34 results on '"Klampfl, T."'
Search Results
2. P325: A KINASE-DEAD VARIANT OF CDK6 IS ASSOCIATED WITH REDUCED TUMORIGENIC POTENTIAL AND A DISTINCT TRANSCRIPTIONAL REWIRING IN AN ALL MODEL
- Author
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Klampfl, T., primary, Nebenführ, S., additional, Zojer, M., additional, Bellutti, F., additional, Sexl, V., additional, and Kollmann, K., additional
- Published
- 2022
- Full Text
- View/download PDF
3. P1407: CDK6 IN HEMATOPOIETIC STEM CELLS: MORE THAN A CELL CYCLE KINASE
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Mayer, I., primary, Doma, E., additional, Prchal-Murphy, M., additional, Klampfl, T., additional, Kollmann, K., additional, and Sexl, V., additional
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- 2022
- Full Text
- View/download PDF
4. Supplement to: Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med 2013;369:2379-90. DOI: 10.1056/NEJMoa1311347
- Author
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Klampfl, T, Gisslinger, H, and Harutyunyan, A S
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- 2013
5. Deletions of the transcription factor Ikaros in myeloproliferative neoplasms
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Jäger, R, Gisslinger, H, Passamonti, F, Rumi, E, Berg, T, Gisslinger, B, Pietra, D, Harutyunyan, A, Klampfl, T, Olcaydu, D, Cazzola, M, and Kralovics, R
- Published
- 2010
- Full Text
- View/download PDF
6. Rare germline variants in regions of loss of heterozygosity may influence clinical course of hematological malignancies
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Harutyunyan, A, Gisslinger, B, Klampfl, T, Berg, T, Bagienski, K, Gisslinger, H, and Kralovics, R
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- 2011
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- View/download PDF
7. Up-regulation of 12(S)-lipoxygenase induces a migratory phenotype in colorectal cancer cells
- Author
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Klampfl, T., Bogner, E., Bednar, W., Mager, L., Massudom, D., Kalny, I., Heinzle, C., Berger, W., Stättner, S., Karner, J., Klimpfinger, M., Fürstenberger, G., Krieg, P., and Marian, B.
- Published
- 2012
- Full Text
- View/download PDF
8. Germline RBBP6 mutations in familial myeloproliferative neoplasms
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Harutyunyan, A. S., Giambruno, R., Krendl, C., Stukalov, A., Klampfl, T., Berg, T., Chen, D., Feenstra, J. D. M., Jäger, R., Gisslinger, B., Gisslinger, H., Rumi, E., Passamonti, Francesco, Pietra, D., Müller, A. C., Parapatics, K., Breitwieser, F. P., Herrmann, R., Colinge, J., Bennett, K. L., Superti Furga, G., Cazzola, M., Hammond, E., Kralovics, R., Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), and CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)
- Subjects
Male ,Myeloproliferative Disorders ,MESH: Humans ,MESH: Pedigree ,Ubiquitin-Protein Ligases ,DNA Mutational Analysis ,Letters to Blood ,MESH: Carrier Proteins ,MESH: Male ,Pedigree ,DNA-Binding Proteins ,MESH: Germ-Line Mutation ,Humans ,Family ,Female ,MESH: DNA Mutational Analysis ,Carrier Proteins ,MESH: Family ,MESH: Female ,Germ-Line Mutation ,ComputingMilieux_MISCELLANEOUS ,MESH: DNA-Binding Proteins ,[SDV.MHEP]Life Sciences [q-bio]/Human health and pathology ,MESH: Myeloproliferative Disorders - Abstract
International audience
- Published
- 2016
9. Germline RBBP6 mutations in familial myeloproliferative neoplasm
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Harutyunyan, A.S., Giambruno, R., Krendl, C., Stukalov, A., Klampfl, T., Berg, T., Chen, D., Milosevic Feenstra, J.D., Jager, R., Gisslinger, B., Gisslinger, H., Rumi, E., Passamonti, F., Pietra, D., Muller, A.C., Parapatics, K., Breitwieser, F.P., Herrmann, R., Colinge, J., Bennett, K.L., Superti-Furga, G., Cazzola, M., Hammond, E., Kralovics, R., Harutyunyan, A.S., Giambruno, R., Krendl, C., Stukalov, A., Klampfl, T., Berg, T., Chen, D., Milosevic Feenstra, J.D., Jager, R., Gisslinger, B., Gisslinger, H., Rumi, E., Passamonti, F., Pietra, D., Muller, A.C., Parapatics, K., Breitwieser, F.P., Herrmann, R., Colinge, J., Bennett, K.L., Superti-Furga, G., Cazzola, M., Hammond, E., and Kralovics, R.
- Abstract
Letter to the Editor
- Published
- 2016
10. High resolution cytogenetic mapping and whole exome sequencing reveal a complex pattern of chromosome 6p aberrations in patients with myeloid malignancies
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Pudja, A., Milosević, J. D., Klampfl, T., Harutyunyan, A., Berg, T., Bagienski, K., Chen, D., Gisslinger, B., Rumi, E., Malcovati, L., Pietra, Daniela, Elena, C., Della Porta, M. G., Pieri, Lisa, Guglielmelli, Paola, Doubek, M., Dvorakova, D., Suvajdžić, Nada, Tomin, D., Tošić, Nataša, Racil, Z., Steurer, M., Pavlović, Sonja, Vannucchi, A. M., Cazzola, M., Gisslinger, H., Kralovics, R., Pudja, A., Milosević, J. D., Klampfl, T., Harutyunyan, A., Berg, T., Bagienski, K., Chen, D., Gisslinger, B., Rumi, E., Malcovati, L., Pietra, Daniela, Elena, C., Della Porta, M. G., Pieri, Lisa, Guglielmelli, Paola, Doubek, M., Dvorakova, D., Suvajdžić, Nada, Tomin, D., Tošić, Nataša, Racil, Z., Steurer, M., Pavlović, Sonja, Vannucchi, A. M., Cazzola, M., Gisslinger, H., and Kralovics, R.
- Published
- 2015
11. Prognostic significance of genetic aberrations in secondary acute myeloid leukemia
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Milosević, J., Puda, Ana, Berg, T., Klampfl, T., Harutyunyan, A., Hofbauer, M., Stukalov, A., Gisslinger, H., Gisslinger, B., Burjanivova, T., Rumi, E., Pietra, Daniela, Malcovati, L., Elena, C., Cazzola, M., Vannucchi, A., Doubek, M., Penka, M., Racil, Z., Dvorakova, D., Wieser, R., Koller, E., Steurer, M., Tošić, Nataša, Pavlović, Sonja, Kralovics, R., Milosević, J., Puda, Ana, Berg, T., Klampfl, T., Harutyunyan, A., Hofbauer, M., Stukalov, A., Gisslinger, H., Gisslinger, B., Burjanivova, T., Rumi, E., Pietra, Daniela, Malcovati, L., Elena, C., Cazzola, M., Vannucchi, A., Doubek, M., Penka, M., Racil, Z., Dvorakova, D., Wieser, R., Koller, E., Steurer, M., Tošić, Nataša, Pavlović, Sonja, and Kralovics, R.
- Published
- 2012
12. An RNA-Seq Strategy to Detect the Complete Coding and Non-Coding Transcriptome Including Full-Length Imprinted Macro ncRNAs
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Huang, R., Jaritz, M., Guenzl, P., Vlatkovic, I., Sommer, A., Tamir, I.M., Marks, H., Klampfl, T., Kralovics, R., Stunnenberg, H.G., Barlow, D. P., Pauler, F. M., Huang, R., Jaritz, M., Guenzl, P., Vlatkovic, I., Sommer, A., Tamir, I.M., Marks, H., Klampfl, T., Kralovics, R., Stunnenberg, H.G., Barlow, D. P., and Pauler, F. M.
- Abstract
Contains fulltext : 91551.pdf (publisher's version ) (Open Access)
- Published
- 2011
13. Plasma decontamination of space equipment using cold atmospheric plasmas
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Thomas, H. M., primary, Shimizu, S., additional, Shimizu, T., additional, Klampfl, T., additional, Zimmermann, J. L., additional, Morfill, G. E., additional, Barczyk, S., additional, Rettberg, P., additional, and Weber, P. K., additional
- Published
- 2012
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14. Genome integrity of myeloproliferative neoplasms in chronic phase and during disease progression
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Ilaria Iacobucci, Ashot S. Harutyunyan, Roland Jäger, Lisa Pieri, Tiina Berg, Paola Guglielmelli, Alessandro M. Vannucchi, Robert Kralovics, Klaudia Bagienski, Elisa Rumi, Daniela Pietra, Bettina Gisslinger, Giovanni Martinelli, Francesco Passamonti, Martin Schalling, Thorsten Klampfl, Heinz Gisslinger, Damla Olcaydu, Mario Cazzola, Klampfl T, Harutyunyan A, Berg T, Gisslinger B, Schalling M, Bagienski K, Olcaydu D, Passamonti F, Rumi E, Pietra D, Jäger R, Pieri L, Guglielmelli P, Iacobucci I, Martinelli G, Cazzola M, Vannucchi AM, Gisslinger H, and Kralovics R.
- Subjects
Myeloid ,Immunology ,Myeloproliferative neoplasm ,Polymorphism, Single Nucleotide ,Biochemistry ,Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative ,medicine ,Humans ,Point Mutation ,acute leukemia ,genome ,mutation ,Oligonucleotide Array Sequence Analysis ,Acute leukemia ,Myeloproliferative Disorders ,Janus kinase 2 ,biology ,Gene Expression Regulation, Leukemic ,Myeloid leukemia ,Karyotype ,Cell Biology ,Hematology ,Janus Kinase 2 ,medicine.disease ,ETV6 ,Leukemia ,medicine.anatomical_structure ,Karyotyping ,Chronic Disease ,Disease Progression ,Cancer research ,biology.protein ,CHRONIC PHASE ,Genome-Wide Association Study - Abstract
Philadelphia chromosome–negative myeloproliferative neoplasms (MPNs) are clonal myeloid disorders with increased production of terminally differentiated cells. The disease course is generally chronic, but some patients show disease progression (secondary myelofibrosis or accelerated phase) and/or leukemic transformation. We investigated chromosomal aberrations in 408 MPN samples using high-resolution single-nucleotide polymorphism microarrays to identify disease-associated somatic lesions. Of 408 samples, 37.5% had a wild-type karyotype and 62.5% harbored at least 1 chromosomal aberration. We identified 25 recurrent aberrations that were found in 3 or more samples. An increased number of chromosomal lesions was significantly associated with patient age, as well as with disease progression and leukemic transformation, but no association was observed with MPN subtypes, Janus kinase 2 (JAK2) mutational status, or disease duration. Aberrations of chromosomes 1q and 9p were positively associated with disease progression to secondary myelofibrosis or accelerated phase. Changes of chromosomes 1q, 7q, 5q, 6p, 7p, 19q, 22q, and 3q were positively associated with post-MPN acute myeloid leukemia. We mapped commonly affected regions to single target genes on chromosomes 3p (forkhead box P1 [FOXP1]), 4q (tet oncogene family member 2 [TET2]), 7p (IKAROS family zinc finger 1 [IKZF1]), 7q (cut-like homeobox 1 [CUX1]), 12p (ets variant 6 [ETV6]), and 21q (runt-related transcription factor 1 [RUNX1]). Our data provide insight into the genetic complexity of MPNs and implicate new genes involved in disease progression.
- Published
- 2011
15. Cyclin C promotes development and progression of B-cell acute lymphoblastic leukemia by counteracting p53-mediated stress responses.
- Author
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Trifinopoulos J, List J, Klampfl T, Klein K, Prchal-Murphy M, Witalisz-Siepracka A, Bellutti F, Fava LL, Heller G, Stummer S, Testori P, Den Boer ML, Boer JM, Marinovic S, Hoermann G, Walter W, Villunger A, Sicinski P, Sexl V, and Gotthardt D
- Abstract
Despite major therapeutic advances in the treatment of acute lymphoblastic leukemia (ALL), resistances and long-term toxicities still pose significant challenges. Cyclins and their associated cyclin-dependent kinases are one focus of cancer research when looking for targeted therapies. We discovered cyclin C as a key factor for B-ALL development and maintenance. While cyclin C is non-essential for normal hematopoiesis, CcncΔ/Δ BCR::ABL1+ B-ALL cells fail to elicit leukemia in mice. RNA sequencing experiments revealed a p53 pathway deregulation in CcncΔ/Δ BCR::ABL1+ cells resulting in the incapability of the leukemic cells to adequately respond to stress. A genome-wide CRISPR/Cas9 loss-of-function screen supplemented with additional knock-outs unveiled a dependency of human B-lymphoid cell lines on CCNC. High cyclin C levels in B-cell precursor (BCP) ALL patients were associated with poor event-free survival and increased risk of early disease recurrence after remission. Our findings highlight cyclin C as potential therapeutic target for B-ALL, particularly to enhance cancer cell sensitivity to stress and chemotherapy.
- Published
- 2024
- Full Text
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16. Kinase-inactivated CDK6 preserves the long-term functionality of adult hematopoietic stem cells.
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Mayer IM, Doma E, Klampfl T, Prchal-Murphy M, Kollmann S, Schirripa A, Scheiblecker L, Zojer M, Kunowska N, Gebrail L, Shaw LE, Mann U, Farr A, Grausenburger R, Heller G, Zebedin-Brandl E, Farlik M, Malumbres M, Sexl V, and Kollmann K
- Subjects
- Animals, Mice, Humans, Adult Stem Cells metabolism, Adult Stem Cells cytology, Cell Proliferation, Cell Differentiation, Mice, Inbred C57BL, Hematopoietic Stem Cell Transplantation, Cell Self Renewal drug effects, Cyclin-Dependent Kinase 6 metabolism, Cyclin-Dependent Kinase 6 genetics, Hematopoietic Stem Cells metabolism, Hematopoietic Stem Cells cytology
- Abstract
Abstract: Hematopoietic stem cells (HSCs) are characterized by the ability to self-renew and to replenish the hematopoietic system. The cell-cycle kinase cyclin-dependent kinase 6 (CDK6) regulates transcription, whereby it has both kinase-dependent and kinase-independent functions. Herein, we describe the complex role of CDK6, balancing quiescence, proliferation, self-renewal, and differentiation in activated HSCs. Mouse HSCs expressing kinase-inactivated CDK6 show enhanced long-term repopulation and homing, whereas HSCs lacking CDK6 have impaired functionality. The transcriptomes of basal and serially transplanted HSCs expressing kinase-inactivated CDK6 exhibit an expression pattern dominated by HSC quiescence and self-renewal, supporting a concept, in which myc-associated zinc finger protein (MAZ) and nuclear transcription factor Y subunit alpha (NFY-A) are critical CDK6 interactors. Pharmacologic kinase inhibition with a clinically used CDK4/6 inhibitor in murine and human HSCs validated our findings and resulted in increased repopulation capability and enhanced stemness. Our findings highlight a kinase-independent role of CDK6 in long-term HSC functionality. CDK6 kinase inhibition represents a possible strategy to improve HSC fitness., (© 2024 American Society of Hematology. Published by Elsevier Inc. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
17. A lineage-specific STAT5BN642H mouse model to study NK-cell leukemia.
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Klein K, Kollmann S, Hiesinger A, List J, Kendler J, Klampfl T, Rhandawa M, Trifinopoulos J, Maurer B, Grausenburger R, Betram CA, Moriggl R, Rülicke T, Mullighan CG, Witalisz-Siepracka A, Walter W, Hoermann G, Sexl V, and Gotthardt D
- Subjects
- Animals, Mice, Humans, Disease Models, Animal, Cell Lineage genetics, Mutation, Mice, Transgenic, STAT5 Transcription Factor genetics, STAT5 Transcription Factor metabolism, Killer Cells, Natural metabolism, Killer Cells, Natural immunology, Killer Cells, Natural pathology, Leukemia, Large Granular Lymphocytic genetics, Leukemia, Large Granular Lymphocytic pathology
- Abstract
Abstract: Patients with T- and natural killer (NK)-cell neoplasms frequently have somatic STAT5B gain-of-function mutations. The most frequent STAT5B mutation is STAT5BN642H, which is known to drive murine T-cell leukemia, although its role in NK-cell malignancies is unclear. Introduction of the STAT5BN642H mutation into human NK-cell lines enhances their potential to induce leukemia in mice. We have generated a mouse model that enables tissue-specific expression of STAT5BN642H and have selectively expressed the mutated STAT5B in hematopoietic cells (N642Hvav/+) or exclusively in NK cells (N642HNK/NK). All N642Hvav/+ mice rapidly develop an aggressive T/NKT-cell leukemia, whereas N642HNK/NK mice display an indolent NK-large granular lymphocytic leukemia (NK-LGLL) that progresses to an aggressive leukemia with age. Samples from patients with NK-cell leukemia have a distinctive transcriptional signature driven by mutant STAT5B, which overlaps with that of murine leukemic N642HNK/NK NK cells. To our knowledge, we have generated the first reliable STAT5BN642H-driven preclinical mouse model that displays an indolent NK-LGLL progressing to aggressive NK-cell leukemia. This novel in vivo tool will enable us to explore the transition from an indolent to an aggressive disease and will thus permit the study of prevention and treatment options for NK-cell malignancies., (© 2024 American Society of Hematology. Published by Elsevier Inc. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
18. SBNO2 is a critical mediator of STAT3-driven hematological malignancies.
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Brandstoetter T, Schmoellerl J, Grausenburger R, Kollmann S, Doma E, Huuhtanen J, Klampfl T, Eder T, Grebien F, Hoermann G, Zuber J, Mustjoki S, Maurer B, and Sexl V
- Subjects
- Animals, Humans, Mice, Anaplastic Lymphoma Kinase metabolism, Cell Line, Tumor, Lymphoma, Large-Cell, Anaplastic genetics, Hematologic Neoplasms genetics, STAT3 Transcription Factor genetics, STAT3 Transcription Factor metabolism
- Abstract
Gain-of-function mutations in the signal transducer and activator of transcription 3 (STAT3) gene are recurrently identified in patients with large granular lymphocytic leukemia (LGLL) and in some cases of natural killer (NK)/T-cell and adult T-cell leukemia/lymphoma. To understand the consequences and molecular mechanisms contributing to disease development and oncogenic transformation, we developed murine hematopoietic stem and progenitor cell models that express mutated STAT3Y640F. These cells show accelerated proliferation and enhanced self-renewal potential. We integrated gene expression analyses and chromatin occupancy profiling of STAT3Y640F-transformed cells with data from patients with T-LGLL. This approach uncovered a conserved set of direct transcriptional targets of STAT3Y640F. Among these, strawberry notch homolog 2 (SBNO2) represents an essential transcriptional target, which was identified by a comparative genome-wide CRISPR/Cas9-based loss-of-function screen. The STAT3-SBNO2 axis is also present in NK-cell leukemia, T-cell non-Hodgkin lymphoma, and NPM-ALK-rearranged T-cell anaplastic large cell lymphoma (T-ALCL), which are driven by STAT3-hyperactivation/mutation. In patients with NPM-ALK+ T-ALCL, high SBNO2 expression correlates with shorter relapse-free and overall survival. Our findings identify SBNO2 as a potential therapeutic intervention site for STAT3-driven hematopoietic malignancies., (© 2023 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2023
- Full Text
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19. CDK6 Degradation Is Counteracted by p16 INK4A and p18 INK4C in AML.
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Schmalzbauer BS, Thondanpallil T, Heller G, Schirripa A, Sperl CM, Mayer IM, Knab VM, Nebenfuehr S, Zojer M, Mueller AC, Fontaine F, Klampfl T, Sexl V, and Kollmann K
- Abstract
Cyclin-dependent kinase 6 (CDK6) represents a novel therapeutic target for the treatment of certain subtypes of acute myeloid leukaemia (AML). CDK4/6 kinase inhibitors have been widely studied in many cancer types and their effects may be limited by primary and secondary resistance mechanisms. CDK4/6 degraders, which eliminate kinase-dependent and kinase-independent effects, have been suggested as an alternative therapeutic option. We show that the efficacy of the CDK6-specific protein degrader BSJ-03-123 varies among AML subtypes and depends on the low expression of the INK4 proteins p16
INK4A and p18INK4C . INK4 protein levels are significantly elevated in KMT2A-MLLT3+ cells compared to RUNX1-RUNX1T1+ cells, contributing to the different CDK6 degradation efficacy. We demonstrate that CDK6 complexes containing p16INK4A or p18INK4C are protected from BSJ-mediated degradation and that INK4 levels define the proliferative response to CDK6 degradation. These findings define INK4 proteins as predictive markers for CDK6 degradation-targeted therapies in AML.- Published
- 2022
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20. A STAT5B-CD9 axis determines self-renewal in hematopoietic and leukemic stem cells.
- Author
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Kollmann S, Grausenburger R, Klampfl T, Prchal-Murphy M, Bastl K, Pisa H, Knab VM, Brandstoetter T, Doma E, Sperr WR, Lagger S, Farlik M, Moriggl R, Valent P, Halbritter F, Kollmann K, Heller G, Maurer B, and Sexl V
- Subjects
- Animals, Cell Self Renewal, Hematopoiesis, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells metabolism, Humans, Leukemia metabolism, Mice, Mice, Inbred C57BL, Neoplastic Stem Cells cytology, Neoplastic Stem Cells metabolism, Tumor Cells, Cultured, Hematopoietic Stem Cells pathology, Leukemia pathology, Neoplastic Stem Cells pathology, STAT5 Transcription Factor metabolism, Signal Transduction, Tetraspanin 29 metabolism
- Abstract
The transcription factors signal transducer and activator of transcription 5A (STAT5A) and STAT5B are critical in hematopoiesis and leukemia. They are widely believed to have redundant functions, but we describe a unique role for STAT5B in driving the self-renewal of hematopoietic and leukemic stem cells (HSCs/LSCs). We find STAT5B to be specifically activated in HSCs and LSCs, where it induces many genes associated with quiescence and self-renewal, including the surface marker CD9. Levels of CD9 represent a prognostic marker for patients with STAT5-driven leukemia, and our findings suggest that anti-CD9 antibodies may be useful in their treatment to target and eliminate LSCs. We show that it is vital to consider STAT5A and STAT5B as distinct entities in normal and malignant hematopoiesis., (© 2021 by The American Society of Hematology.)
- Published
- 2021
- Full Text
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21. Mutant Calreticulin in the Myeloproliferative Neoplasms.
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Prins D, González Arias C, Klampfl T, Grinfeld J, and Green AR
- Abstract
Mutations in the gene for calreticulin ( CALR ) were identified in the myeloproliferative neoplasms (MPNs) essential thrombocythaemia (ET) and primary myelofibrosis (MF) in 2013; in combination with previously described mutations in JAK2 and MPL , driver mutations have now been described for the majority of MPN patients. In subsequent years, researchers have begun to unravel the mechanisms by which mutant CALR drives transformation and to understand their clinical implications. Mutant CALR activates the thrombopoietin receptor (MPL), causing constitutive activation of Janus kinase 2 (JAK2) signaling and cytokine independent growth in vitro. Mouse models show increased numbers of hematopoietic stem cells (HSCs) and overproduction of megakaryocytic lineage cells with associated thrombocytosis. In the clinic, detection of CALR mutations has been embedded in World Health Organization and other international diagnostic guidelines. Distinct clinical and laboratory associations of CALR mutations have been identified together with their prognostic significance, with CALR mutant patients showing increased overall survival. The discovery and subsequent study of CALR mutations have illuminated novel aspects of megakaryopoiesis and raised the possibility of new therapeutic approaches., Competing Interests: The authors have indicated they have no potential conflicts of interest to disclose., (Copyright © 2020 the Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the European Hematology Association.)
- Published
- 2020
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22. Mutant calreticulin knockin mice develop thrombocytosis and myelofibrosis without a stem cell self-renewal advantage.
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Li J, Prins D, Park HJ, Grinfeld J, Gonzalez-Arias C, Loughran S, Dovey OM, Klampfl T, Bennett C, Hamilton TL, Pask DC, Sneade R, Williams M, Aungier J, Ghevaert C, Vassiliou GS, Kent DG, and Green AR
- Subjects
- Animals, Cells, Cultured, Homozygote, Leukocytosis genetics, Leukocytosis pathology, Mice, Mice, Inbred C57BL, Mice, Transgenic, Mutation, Missense, Splenomegaly genetics, Splenomegaly pathology, Thrombocythemia, Essential genetics, Thrombocythemia, Essential pathology, Calreticulin genetics, Cell Self Renewal genetics, Hematopoietic Stem Cells physiology, Primary Myelofibrosis genetics, Thrombocytosis genetics
- Abstract
Somatic mutations in the endoplasmic reticulum chaperone calreticulin (CALR) are detected in approximately 40% of patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF). Multiple different mutations have been reported, but all result in a +1-bp frameshift and generate a novel protein C terminus. In this study, we generated a conditional mouse knockin model of the most common CALR mutation, a 52-bp deletion. The mutant novel human C-terminal sequence is integrated into the otherwise intact mouse CALR gene and results in mutant CALR expression under the control of the endogenous mouse locus. CALR
del/+ mice develop a transplantable ET-like disease with marked thrombocytosis, which is associated with increased and morphologically abnormal megakaryocytes and increased numbers of phenotypically defined hematopoietic stem cells (HSCs). Homozygous CALRdel/del mice developed extreme thrombocytosis accompanied by features of MF, including leukocytosis, reduced hematocrit, splenomegaly, and increased bone marrow reticulin. CALRdel/+ HSCs were more proliferative in vitro, but neither CALRdel/+ nor CALRdel/del displayed a competitive transplantation advantage in primary or secondary recipient mice. These results demonstrate the consequences of heterozygous and homozygous CALR mutations and provide a powerful model for dissecting the pathogenesis of CALR-mutant ET and PMF., (© 2018 by The American Society of Hematology.)- Published
- 2018
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23. Determination of complex subclonal structures of hematological malignancies by multiplexed genotyping of blood progenitor colonies.
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Nice FL, Massie CE, Klampfl T, and Green AR
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- Alleles, Clone Cells, Cluster Analysis, Granulocytes chemistry, Hematologic Neoplasms pathology, Humans, Janus Kinase 2 genetics, Mutation, Missense, Polycythemia Vera pathology, Polymorphism, Single Nucleotide, DNA Mutational Analysis methods, DNA, Neoplasm genetics, Erythroid Precursor Cells chemistry, Genotyping Techniques methods, Hematologic Neoplasms genetics, High-Throughput Nucleotide Sequencing methods, Mutation, Polycythemia Vera genetics, Sequence Analysis, DNA methods
- Abstract
Current next-generation sequencing (NGS) technologies allow unprecedented insights into the mutational profiles of tumors. Recent studies in myeloproliferative neoplasms have further demonstrated that, not only the mutational profile, but also the order in which these mutations are acquired is relevant for our understanding of the disease. Our ability to assign mutation order from NGS data alone is, however, limited. Here, we present a strategy of highly multiplexed genotyping of burst forming unit-erythroid colonies based on NGS results to assess subclonal tumor structure. This allowed for the generation of complex clonal hierarchies and determination of order of mutation acquisition far more accurately than was possible from NGS data alone., (Copyright © 2018 ISEH – Society for Hematology and Stem Cells. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
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24. Germline RBBP6 mutations in familial myeloproliferative neoplasms.
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Harutyunyan AS, Giambruno R, Krendl C, Stukalov A, Klampfl T, Berg T, Chen D, Milosevic Feenstra JD, Jäger R, Gisslinger B, Gisslinger H, Rumi E, Passamonti F, Pietra D, Müller AC, Parapatics K, Breitwieser FP, Herrmann R, Colinge J, Bennett KL, Superti-Furga G, Cazzola M, Hammond E, and Kralovics R
- Subjects
- DNA Mutational Analysis, Family, Female, Humans, Male, Myeloproliferative Disorders diagnosis, Pedigree, Ubiquitin-Protein Ligases, Carrier Proteins genetics, DNA-Binding Proteins genetics, Germ-Line Mutation, Myeloproliferative Disorders genetics
- Published
- 2016
- Full Text
- View/download PDF
25. The solute carrier SLC35F2 enables YM155-mediated DNA damage toxicity.
- Author
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Winter GE, Radic B, Mayor-Ruiz C, Blomen VA, Trefzer C, Kandasamy RK, Huber KVM, Gridling M, Chen D, Klampfl T, Kralovics R, Kubicek S, Fernandez-Capetillo O, Brummelkamp TR, and Superti-Furga G
- Subjects
- Animals, Apoptosis drug effects, Cell Division drug effects, Cell Division physiology, Cell Line, Tumor, Cell Survival, Cloning, Molecular, Comet Assay, Genome, Human drug effects, Genome, Human genetics, Haploidy, Humans, Imidazoles metabolism, Immunohistochemistry, Mice, Mice, SCID, Naphthoquinones metabolism, RNA, Neoplasm chemistry, RNA, Neoplasm genetics, DNA Damage drug effects, Imidazoles pharmacology, Intercalating Agents pharmacology, Membrane Transport Proteins genetics, Membrane Transport Proteins metabolism, Naphthoquinones pharmacology
- Abstract
Genotoxic chemotherapy is the most common cancer treatment strategy. However, its untargeted generic DNA-damaging nature and associated systemic cytotoxicity greatly limit its therapeutic applications. Here, we used a haploid genetic screen in human cells to discover an absolute dependency of the clinically evaluated anticancer compound YM155 on solute carrier family member 35 F2 (SLC35F2), an uncharacterized member of the solute carrier protein family that is highly expressed in a variety of human cancers. YM155 generated DNA damage through intercalation, which was contingent on the expression of SLC35F2 and its drug-importing activity. SLC35F2 expression and YM155 sensitivity correlated across a panel of cancer cell lines, and targeted genome editing verified SLC35F2 as the main determinant of YM155-mediated DNA damage toxicity in vitro and in vivo. These findings suggest a new route to targeted DNA damage by exploiting tumor and patient-specific import of YM155.
- Published
- 2014
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26. JAK2 or CALR mutation status defines subtypes of essential thrombocythemia with substantially different clinical course and outcomes.
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Rumi E, Pietra D, Ferretti V, Klampfl T, Harutyunyan AS, Milosevic JD, Them NC, Berg T, Elena C, Casetti IC, Milanesi C, Sant'antonio E, Bellini M, Fugazza E, Renna MC, Boveri E, Astori C, Pascutto C, Kralovics R, and Cazzola M
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Alleles, Cell Transformation, Neoplastic genetics, Codon, Exons, Female, Granulocytes, Humans, Male, Middle Aged, Myeloproliferative Disorders genetics, Polycythemia Vera genetics, Primary Myelofibrosis genetics, Prognosis, Receptors, Thrombopoietin genetics, Thrombocythemia, Essential mortality, Thrombosis genetics, Young Adult, Calreticulin genetics, Janus Kinase 2 genetics, Mutation, Thrombocythemia, Essential diagnosis, Thrombocythemia, Essential genetics
- Abstract
Patients with essential thrombocythemia may carry JAK2 (V617F), an MPL substitution, or a calreticulin gene (CALR) mutation. We studied biologic and clinical features of essential thrombocythemia according to JAK2 or CALR mutation status and in relation to those of polycythemia vera. The mutant allele burden was lower in JAK2-mutated than in CALR-mutated essential thrombocythemia. Patients with JAK2 (V617F) were older, had a higher hemoglobin level and white blood cell count, and lower platelet count and serum erythropoietin than those with CALR mutation. Hematologic parameters of patients with JAK2-mutated essential thrombocythemia or polycythemia vera were related to the mutant allele burden. While no polycythemic transformation was observed in CALR-mutated patients, the cumulative risk was 29% at 15 years in those with JAK2-mutated essential thrombocythemia. There was no significant difference in myelofibrotic transformation between the 2 subtypes of essential thrombocythemia. Patients with JAK2-mutated essential thrombocythemia and those with polycythemia vera had a similar risk of thrombosis, which was twice that of patients with the CALR mutation. These observations are consistent with the notion that JAK2-mutated essential thrombocythemia and polycythemia vera represent different phenotypes of a single myeloproliferative neoplasm, whereas CALR-mutated essential thrombocythemia is a distinct disease entity.
- Published
- 2014
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27. Somatic mutations of calreticulin in myeloproliferative neoplasms.
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Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, Them NC, Berg T, Gisslinger B, Pietra D, Chen D, Vladimer GI, Bagienski K, Milanesi C, Casetti IC, Sant'Antonio E, Ferretti V, Elena C, Schischlik F, Cleary C, Six M, Schalling M, Schönegger A, Bock C, Malcovati L, Pascutto C, Superti-Furga G, Cazzola M, and Kralovics R
- Subjects
- Bone Marrow Diseases genetics, Exons, Humans, Janus Kinase 2 genetics, Leukemia, Myeloid genetics, Polymerase Chain Reaction, Primary Myelofibrosis mortality, Proportional Hazards Models, Receptors, Thrombopoietin genetics, Sequence Analysis, DNA, Thrombocythemia, Essential complications, Thrombocythemia, Essential mortality, Thrombosis etiology, Calreticulin genetics, Mutation, Primary Myelofibrosis genetics, Thrombocythemia, Essential genetics
- Abstract
Background: Approximately 50 to 60% of patients with essential thrombocythemia or primary myelofibrosis carry a mutation in the Janus kinase 2 gene (JAK2), and an additional 5 to 10% have activating mutations in the thrombopoietin receptor gene (MPL). So far, no specific molecular marker has been identified in the remaining 30 to 45% of patients., Methods: We performed whole-exome sequencing to identify somatically acquired mutations in six patients who had primary myelofibrosis without mutations in JAK2 or MPL. Resequencing of CALR, encoding calreticulin, was then performed in cohorts of patients with myeloid neoplasms., Results: Somatic insertions or deletions in exon 9 of CALR were detected in all patients who underwent whole-exome sequencing. Resequencing in 1107 samples from patients with myeloproliferative neoplasms showed that CALR mutations were absent in polycythemia vera. In essential thrombocythemia and primary myelofibrosis, CALR mutations and JAK2 and MPL mutations were mutually exclusive. Among patients with essential thrombocythemia or primary myelofibrosis with nonmutated JAK2 or MPL, CALR mutations were detected in 67% of those with essential thrombocythemia and 88% of those with primary myelofibrosis. A total of 36 types of insertions or deletions were identified that all cause a frameshift to the same alternative reading frame and generate a novel C-terminal peptide in the mutant calreticulin. Overexpression of the most frequent CALR deletion caused cytokine-independent growth in vitro owing to the activation of signal transducer and activator of transcription 5 (STAT5) by means of an unknown mechanism. Patients with mutated CALR had a lower risk of thrombosis and longer overall survival than patients with mutated JAK2., Conclusions: Most patients with essential thrombocythemia or primary myelofibrosis that was not associated with a JAK2 or MPL alteration carried a somatic mutation in CALR. The clinical course in these patients was more indolent than that in patients with the JAK2 V617F mutation. (Funded by the MPN Research Foundation and Associazione Italiana per la Ricerca sul Cancro.).
- Published
- 2013
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28. Complex patterns of chromosome 11 aberrations in myeloid malignancies target CBL, MLL, DDB1 and LMO2.
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Klampfl T, Milosevic JD, Puda A, Schönegger A, Bagienski K, Berg T, Harutyunyan AS, Gisslinger B, Rumi E, Malcovati L, Pietra D, Elena C, Della Porta MG, Pieri L, Guglielmelli P, Bock C, Doubek M, Dvorakova D, Suvajdzic N, Tomin D, Tosic N, Racil Z, Steurer M, Pavlovic S, Vannucchi AM, Cazzola M, Gisslinger H, and Kralovics R
- Subjects
- Exome genetics, Histone-Lysine N-Methyltransferase, Humans, Leukemia, Myeloid, Acute genetics, Myelodysplastic Syndromes genetics, Myeloid-Lymphoid Leukemia Protein genetics, Polycythemia Vera genetics, Polymerase Chain Reaction, Primary Myelofibrosis genetics, Proto-Oncogene Proteins c-cbl genetics, Thrombocytosis genetics, Adaptor Proteins, Signal Transducing genetics, Chromosome Aberrations, Chromosomes, Human, Pair 11 genetics, DNA-Binding Proteins genetics, LIM Domain Proteins genetics, Proto-Oncogene Proteins genetics
- Abstract
Exome sequencing of primary tumors identifies complex somatic mutation patterns. Assignment of relevance of individual somatic mutations is difficult and poses the next challenge for interpretation of next generation sequencing data. Here we present an approach how exome sequencing in combination with SNP microarray data may identify targets of chromosomal aberrations in myeloid malignancies. The rationale of this approach is that hotspots of chromosomal aberrations might also harbor point mutations in the target genes of deletions, gains or uniparental disomies (UPDs). Chromosome 11 is a frequent target of lesions in myeloid malignancies. Therefore, we studied chromosome 11 in a total of 813 samples from 773 individual patients with different myeloid malignancies by SNP microarrays and complemented the data with exome sequencing in selected cases exhibiting chromosome 11 defects. We found gains, losses and UPDs of chromosome 11 in 52 of the 813 samples (6.4%). Chromosome 11q UPDs frequently associated with mutations of CBL. In one patient the 11qUPD amplified somatic mutations in both CBL and the DNA repair gene DDB1. A duplication within MLL exon 3 was detected in another patient with 11qUPD. We identified several common deleted regions (CDR) on chromosome 11. One of the CDRs associated with de novo acute myeloid leukemia (P=0.013). One patient with a deletion at the LMO2 locus harbored an additional point mutation on the other allele indicating that LMO2 might be a tumor suppressor frequently targeted by 11p deletions. Our chromosome-centered analysis indicates that chromosome 11 contains a number of tumor suppressor genes and that the role of this chromosome in myeloid malignancies is more complex than previously recognized.
- Published
- 2013
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29. Clinical significance of genetic aberrations in secondary acute myeloid leukemia.
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Milosevic JD, Puda A, Malcovati L, Berg T, Hofbauer M, Stukalov A, Klampfl T, Harutyunyan AS, Gisslinger H, Gisslinger B, Burjanivova T, Rumi E, Pietra D, Elena C, Vannucchi AM, Doubek M, Dvorakova D, Robesova B, Wieser R, Koller E, Suvajdzic N, Tomin D, Tosic N, Colinge J, Racil Z, Steurer M, Pavlovic S, Cazzola M, and Kralovics R
- Subjects
- DNA genetics, Gene Expression Profiling, Genome-Wide Association Study, Humans, Kaplan-Meier Estimate, Karyotyping, Leukemia, Myeloid, Acute mortality, Loss of Heterozygosity, Multivariate Analysis, Neoplasms, Second Primary mortality, Nucleophosmin, Oligonucleotide Array Sequence Analysis, Polymorphism, Single Nucleotide, Prognosis, Chromosome Aberrations statistics & numerical data, Leukemia, Myeloid, Acute genetics, Neoplasms, Second Primary genetics
- Abstract
The study aimed to identify genetic lesions associated with secondary acute myeloid leukemia (sAML) in comparison with AML arising de novo (dnAML) and assess their impact on patients' overall survival (OS). High-resolution genotyping and loss of heterozygosity mapping was performed on DNA samples from 86 sAML and 117 dnAML patients, using Affymetrix Genome-Wide Human SNP 6.0 arrays. Genes TP53, RUNX1, CBL, IDH1/2, NRAS, NPM1, and FLT3 were analyzed for mutations in all patients. We identified 36 recurrent cytogenetic aberrations (more than five events). Mutations in TP53, 9pUPD, and del7q (targeting CUX1 locus) were significantly associated with sAML, while NPM1 and FLT3 mutations associated with dnAML. Patients with sAML carrying TP53 mutations demonstrated lower 1-year OS rate than those with wild-type TP53 (14.3% ± 9.4% vs. 35.4% ± 7.2%; P = 0.002), while complex karyotype, del7q (CUX1) and del7p (IKZF1) showed no significant effect on OS. Multivariate analysis confirmed that mutant TP53 was the only independent adverse prognostic factor for OS in sAML (hazard ratio 2.67; 95% CI: 1.33-5.37; P = 0.006). Patients with dnAML and complex karyotype carried sAML-associated defects (TP53 defects in 54.5%, deletions targeting FOXP1 and ETV6 loci in 45.4% of the cases). We identified several co-occurring lesions associated with either sAML or dnAML diagnosis. Our data suggest that distinct genetic lesions drive leukemogenesis in sAML. High karyotype complexity of sAML patients does not influence OS. Somatic mutations in TP53 are the only independent adverse prognostic factor in sAML. Patients with dnAML and complex karyotype show genetic features associated with sAML and myeloproliferative neoplasms., (Copyright © 2012 Wiley Periodicals, Inc.)
- Published
- 2012
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30. Frequent deletions of JARID2 in leukemic transformation of chronic myeloid malignancies.
- Author
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Puda A, Milosevic JD, Berg T, Klampfl T, Harutyunyan AS, Gisslinger B, Rumi E, Pietra D, Malcovati L, Elena C, Doubek M, Steurer M, Tosic N, Pavlovic S, Guglielmelli P, Pieri L, Vannucchi AM, Gisslinger H, Cazzola M, and Kralovics R
- Subjects
- Acute Disease, Aged, Carrier Proteins genetics, Chromosome Aberrations, Chromosome Mapping, Chromosomes, Human, Pair 6 genetics, DNA-Binding Proteins deficiency, DNA-Binding Proteins genetics, Disease Progression, Enhancer of Zeste Homolog 2 Protein, Female, Genotype, Humans, Leukemia, Myeloid genetics, Male, Neoplasm Proteins deficiency, Neoplasm Proteins genetics, Nerve Tissue Proteins deficiency, Nerve Tissue Proteins genetics, Nuclear Proteins deficiency, Nuclear Proteins genetics, Oligonucleotide Array Sequence Analysis, Polycomb Repressive Complex 2, Polycomb-Group Proteins, Repressor Proteins deficiency, Repressor Proteins genetics, Repressor Proteins physiology, Sequence Analysis, DNA, Transcription Factors deficiency, Transcription Factors genetics, Tumor Suppressor Proteins deficiency, Tumor Suppressor Proteins genetics, Cell Transformation, Neoplastic genetics, Chromosome Deletion, Chromosomes, Human, Pair 6 ultrastructure, Genes, Tumor Suppressor, Myelodysplastic Syndromes genetics, Myeloproliferative Disorders genetics, Neoplasm Proteins physiology, Nerve Tissue Proteins physiology, Tumor Suppressor Proteins physiology
- Abstract
Chronic myeloproliferative neoplasms (MPN) and myelodysplastic syndromes (MDS) have an inherent tendency to progress to acute myeloid leukemia (AML). Using high-resolution SNP microarrays, we studied a total of 517 MPN and MDS patients in different disease stages, including 77 AML cases with previous history of MPN (N = 46) or MDS (N = 31). Frequent chromosomal deletions of variable sizes were detected, allowing the mapping of putative tumor suppressor genes involved in the leukemic transformation process. We detected frequent deletions on the short arm of chromosome 6 (del6p). The common deleted region on 6p mapped to a 1.1-Mb region and contained only the JARID2 gene--member of the polycomb repressive complex 2 (PRC2). When we compared the frequency of del6p between chronic and leukemic phase, we observed a strong association of del6p with leukemic transformation (P = 0.0033). Subsequently, analysis of deletion profiles of other PRC2 members revealed frequent losses of genes such as EZH2, AEBP2, and SUZ12; however, the deletions targeting these genes were large. We also identified two patients with homozygous losses of JARID2 and AEBP2. We observed frequent codeletion of AEBP2 and ETV6, and similarly, SUZ12 and NF1. Using next generation exome sequencing of 40 patients, we identified only one somatic mutation in the PRC2 complex member SUZ12. As the frequency of point mutations in PRC2 members was found to be low, deletions were the main type of lesions targeting PRC2 complex members. Our study suggests an essential role of the PRC2 complex in the leukemic transformation of chronic myeloid disorders., (Copyright © 2011 Wiley Periodicals, Inc.)
- Published
- 2012
- Full Text
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31. Identification of genomic aberrations associated with disease transformation by means of high-resolution SNP array analysis in patients with myeloproliferative neoplasm.
- Author
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Rumi E, Harutyunyan A, Elena C, Pietra D, Klampfl T, Bagienski K, Berg T, Casetti I, Pascutto C, Passamonti F, Kralovics R, and Cazzola M
- Subjects
- Blast Crisis etiology, Blast Crisis genetics, Blast Crisis metabolism, DNA chemistry, DNA metabolism, Disease Progression, Female, Genome-Wide Association Study, Granulocytes metabolism, Humans, Italy, Janus Kinase 2 genetics, Janus Kinase 2 metabolism, Leukemia, Myeloid genetics, Leukemia, Myeloid metabolism, Leukemia, Myeloid physiopathology, Male, Mutation, Myeloproliferative Disorders metabolism, Myeloproliferative Disorders pathology, Myeloproliferative Disorders physiopathology, Oligonucleotide Array Sequence Analysis, Polycythemia Vera etiology, Polycythemia Vera genetics, Polycythemia Vera metabolism, Primary Myelofibrosis etiology, Primary Myelofibrosis genetics, Primary Myelofibrosis metabolism, Receptors, Thrombopoietin genetics, Receptors, Thrombopoietin metabolism, Survival Analysis, Thrombocythemia, Essential etiology, Thrombocythemia, Essential genetics, Thrombocythemia, Essential metabolism, Cell Transformation, Neoplastic genetics, Chromosome Aberrations, Myeloproliferative Disorders genetics, Polymorphism, Single Nucleotide
- Abstract
Myeloproliferative neoplasms (MPN) include polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). These disorders may undergo phenotypic shifts, and may specifically evolve into secondary myelofibrosis (MF) or acute myeloid leukemia (AML). We studied genomic changes associated with these transformations in 29 patients who had serial samples collected in different phases of disease. Genomic DNA from granulocytes, i.e., the myeloproliferative genome, was processed and hybridized to genome-wide human SNP 6.0 arrays. Most patients in chronic phase had chromosomal regions with uniparental disomy (UPD) and/or copy number changes. Disease progression to secondary MF or AML was associated with the acquisition of additional chromosomal aberrations in granulocytes (P = 0.002). A close relationship was observed between aberrations of chromosome 9p (UPD and/or gain) and progression from PV to post-PV MF (P = 0.002). The acquisition of one or more aberrations involving chromosome 5, 7, or 17p was specifically associated with progression to AML (OR 5.9, 95% CI 1.2-27.7, P = 0.006), and significantly affected overall survival (HR 18, 95% CI 1.9-164, P = 0.01). These observations indicate that disease progression from chronic-phase MPN to secondary MF or AML is associated with specific chromosomal aberrations that can be detected by means of high-resolution SNP array analysis of granulocyte DNA., (Copyright © 2011 Wiley-Liss, Inc.)
- Published
- 2011
- Full Text
- View/download PDF
32. Genome integrity of myeloproliferative neoplasms in chronic phase and during disease progression.
- Author
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Klampfl T, Harutyunyan A, Berg T, Gisslinger B, Schalling M, Bagienski K, Olcaydu D, Passamonti F, Rumi E, Pietra D, Jäger R, Pieri L, Guglielmelli P, Iacobucci I, Martinelli G, Cazzola M, Vannucchi AM, Gisslinger H, and Kralovics R
- Subjects
- Chronic Disease, Disease Progression, Gene Expression Regulation, Leukemic genetics, Humans, Janus Kinase 2 genetics, Karyotyping, Point Mutation, Polymorphism, Single Nucleotide, Genome-Wide Association Study methods, Leukemia, Myeloid, Chronic, Atypical, BCR-ABL Negative genetics, Myeloproliferative Disorders genetics, Oligonucleotide Array Sequence Analysis methods
- Abstract
Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs) are clonal myeloid disorders with increased production of terminally differentiated cells. The disease course is generally chronic, but some patients show disease progression (secondary myelofibrosis or accelerated phase) and/or leukemic transformation. We investigated chromosomal aberrations in 408 MPN samples using high-resolution single-nucleotide polymorphism microarrays to identify disease-associated somatic lesions. Of 408 samples, 37.5% had a wild-type karyotype and 62.5% harbored at least 1 chromosomal aberration. We identified 25 recurrent aberrations that were found in 3 or more samples. An increased number of chromosomal lesions was significantly associated with patient age, as well as with disease progression and leukemic transformation, but no association was observed with MPN subtypes, Janus kinase 2 (JAK2) mutational status, or disease duration. Aberrations of chromosomes 1q and 9p were positively associated with disease progression to secondary myelofibrosis or accelerated phase. Changes of chromosomes 1q, 7q, 5q, 6p, 7p, 19q, 22q, and 3q were positively associated with post-MPN acute myeloid leukemia. We mapped commonly affected regions to single target genes on chromosomes 3p (forkhead box P1 [FOXP1]), 4q (tet oncogene family member 2 [TET2]), 7p (IKAROS family zinc finger 1 [IKZF1]), 7q (cut-like homeobox 1 [CUX1]), 12p (ets variant 6 [ETV6]), and 21q (runt-related transcription factor 1 [RUNX1]). Our data provide insight into the genetic complexity of MPNs and implicate new genes involved in disease progression.
- Published
- 2011
- Full Text
- View/download PDF
33. p53 lesions in leukemic transformation.
- Author
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Harutyunyan A, Klampfl T, Cazzola M, and Kralovics R
- Subjects
- Chromosomes, Human, Pair 1, Humans, Cell Transformation, Neoplastic genetics, Genes, p53, Leukemia, Myeloid, Acute genetics, Mutation, Myeloproliferative Disorders genetics
- Published
- 2011
- Full Text
- View/download PDF
34. An RNA-Seq strategy to detect the complete coding and non-coding transcriptome including full-length imprinted macro ncRNAs.
- Author
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Huang R, Jaritz M, Guenzl P, Vlatkovic I, Sommer A, Tamir IM, Marks H, Klampfl T, Kralovics R, Stunnenberg HG, Barlow DP, and Pauler FM
- Subjects
- Animals, Biomarkers, Tumor genetics, Biomarkers, Tumor metabolism, Fetus, Gene Expression Profiling, Genome, Mice, Oligonucleotide Array Sequence Analysis, RNA, Messenger genetics, Real-Time Polymerase Chain Reaction, Cell Differentiation, Embryonic Stem Cells cytology, Embryonic Stem Cells metabolism, Genomic Imprinting, Head physiology, RNA, Untranslated genetics
- Abstract
Imprinted macro non-protein-coding (nc) RNAs are cis-repressor transcripts that silence multiple genes in at least three imprinted gene clusters in the mouse genome. Similar macro or long ncRNAs are abundant in the mammalian genome. Here we present the full coding and non-coding transcriptome of two mouse tissues: differentiated ES cells and fetal head using an optimized RNA-Seq strategy. The data produced is highly reproducible in different sequencing locations and is able to detect the full length of imprinted macro ncRNAs such as Airn and Kcnq1ot1, whose length ranges between 80-118 kb. Transcripts show a more uniform read coverage when RNA is fragmented with RNA hydrolysis compared with cDNA fragmentation by shearing. Irrespective of the fragmentation method, all coding and non-coding transcripts longer than 8 kb show a gradual loss of sequencing tags towards the 3' end. Comparisons to published RNA-Seq datasets show that the strategy presented here is more efficient in detecting known functional imprinted macro ncRNAs and also indicate that standardization of RNA preparation protocols would increase the comparability of the transcriptome between different RNA-Seq datasets.
- Published
- 2011
- Full Text
- View/download PDF
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