzidimitriou, Zadie Davis, Fie Juhl Vojdeman, Uppsala Universitet [Uppsala], Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden, Institute of Applied Biosciences, Centre for Research and Technology-Hellas, Thessaloniki, Greece, Department of Immunology, Genetics and Pathology [Uppsala, Sueden] (IGP), Uppsala University, Department of Hematology [Uppsala], Università Vita-Salute San Raffaele, Milan, Italy., Strategic Research Program in CLL, Division of Experimental Oncology, IRCCS San Raffaele Scientific Institute, Milan, Italy, Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden, Università Vita e Salute, San Raffaele, Milano, Italy, Strategic Research Program in CLL, Division of Experimental Oncology, IRCCS San Raffaele Scientific Institute, Milan, Italy., Department of Haematology, Royal Bournemouth Hospital, Bournemouth, UK., The Feinstein Institute for Medical Research, CEITEC-Central European Institute of Technology, MasarykBrno, Czech Republic., Central European Institute of Technology [Brno] (CEITEC MU), Brno University of Technology [Brno] (BUT), Erasmus University Medical Center [Rotterdam] (Erasmus MC), Department of Hematology, Rigshospitalet, Copenhagen, Denmark, First Department of Propaedeutic Medicine, University of Athens, Athens, Greece, Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico, Department of Medicine Solna, Clinical Epidemiology Unit, Karolinska Institutet, and Hematology Center, Karolinska University Hospital, Stockholm, Sweden, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Service d'Hématologie clinique [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Hematology and Transplantation, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, Lund University and Hospital Department of Hematology, Lund Stem Cell Center, Lund, Sweden, Hematology Department and HCT Unit, G. Papanicolaou Hospital, Thessaloniki, Greece, Universita degli Studi di Padova, Venetian Institute Molecular Medicine (VIMM), Department of Hemato-Oncology, Belfast City Hospital, Belfast, UK, University Hospital Brno, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Department of Immunology, Mayo Clinic, Rochester, MV, USA, Mayo Clinic [Rochester], Hematology Department, Nikea General Hospital, Piraeus, Greece, Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden., Department of Molecular Medicine and Surgery, Karolinska Institutet, Stockholm, Sweden, Hematology Department and HCT Unit, G. Papanicolaou Hospital, Thessaloniki, Greece., Institute of Applied Biosciences, Thessaloniki, Greece., Department of Immunology, Baliakas, Panagioti, Mattsson, Mattia, Hadzidimitriou, Anastasia, Minga, Eva, Agathangelidis, Andrea, Sutton, Lesley-Ann, Scarfo, Lydia, Davis, Zadie, Yan, Xiao-Jie, Plevova, Karla, Sandberg, Yorick, Vojdeman, Fie J, Tzenou, Tatiana, Chu, Charles C, Veronese, Silvio, Mansouri, Larry, Smedby, Karin E, Giudicelli, Véronique, Nguyen-Khac, Florence, Panagiotidis, Panagioti, Juliusson, Gunnar, Anagnostopoulos, Achille, Lefranc, Marie-Paule, Trentin, Livio, Catherwood, Mark, Montillo, Marco, Niemann, Carsten U, Langerak, Anton W, Pospisilova, Sarka, Stavroyianni, Niki, Chiorazzi, Nichola, Oscier, David, Jelinek, Diane F, Shanafelt, Tait, Darzentas, Niko, Belessi, Chrysoula, Davi, Frederic, Ghia, Paolo, Rosenquist, Richard, Stamatopoulos, Kostas, and Immunology
- Subjects
Adult ,Male ,chemorefractorine ,Oncology ,medicine.medical_specialty ,Cyclophosphamide ,Chronic lymphocytic leukemia ,[SDV]Life Sciences [q-bio] ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Drug Therapy ,Chemoimmunotherapy ,Internal medicine ,medicine ,Humans ,Chronic Lymphocytic Leukemia ,[INFO]Computer Science [cs] ,stereotyped subsets ,Online Only Articles ,Survival rate ,ComputingMilieux_MISCELLANEOUS ,Aged ,Retrospective Studies ,Aged, 80 and over ,[SDV.GEN]Life Sciences [q-bio]/Genetics ,Hematology ,business.industry ,Middle Aged ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,3. Good health ,Fludarabine ,Survival Rate ,Leukemia ,030220 oncology & carcinogenesis ,Female ,Rituximab ,Immunotherapy ,business ,030215 immunology ,medicine.drug - Abstract
The overall survival (OS) of patients with chronic lymphocytic leukemia (CLL) has improved over the last decades mainly due to advances in the understanding of the disease biology and the introduction of novel therapeutic approaches(1). In the present retrospective study we investigated trends in OS in subgroups of cases defined by genetic and immunogenetic features aiming at addressing the question whether advances in chemoimmunotherapy had a uniform impact across all CLL patients. We found that such advances have translated into prolonged OS in all prognostic subgroups examined except those carrying TP53 abnormalities, as expected, but also those assigned to stereotyped subsets #1 and #2, that are generally devoid of such gene aberrations. This latter finding, reported here for the first time, indicates the need for alternative treatment options for these patients.
- Published
- 2018
16. Clinical and functional impact of recurrent S1PR1 mutations in mantle cell lymphoma
- Author
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Agata M. Wasik, Larry Mansouri, Birgitta Sander, Chenglin Wu, Richard Rosenquist, and Qiang Pan-Hammarström
- Subjects
0301 basic medicine ,medicine.medical_specialty ,Gene Expression ,Lymphoma, Mantle-Cell ,medicine.disease_cause ,03 medical and health sciences ,Exon ,immune system diseases ,Recurrence ,Internal medicine ,hemic and lymphatic diseases ,Medicine ,Humans ,Genetic Predisposition to Disease ,Receptor ,neoplasms ,Sphingosine-1-Phosphate Receptors ,S1PR1 ,Genetic Association Studies ,Mutation ,Hematology ,business.industry ,Exons ,medicine.disease ,Prognosis ,Stimulus Report ,Immunohistochemistry ,Lymphoma ,Receptors, Lysosphingolipid ,030104 developmental biology ,Cancer research ,Mantle cell lymphoma ,business - Abstract
Key Points S1PR1 mutations are present in 7.8% of patients with MCL and are significantly more frequent at relapse. S1PR1 mutations reduce expression of the S1PR1 receptor, which mediates migration towards the tissue-to-blood egress factor S1P in MCL.
- Published
- 2018
17. Not all IGHV3-21 chronic lymphocytic leukemias are equal: prognostic considerations
- Author
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Eva Minga, Karin E. Smedby, Lesley-Ann Sutton, Nicholas Chiorazzi, Myriam Boudjogra, Kostas Stamatopoulos, Karla Plevová, Lone Bredo Pedersen, Zadie Davis, Lydia Scarfò, Andreas Agathangelidis, Monica Facco, Achilles Anagnostopoulos, Maria Chatzouli, Chrysoula Belessi, Athina Tsanousa, Panagiotis Baliakas, Kirsten van Lom, Lefteris Angelis, Yorick Sandberg, Gunnar Juliusson, Diane F. Jelinek, Fie Juhl Vojdeman, Anne Gardiner, Panagiotis Panagiotidis, Anton W. Langerak, Florence Nguyen-Khac, Hana Skuhrová Francová, Frederic Davi, Denis Moreno, Silvio Veronese, Richard Rosenquist, Marie-Paule Lefranc, Nikos Darzentas, Šárka Pospíšilová, Véronique Giudicelli, Xiao-Jie Yan, Charles C. Chu, Christian H. Geisler, Larry Mansouri, David Oscier, Mark Catherwood, Marco Montillo, Anastasia Hadzidimitriou, Livio Trentin, Paolo Ghia, Tait D. Shanafelt, Tatiana Tzenou, Baliakas, P, Agathangelidis, A, Hadzidimitriou, A, Sutton, La, Minga, E, Tsanousa, A, Scarfò, L, Davis, Z, Yan, Xj, Shanafelt, T, Plevova, K, Sandberg, Y, Vojdeman, Fj, Boudjogra, M, Tzenou, T, Chatzouli, M, Chu, Cc, Veronese, S, Gardiner, A, Mansouri, L, Smedby, Ke, Pedersen, Lb, Moreno, D, Van Lom, K, Giudicelli, V, Francova, H, Nguyen Khac, F, Panagiotidis, P, Juliusson, G, Angelis, L, Anagnostopoulos, A, Lefranc, Mp, Facco, M, Trentin, L, Catherwood, M, Montillo, M, Geisler, Ch, Langerak, Aw, Pospisilova, S, Chiorazzi, N, Oscier, D, Jelinek, Df, Darzentas, N, Belessi, C, Davi, F, Ghia, PAOLO PROSPERO, Rosenquist, R, Stamatopoulos K. Ghia P., is Co senior author, Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden, Università Vita-Salute San Raffaele, Milan, Italy, Division of Molecular Oncology and Department of Onco-Hematology, Istituto di Ricovero e Cura a Carattere Scientifico, San Raffaele Scientific Institute, Milan, Italy, Institute of Applied Biosciences, Centre for Research and Technology-Hellas, Thessaloniki, Greece, Department of Informatics, Aristotle University of Thessaloniki, Thessaloniki, Greece, Department of Haematology, Royal Bournemouth Hospital, Bournemouth, United Kingdom, The Feinstein Institute for Medical Research, Mayo Clinic [Rochester], Central European Institute of Technology [Brno] (CEITEC MU), Brno University of Technology [Brno] (BUT), Erasmus University Medical Center [Rotterdam] (Erasmus MC), Department of Hematology, Rigshospitalet, Copenhagen, Denmark, Service d'Hématologie Biologique [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), First Department of Propaedeutic Medicine, University of Athens, Athens, Greece, Hematology Department, Nikea General Hospital, Piraeus, Greece, Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico, Department of Medicine, Solna, Clinical Epidemiology Unit, Karolinska Institutet, Stockholm, Sweden, Institut de génétique humaine (IGH), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Lund University and Hospital Department of Hematology, Lund Stem Cell Center, Lund, Sweden, Hematology Department and Hematopoietic Cell Transplantation Unit, Georgios Papanicolaou Hospital, Thessaloniki, Greece, Universita degli Studi di Padova, Department of Hemato-Oncology, Belfast City Hospital, Belfast, United Kingdom, Immunology, and Hematology
- Subjects
Male ,Oncology ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Immunology ,B-cell receptor ,Gene Rearrangement, B-Lymphocyte, Heavy Chain ,Antineoplastic Agents ,Biology ,Biochemistry ,Time-to-Treatment ,Genetic Heterogeneity ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Humans ,10. No inequality ,ComputingMilieux_MISCELLANEOUS ,Survival analysis ,Aged ,030304 developmental biology ,[SDV.GEN]Life Sciences [q-bio]/Genetics ,B-Lymphocytes ,0303 health sciences ,Lymphoid Neoplasia ,Hematology ,Gene Expression Regulation, Leukemic ,Genetic heterogeneity ,Cell Biology ,Middle Aged ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,Treatment Outcome ,030220 oncology & carcinogenesis ,biology.protein ,Immunoglobulin heavy chain ,Female ,Somatic Hypermutation, Immunoglobulin ,Antibody ,Immunoglobulin Heavy Chains ,IGHV@ - Abstract
An unresolved issue in chronic lymphocytic leukemia (CLL) is whether IGHV3-21 gene usage, in general, or the expression of stereotyped B-cell receptor immunoglobulin defining subset # 2 (IGHV3-21/IGLV3-21), in particular, determines outcome for IGHV3-21-utilizing cases. We reappraised this issue in 8593 CLL patients of whom 437 (5%) used the IGHV3-21 gene with 254/437 (58%) classified as subset # 2. Within subset # 2, immunoglobulin heavy variable (IGHV)-mutated cases predominated, whereas non-subset # 2/IGHV3-21 was enriched for IGHV-unmutated cases (P =.002). Subset # 2 exhibited significantly shorter time-to-first-treatment (TTFT) compared with non-subset # 2/IGHV3-21 (22 vs 60 months, P =.001). No such difference was observed between non-subset # 2/IGHV3-21 vs the remaining CLL with similar IGHV mutational status. In conclusion, IGHV3-21 CLL should not be axiomatically considered a homogeneous entity with adverse prognosis, given that only subset # 2 emerges as uniformly aggressive, contrasting non-subset # 2/IGVH3-21 patients whose prognosis depends on IGHV mutational status as the remaining CLL.
- Published
- 2015
18. Targeted next-generation sequencing in chronic lymphocytic leukemia: a high-throughput yet tailored approach will facilitate implementation in a clinical setting
- Author
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Jitka Malčíková, Emma Young, Veronika Navrkalová, Šárka Pospíšilová, Martin Trbušek, Kostas Stamatopoulos, Frederic Davi, Diego Cortese, Panagiotis Panagiotidis, Chrysoula Belessi, Alice F. Muggen, Anton W. Langerak, Richard Rosenquist, Lesley-Ann Sutton, Viktor Ljungström, Paolo Ghia, Larry Mansouri, Immunology, Sutton, La, Ljungström, V, Mansouri, L, Young, E, Cortese, D, Navrkalova, V, Malcikova, J, Muggen, Af, Trbusek, M, Panagiotidis, P, Davi, F, Belessi, C, Langerak, Aw, Ghia, PAOLO PROSPERO, Pospisilova, S, Stamatopoulos, K, and Rosenquist, R.
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Chronic lymphocytic leukemia ,Gene Expression ,Computational biology ,Biology ,medicine.disease_cause ,DNA sequencing ,symbols.namesake ,Gene Frequency ,medicine ,Humans ,Receptor, Notch1 ,Allele ,Allele frequency ,Alleles ,Sanger sequencing ,Genetics ,Mutation ,High-Throughput Nucleotide Sequencing ,Articles ,Hematology ,Ribonucleoprotein, U2 Small Nuclear ,Phosphoproteins ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Neoplasm Proteins ,Leukemia ,Myeloid Differentiation Factor 88 ,symbols ,RNA Splicing Factors ,Tumor Suppressor Protein p53 ,IGHV@ - Abstract
Next- generation sequencing has revealed novel recurrent mutations in chronic lymphocytic leukemia, particularly in patients with aggressive disease. Here, we explored targeted re- sequencing as a novel strategy to assess the mutation status of genes with prognostic potential. To this end, we utilized HaloPlex targeted enrichment technology and designed a panel including nine genes: ATM, BIRC3, MYD88, NOTCH1, SF3B1 and TP53, which have been linked to the prognosis of chronic lymphocytic leukemia, and KLHL6, POT1 and XPO1, which are less characterized but were found to be recurrently mutated in various sequencing studies. A total of 188 chronic lymphocytic leukemia patients with poor prognostic features ( unmutated IGHV, n= 137; IGHV3- 21 subset # 2, n= 51) were sequenced on the HiSeq 2000 and data were analyzed using well- established bioinformatics tools. Using a conservative cutoff of 10% for the mutant allele, we found that 114/ 180 ( 63%) patients carried at least one mutation, with mutations in ATM, BIRC3, NOTCH1, SF3B1 and TP53 accounting for 149/ 177 ( 84%) of all mutations. We selected 155 mutations for Sanger validation ( variant allele frequency, 10- 99%) and 93% ( 144/ 155) of mutations were confirmed; notably, all 11 discordant variants had a variant allele frequency between 11- 27%, hence at the detection limit of conventional Sanger sequencing. Technical precision was assessed by repeating the entire HaloPlex procedure for 63 patients; concordance was found for 77/ 82 ( 94%) mutations. In summary, this study demonstrates that targeted next- generation sequencing is an accurate and reproducible technique potentially suitable for routine screening, eventually as a stand- alone test without the need for confirmation by Sanger sequencing.
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- 2015
19. UGT2B17 expression: a novel prognostic marker within IGHV-mutated chronic lymphocytic leukemia?
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Sujata Bhoi, Karin E. Smedby, Panagiotis Baliakas, Lesley-Ann Sutton, Larry Mansouri, Gunnar Juliusson, Marie Engvall, Mattias Mattsson, and Diego Cortese
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medicine.medical_specialty ,Chronic lymphocytic leukemia ,DNA Mutational Analysis ,Population ,Immunoglobulin Variable Region ,Gene Expression ,Minor Histocompatibility Antigens ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,immune system diseases ,hemic and lymphatic diseases ,Molecular marker ,Internal medicine ,Gene expression ,Humans ,Medicine ,RNA, Messenger ,Glucuronosyltransferase ,Receptor, Notch1 ,Online Only Articles ,education ,Survival analysis ,education.field_of_study ,Hematology ,business.industry ,Phosphoproteins ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,Lipoprotein Lipase ,Leukemia ,ROC Curve ,chemistry ,030220 oncology & carcinogenesis ,Multivariate Analysis ,Mutation ,Immunology ,Cancer research ,RNA Splicing Factors ,Tumor Suppressor Protein p53 ,Immunoglobulin Heavy Chains ,business ,IGHV@ ,Follow-Up Studies ,030215 immunology - Abstract
High UGT2B17 mRNA expression has recently been correlated with poor prognosis in chronic lymphocytic leukemia (CLL).1 In the present study, we investigated the expression of UGT2B17 in a Scandinavian population-based CLL cohort (n=253) and can confirm that high expression of UGT2B17 is associated with advanced clinical stage at diagnosis, unmutated IGHV genes (U-CLL) and poor clinical outcome. That said, we discovered a notable and novel finding based on the expression of UGT2B17, that of identifying patients with a poor prognosis within the IGHV-mutated group (M-CLL) (31/120, 26%), which previously could not be discriminated by any other established molecular marker, including recurrent genomic aberrations, novel mutations (SF3B1, NOTCH1 and TP53) and CD38 expression. Interestingly, high UGT2B17 expression arose as the strongest independent molecular prognostic marker of overall survival (OS) in multivariate analysis within M-CLL. The incorporation of LPL into our expression analysis enabled the further stratification of M-CLL, thus highlighting the potential use of RNA-based markers in the prognostic stratification of CLL, particularly for cases exhibiting an otherwise favorable clinicobiological profile.
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- 2015
20. SHP1 Deficiency Is Responsible for the Constitutive Activation of the BCR Pathway in GCB DLBCL
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Fabiola Porro, Engin Bojnik, Francesco Bertoni, Binu K. Sasi, Beata Pyrzynska, Abdessamad Zerrouqi, Richard Rosenquist, Dimitar G. Efremov, Magdalena Winiarska, Malgorzata Bobrowicz, Sven Turkalj, Valdemar Priebe, Hilal Kalkan, and Larry Mansouri
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medicine.medical_specialty ,Hematology ,Protein Tyrosine Phosphatase SHP-1 ,Venetoclax ,Btk inhibitors ,Immunology ,breakpoint cluster region ,Cell Biology ,NFKB1 ,medicine.disease ,Biochemistry ,chemistry.chemical_compound ,chemistry ,hemic and lymphatic diseases ,Internal medicine ,Ibrutinib ,medicine ,Cancer research ,Diffuse large B-cell lymphoma - Abstract
Functional and transcriptional profiling studies have identified a subset of diffuse large B cell lymphoma (DLBCL) tumors that rely on B cell receptor (BCR) signals for proliferation and survival. These BCR-dependent tumors have been identified among both the ABC and GCB DLBCL subtype. However, differences have been observed between the two subtypes in terms of the mechanism of BCR pathway activation and activity of downstream signaling molecules. BCR-dependent ABC DLBCL tumors are characterized by high basal NF-kB and PI3K/AKT activity and the presence of gain-of-function mutations in the CD79A or CD79B subunit of the BCR that are responsible for the chronic activation of the BCR pathway. In contrast, BCR-dependent GCB DLBCL tumors have low baseline NF-kB activity and lack CD79A/CD79B mutations, suggesting a different mechanism of BCR pathway activation. In this study, we investigated whether activation of the BCR pathway in GCB DLBCL is potentially caused by deficiency of the phosphatase SHP1, which is an important negative regulator of the BCR pathway and has been reported to be downregulated in approximately 40% of primary DLBCL tumors. For this purpose, we first correlated SHP1 expression with the presence of phosphorylated SYK and BLNK in the GCB DLBCL cell lines OCI-Ly1, OCI-Ly7, OCI-Ly18, SU-DHL-4, SU-DHL-6, WSU-NHL, SU-DHL-8, Toledo, BJAB, OCI-Ly19, OCI-Ly4, and Farage. Immunoblotting analysis revealed that SHP1 is expressed in SU-DHL-8, Toledo, BJAB, OCI-Ly19, OCI-Ly4, and Farage but not in the remaining 6 cell lines. Expression of SHP1 inversely correlated with expression of phosphorylated SYK (pSYK-Y352) and BLNK (pBLNK-Y84), suggesting that these two BCR-proximal signaling molecules are activated because of SHP1 deficiency. To further evaluate this possibility, we re-expressed SHP1 in OCI-Ly1 and SU-DHL-4 cells by transient transfection with a plasmid or a lentiviral SHP1 expression vector. Both approaches resulted in a substantial reduction of pSYK-Y352 and pBLNK-Y84 levels, confirming that SHP1 deficiency leads to activation of the BCR pathway. To determine whether activation of the BCR pathway contributes to tumor cell survival, we investigated the effects of the SYK inhibitor R406 (active substance of the drug fostamatinib) on the viability of the 6 SHP1-positive and 6 SHP1-negative GCB DLBCL cell lines. At concentrations up to 2 μM, R406 displayed only minimal toxicity against each of the SHP1-positive and SHP1-negative GCB DLBCL cell lines ( Disclosures No relevant conflicts of interest to declare.
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- 2018
21. RPS15 mutations Repress mRNA Translation in Chronic Lymphocytic Leukemia Cells
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Stavroula Ntoufa, Niki Stavroyianni, Georgios Tsiolas, Antonios M. Makris, Kostas Stamatopoulos, Achilles Anagnostopoulos, Richard Rosenquist, Marina Gerousi, Fotis Psomopoulos, Nikos Papakonstantinou, Stamatia Laidou, Larry Mansouri, and Anastasia Hadzidimitriou
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0301 basic medicine ,03 medical and health sciences ,030104 developmental biology ,business.industry ,Chronic lymphocytic leukemia ,Immunology ,Cancer research ,Medicine ,Cell Biology ,Hematology ,business ,medicine.disease ,Biochemistry - Abstract
We and others recently reported mutations within the RPS15 gene, encoding a component of the 40S ribosomal subunit, in clinically aggressive chronic lymphocytic leukemia (CLL). RPS15 mutations resided within an evolutionary conserved region, alluding to an oncogenic rather than a tumor-suppressor role. Our pilot functional analysis revealed that, similar to other ribosomal proteins (RPs), RPS15 also binds MDM2 and may impact on the p53 response. Here, we performed ribosome profiling in order to gain global insight into changes in translation induced by RPS15 mutations in CLL cells. This technique involves measuring translational efficiency (TE), by comparing the levels of ribosome-associated mRNA footprints against the total mRNA for each gene. For 6 CLL cases bearing mutant (mut, n=3) or wildtype (wt, n=3) RPS15, we obtained both ribosome-protected footprints (RPFs) and matching mRNA sequencing data. In parallel, we created stable MEC1 CLL cell lines expressing an additional copy of wt or mut RPS15 (131S) by lentiviral transduction; validation of the transgene expression was performed by Sanger sequencing of amplified cDNAs. Ribosome footprinting and subsequent library preparation of RPFs and total mRNA for all samples was performed with the Illumina Truseq Ribo Profile Kit and all libraries were sequenced on a NextSeq500 instrument. Reads were aligned to the human hg19 genome using Bowtie2. SystemPipeR was used to determine the percentage of reads mapping to 5' UTRs, CDS, and 3' UTRs and triplet periodicity was assessed using RibORF. The RPFs were of high quality, as assessed by expected RPF size (28-30nt), CDS enrichment, and triplet periodicity. To determine differentially expressed genes between RPS15-mut vs RPS15-wt cases we used DESeq2 while, for differentially translated genes we used Xtail. Changes in transcription and translation between PRS15-wt vs RPS15-mut cases showed limited overlap in both primary CLL cells and cell lines (12.8% and 12.9%, respectively), indicating the potential of ribosome profiling to reveal additional information compared to RNA sequencing alone. In primary CLL cells, 474 genes showed differences only at the transcription level (log2FC mRNA>I1I, pI1I, pI1I), while 1859 genes were regulated only at the translation level (log2FC RPF>I1I). Overall, 771 genes displayed differential TE (log2FC TE Disclosures Hadzidimitriou: Abbvie: Research Funding; Gilead: Research Funding; Janssen: Honoraria, Research Funding. Stamatopoulos:Janssen: Honoraria, Research Funding; Abbvie: Honoraria, Research Funding; Gilead: Honoraria, Research Funding.
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- 2018
22. Short telomere length is associated withNOTCH1/SF3B1/TP53aberrations and poor outcome in newly diagnosed chronic lymphocytic leukemia patients
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Richard Rosenquist, Karin E. Smedby, Ulrika Svenson, Christian H. Geisler, Göran Roos, Gunnar Juliusson, Pawel Grabowski, Nicola Cahill, Larry Mansouri, Rebeqa Gunnarsson, and Sofie Degerman
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Male ,Oncology ,medicine.medical_specialty ,Pathology ,Chronic lymphocytic leukemia ,Disease-Free Survival ,03 medical and health sciences ,0302 clinical medicine ,Predictive Value of Tests ,hemic and lymphatic diseases ,Internal medicine ,Biomarkers, Tumor ,medicine ,Humans ,Receptor, Notch1 ,Survival rate ,Aged ,Retrospective Studies ,030304 developmental biology ,0303 health sciences ,Hematology ,business.industry ,Retrospective cohort study ,Middle Aged ,Ribonucleoprotein, U2 Small Nuclear ,Telomere ,Phosphoproteins ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Rate ,Clinical trial ,Leukemia ,030220 oncology & carcinogenesis ,Predictive value of tests ,Mutation ,Female ,RNA Splicing Factors ,Tumor Suppressor Protein p53 ,business ,Follow-Up Studies - Abstract
Most previous studies on telomere length (TL) in chronic lymphocytic leukemia (CLL) are based on referral cohorts including a high proportion of aggressive cases. Here, the impact of TL was analyzed in a population-based cohort of newly diagnosed CLL (n = 265) and in relation to other prognostic markers. Short telomeres were particularly associated with high-risk genetic markers, such as NOTCH1, SF3B1, or TP53 aberrations, and predicted a short time to treatment (TTT) and overall survival (OS) (both P0.0001). TL was an independent prognostic factor and subdivided patients with otherwise good-prognostic features (e.g., mutated IGHV genes, favorable cytogenetics) into subgroups with different outcome. Furthermore, in follow-up samples (n = 119) taken 5-8 years after diagnosis, TL correlated well with TL at diagnosis and remained unaffected by treatment. Altogether, these novel data indicate that short TL already at diagnosis is associated with poor outcome in CLL and that TL can be measured at later stages of the disease.
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- 2013
23. rs2072135, a low-penetrance variant for chronic lymphocytic leukaemia?
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Helen E. Speedy, Daniel Catovsky, Dariusz Woszczyk, Davide Rossi, Richard Rosenquist, Irena Frydecka, Gianluca Gaidano, Silvia Deaglio, Lidia Karabon, Larry Mansouri, Maria Chiara Di Bernardo, Gunnar Juliusson, Karin E. Smedby, Georgina P. Sava, and Richard S. Houlston
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Adult ,Male ,Genotype ,Quantitative Trait Loci ,Gene Expression ,Penetrance ,Locus (genetics) ,Single-nucleotide polymorphism ,Genome-wide association study ,Biology ,Polymorphism, Single Nucleotide ,Genome ,Humans ,Genetic Predisposition to Disease ,Aged ,Genetic association ,Genetics ,Lymphocytic leukaemia ,Hematology ,Middle Aged ,Leukemia, Lymphocytic, Chronic, B-Cell ,Case-Control Studies ,Expression quantitative trait loci ,Female ,Genome-Wide Association Study - Abstract
Recent multi-stage genome-wide association studies (GWAS) have identified single nucleotide polymorphisms (SNPs) that are robustly associated with chronic lymphocytic leukaemia (CLL) risk. Given that most of these SNPs map to non-coding regions of the genome, it suggests that the functional basis of many GWAS signals will be through differential gene expression. By referencing publically accessible expression quantitative trait loci (eQTL) data on lymphoblastoid cells lines (LCLs) we have globally demonstrated an association between GWAS P-values and eQTLs, consistent with much of the variation in CLL risk being defined by variants impacting on gene expression. To explore using eQTL data to select GWAS SNPs for replication, we genotyped rs2072135 (GWAS P-value = 0·0024, eQTL P-value = 1·510(-19)) in five independent case-control series totalling 1968 cases and 3538 controls. While not attaining statistical significance (combined P-value = 1 × 10(-4)), rs2072135 defines a promising risk locus for CLL. Incorporating eQTL information offers an attractive strategy for selecting SNPs from GWAS for validation.
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- 2013
24. Prognostic markers and their clinical applicability in chronic lymphocytic leukemia: where do we stand?
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Diego Cortese, Rebeqa Gunnarsson, Richard Rosenquist, Larry Mansouri, and Sujata Bhoi
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Cancer Research ,medicine.diagnostic_test ,business.industry ,ZAP70 ,Chronic lymphocytic leukemia ,Hematology ,Disease ,Gene mutation ,CD38 ,Prognosis ,medicine.disease ,Bioinformatics ,Leukemia, Lymphocytic, Chronic, B-Cell ,Leukemia ,Oncology ,Biomarkers, Tumor ,medicine ,Humans ,IGHV@ ,business ,Neoplasm Staging ,Fluorescence in situ hybridization - Abstract
Chronic lymphocytic leukemia (CLL) is a clinically and biologically heterogeneous disease where the majority of patients have an indolent disease course, while others may experience a far more aggressive disease, treatment failure and poor overall survival. During the last two decades, there has been an intense search to find novel biomarkers that can predict prognosis as well as guide treatment decisions. Two of the most reliable molecular prognostic markers, both of which are offered in routine diagnostics, are the immunoglobulin heavy chain variable (IGHV) gene mutational status and fluorescence in situ hybridization (FISH) detection of prognostically relevant genomic aberrations (e.g. 11q-, 13q-, +12 and 17p-). In addition to these markers, a myriad of additional biomarkers have been postulated as potential prognosticators in CLL, on the protein (e.g. CD38, ZAP70, TCL1), the RNA (e.g. LPL, CLLU1, micro-RNAs) and the genomic (e.g. TP53, NOTCH1, SF3B1 and BIRC3 mutations) level. Efforts are now being made to test these novel markers in larger patient cohorts as well as in prospective trials, with the ultimate goal to combine the "best" markers in a "CLL prognostic index" applicable for the individual patient. Although it is clear that these studies have significantly improved our knowledge regarding both prognostication and the biology of the disease, there is still an immediate need for recognizing biomarkers that can predict therapy response, and efforts should now focus on addressing this pertinent issue. In the present article, we review the extensive literature in the field of prognostic markers in CLL, focus on the most clinically relevant markers and discuss future directions regarding biomarkers in CLL.
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- 2013
25. Distinct patterns of novel gene mutations in poor-prognostic stereotyped subsets of chronic lymphocytic leukemia: the case of SF3B1 and subset #2
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Jitka Malčíková, Richard Rosenquist, Panagiotis Panagiotidis, C. Belessi, Tatiana Tzenou, P. F. Di Celle, Karla Plevová, Šárka Pospíšilová, Nicola Cahill, Anthonie Willem Langerak, Larry Mansouri, Nicholas Chiorazzi, Christian H. Geisler, Kostas Stamatopoulos, Lone B. Pedersen, Zadie Davis, Lesley-Ann Sutton, Diego Cortese, David Oscier, Panagiotis Baliakas, Evangelia Stalika, Paolo Ghia, Lydia Scarfò, Andreas Agathangelidis, Frederic Davi, Jonathan C. Strefford, Strefford, Jc, Sutton, La, Baliakas, P, Agathangelidis, A, Malcikova, J, Plevova, K, Scarfo, L, Davis, Z, Stalika, E, Cortese, D, Cahill, N, Pedersen, Lb, di Celle, Pf, Tzenou, T, Geisler, C, Panagiotidis, P, Langerak, Aw, Chiorazzi, N, Pospisilova, S, Oscier, D, Davi, F, Belessi, C, Mansouri, L, Ghia, PAOLO PROSPERO, Stamatopoulos, K, Rosenquist R. P., Ghia is joint last author, and Immunology
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Cancer Research ,Chronic lymphocytic leukemia ,Ubiquitin-Protein Ligases ,Biology ,medicine.disease_cause ,Somatic evolution in cancer ,Polymerase Chain Reaction ,law.invention ,Inhibitor of Apoptosis Proteins ,Cohort Studies ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,law ,medicine ,Biomarkers, Tumor ,Humans ,Receptor, Notch1 ,Survival rate ,Gene ,Polymerase chain reaction ,030304 developmental biology ,0303 health sciences ,Mutation ,Hematology ,DNA, Neoplasm ,Ribonucleoprotein, U2 Small Nuclear ,medicine.disease ,Phosphoproteins ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,Baculoviral IAP Repeat-Containing 3 Protein ,Survival Rate ,Leukemia ,Oncology ,030220 oncology & carcinogenesis ,Immunology ,RNA Splicing Factors ,Follow-Up Studies - Abstract
Recent studies have revealed recurrent mutations of the NOTCH1, SF3B1 and BIRC3 genes in chronic lymphocytic leukemia (CLL), especially among aggressive, chemorefractory cases. Nevertheless, it is currently unknown whether their presence may differ in subsets of patients carrying stereotyped B-cell receptors and also exhibiting distinct prognoses. Here, we analyzed the mutation status of NOTCH1, SF3B1 and BIRC3 in three subsets with particularly poor prognosis, that is, subset # 1, # 2 and # 8, aiming to explore links between genetic aberrations and immune signaling. A remarkably higher frequency of SF3B1 mutations was revealed in subset # 2 (44%) versus subset # 1 and # 8 (4.6% and 0%, respectively; P
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- 2013
26. Tailored Approaches for Refined Prognostication in Chronic Lymphocytic Leukemia Patients with Mutated Versus Unmutated Immunoglobulin Receptors
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Panagiotis Baliakas, Theodoros Moysiadis, Anastasia Hadzidimitriou, Aliki Xochelli, Mattias Mattsson, Lesley-Ann Sutton, Eva Minga, Lydia Scarfo, Davide Rossi, Zadie Davis, Andreas Agathangelidis, Neus Villamor, Helen Parker, Jana Kotaskova, Evangelia Stalika, Karla Plevova, Larry Mansouri, Diego Cortese, Alba Navaro Lopez, Julio Delgado, Marta Larrayoz, Emma Young, Achilles Anagnostopoulos, Karin E Smedby, Gunnar Juliusson, Mark Catherwood, Jonathan C Strefford, Niki Stavroyianni, Chrysoula Belessi, Sarka Pospisilova, David Oscier, Gianluca Gaidano, Elias Campo, Paolo Ghia, Richard Rosenquist, and Kostas Stamatopoulos, on behalf of ERIC
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Oncology ,medicine.medical_specialty ,Proportional hazards model ,business.industry ,Chronic lymphocytic leukemia ,Immunology ,Somatic hypermutation ,Recursive partitioning ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Log-rank test ,03 medical and health sciences ,0302 clinical medicine ,hemic and lymphatic diseases ,Internal medicine ,Cohort ,medicine ,Clinical endpoint ,IGHV@ ,business ,030215 immunology - Abstract
The classification of CLL patients according to the somatic hypermutation status (SHM) of the immunoglobulin heavy variable (IGHV) genes, namely mutated (M-CLL) versus unmutated (U-CLL), reflects fundamental differences in disease biology and clinical course. Realizing this, here we followed a compartmentalized approach and addressed the issue of prognostication separately for M-CLL and U-CLL. In a multi-institutional cohort of 2366 patients [M-CLL, n=1364 (58%); U-CLL, n=1002 (42%)] consolidated within ERIC, the European Initiative in CLL, we assessed the clinical impact of 'traditional' (age and clinical stage at the time of diagnosis, gender, CD38 expression, FISH detected abnormalities included in the Döhner hierarchical model of cytogenetic aberrations), and novel prognosticators (recurrent mutations within the TP53, SF3B1, NOTCH1, MYD88, and BIRC3 genes; IGHV gene usage; membership in stereotyped subsets) within M-CLL and U-CLL. Our statistical approach was based both on Cox regression models and recursive partitioning algorithms; internal validation was performed via bootstrapping procedures. Given the retrospective nature of our study, time-to-first-treatment (TTFT) was the primary endpoint. As expected, M-CLL exhibited significantly longer TTFT compared to U-CLL [median TTFT: not yet reached (M-CLL) vs 1.9 years (95% CI: 0.01-12.3 years, U-CLL), p PB and TM contributed equally as first authors Figure 1 Figure 1. Disclosures Rosenquist: Gilead Sciences: Speakers Bureau.
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- 2016
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27. Different spectra of recurrent gene mutations in subsets of chronic lymphocytic leukemia harboring stereotyped B-cell receptors
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Marta Larrayoz, Panagiotis Panagiotidis, Theodoros Moysiadis, Evangelia Stalika, Kostas Stamatopoulos, Alba Navarro, Šárka Pospíšilová, Xiao-Jie Yan, Chrysoula Belessi, Jitka Malčíková, Florence Nguyen-Khac, Frederic Davi, Richard Rosenquist, Lesley-Ann Sutton, Lone Bredo Pedersen, Nicholas Chiorazzi, Karla Plevová, Gianluca Gaidano, Panagiotis Baliakas, Lydia Scarfò, Emma Young, Andreas Agathangelidis, Paolo Ghia, Anton W. Langerak, Carsten Utoft Niemann, Davide Rossi, Jonathan C. Strefford, Elias Campo, Alice F. Muggen, Myriam Boudjoghra, Larry Mansouri, Jana Kminkova, Anastasia Hadzidimitriou, Zadie Davis, David Oscier, Sutton, Lesley Ann, Young, Emma, Baliakas, Panagioti, Hadzidimitriou, Anastasia, Moysiadis, Theodoro, Plevova, Karla, Rossi, Davide, Kminkova, Jana, Stalika, Evangelia, Pedersen, Lone Bredo, Malcikova, Jitka, Agathangelidis, Andrea, Davis, Zadie, Mansouri, Larry, Scarfò, Lydia, Boudjoghra, Myriam, Navarro, Alba, Muggen, Alice F., Yan, Xiao Jie, Nguyen Khac, Florence, Larrayoz, Marta, Panagiotidis, Panagioti, Chiorazzi, Nichola, Niemann, Carsten Utoft, Belessi, Chrysoula, Campo, Elia, Strefford, Jonathan C., Langerak, Anton W., Oscier, David, Gaidano, Gianluca, Pospisilova, Sarka, Davi, Frederic, Ghia, PAOLO PROSPERO, Stamatopoulos, Kosta, Rosenquist, Richard, Immunology, Uppsala Universitet [Uppsala], Centre for Research and Technology Hellas (CERTH), Masaryk University [Brno] (MUNI), Università degli Studi del Piemonte Orientale - Amedeo Avogadro (UPO), Rigshospitalet [Copenhagen], Copenhagen University Hospital, Università Vita e Salute, San Raffaele, Milano, Italy, Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS San Raffaele Pisana), Royal Bournemouth Hospital, Université Pierre et Marie Curie - Paris 6 (UPMC), Service d'Hématologie clinique [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona (UB), Erasmus University Medical Center [Rotterdam] (Erasmus MC), The Feinstein Institute for Medical Research, University of Southampton, National and Kapodistrian University of Athens (NKUA), Hematology Department, Nikea General Hospital, Piraeus, Greece, George Papanicolau Hospital, Universitat de Barcelona, and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)
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Male ,Chronic lymphocytic leukemia ,Immunoglobulin Variable Region ,Receptors, Antigen, B-Cell ,Somatic hypermutation ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Gene mutation ,Biology ,medicine.disease_cause ,Polymorphism, Single Nucleotide ,Somatic evolution in cancer ,03 medical and health sciences ,Genètica mèdica ,0302 clinical medicine ,Gene Frequency ,Biomarkers, Tumor ,medicine ,Humans ,Leucèmia limfocítica crònica ,Gene Rearrangement, B-Lymphocyte ,Genetics ,Mutation ,Genes, Immunoglobulin ,Mutació (Biologia) ,Medical genetics ,[SDV.MHEP.HEM]Life Sciences [q-bio]/Human health and pathology/Hematology ,Articles ,Gene rearrangement ,Hematology ,Mutation (Biology) ,Prognosis ,medicine.disease ,Complementarity Determining Regions ,Leukemia, Lymphocytic, Chronic, B-Cell ,3. Good health ,Leukemia ,030220 oncology & carcinogenesis ,Cytogenetic Analysis ,Immunoglobulin Joining Region ,Immunoglobulin heavy chain ,Female ,Immunoglobulin Heavy Chains ,030215 immunology - Abstract
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
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- 2016
28. ATM mutations in major stereotyped subsets of chronic lymphocytic leukemia: Enrichment in subset #2 is associated with markedly short telomeres
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Panagiotis Baliakas, Larry Mansouri, Viktor Ljungström, Richard Rosenquist, Šárka Pospíšilová, Gunnar Juliusson, Frederic Davi, Martin Trbušek, Stavroula Ntoufa, Paolo Ghia, Karin E. Smedby, Chrysoula Belessi, Anton W. Langerak, Lesley-Ann Sutton, Jiri Mayer, Karla Plevová, Kostas Stamatopoulos, Panagiotis Panagiotidis, Tasoula Touloumenidou, Lenka Radová, Emma Young, Jonathan C. Strefford, Zadie Davis, David Oscier, Veronika Navrkalová, Navrkalova, Veronika, Young, Emma, Baliakas, Panagioti, Radova, Lenka, Sutton, Lesley Ann, Plevova, Karla, Mansouri, Larry, Ljungström, Viktor, Ntoufa, Stavroula, Davis, Zadie, Juliusson, Gunnar, Smedby, Karin E., Belessi, Chrysoula, Panagiotidis, Panagioti, Touloumenidou, Tasoula, Davi, Frederic, Langerak, Anton W., Ghia, PAOLO PROSPERO, Strefford, Jonathan C., Oscier, David, Mayer, Jiri, Stamatopoulos, Kosta, Pospisilova, Sarka, Rosenquist, Richard, Trbusek, Martin, and Immunology
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0301 basic medicine ,medicine.medical_specialty ,Antigenic selection ,Chronic lymphocytic leukemia ,Ataxia Telangiectasia Mutated Proteins ,Biology ,Cohort Studies ,03 medical and health sciences ,0302 clinical medicine ,Internal medicine ,medicine ,Humans ,Stereotyped subset ,Online Only Articles ,Telomere Shortening ,Hematology ,Atm mutation ,Sequence Analysis, DNA ,Telomere ,medicine.disease ,Short telomere ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,030104 developmental biology ,030220 oncology & carcinogenesis ,Immunology ,Mutation - Abstract
ATM mutations in major stereotyped subsets of chronic lymphocytic leukemia: enrichment in subset 2 is associated with markedly short telomeres, ATM mutations in major stereotyped subsets of chronic lymphocytic leukemia: enrichment in subset 2 is associated with markedly short telomeres, ATM mutations in major stereotyped subsets of chronic lymphocytic leukemia: enrichment in subset 2 is associated with markedly short telomeres.
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- 2016
29. Whole-exome sequencing in relapsing chronic lymphocytic leukemia: clinical impact of recurrent RPS15 mutations
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Larry Mansouri, Anton W. Langerak, Anna Schuh, Tatjana Pandzic, Martin Höglund, Lesley-Ann Sutton, David Oscier, Stuart Blakemore, Karla Plevová, Christiane Pott, Karin E. Smedby, Emma Young, R Clifford, Andreas Agathangelidis, Livio Trentin, Davide Rossi, Viktor Ljungström, Jonathan C. Strefford, Panagiotis Panagiotidis, Jana Kotašková, Šárka Pospíšilová, Paolo Ghia, Tobias Sjöblom, Kostas Stamatopoulos, Stavroula Ntoufa, Niki Stavroyianni, Diego Cortese, Richard Rosenquist, Gianluca Gaidano, Nicholas Chiorazzi, Gunnar Juliusson, Frederic Davi, Panagiotis Baliakas, Chrysoula Belessi, Immunology, Ljungström, V, Cortese, D, Young, E, Pandzic, T, Mansouri, L, Plevova, K, Ntoufa, S, Baliakas, P, Clifford, R, Sutton, La, Blakemore, S, Stavroyianni, N, Agathangelidis, A, Rossi, D, Höglund, M, Kotaskova, J, Juliusson, G, Belessi, C, Chiorazzi, N, Panagiotidis, P, Langerak, Aw, Smedby, Ke, Oscier, D, Gaidano, G, Schuh, A, Davi, F, Pott, C, Strefford, Jc, Trentin, L, Pospisilova, S, Ghia, PAOLO PROSPERO, Stamatopoulos, K, Sjöblom, T, and Rosenquist, R.
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Ribosomal Proteins ,0301 basic medicine ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Blotting, Western ,DNA Mutational Analysis ,Immunology ,Mutation, Missense ,Cell Separation ,Kaplan-Meier Estimate ,Biology ,Transfection ,medicine.disease_cause ,Biochemistry ,Somatic evolution in cancer ,03 medical and health sciences ,Internal medicine ,Antineoplastic Combined Chemotherapy Protocols ,medicine ,Humans ,Immunoprecipitation ,Exome ,Cyclophosphamide ,Exome sequencing ,Mutation ,Lymphoid Neoplasia ,Hematology ,Cell Biology ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,NFKBIE ,3. Good health ,Fludarabine ,030104 developmental biology ,Drug Resistance, Neoplasm ,Cancer research ,Neoplasm Recurrence, Local ,Tumor Suppressor Protein p53 ,Rituximab ,Vidarabine ,medicine.drug - Abstract
Fludarabine, cyclophosphamide, and rituximab (FCR) is first-line treatment of medically fit chronic lymphocytic leukemia (CLL) patients; however, despite good response rates, many patients eventually relapse. Although recent high-throughput studies have identified novel recurrent genetic lesions in adverse prognostic CLL, the mechanisms leading to relapse after FCR therapy are not completely understood. To gain insight into this issue, we performed whole-exome sequencing of sequential samples from 41 CLL patients who were uniformly treated with FCR but relapsed after a median of 2 years. In addition to mutations with known adverse-prognostic impact (TP53, NOTCH1, ATM, SF3B1, NFKBIE, and BIRC3), a large proportion of cases (19.5%) harbored mutations in RPS15, a gene encoding a component of the 40S ribosomal subunit. Extended screening, totaling 1119 patients, supported a role for RPS15 mutations in aggressive CLL, with one-third of RPS15-mutant cases also carrying TP53 aberrations. In most cases, selection of dominant, relapse-specific subclones was observed over time. However, RPS15 mutations were clonal before treatment and remained stable at relapse. Notably, all RPS15 mutations represented somatic missense variants and resided within a 7 amino-acid, evolutionarily conserved region. We confirmed the recently postulated direct interaction between RPS15 and MDM2/MDMX and transient expression of mutant RPS15 revealed defective regulation of endogenous p53 compared with wild-type RPS15. In summary, we provide novel insights into the heterogeneous genetic landscape of CLL relapsing after FCR treatment and highlight a novel mechanism underlying clinical aggressiveness involving a mutated ribosomal protein, potentially representing an early genetic lesion in CLL pathobiology.
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- 2016
30. 450K-array analysis of chronic lymphocytic leukemia cells reveals global DNA methylation to be relatively stable over time and similar in resting and proliferative compartments
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Karin E. Smedby, Richard Rosenquist, Anders Rosén, Meena Kanduri, Hanna Göransson-Kultima, A-C Bergh, Anders Isaksson, Gunnar Juliusson, Fergus Ryan, Christer Sundström, Larry Mansouri, and Nicola Cahill
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Male ,Cancer Research ,Chronic lymphocytic leukemia ,Immunoglobulin Variable Region ,Biology ,Cohort Studies ,Biomarkers, Tumor ,medicine ,Humans ,Epigenetics ,Aged ,Cell Proliferation ,Oligonucleotide Array Sequence Analysis ,B-Lymphocytes ,Gene Expression Profiling ,ZAP70 ,DNA, Neoplasm ,Hematology ,Methylation ,DNA Methylation ,Middle Aged ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Gene expression profiling ,Leukemia ,Oncology ,CpG site ,Case-Control Studies ,DNA methylation ,Immunology ,Disease Progression ,Cancer research ,CpG Islands ,Female ,Immunoglobulin Heavy Chains ,Genome-Wide Association Study - Abstract
In chronic lymphocytic leukemia (CLL), the microenvironment influences gene expression patterns; however, knowledge is limited regarding the extent to which methylation changes with time and exposure to specific microenvironments. Using high-resolution 450K arrays, we provide the most comprehensive DNA methylation study of CLL to date, analyzing paired diagnostic/follow-up samples from IGHV-mutated/untreated and IGHV-unmutated/treated patients (n=36) and patient-matched peripheral blood and lymph node samples (n=20). On an unprecedented scale, we revealed 2239 differentially methylated CpG sites between IGHV-mutated and unmutated patients, with the majority of sites positioned outside annotated CpG islands. Intriguingly, CLL prognostic genes (for example, CLLU1, LPL, ZAP70 and NOTCH1), epigenetic regulator (for example, HDAC9, HDAC4 and DNMT3B), B-cell signaling (for example, IBTK) and numerous TGF-β and NF-κB/TNF pathway genes were alternatively methylated between subgroups. Contrary, DNA methylation over time was deemed rather stable with few recurrent changes noted within subgroups. Although a larger number of non-recurrent changes were identified among IGHV-unmutated relative to mutated cases over time, these equated to a low global change. Similarly, few changes were identified between compartment cases. Altogether, we reveal CLL subgroups to display unique methylation profiles and unveil methylation as relatively stable over time and similar within different CLL compartments, implying aberrant methylation as an early leukemogenic event.
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- 2012
31. IGHV3-21 Gene Frequency in a Swedish Cohort of Patients With Newly Diagnosed Chronic Lymphocytic Leukemia
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Fergus Ryan, Karin E. Smedby, Rebeqa Gunnarsson, Richard Rosenquist, Gunnar Juliusson, Mattias Jansson, Fiona Murray, Larry Mansouri, Lesley-Ann Sutton, Christian H. Geisler, and Nicola Cahill
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Adult ,Male ,Cancer Research ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Population ,Immunoglobulin Variable Region ,White People ,Cohort Studies ,Gene Frequency ,Internal medicine ,medicine ,Humans ,education ,Allele frequency ,Aged ,Neoplasm Staging ,Sweden ,education.field_of_study ,Hematology ,business.industry ,Middle Aged ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Stereotypy (non-human) ,Oncology ,Mutation ,Cohort ,Immunology ,Medical genetics ,Female ,Immunoglobulin Heavy Chains ,business ,Cohort study - Abstract
The IGHV3-21 gene has been shown to be overrepresented in Scandinavian patients with chronic lymphocytic leukemia (CLL). By investigating a population-based cohort of 337 Swedish patients with CLL, a lower (6.5%) IGHV3-21 frequency was determined relative to our previous hospital-based studies (10.1%-12.7%), yet this frequency remained higher compared to other Western CLL cohorts (2.6%-4.1%). Furthermore, we confirmed the poor outcome for patients with IGHV3-21 to be independent of mutational and stereotypy status. Background: Scandinavian patients with CLL have shown an overrepresentation of the poor-prognostic IGHV3-21 gene. Furthermore, approximately 50% of patients with IGHV3-21 carry stereotyped B-cell receptors, which implicate antigen selection in leukemogenesis. These patients have also been reported to have shorter time to progression than patients with nonstereotyped IGHV3-21. Materials and Methods: To investigate the IGHV3-21 frequency and the clinical impact of IGHV3-21 stereotypy, 337 newly diagnosed Swedish CLL patients from a population-based cohort were analyzed. Results: Interestingly, the IGHV3-21 frequency was indeed lower (6.5%) in this indolent patient cohort than in our previous hospital-based cohort studies (10.1%-12.7%). Hence, a selection bias of more-aggressive cases rendered a higher proportion of IGHV3-21 cases in our original studies. Nevertheless, the Swedish IGHV3-21 frequency still remained higher when compared with other larger European or American studies (2.6%-4.1%). Finally, we confirmed the poor outcome for IGHV3-21 patients to be independent of mutational status and found stereotypy to have no impact on survival or time to treatment. Conclusion: The Swedish geographic bias in IGHV3-21 gene frequency was validated albeit at a lower frequency than previously reported. Moreover, no prognostic value could be attributed to IGHV3-21 stereotype status.
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- 2012
32. Impact of TP53 mutation and 17p deletion in mantle cell lymphoma
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Fiona Murray, Anna Margrét Halldórsdóttir, Anna Laurell, A Lundin, Sakari Knuutila, Birgitta Sander, Hans Ehrencrona, Larry Mansouri, Christer Sundström, and Richard Rosenquist
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Male ,Cancer Research ,Chronic lymphocytic leukemia ,Lymphoma, Mantle-Cell ,Biology ,Cohort Studies ,03 medical and health sciences ,Exon ,0302 clinical medicine ,immune system diseases ,hemic and lymphatic diseases ,medicine ,Humans ,neoplasms ,Gene ,Aged ,030304 developmental biology ,Aged, 80 and over ,Chromosome Aberrations ,Genetics ,0303 health sciences ,Hematology ,Methylation ,Middle Aged ,Prognosis ,medicine.disease ,Candidate Tumor Suppressor Gene ,3. Good health ,Survival Rate ,Oncology ,030220 oncology & carcinogenesis ,Mutation ,DNA methylation ,Cancer research ,Female ,Mantle cell lymphoma ,Tumor Suppressor Protein p53 ,IGHV@ ,Gene Deletion ,Chromosomes, Human, Pair 17 - Abstract
Mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) both belong to the group of mature B-cell malignancies. However, MCL is typically clinically aggressive while the clinical course of CLL varies. CLL can be divided into prognostic subgroups based on IGHV mutational status and into multiple subsets based on closely homologous (stereotyped) B-cell receptors. In paper I we investigated 31 MCL cases using high-density 250K single-nucleotide polymorphism arrays and gene expression arrays. Although most copy-number aberrations (CNAs) were previously reported in MCL, a novel deletion was identified at 20q (16%) containing the candidate tumor suppressor gene ZFP64. A high proliferation gene expression signature was associated with poor prognosis, large CNAs, 7p gains and 9q losses. Losses at 1p/8p/13q/17p were associated with increased genomic complexity. In paper II we sequenced exons 4 to 8 of the TP53 gene in 119 MCL cases. 17p copy-number status was known from previous studies or determined by real-time quantitative polymerase chain reaction. TP53 mutations were detected in 14% of cases and were strongly associated with poor survival while 17p deletions were more common (32%) but did not predict survival. In papers III and IV we applied high-resolution genomic 27K methylation arrays to 20 MCL and 39 CLL samples. In paper III MCL displayed a homogenous methylation profile without correlation with the proliferation signature whereas MCL was clearly separated from CLL. Gene ontology analysis revealed enrichment of developmental genes, in particular homeobox transcription factor genes, among targets methylated in MCL. In paper IV we compared three different stereotyped CLL subsets: #1 (IGHV unmutated), #2 (IGHV3-21) and #4 (IGHV mutated). Many genes were differentially methylated between each two subsets and immune response genes (e.g. CD80 and CD86) were enriched among genes methylated in subset #1 but not in subsets #2/#4. In summary, CNAs were frequent and not random in MCL. Specific CNAs correlated with a high proliferation gene expression signature or genomic complexity. TP53 mutations predicted short survival whereas 17p deletions did not. A high proliferation signature was not associated with differential DNA methylation in MCL, which demonstrated a homogeneous methylation pattern. In contrast, genomic methylation patterns differed between MCL and CLL and between stereotyped CLL subsets.
- Published
- 2011
33. LPL is the strongest prognostic factor in a comparative analysis of RNA-based markers in early chronic lymphocytic leukemia
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Henrik Hjalgrim, Anne Mette Buhl, Rebeqa Gunnarsson, Marie Sevov, Gunnar Juliusson, Karin E. Smedby, Richard Rosenquist, Mohd Arifin Kaderi, Meena Kanduri, Mattias Jansson, Jesper Jurlander, Nicola Cahill, and Larry Mansouri
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Male ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Internal medicine ,Biomarkers, Tumor ,Humans ,Medicine ,MCL1 ,RNA, Messenger ,Survival analysis ,Aged ,Lipoprotein lipase ,Hematology ,business.industry ,ZAP70 ,RNA ,Original Articles ,Middle Aged ,Prognosis ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,Lipoprotein Lipase ,Treatment Outcome ,Multivariate Analysis ,Mutation ,Immunology ,Cancer research ,Medical genetics ,Female ,Immunoglobulin Heavy Chains ,business - Abstract
The expression levels of LPL, ZAP70, TCL1A, CLLU1 and MCL1 have recently been proposed as prognostic factors in chronic lymphocytic leukemia. However, few studies have systematically compared these different RNA-based markers.Using real-time quantitative PCR, we measured the mRNA expression levels of these genes in unsorted samples from 252 newly diagnosed chronic lymphocytic leukemia patients and correlated our data with established prognostic markers (for example Binet stage, CD38, IGHV gene mutational status and genomic aberrations) and clinical outcome.High expression levels of all RNA-based markers, except MCL1, predicted shorter overall survival and time to treatment, with LPL being the most significant. In multivariate analysis including the RNA-based markers, LPL expression was the only independent prognostic marker for overall survival and time to treatment. When studying LPL expression and the established markers, LPL expression retained its independent prognostic strength for overall survival. All of the RNA-based markers, albeit with varying ability, added prognostic information to established markers, with LPL expression giving the most significant results. Notably, high LPL expression predicted a worse outcome in good-prognosis subgroups, such as patients with mutated IGHV genes, Binet stage A, CD38 negativity or favorable cytogenetics. In particular, the combination of LPL expression and CD38 could further stratify Binet stage A patients.LPL expression is the strongest RNA-based prognostic marker in chronic lymphocytic leukemia that could potentially be applied to predict outcome in the clinical setting, particularly in the large group of patients with favorable prognosis.
- Published
- 2011
34. Functional loss of IκBε leads to NF-κB deregulation in aggressive chronic lymphocytic leukemia
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Viktor Ljungström, Frederic Davi, Kostas Stamatopoulos, Diego Cortese, Nicholas Chiorazzi, Johan Rung, Šárka Pospíšilová, Xiao-Jie Yan, Antonia Kalushkova, Gunnar Juliusson, Karin E. Smedby, Gunilla Enblad, Jonathan C. Strefford, Karla Plevová, Sina Bondza, Lesley-Ann Sutton, Brigitta Sander, Linda Arngården, Rebeqa Gunnarsson, Anton W. Langerak, Larry Mansouri, Jimmy Larsson, Erin Young, Richard Rosenquist, Elin Falk-Sörqvist, Helena Jernberg-Wiklund, Marcus Lars Vittorio Nilsson, Alice F. Muggen, Paolo Ghia, Sujata Bhoi, Ola Söderberg, Chrysoula Belessi, Mats Hellström, Peter Lönn, Mansouri, L, Sutton, La, Ljungström, V, Bondza, S, Arngården, L, Bhoi, S, Larsson, J, Cortese, D, Kalushkova, A, Plevova, K, Young, E, Gunnarsson, R, Falk Sörqvist, E, Lönn, P, Muggen, Af, Yan, Xj, Sander, B, Enblad, G, Smedby, Ke, Juliusson, G, Belessi, C, Rung, J, Chiorazzi, N, Strefford, Jc, Langerak, Aw, Pospisilova, S, Davi, F, Hellström, M, Jernberg Wiklund, H, Ghia, PAOLO PROSPERO, Söderberg, O, Stamatopoulos, K, Nilsson, M, and Rosenquist, R.
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Cell- och molekylärbiologi ,Chronic lymphocytic leukemia ,Immunology ,B-cell receptor ,Biology ,Frameshift mutation ,03 medical and health sciences ,NFKBIE Gene ,0302 clinical medicine ,hemic and lymphatic diseases ,medicine ,Humans ,Immunology and Allergy ,B cell ,030304 developmental biology ,0303 health sciences ,Gene Expression Regulation, Leukemic ,Brief Definitive Report ,NF-kappa B ,breakpoint cluster region ,Hematology ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,NFKBIE ,I-kappa B Kinase ,3. Good health ,Leukemia ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Cancer research ,Cell and Molecular Biology - Abstract
Mansouri et al. applied targeted deep sequencing to identify mutations within NF-κB core complex genes in CLL. NFKBIE, the gene encoding the inhibitory IκBε molecule, was most frequently mutated, especially in poor-prognostic subgroups of CLL. The authors show that NFKBIE mutations were associated with significantly reduced IkBε expression and p65 inhibition, ultimately leading to NF-κB activation and a more aggressive disease., NF-κB is constitutively activated in chronic lymphocytic leukemia (CLL); however, the implicated molecular mechanisms remain largely unknown. Thus, we performed targeted deep sequencing of 18 core complex genes within the NF-κB pathway in a discovery and validation CLL cohort totaling 315 cases. The most frequently mutated gene was NFKBIE (21/315 cases; 7%), which encodes IκBε, a negative regulator of NF-κB in normal B cells. Strikingly, 13 of these cases carried an identical 4-bp frameshift deletion, resulting in a truncated protein. Screening of an additional 377 CLL cases revealed that NFKBIE aberrations predominated in poor-prognostic patients and were associated with inferior outcome. Minor subclones and/or clonal evolution were also observed, thus potentially linking this recurrent event to disease progression. Compared with wild-type patients, NFKBIE-deleted cases showed reduced IκBε protein levels and decreased p65 inhibition, along with increased phosphorylation and nuclear translocation of p65. Considering the central role of B cell receptor (BcR) signaling in CLL pathobiology, it is notable that IκBε loss was enriched in aggressive cases with distinctive stereotyped BcR, likely contributing to their poor prognosis, and leading to an altered response to BcR inhibitors. Because NFKBIE deletions were observed in several other B cell lymphomas, our findings suggest a novel common mechanism of NF-κB deregulation during lymphomagenesis.
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- 2015
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35. 4.23 MDM2 Promotor Polymorphism and Disease Characteristics in CLL: Individual Patient Data Meta-Analysis of 2598 CLL Patients
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Olaf Merkel, Hartmut Döhner, Šárka Pavlová, Šárka Pospíšilová, Mohd Arifin Kaderi, Peter Söderkvist, Richard Greil, Thorsten Zenz, Larry Mansouri, Axel Benner, Davide Rossi, Stephan Stilgenbauer, Ulrich Dührsen, Aneela Majid, Gareth L. Bond, David Oscier, Holger Nückel, Gianluca Gaidano, Mats Linderholm, Kerstin Willander, Martin J. S. Dyer, Richard Rosenquist, and Anton Parker
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Oncology ,Cancer Research ,medicine.medical_specialty ,biology ,business.industry ,Promoter ,Hematology ,Patient data ,03 medical and health sciences ,0302 clinical medicine ,030220 oncology & carcinogenesis ,Meta-analysis ,Internal medicine ,medicine ,biology.protein ,Mdm2 ,Disease characteristics ,business ,030215 immunology - Published
- 2011
36. Recurrent mutations refine prognosis in chronic lymphocytic leukemia
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Kostas Stamatopoulos, Achilles Anagnostopoulos, Karin E. Smedby, Diego Cortese, Neus Villamor, Alba Navarro, C. Belessi, La. Sutton, E. Minga, Larry Mansouri, Gianluca Gaidano, Eugen Tausch, Paolo Ghia, Richard Rosenquist, Veronika Navrkalová, Šárka Pospíšilová, Jana Kminkova, Julio Delgado, Anastasia Hadzidimitriou, Lydia Scarfò, Gunnar Juliusson, Andreas Agathangelidis, Davide Rossi, Jonathan C. Strefford, Zadie Davis, Antonios M. Makris, Matthew J. J. Rose-Zerilli, David Oscier, Stephan Stilgenbauer, Panagiotis Baliakas, Elias Campo, Barbara Kantorová, Marta Larrayoz, Evangelia Stalika, Baliakas, P., Hadzidimitriou, A., Sutton, L. -A., Rossi, D., Minga, E., Villamor, N., Larrayoz, M., Kminkova, J., Agathangelidis, A., Davis, Z., Tausch, E., Stalika, E., Kantorova, B., Mansouri, L., Scarfo', L., Cortese, D., Navrkalova, V., Rose-Zerilli, M. J. J., Smedby, K. E., Juliusson, G., Anagnostopoulos, A., Makris, A. M., Navarro, A., Delgado, J., Oscier, D., Belessi, C., Stilgenbauer, S., Ghia, P., Pospisilova, S., Gaidano, G., Campo, E., Strefford, J. C., Stamatopoulos, K., and Rosenquist, R.
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Oncology ,Male ,Cancer Research ,Time Factors ,Chronic lymphocytic leukemia ,DNA Mutational Analysis ,medicine.disease_cause ,0302 clinical medicine ,Recurrence ,hemic and lymphatic diseases ,Receptor, Notch1 ,0303 health sciences ,Mutation ,Hematology ,Middle Aged ,Prognosis ,3. Good health ,Europe ,Leukemia ,030220 oncology & carcinogenesis ,Female ,RNA Splicing Factors ,IGHV@ ,medicine.medical_specialty ,Polymorphism, Single Nucleotide ,03 medical and health sciences ,Cytogenetics ,Internal medicine ,medicine ,Humans ,Clinical significance ,neoplasms ,030304 developmental biology ,Aged ,business.industry ,Ribonucleoprotein, U2 Small Nuclear ,medicine.disease ,Phosphoproteins ,Leukemia, Lymphocytic, Chronic, B-Cell ,Lymphoma ,Immunology ,Multivariate Analysis ,Tumor Suppressor Protein p53 ,business ,Trisomy ,Gene Deletion - Abstract
Through the European Research Initiative on chronic lymphocytic leukemia (CLL) (ERIC), we screened 3490 patients with CLL for mutations within the NOTCH1 (n=3334), SF3B1 (n=2322), TP53 (n=2309), MYD88 (n=1080) and BIRC3 (n=919) genes, mainly at diagnosis (75%) and before treatment (>90%). BIRC3 mutations (2.5%) were associated with unmutated IGHV genes (U-CLL), del(11q) and trisomy 12, whereas MYD88 mutations (2.2%) were exclusively found among M-CLL. NOTCH1, SF3B1 and TP53 exhibited variable frequencies and were mostly enriched within clinically aggressive cases. Interestingly, as the timespan between diagnosis and mutational screening increased, so too did the incidence of SF3B1 mutations; no such increase was observed for NOTCH1 mutations. Regarding the clinical impact, NOTCH1 mutations, SF3B1 mutations and TP53 aberrations (deletion/mutation, TP53ab) correlated with shorter time-to-first-treatment (P
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- 2014
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37. ΝFΚΒΙΕ Deletions: A Novel Marker of Clinical Aggressiveness in Primary Mediastinal B-Cell Lymphoma
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Jude Fitzgibbon, Theodoros P. Vassilakopoulos, Richard Rosenquist, Frederik Damm, Gunilla Enblad, Elias Campo, Hans G. Drexler, Emma Young, Niki Stavroyianni, German Ott, Elisabeth Ralfkiaer, Maria K. Angelopoulou, Frick Mareike, Lucile Couronné, Marita Ziepert, Viktor Ljungström, Magnus Hultdin, Christopher Maximilian Arends, Rose-Marie Amini, Theodora Papadaki, Kirsten Grønbæk, Blanca Gonzalez-Farre, Jonathan C. Strefford, Tasidou Anna, Claudia D. Baldus, Thorsten Zenz, Larry Mansouri, Armando López-Guillermo, Maysaa Abdulla, George Kanellis, Christian Bastard, Dido Lenze, Kostas Stamatopoulos, Damien Roos-Weil, Penelope Korkolopoulou, Nils Waldhueter, Bernd Dörken, Alfredo Rivas, Jessica Okosun, Clemens A. Schmitt, Daniel Noerenberg, Helga D. Munch-Petersen, Olivier A. Bernard, Andreas Rosenwald, Michael Hummel, and Fazila Asmar
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Oncology ,medicine.medical_specialty ,business.industry ,Immunology ,Follicular lymphoma ,Primary central nervous system lymphoma ,Cell Biology ,Hematology ,Gene mutation ,medicine.disease ,Biochemistry ,NFKBIE ,NFKBIE Gene ,Internal medicine ,medicine ,Mantle cell lymphoma ,Splenic marginal zone lymphoma ,Primary mediastinal B-cell lymphoma ,business - Abstract
Deregulated NF-κB signaling is a hallmark of most, if not all, lymphoid malignancies, and recurrent gene mutations in both the canonical and non-canonical NF-κB pathway are known to lead to NF-κB activation. However, the full compendium of NF-κB gene mutations in lymphoid malignancies remains to be elucidated. Recently, we reported a 4-bp truncating mutation in the NFKBIE gene, which encodes IκBε, a negative regulator of NF-κB, in patients with chronic lymphocytic leukemia (CLL). The NFKBIE deletion was enriched in clinically aggressive CLL patients (7-8%) and associated with a worse clinical outcome. At the functional level, NFKBIE-deleted CLL showed reduced IκBε levels and decreased p65 inhibition, along with increased phosphorylation and nuclear translocation of p65, compared to wildtype patients. Preliminary data has indicated an increased frequency of NFKBIE aberrations in other lymphoid malignancies as well. To explore this further, we screened for NFKBIE deletions in a large cohort of patients diagnosed with a range of different lymphoid neoplasms. Overall, NFKBIE deletions were identified in 76 of 1414 patients (5.4%). While NFKBIE deletions were relatively infrequent in patients diagnosed with follicular lymphoma (3/225, 1.3%), splenic marginal zone lymphoma (3/175, 1.7%), and T-cell acute lymphoblastic leukemia (1/94, 1.1%), moderate frequencies were observed among diffuse large B-cell lymphoma (18/521, 3.5%), mantle cell lymphoma (8/189, 4.2%), and primary CNS lymphoma (1/34, 2.9%) patients. In contrast, a remarkably high frequency of NFKBIE deletions (41/176 cases, 23%) was observed among primary mediastinal B-cell lymphoma (PMBL) patients. Noteworthy, the prevalence of NFKBIE-deleted PMBL cases was similar in the different contributing centers. All PMBL patients in the present series received a CHOP based treatment regime; in ~75% of cases rituximab was added and ~25% were treated with dose intensified schemes. For the latter, the vast majority of patients received CHOEP, while individual cases were treated with MegaCHOEP, DA-EPOCH or ACVBP. Regarding clinicobiological associations, there were no significant differences between NFKBIE-deleted and wildtype PMBL patients with respect to age, sex, Ann Arbor stage, IPI risk-groups, extranodal or bone marrow involvement, bulky disease, and LDH elevation. However, NFKBIE-deleted patients were more likely to be refractory to primary chemotherapy (31% vs. 3%, P=.001) and had a shorter overall survival compared to wildtype patients (5-year overall survival: 63% vs 84%, P=.013). In multivariate analysis (including age, gender, Ann Arbor stage, IPI, and NFKBIE mutation status), NFKBIE mutation status (95% CI: 1.23-10.61; HR: 3.61; P=0.020) remained an independent factor for poor prognosis. In summary, we document NFKBIE deletions as a common genetic event across B-cell malignancies, albeit at varying frequencies. The high frequency of NFKBIE deletions in PMBL alludes to the critical role of this aberration in the pathophysiology of the disease. NFKBIE deletions were associated witha worse clinical outcome, hence potentially representing a novel poor-prognostic marker in PMBL. *Contributed equally as first authors. **Contributed equally as senior authors. Disclosures Stamatopoulos: Gilead: Consultancy, Honoraria, Research Funding; Abbvie: Honoraria, Other: Travel expenses; Novartis: Honoraria, Research Funding; Janssen: Honoraria, Other: Travel expenses, Research Funding.
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- 2016
38. On the way towards a 'CLL prognostic index': focus on TP53, BIRC3, SF3B1, NOTCH1 and MYD88 in a population-based cohort
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Gunnar Juliusson, Lesley-Ann Sutton, Christian H. Geisler, Larry Mansouri, Karin E. Smedby, Diego Cortese, Richard Rosenquist, Nicola Cahill, and Rebeqa Gunnarsson
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Gerontology ,Cancer Research ,Index (economics) ,Ubiquitin-Protein Ligases ,Population ,Inhibitor of Apoptosis Proteins ,Cohort Studies ,Population based cohort ,Medicine ,Humans ,Receptor, Notch1 ,education ,Focus (computing) ,education.field_of_study ,business.industry ,Hematology ,Ribonucleoprotein, U2 Small Nuclear ,Phosphoproteins ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,Baculoviral IAP Repeat-Containing 3 Protein ,Oncology ,Cohort ,Immunology ,Myeloid Differentiation Factor 88 ,RNA Splicing Factors ,Tumor Suppressor Protein p53 ,business - Abstract
On the way towards a 'CLL prognostic index' : focus on TP53, BIRC3, SF3B1, NOTCH1 and MYD88 in a population-based cohort.
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- 2013
39. Feasibility of targeted next-generation sequencing of the TP53 and ATM genes in chronic lymphocytic leukemia
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Elin Falk Sorqvist, Larry Mansouri, Lesley-Ann Sutton, Kostas Stamatopoulos, Mats Nilsson, Karin E. Smedby, Viktor Ljungström, Rebeqa Gunnarsson, Gunnar Juliusson, and Richard Rosenquist
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Cancer Research ,endocrine system diseases ,business.industry ,Chronic lymphocytic leukemia ,Hematology ,Ataxia Telangiectasia Mutated Proteins ,medicine.disease ,Genes, p53 ,Leukemia, Lymphocytic, Chronic, B-Cell ,DNA sequencing ,stomatognathic system ,Oncology ,hemic and lymphatic diseases ,Immunology ,Cancer research ,Medicine ,Feasibility Studies ,Humans ,business ,neoplasms ,Gene - Abstract
Feasibility of targeted next-generation sequencing of the TP53 and ATM genes in chronic lymphocytic leukemia
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- 2013
40. Exploring the genetic landscape in chronic lymphocytic leukemia using high-resolution technologies
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Richard Rosenquist, Rebeqa Gunnarsson, and Larry Mansouri
- Subjects
Genetics ,Chromosome Aberrations ,Cancer Research ,Microarray ,Chronic lymphocytic leukemia ,High-Throughput Nucleotide Sequencing ,Hematology ,Disease ,Genomics ,Biology ,medicine.disease ,Genome ,Leukemia, Lymphocytic, Chronic, B-Cell ,DNA sequencing ,Leukemia ,Oncology ,medicine ,Humans ,Gene ,Exome sequencing - Abstract
During recent years, microarray-based technologies and next-generation sequencing (NGS) have been applied in chronic lymphocytic leukemia (CLL) in order to identify novel genomic aberrations that may contribute to the pathogenesis of the disease. Even though high-resolution microarray studies have confirmed the importance of the known recurrent aberrations, i.e. del(11q), trisomy 12, del(13q) and del(17p), and have more precisely delineated the genomic borders of these aberrations, only a few novel aberrations, found at a low frequency, have been detected with these techniques. In contrast to this, the application of NGS technology of the coding genome (exome sequencing) or the entire genome (whole-genome sequencing) has unveiled a number of novel recurrent mutations in e.g. the NOTCH1, SF3B1 and BIRC3 genes. Importantly, mutations in these latter genes were reported to be associated with a particularly poor outcome, similar to TP53 aberrations, and may play key roles in tumor development, treatment resistance and prognosis. In this review, we not only summarize the latest achievements using array-based or NGS technologies, but also point to new directions for research aiming to unravel the complex genetic "map" in CLL and its prognostic subsets.
- Published
- 2012
41. Next generation RNA-sequencing in prognostic subsets of chronic lymphocytic leukemia
- Author
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Sean D. Hooper, Richard Rosenquist, Markus Mayrhofer, Gunnar Juliusson, Ulf Gyllensten, Larry Mansouri, Adam Ameur, Anders Isaksson, Rebeqa Gunnarsson, and Lesley-Ann Sutton
- Subjects
Male ,RNA, Untranslated ,Time Factors ,Sequence analysis ,Chronic lymphocytic leukemia ,Mutation, Missense ,Pilot Projects ,Biology ,medicine.disease_cause ,Exon ,medicine ,Humans ,RNA, Neoplasm ,Gene ,Aged ,Genetics ,Regulation of gene expression ,Sweden ,Mutation ,Base Sequence ,Sequence Analysis, RNA ,Gene Expression Profiling ,Alternative splicing ,Hematology ,Middle Aged ,medicine.disease ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Alternative Splicing ,RNA splicing ,Female - Abstract
Advances in next-generation RNA-sequencing have revealed the complexity of transcriptomes by allowing both coding and noncoding (nc) RNAs to be analyzed. However, limited data exist regarding the whole transcriptional landscape of chronic lymphocytic leukemia (CLL). In this pilot-study, we evaluated RNA-sequencing in CLL by comparing two subsets which carry almost identical or `` stereotyped'' B-cell receptors with distinct clinical outcome, that is the poor-prognostic subset # 1 (n = 4) and the more favorable-prognostic subset # 4 (n = 4). Our analysis revealed that 156 genes (e.g. LPL, WNT9A) and 76 ncRNAs, (e. g. SNORD48, SNORD115) were differentially expressed between the subsets. This technology also enabled us to identify numerous subset-specific splice variants (n = 406), which were predominantly expressed in subset # 1, including a splice-isoform of MSI2 with a novel start exon. A further important application of RNA-sequencing was for mutation detection and revealed 16-30 missense mutations per sample; notably many of these changes were found in genes with a strong potential for involvement in CLL pathogenesis, e. g., ATM and NOTCH2. This study not only demonstrates the effectiveness of RNA-sequencing for identifying mutations, quantifying gene expression and detecting splicing events, but also highlights the potential such global approaches have to significantly advance our understanding of the molecular mechanisms behind CLL development.
- Published
- 2012
42. RNA-based markers as prognostic factors in chronic lymphocytic leukemia
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Richard Rosenquist, Marie Sevov, and Larry Mansouri
- Subjects
business.industry ,ZAP70 ,Chronic lymphocytic leukemia ,Mrna expression ,Gene Expression Profiling ,Clinical course ,RNA ,Hematology ,Bioinformatics ,medicine.disease ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,Lipoprotein Lipase ,hemic and lymphatic diseases ,microRNA ,medicine ,Biomarkers, Tumor ,Humans ,RNA, Messenger ,RNA, Neoplasm ,business ,Gene ,Progressive disease - Abstract
Chronic lymphocytic leukemia (CLL) is most often indolent at diagnosis but has a highly variable clinical course, and many patients will eventually progress and require treatment. Currently, there are a number of clinical and molecular markers known to be predictive of prognosis in CLL that can be applied to discriminate patients that are more likely to develop a progressive disease. Gene-expression profiling studies have identified genes with differential expression between prognostic subgroups in CLL, and research on these RNA-based prognostic markers has expanded during recent years. For example, high lipoprotein lipase and CLLU1 mRNA expression have recently been shown to be strong markers of poor clinical outcome. In this review we will provide a summary of the most significant prognostic markers in CLL, focusing on the recent category of RNA-based markers in particular.
- Published
- 2012
43. EGR2 Mutations in Chronic Lymphocytic Leukemia: A New Bad Player
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Panagiotis Panagiotidis, Nicholas Chiorazzi, Ruth Clifford, Šárka Pospíšilová, Anton W. Langerak, Richard Rosenquist, Gunnar Juliusson, Paolo Ghia, Emma Young, Viktor Ljungström, Anna Schuh, Gianluca Gaidano, Lesley-Ann Sutton, Frederic Davi, Veronika Navrkalová, Kostas Stamatopoulos, Daniel Noerenberg, Karin E. Smedby, Panagiotis Baliakas, Davide Rossi, Stuart Blakemore, Jonathan C. Strefford, Frederik Damm, Thorsten Zenz, Larry Mansouri, Karla Plevová, and Chrysoula Belessi
- Subjects
Oncology ,Sanger sequencing ,medicine.medical_specialty ,Hematology ,business.industry ,Chronic lymphocytic leukemia ,Immunology ,breakpoint cluster region ,Cell Biology ,Bioinformatics ,medicine.disease ,Biochemistry ,symbols.namesake ,Internal medicine ,medicine ,symbols ,Mutation testing ,Missense mutation ,IGHV@ ,business ,Progressive disease - Abstract
*Contributed equally as first authors. **Contributed equally as senior authors. Recurrent mutations within EGR2, a versatile transcription factor involved in differentiation of hematopoietic cells, were recently reported in 8% of advanced-stage chronic lymphocytic leukemia (CLL) patients, where they appear to be associated with a worse outcome. EGR2 is activated through ERK phosphorylation upon B-cell receptor (BcR) stimulation, and we have previously shown that EGR2 -mutated CLL patients display altered expression of EGR2 down-stream target genes compared to wildtype (wt) patients, thereby pointing to a pathogenic role for EGR2 mutations in dysregulating BcR signaling. To gain further insight into the incidence and prognostic impact of EGR2 mutations in CLL, we screened samples from a well-characterized series of 1430 patients, either by Sanger sequencing (n=1019) or targeted deep-sequencing (n=370), both covering the recently reported EGR2 hotspot in exon 2. In addition, whole-exome data was available for an additional 43 patients. Different cohorts were included in our analysis ranging from 'general practice' CLL (33% IGHV-unmutated (U-CLL), 6% TP53 -aberrant (TP53abn), n=693), to adverse-prognostic CLL (89% U-CLL, 26% TP53abn, n=325), patients belonging to clinically aggressive stereotyped subsets #1-3 & #5-8 (n=342), patients relapsing after FCR therapy (n=41) and Richter transformed cases (n=31), thus reflecting the heterogeneous nature of CLL. Nineteen EGR2 mutations were detected by Sanger sequencing, while 22 additional mutations were identified with deep-sequencing using a 5% variant allele frequency (VAF) cutoff (median 39%, range 5.6-63.9%, median coverage 43,000X). With the exception of one in-frame deletion, all mutations were missense alterations located within the three zinc-finger domains. Significant enrichment of EGR2 mutations was observed in adverse-prognostic (18/325, 5.5%) and FCR-relapsing (4/41, 9.8%) CLL compared to the 'general practice' cohort (18/693, 2.6%, Figure 1A). A surprisingly low frequency was observed among clinically aggressive stereotyped subsets (5/342, 1.5%), although the cause for this observation is currently unknown. Finally, 2/31 (6.5%) cases with Richter transformation carried an EGR2 mutation. Of the 4 FCR-relapsing, EGR2 -mutated cases with available overtime samples, all demonstrated a significant expansion of the EGR2 -mutated clone at relapse (VAF-increase between 15-41%). In addition, subclonal levels of EGR2 hotspot mutations (VAF 0.5-5%) were detected in an additional 13/370 (3.5%) cases by deep-sequencing. The majority of EGR2 -mutated CLL patients (32/39, 82%) concerned U-CLL and the following aberrations co-occurred: 11q-deletions (n=10), TP53abn (n=6), NOTCH1 (n=3)or SF3B1 (n=3) mutations. EGR2 -mutated patients displayed a significantly worse overall survival compared to wt patients (median survival 59 vs. 141 months, p=0.003, using a conservative 10% VAF cutoff), and a poor outcome similar to cases with TP53abn (Figure 1B). In multivariate analysis (n=583), EGR2 status remained an independent factor (p=0.038), along with stage (p=0.048) and IGHV status (p In summary, EGR2 -mutant cases appear to constitute a novel poor-prognostic subgroup of CLL, with mutations occurring either as disease-initiating aberrations, i.e. in cases where mutations were found in the entire clone, or as subclonal driver events linked to progressive disease. The latter is reflected by the enrichment of EGR2 mutations in aggressive CLL and the association of EGR2 mutations with an overall dismal prognosis. Considering the potential role of mutated EGR2 in altering BcR signaling, it will be particularly relevant to study the efficacy of BcR inhibitors in this patient group. Figure 1. Figure 1. Figure 2. Figure 2. Disclosures Langerak: Roche: Other: Lab services in the field of MRD diagnostics provided by Dept of Immunology, Erasmus MC (Rotterdam); InVivoScribe: Patents & Royalties: Licensing of IP and Patent on BIOMED-2-based methods for PCR-based Clonality Diagnostics.; DAKO: Patents & Royalties: Licensing of IP and Patent on Split-Signal FISH. Royalties for Dept. of Immunology, Erasmus MC, Rotterdam, NL. Schuh:Acerta Pharma BV: Research Funding. Strefford:Roche: Research Funding.
- Published
- 2015
44. ATM Mutations in Major Stereotyped CLL Subsets: Enrichment in Subset #2 is Associated with Unfavourable Outcome
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Panagiotis Panagiotidis, Viktor Ljungström, Šárka Pospíšilová, Kostas Stamatopoulos, Karla Plevová, Anton W. Langerak, Emma Young, Chrysoula Belessi, Veronika Navrkalová, Richard Rosenquist, Lenka Radová, Martin Trbušek, Paolo Ghia, Panagiotis Baliakas, Lesley-Ann Sutton, Larry Mansouri, Gunnar Juliusson, Frederic Davi, Karin E. Smedby, Jonathan C. Strefford, Stavroula Ntoufa, Zadie Davis, and David Oscier
- Subjects
Pediatrics ,medicine.medical_specialty ,Chronic lymphocytic leukemia ,Immunology ,Biochemistry ,law.invention ,03 medical and health sciences ,Exon ,0302 clinical medicine ,law ,Internal medicine ,medicine ,Gene ,Polymerase chain reaction ,030304 developmental biology ,0303 health sciences ,Hematology ,business.industry ,Cancer ,Cell Biology ,medicine.disease ,3. Good health ,RNA splicing ,Cancer research ,business ,030215 immunology - Abstract
Chronic lymphocytic leukemia (CLL) is a paradigmatic malignancy where the interplay of cell-extrinsic and cell-intrinsic factors has a major impact on disease evolution. Indeed, extrinsic triggering, e.g. antigenic stimulation through the B-cell receptor (BcR), together with intrinsic aberrations, e.g. accumulation of genetic defects, play a major role throughout the natural history of CLL. The importance of antigen involvement is underscored by the existence of 'stereotyped' BcR in up to 30% of CLL patients. Notably, CLL patients with stereotyped BcR can be grouped into different subsets, each with a subset-biased biological and clinical profile. For instance, while the clinically aggressive subset #2 (IGHV3-21/IGLV3-21, comprising both mutated (M-CLL) and unmutated (U-CLL) IGHV genes) displays a remarkably high frequency of SF3B1 mutations, subset #8, a subset with the highest risk of Richter transformation, shows a strong association with trisomy 12 and NOTCH1 mutations. ATM defects are implicated in the evolution of CLL and are associated with a dismal prognosis, however the extent to which they contribute to the genetic landscape in stereotyped subsets remains unexplored. To gain insight into this issue, we assessed the frequency of ATM mutations in 249 well-characterized CLL patients assigned to major subsets #1-8. The entire coding region of ATM (62 exons) was investigated with two different targeted deep-sequencing approaches, i.e. Haloplex technology (HiSeq, coverage ~1500X) or the Nextera XT kit (MiSeq, coverage ~4000X). A conservative variant allele frequency cut-off of 10% was selected, and mutations were validated by Sanger sequencing. Altogether, we identified 61 ATM mutations in 47/249 (19%) patients across all major subsets (Fig. 1). As expected, the majority of identified ATM mutations (n=43, 70%) have not yet been reported while the remaining 30% were listed in the HGMD or COSMIC mutation databases. The spectrum of ATM mutations included missense (n=31), nonsense (n=9), splicing (n=6), and frame-shift (n=14) mutations, and one in-frame deletion. Missense substitutions were distributed along the entire gene without any 'hotspot' region or preferred domain. The highest mutation frequency was detected in subset #2 (26%), with a significant enrichment in U-CLL vs. M-CLL cases, (13/33 vs. 8/48 subset #2 cases, respectively; p=0.021). Within poor-prognostic U-CLL subsets, ATM mutations were also frequent in subsets #6 (25%) and #7 (23%), while subsets #3, #5, #1, and #8 showed lower frequencies (17%, 17%, 13%, and 7%, respectively). The favorable prognostic M-CLL subset #4 exhibited a low frequency (7%) of ATM mutations. Notably, when comparing the two most populated subsets, i.e. #1 and #2, ATM mutations were overrepresented in the latter with a borderline significance value (p=0.086); when restricting the analysis to U-CLL #2 cases a significantly higher frequency was observed compared to #1 (13/33 vs 9/68; p=0.0045). Regarding the clinical impact of ATM defects in subset #2, we divided patients into subgroups with biallelic inactivation (def-ATM), sole 11q-, sole ATM -mutation, TP53 -aberrations and WT. While both groups with mono-allelic ATM disruption showed a significantly reduced overall survival compared to WT (median survival sole ATM -mutation, 71 months, sole 11q-, 40 months, vs. 123 months in the WT group; p=0.002 and 0.02, respectively), a non-significant reduction of overall survival was observed for patients with bi-allelic ATM aberrations (70 months, p=0.29) (Fig. 2). The few subset #2 patients with TP53 defects showed a similar survival as WT group, underscoring previous observations that TP53 dysfunction per se plays a minor role in this subset. In summary, we demonstrate that ATM mutations can be added to the list of genetic defects with a biased distribution in stereotyped subsets. The enrichment of ATM defects in subset #2 was associated with a negative impact on overall survival, suggesting a role for ATM inactivation in shaping the aggressive phenotype of this subset. This study further supports the recent suggestion that CLL development is driven by antigenic selection, coupled with preferential acquisition of specific genetic defects. The work was supported by the projects MSMT CR CZ.1.05/1.1.00/02.0068, IGA NT13493-4/2012 and TACR TE02000058. Figure 1. Figure 1. Figure 2. Figure 2. Disclosures Langerak: Roche: Other: Lab services in the field of MRD diagnostics provided by Dept of Immunology, Erasmus MC (Rotterdam); DAKO: Patents & Royalties: Licensing of IP and Patent on Split-Signal FISH. Royalties for Dept. of Immunology, Erasmus MC, Rotterdam, NL; InVivoScribe: Patents & Royalties: Licensing of IP and Patent on BIOMED-2-based methods for PCR-based Clonality Diagnostics. . Strefford:Roche: Research Funding. Stamatopoulos:Gilead Sciences: Research Funding; Janssen Pharmaceuticals: Research Funding.
- Published
- 2015
45. CLL with Mutated IGHV4-34 Antigen Receptors Is Clinically Heterogeneous: Antigen Receptor Stereotypy Makes the Difference
- Author
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Alba Navarro, Panagiotis Baliakas, Lone Bredo Pedersen, Richard Rosenquist, Stephan Stilgenbauer, Véronique Giudicelli, Yorick Sandberg, Elias Campo, Tatiana Tzenou, Gunnar Juliusson, Monica Facco, Andrey Sudarikov, Niki Stavroyianni, Teodora Karan-Djurasevic, Nikos Darzentas, Charles C. Chu, Eva Minga, Karla Plevová, Frederic Davi, Maria Chatzouli, Chrysoula Belessi, Livio Trentin, Paolo Ghia, Marie-Paule Lefranc, Davide Rossi, Xiao J. Yan, Šárka Pospíšilová, Diane F. Jelinek, Carsten Utoft Niemann, Jasmin Bahlo, Karin E. Smedby, Myriam Boudjogra, Mark Catherwood, Larry Mansouri, Darko Antic, Silvio Veronese, Lydia Scarfò, Andreas Agathangelidis, Marco Montillo, Zadie Davis, Anastasia Hadzidimitriou, Michael Hallek, Tait D. Shanafelt, Hartmut Döhner, Kostas Stamatopoulos, Achilles Anagnostopoulos, David Oscier, Nicholas Chiorazzi, Gianluca Gaidano, Eugen Tausch, Fie Juhl Vojdeman, Panagiotis Panagiotidis, Dirk Kienle, Aliki Xochelli, Lesley-Ann Sutton, and Anton W. Langerak
- Subjects
0303 health sciences ,medicine.medical_specialty ,Mutation ,Hematology ,biology ,Immunology ,Cell Biology ,CD38 ,medicine.disease_cause ,Biochemistry ,3. Good health ,03 medical and health sciences ,Stereotypy (non-human) ,0302 clinical medicine ,Antigen ,Immunoglobulin M ,Internal medicine ,biology.protein ,medicine ,Gene ,Antigen receptors ,030304 developmental biology ,030215 immunology - Abstract
The IGHV4-34 gene is very frequent (~10%) in the B cell receptor immunoglobulin (BcR IG) gene repertoire of chronic lymphocytic leukemia (CLL). Over 30% of IGHV4-34 CLL cases can be assigned to different subsets with stereotyped BcR IG. The largest is subset #4 which represents ~1% of all CLL and ~10% of IGHV4-34 CLL and is considered a prototype for indolent disease. The BcR IG of a great majority (~85%) of IGHV4-34 CLL cases carry a significant load of somatic hypermutation (SHM), often with distinctive SHM patterns. This holds especially true for stereotyped subsets and is suggestive of particular modes of interactions with the selecting antigen(s). In detail, subsets #4 and #16, both involving IgG-switched cases (IgG-CLL), exhibit the greatest sequence similarity in SHM profiles, whereas they differ in this respect from IgM/D subsets #29 and #201. Prompted by these observations, here we explored the extent that these subset-biased SHM profiles in different IGHV4-34 stereotyped subsets were reflected in distinct demographics, clinical presentation, genomic aberrations and outcomes. Within a multi-institutional series of 20,331 CLL patients, 1790 (8.8%) expressed IGHV4-34 BcR IG. Following established bioinformatics approaches for the identification of BcR IG stereotypy, 573/1790 IGHV4-34 CLL cases (32%) were assigned to stereotyped subsets; of these, 340 cases (19% of all IGHV4-34 CLL and 60% of stereotyped IGHV4-34 cases) belonged to subsets #4, #16, #29 and #201, all concerning IGHV-mutated CLL (M-CLL). Clinicobiological information was available for 275/340 patients: #4, n=150; #16, n=44; #29, n=39; and #201, n=42. Comparisons between subsets revealed no differences in gender and age distribution. Interestingly, however, 36-43% of each subset cases were young for CLL (defined as patients aged ≤55 years), which is higher compared to general CLL cohorts, where young patients generally account for ~25% of cases. In contrast, significant differences were identified between subsets regarding: (i) disease stage at diagnosis, with >90% of IgG subsets #4 and #16 diagnosed at Binet stage A versus 83% in subset #201 and 74% in subset #29 (p=0.029); (ii) CD38 expression, ranging from 1% in subset #4 to 10% in subset #201 (p=0.013); (iii) the distribution of del(13q), peaking at a remarkable 92% in subset #29 versus only 37% in subset #16 (p Disclosures Tausch: Gilead: Other: Travel support. Shanafelt:Glaxo-Smith_Kline: Research Funding; Genentech: Research Funding; Celgene: Research Funding; Polyphenon E Int'l: Research Funding; Hospira: Research Funding; Janssen: Research Funding; Pharmactckucs: Research Funding; Cephalon: Research Funding. Niemann:Gilead: Consultancy; Janssen: Consultancy; Roche: Consultancy; Novartis: Other: Travel grant. Langerak:InVivoScribe: Patents & Royalties: Licensing of IP and Patent on BIOMED-2-based methods for PCR-based Clonality Diagnostics.; DAKO: Patents & Royalties: Licensing of IP and Patent on Split-Signal FISH. Royalties for Dept. of Immunology, Erasmus MC, Rotterdam, NL; Roche: Other: Lab services in the field of MRD diagnostics provided by Dept of Immunology, Erasmus MC (Rotterdam). Hallek:Celgene: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; AbbVie: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; Roche: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; Boehringher Ingelheim: Honoraria, Other: Speakers Bureau and/or Advisory Boards; Pharmacyclics: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; Mundipharma: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; Janssen: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding; Gilead: Honoraria, Other: Speakers Bureau and/or Advisory Boards, Research Funding. Ghia:Janssen Pharmaceuticals: Research Funding.
- Published
- 2015
46. Mantle cell lymphoma displays a homogenous methylation profile: a comparative analysis with chronic lymphocytic leukemia
- Author
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Millaray Marincevic, Larry Mansouri, Prasoon Agarwal, Hanna Göransson, Tomas Axelsson, Helena Jernberg-Wiklund, Kostas Stamatopoulos, Birgitta Sander, Richard Rosenquist, Anna Margrét Halldórsdóttir, Anders Isaksson, Meena Kanduri, and Hans Ehrencrona
- Subjects
DNA (Cytosine-5-)-Methyltransferase 1 ,Male ,Chronic lymphocytic leukemia ,Lymphoma, Mantle-Cell ,Biology ,Decitabine ,Hydroxamic Acids ,immune system diseases ,hemic and lymphatic diseases ,Cell Line, Tumor ,medicine ,Humans ,DNA (Cytosine-5-)-Methyltransferases ,neoplasms ,Gene ,Genetics ,Genes, Immunoglobulin ,Gene Expression Regulation, Leukemic ,Genes, Homeobox ,Hematology ,Methylation ,DNA, Neoplasm ,Sequence Analysis, DNA ,DNA Methylation ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Neoplasm Proteins ,Gene Expression Regulation, Neoplastic ,Histone Deacetylase Inhibitors ,CpG site ,DNA methylation ,Cancer research ,Azacitidine ,Mantle cell lymphoma ,CpG Islands ,Female ,CD5 ,IGHV@ ,Apoptosis Regulatory Proteins ,Immunoglobulin Heavy Chains ,Cell Division ,Transcription Factors - Abstract
Mantle cell lymphoma (MCL) and chronic lymphocytic leukemia (CLL) are mature CD5(+) B-cell malignancies with different biological/clinical characteristics. We recently reported an association between different prognostic subgroups of CLL (i.e., IGHV mutated and unmutated) and genomic methylation pattern. However, the relationship between DNA methylation and prognostic markers, such as the proliferation gene expression signature, has not been investigated in MCL. We applied high-resolution methylation microarrays (27,578 CpG sites) to assess the global DNA methylation profiles in 20 MCL (10 each with high/low proliferation signature) and 30 CLL (15 poor-prognostic IGHV unmutated subset #1 and 15 good-prognostic IGHV mutated subset #4) samples. Notably, MCL and each CLL subset displayed distinct genomic methylation profiles. After unsupervised hierarchical clustering, 17/20 MCL cases formed a cluster separate from CLL, while CLL subsets #1 and #4 formed subclusters. Surprisingly, few differentially methylated genes (n = 6) were identified between high vs. low proliferation MCL. In contrast, distinct methylation profiles were demonstrated for MCL and CLL. Importantly, certain functional classes of genes were preferentially methylated in either disease. For instance, developmental genes, in particular homeobox transcription factor genes (e.g., HLXB9, HOXA13), were more highly methylated in MCL, whereas apoptosis-related genes were enriched among targets methylated in CLL (e.g., CYFIP2, NR4A1). Results were validated using pyrosequencing, RQ-PCR and reexpression of specific genes. In summary, the methylation profile of MCL was homogeneous and no correlation with the proliferation signature was observed. Compared to CLL, however, marked differences were discovered such as the preferential methylation of homeobox genes in MCL.
- Published
- 2011
47. Array-based genomic screening at diagnosis and during follow-up in chronic lymphocytic leukemia
- Author
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Karin E. Smedby, Mattias Jansson, Nicola Cahill, Henrik Hjalgrim, Karin Karlsson, Rebeqa Gunnarsson, Jeanette Lundin, Christian H. Geisler, Markus Rasmussen, Gunnar Juliusson, Hanna Göransson, Stefan Norin, Larry Mansouri, Anders Isaksson, Anne Mette Buhl, Jesper Jurlander, and Richard Rosenquist
- Subjects
Oncology ,Adult ,medicine.medical_specialty ,Pathology ,Adolescent ,DNA Copy Number Variations ,Chronic lymphocytic leukemia ,Population ,Loss of Heterozygosity ,Biology ,Somatic evolution in cancer ,Polymorphism, Single Nucleotide ,Loss of heterozygosity ,Young Adult ,Internal medicine ,medicine ,Humans ,education ,Aged ,Oligonucleotide Array Sequence Analysis ,Chromosome Aberrations ,education.field_of_study ,Chromosomes, Human, Pair 13 ,Genome, Human ,Hematology ,Original Articles ,Middle Aged ,medicine.disease ,Prognosis ,Leukemia, Lymphocytic, Chronic, B-Cell ,Survival Analysis ,Leukemia ,IGHV@ ,Trisomy ,SNP array ,Follow-Up Studies - Abstract
Background High-resolution genomic microarrays enable simultaneous detection of copy-number aberrations such as the known recurrent aberrations in chronic lymphocytic leukemia [del(11q), del(13q), del(17p) and trisomy 12], and copy-number neutral loss of heterozygosity. Moreover, comparison of genomic profiles from sequential patients' samples allows detection of clonal evolution. Design and Methods We screened samples from 369 patients with newly diagnosed chronic lymphocytic leukemia from a population-based cohort using 250K single nucleotide polymorphism-arrays. Clonal evolution was evaluated in 59 follow-up samples obtained after 5-9 years. Results At diagnosis, copy-number aberrations were identified in 90% of patients; 70% carried known recurrent alterations, including del(13q) (55%), trisomy 12 (10.5%), del(11q) (10%), and del(17p) (4%). Additional recurrent aberrations were detected on chromosomes 2 (1.9%), 4 (1.4%), 8 (1.6%) and 14 (1.6%). Thirteen patients (3.5%) displayed recurrent copy-number neutral loss of heterozygosity on 13q, of whom 11 had concurrent homozygous del(13q). Genomic complexity and large 13q deletions correlated with inferior outcome, while the former was linked to poor-prognostic aberrations. In the follow-up study, clonal evolution developed in 8/24 (33%) patients with unmutated IGHV, and in 4/25 (16%) IGHV-mutated and treated patients. In contrast, untreated patients with mutated IGHV (n=10) did not acquire additional aberrations. The most common secondary event, del(13q), was detected in 6/12 (50%) of all patients with acquired alterations. Interestingly, aberrations on, for example, chromosome 6q, 8p, 9p and 10q developed exclusively in patients with unmutated IGHV. Conclusions Whole-genome screening revealed a high frequency of genomic aberrations in newly diagnosed chronic lymphocytic leukemia. Clonal evolution was associated with other markers of aggressive disease and commonly included the known recurrent aberrations.
- Published
- 2011
48. Recurrent Mutations within the Nfkbie gene: A Novel Mechanism for NF-κB Deregulation in Aggressive Chronic Lymphocytic Leukemia
- Author
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Frederic Davi, Antonia Kalushkova, Chrysoula Belessi, Paolo Ghia, Mats Hellström, Karin E. Smedby, Anton W. Langerak, Mats Nilsson, Jonathan C. Strefford, Viktor Ljungström, Jimmy Larsson, Nicholas Chiorazzi, Sina Bondza, Larry Mansouri, Rebeqa Gunnarsson, Richard Rosenquist, Ola Söderberg, Šárka Pospíšilová, Xiao-Jie Yan, Emma Young, Elin Falk-Sörqvist, Linda Arngården, Birgitta Sander, Helena Jernberg Wiklund, Karla Plevová, Alice F. Muggen, Gunnar Juliusson, Kostas Stamatopoulos, Diego Cortese, Gunilla Enblad, Sujata Bhoi, and Lesley-Ann Sutton
- Subjects
medicine.medical_specialty ,Chronic lymphocytic leukemia ,Immunology ,medicine.disease_cause ,Biochemistry ,03 medical and health sciences ,chemistry.chemical_compound ,NFKBIE Gene ,0302 clinical medicine ,Antigen ,Internal medicine ,medicine ,030304 developmental biology ,0303 health sciences ,Mutation ,Hematology ,Mechanism (biology) ,business.industry ,NF-κB ,Cell Biology ,medicine.disease ,3. Good health ,chemistry ,Cancer research ,business ,Diffuse large B-cell lymphoma ,030215 immunology - Abstract
Dysregulated NF-κB signaling appears to be particularly important in B-cell malignancies, with recurrent mutations identified within both the canonical and non-canonical NF-κB pathways, as well as in components of the B-cell receptor (BcR) and Toll-like receptor (TLR) signaling pathways. In chronic lymphocytic leukemia (CLL), although recurrent mutations have been identified in MYD88 (TLR signaling) and BIRC3 (non-canonical NF-κB pathway), their frequency is low (10% for the mutant allele, we identified mutations (n=35) within 30/124 (24%) patients in 14/18 NF-κB genes analyzed. IκB genes, which encode for cytoplasmic inhibitor proteins, accounted for 20/35 (57%) mutations, with IκBε (encoded by NFKBIE) mutated in 8 patients; notably, 3/8 cases carried an identical 4bp deletion within exon 1 of NFKBIE. Prompted by these findings, we proceeded to validate our findings in an independent CLL cohort (n=168) using the same methodology as above and primarily focusing on cases with poor-prognostic features. We identified 30 mutations within 28 CLL patients in 11/18 NF-κB genes analyzed. Strikingly, 13/30 mutations were found within IκBε, with 10/13 patients carrying the same 4bp NFKBIE deletion. Notably, investigations into whether additional cases (within both the discovery and validation cohort) may harbor mutations of low clonal abundance ( Disclosures No relevant conflicts of interest to declare.
- Published
- 2014
49. Subset-Specific Spectra of Recurrent Gene Mutations in Chronic Lymphocytic Leukemia with Stereotyped B-Cell Receptors
- Author
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Evangelia Stalika, Richard Rosenquist, Frederic Davi, Lydia Scarfò, Andreas Agathangelidis, Christian H. Geisler, Jitka Malčíková, Anton W. Langerak, Kostas Stamatopoulos, Panagiotis Panagiotidis, Gianluca Gaidano, Karla Plevová, Alice F. Muggen, Lesley-Ann Sutton, Alba Navarro, Myriam Boudjogra, Šárka Pospíšilová, Xiao-Jie Yan, Zadie Davis, David Oscier, Nicholas Chiorazzi, Paolo Ghia, Anastasia Hadzidimitriou, Chrysoula Belessi, Panagiotis Baliakas, Larry Mansouri, Elias Campo, Lone Bredo Pedersen, Emma Young, Davide Rossi, and Jonathan C. Strefford
- Subjects
medicine.medical_specialty ,Hematology ,Chronic lymphocytic leukemia ,Immunology ,B-cell receptor ,Cell Biology ,Biology ,Gene mutation ,medicine.disease ,Biochemistry ,Internal medicine ,medicine ,Cancer research ,biology.protein ,Antibody ,Receptor - Abstract
Preliminary observations from essentially small patient series indicate that certain recurrent gene mutations may be enriched in subsets of chronic lymphocytic leukemia (CLL) with stereotyped B-cell receptors (BcR). On these grounds, it could be argued that differential modes of immune signaling, in the context of subset-biased antigen-immunoglobulin (IG) interactions, may be associated with the acquisition and/or selection of certain genomic aberrations within various stereotyped CLL subsets. With this in mind, we here sought to explore the genetic background of 10 major stereotyped subsets which collectively account for ~11% of all CLL and represent both IGHV unmutated (U-CLL) and/or mutated (M-CLL) cases. We focused on recurrent mutations within the NOTCH1 (entire exon 34 or targeted analysis for del7544-45), TP53 (exons 4-9), SF3B1 (exons 14-16), BIRC3 (exons 6-9) and MYD88 (exon 5) genes. Overall, 647 cases were analyzed, belonging to the following major subsets: (i) U-CLL: #1 (the largest within U-CLL, clinically aggressive), n=139; #3, n=39; #5, n=22; #6, n=48; #7, n=74; #8, n=46; #59, n=19 and #99, n=18; (ii) M-CLL: #4 (the largest within M-CLL, particularly indolent), n=78; and, (iii) subset #2 (the largest overall, variable mutational status and clinically aggressive), n=164. All cases were devoid of MYD88 mutations, which was not surprising given that our cohort was predominantly composed of U-CLL. Mutations within the BIRC3 gene were either absent (#2, #4, #6 and #59) or rare (#1, #3, #5, #7, #8 and #99; frequency 1.5%-7%) with no clear bias to any subset. BIRC3-mutant cases frequently co-existed with either del(11q) or trisomy 12. NOTCH1 mutations were more frequent in subsets #1, #6, #8, #59 and #99 (frequency, 22%-32%), sharply contrasting subsets #2 or #3 (4% and 7%, respectively) (p Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.
- Published
- 2014
50. 1.12 Transcriptome Sequencing Reveals Novel Mutations and Differential Gene Expression in Stereotyped Subsets of Chronic Lymphocytic Leukemia
- Author
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Rebeqa Gunnarsson, Markus Rasmussen, Richard Rosenquist, Adam Ameur, Lesley-Ann Sutton, Ulf Gyllensten, Sean D. Hooper, Anders Isaksson, and Larry Mansouri
- Subjects
Genetics ,Cancer Research ,Oncology ,business.industry ,Chronic lymphocytic leukemia ,Gene expression ,Medicine ,Hematology ,business ,medicine.disease ,Differential (mathematics) ,Transcriptome Sequencing - Published
- 2011
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