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• W2513915644 abstract "Article29 August 2016free access Transparent process Myeloid leukemia with transdifferentiation plasticity developing from T-cell progenitors Pia Riemke Pia Riemke Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Melinda Czeh Melinda Czeh Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Josephine Fischer Josephine Fischer Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Carolin Walter Carolin Walter Institute of Medical Informatics, University of Münster, Münster, Germany Search for more papers by this author Saeed Ghani Saeed Ghani Department of Hematology, Oncology, and Tumor Immunology, Robert-Rössle-Clinic, Berlin, Germany Search for more papers by this author Matthias Zepper Matthias Zepper Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Konstantin Agelopoulos Konstantin Agelopoulos Department of Dermatology, Competence Center Chronic Pruritus, University of Münster, Münster, Germany Search for more papers by this author Stephanie Lettermann Stephanie Lettermann Molecular Hematology and Oncology, Medical Clinics A, University of Münster, Münster, Germany Search for more papers by this author Marie L Gebhardt Marie L Gebhardt Department of Computational Biology and Data Mining, Max-Delbrück-Center for Molecular Medicine, Berlin, Germany Search for more papers by this author Nancy Mah Nancy Mah Berlin-Brandenburger Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin, Berlin, Germany Search for more papers by this author Andre Weilemann Andre Weilemann Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Michael Grau Michael Grau Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Verena Gröning Verena Gröning Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Torsten Haferlach Torsten Haferlach MLL Munich Leukemia Laboratory, Munich, Germany Search for more papers by this author Dido Lenze Dido Lenze Institute of Pathology, Charité - Universitätsmedizin Berlin, Berlin, Germany Search for more papers by this author Ruud Delwel Ruud Delwel Department of Hematology, Erasmus University Medical Center, Rotterdam, the Netherlands Search for more papers by this author Marco Prinz Marco Prinz orcid.org/0000-0002-0349-1955 Institute of Neuropathology, University of Freiburg, Freiburg, Germany BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany Search for more papers by this author Miguel A Andrade-Navarro Miguel A Andrade-Navarro Department of Medical Informatics and Biomathematics, Institute of Molecular Biology, Johannes Gutenberg University of Mainz, Mainz, Germany Search for more papers by this author Georg Lenz Georg Lenz Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Martin Dugas Martin Dugas Institute of Medical Informatics, University of Münster, Münster, Germany Search for more papers by this author Carsten Müller-Tidow Carsten Müller-Tidow Department of Internal Medicine, Hematology and Oncology, University of Halle-Wittenberg, Halle, Germany Search for more papers by this author Frank Rosenbauer Corresponding Author Frank Rosenbauer [email protected] orcid.org/0000-0001-7977-9421 Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Pia Riemke Pia Riemke Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Melinda Czeh Melinda Czeh Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Josephine Fischer Josephine Fischer Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Carolin Walter Carolin Walter Institute of Medical Informatics, University of Münster, Münster, Germany Search for more papers by this author Saeed Ghani Saeed Ghani Department of Hematology, Oncology, and Tumor Immunology, Robert-Rössle-Clinic, Berlin, Germany Search for more papers by this author Matthias Zepper Matthias Zepper Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Konstantin Agelopoulos Konstantin Agelopoulos Department of Dermatology, Competence Center Chronic Pruritus, University of Münster, Münster, Germany Search for more papers by this author Stephanie Lettermann Stephanie Lettermann Molecular Hematology and Oncology, Medical Clinics A, University of Münster, Münster, Germany Search for more papers by this author Marie L Gebhardt Marie L Gebhardt Department of Computational Biology and Data Mining, Max-Delbrück-Center for Molecular Medicine, Berlin, Germany Search for more papers by this author Nancy Mah Nancy Mah Berlin-Brandenburger Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin, Berlin, Germany Search for more papers by this author Andre Weilemann Andre Weilemann Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Michael Grau Michael Grau Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Verena Gröning Verena Gröning Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Torsten Haferlach Torsten Haferlach MLL Munich Leukemia Laboratory, Munich, Germany Search for more papers by this author Dido Lenze Dido Lenze Institute of Pathology, Charité - Universitätsmedizin Berlin, Berlin, Germany Search for more papers by this author Ruud Delwel Ruud Delwel Department of Hematology, Erasmus University Medical Center, Rotterdam, the Netherlands Search for more papers by this author Marco Prinz Marco Prinz orcid.org/0000-0002-0349-1955 Institute of Neuropathology, University of Freiburg, Freiburg, Germany BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany Search for more papers by this author Miguel A Andrade-Navarro Miguel A Andrade-Navarro Department of Medical Informatics and Biomathematics, Institute of Molecular Biology, Johannes Gutenberg University of Mainz, Mainz, Germany Search for more papers by this author Georg Lenz Georg Lenz Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany Search for more papers by this author Martin Dugas Martin Dugas Institute of Medical Informatics, University of Münster, Münster, Germany Search for more papers by this author Carsten Müller-Tidow Carsten Müller-Tidow Department of Internal Medicine, Hematology and Oncology, University of Halle-Wittenberg, Halle, Germany Search for more papers by this author Frank Rosenbauer Corresponding Author Frank Rosenbauer [email protected] orcid.org/0000-0001-7977-9421 Institute of Molecular Tumor Biology, University of Münster, Münster, Germany Search for more papers by this author Author Information Pia Riemke1,‡, Melinda Czeh1,‡, Josephine Fischer1,‡, Carolin Walter2, Saeed Ghani3, Matthias Zepper1, Konstantin Agelopoulos4, Stephanie Lettermann5, Marie L Gebhardt6, Nancy Mah7, Andre Weilemann8,9, Michael Grau8,9, Verena Gröning1, Torsten Haferlach10, Dido Lenze11, Ruud Delwel12, Marco Prinz13,14, Miguel A Andrade-Navarro15, Georg Lenz8,9, Martin Dugas2, Carsten Müller-Tidow16 and Frank Rosenbauer *,1 1Institute of Molecular Tumor Biology, University of Münster, Münster, Germany 2Institute of Medical Informatics, University of Münster, Münster, Germany 3Department of Hematology, Oncology, and Tumor Immunology, Robert-Rössle-Clinic, Berlin, Germany 4Department of Dermatology, Competence Center Chronic Pruritus, University of Münster, Münster, Germany 5Molecular Hematology and Oncology, Medical Clinics A, University of Münster, Münster, Germany 6Department of Computational Biology and Data Mining, Max-Delbrück-Center for Molecular Medicine, Berlin, Germany 7Berlin-Brandenburger Center for Regenerative Therapies, Charité - Universitätsmedizin Berlin, Berlin, Germany 8Translational Oncology, Medical Clinics A, University of Münster, Münster, Germany 9Cluster of Excellence EXC 1003, Cells in Motion, Münster, Germany 10MLL Munich Leukemia Laboratory, Munich, Germany 11Institute of Pathology, Charité - Universitätsmedizin Berlin, Berlin, Germany 12Department of Hematology, Erasmus University Medical Center, Rotterdam, the Netherlands 13Institute of Neuropathology, University of Freiburg, Freiburg, Germany 14BIOSS Centre for Biological Signalling Studies, University of Freiburg, Freiburg, Germany 15Department of Medical Informatics and Biomathematics, Institute of Molecular Biology, Johannes Gutenberg University of Mainz, Mainz, Germany 16Department of Internal Medicine, Hematology and Oncology, University of Halle-Wittenberg, Halle, Germany ‡These authors contributed equally to this work *Corresponding author. Tel: +49 251 83 55312; Fax:+49 251 83 55303; E-mail: [email protected] The EMBO Journal (2016)35:2399-2416https://doi.org/10.15252/embj.201693927 See also: L Bullinger (November 2016) PDFDownload PDF of article text and main figures. Peer ReviewDownload a summary of the editorial decision process including editorial decision letters, reviewer comments and author responses to feedback. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Abstract Unfavorable patient survival coincides with lineage plasticity observed in human acute leukemias. These cases are assumed to arise from hematopoietic stem cells, which have stable multipotent differentiation potential. However, here we report that plasticity in leukemia can result from instable lineage identity states inherited from differentiating progenitor cells. Using mice with enhanced c-Myc expression, we show, at the single-cell level, that T-lymphoid progenitors retain broad malignant lineage potential with a high capacity to differentiate into myeloid leukemia. These T-cell-derived myeloid blasts retain expression of a defined set of T-cell transcription factors, creating a lymphoid epigenetic memory that confers growth and propagates myeloid/T-lymphoid plasticity. Based on these characteristics, we identified a correlating human leukemia cohort and revealed targeting of Jak2/Stat3 signaling as a therapeutic possibility. Collectively, our study suggests the thymus as a source for myeloid leukemia and proposes leukemic plasticity as a driving mechanism. Moreover, our results reveal a pathway-directed therapy option against thymus-derived myeloid leukemogenesis and propose a model in which dynamic progenitor differentiation states shape unique neoplastic identities and therapy responses. Synopsis The inherent lineage plasticity of multipotent hematopoietic stem cells is thought to be causal for the development of human acute myeloid leukemia (AML). Here, early non-committed lymphoid progenitors are shown to give rise to myeloid blasts, which retain epigenetic cell-of-origin memory and are promoted by Jak/Stat signaling. Myc/Bcl2-transformed mouse early T-cell progenitors have broad malignant lineage potential and can generate myeloid leukemic progeny in in vitro colony formation and in vivo transplantation assays. T-cell progenitor-derived myeloid leukemia cells express a lymphoid epigenetic memory that confers growth and propagates myeloid/T-lymphoid plasticity. A cohort of ˜5% human AML cases resembles mouse T-cell-derived leukemia and thus may be of thymic origin. Jak2/Stat3 signaling is required for proliferation of T-cell progenitor-derived myeloid leukemia cells and leukemogenesis in vivo. Introduction Acute leukemia (AL) refers to a heterogeneous group of blood cell disorders in which differentiation-blocked malignant myeloid or lymphoid cells expand uncontrolled. The treatment for AL depends on how the disease is classified according to both cytogenetic and phenotypic markers (Bene et al, 1995; Dohner et al, 2010). However, up to 20% of ALs remain difficult to classify, as they express both lymphoid and myeloid antigens (Matutes et al, 1997; Golemovic et al, 2006). Clinically, these “ambiguous” AL cases are highly relevant because they result in particularly poor patient survival and are frequently resistant to standard therapies (Golemovic et al, 2006; Coustan-Smith et al, 2009; Dorantes-Acosta & Pelayo, 2012). In addition to lineage marker ambiguity, ALs that switch lineages in treatment-associated relapse have been described (Stass et al, 1984; Risueno et al, 2011). These findings imply that hematopoietic malignancies can hold immanent potential to differentiate into alternative lineages. While earlier studies have assumed that leukemias with lineage infidelity and switching capacity can arise upon malignant transformation of a hematopoietic stem cell (HSC; Matutes et al, 1997; Neumann et al, 2012), the underlying cellular origin of such lineage plasticity remains poorly defined. One type of AL, acute myeloid leukemia (AML), is believed to originate from transformed HSCs or myeloid-restricted progenitors (Walter et al, 2012). However, these bone marrow (BM)-resident precursors are not the only cells with myeloid potential: myeloid potential has also been found in thymic CD4/CD8 double negative 1 (DN1) and 2 (DN2) cells, which represent the earliest stages of T-cell development (Bell & Bhandoola, 2008; Wada et al, 2008). However, the contribution of DN1 and DN2 cells to physiological myelopoiesis in vivo has been questioned by lineage-tracing experiments (Schlenner et al, 2010). Nevertheless, the same authors found that approximately 15% of DN1 cells had robust myeloid differentiation capacity in vitro, confirming that thymic progenitors can produce myeloid progeny (Schlenner et al, 2010). In another study, myeloid potential was attributed to thymic progenitors in vivo, and early T cells were suggested as precursors of thymic granulocytes (De Obaldia et al, 2013). However, even if a contribution to myelopoiesis under homeostatic conditions is still unclear, these results demonstrated that the lymphoid fate of early T-cell progenitors is not yet fully stabilized and may compete with a residual option of myeloid differentiation. Therefore, we hypothesized that this myeloid potential may be exploited under pathological conditions so that T-cell precursors could transdifferentiate into leukemia with myeloid identity. To test this hypothesis, we aimed to analyze the lineage potential of DN2 cells upon transformation. We reasoned that ectopic c-Myc (Myc) expression would serve our purpose best, as MYC has been demonstrated to transform cells, at least in part, by amplifying the expression of already transcribed genes without enforcing a lineage bias (Lin et al, 2012; Nie et al, 2012). Moreover, MYC is frequently overexpressed in both human acute myeloid and lymphoid leukemia (Majeti et al, 2009; Haferlach et al, 2010) (Appendix Fig S1A) and can rapidly induce hematopoietic neoplasias in mice (Luo et al, 2005). We therefore overexpressed Myc, together with Bcl2 to prevent apoptosis, in murine thymic and BM-resident precursor cells and compared their lineage potential in leukemogenesis. Results Single DN2 cells possess malignant myeloid and T-lymphoid potential in vitro To evaluate the lineage potential of transformed T-cell precursors, we isolated DN2 cells from the thymus along with BM LSK cells (Lineage−Sca1+c-kit+ as a population enriched for HSCs) and granulocyte–macrophage progenitors (GMPs) from C57BL/6 CD45.2 donor wild-type mice. Cells were transduced with a retrovirus co-expressing Myc and Bcl2 from a bicistronic mRNA (Myc/Bcl2) (Appendix Fig S1B–D). From these populations, we sorted single cells into 96-well plates containing 1:1 mixtures of OP9:OP9-DL1 as feeder cell layers and analyzed the resulting clones via flow cytometry for the expression of CD11b and Gr1 as myeloid markers, and CD25 as an early T-lineage marker which was in line with previous reports (Bell & Bhandoola, 2008; Wada et al, 2008). Myc/Bcl2-transduced DN2 cells generated immortalized clones with 8.6% plating efficiency, all of which had either mixed myeloid/T-cell or pure T-cell phenotypes (Fig 1A and B). From the mixed-lineage clones, flow-sorted myeloid cells or lymphoid cells demonstrated typical blast-like myeloid or lymphoid morphologies, respectively, and produced short-latency leukemias in transplanted mice, indicating their fully transformed states (Fig 1C). In contrast to DN2 cells, single Myc/Bcl2+ GMPs produced exclusively myeloid clones, and single Myc/Bcl2+ LSK cells produced predominantly pure myeloid clones but also mixed myeloid/T-cell and pure T-cell clones at lower frequencies (Fig 1B and D). These results show that at the single-cell level, transformed DN2 cells do not exclusively produce T-lymphoid leukemia, but can also develop into myeloid leukemia. Figure 1. Single DN2 cells possess malignant myeloid and T-lymphoid potential Representative FACS plot (left) showing a mixed-lineage clone with a T-lymphoid (CD25+/CD11b−Gr1−), myeloid (CD11b+Gr1+/CD25−), and biphenotypic (CD11b+Gr1+/CD25+) fraction derived from a single Myc/Bcl2-transformed DN2 cell grown on 1:1 OP9:OP9-Dll1 co-culture with lineage-promiscuous cytokines IL-2, IL-3, IL-6, IL-7, SCF, GM-CSF, Flt3. Morphology of the indicated flow-sorted cell fractions was analyzed by Giemsa staining (right). Phenotype frequencies of clones from Myc/Bcl2-transformed single cells (grown and phenotyped as mentioned in A). Analyzed were 31 clones from LSKs, 107 from GMPs and 28 from DN2 cells. Mice that received 5 × 104 to 5 × 105 T-lymphoid cells or myeloid cells flow-sorted from mixed Myc/Bcl2+ DN2-derived clones showed a cumulative survival of 31 ± 2 and 27 ± 6 days respectively. The summarized data are from three mice injected with the T-lymphoid and four mice injected with the myeloid fraction of two different clones. Representative FACS plots demonstrating predominant outgrowth of myeloid clones from single Myc/Bcl2-transformed LSK or GMP cells on OP9:OP9-Dll1 co-cultures. (top panels) Serial replating of Myc-Bcl2+ DN2 cells in methylcellulose generated myeloid clones with stable genomic reassembly of the Dβ1 TCR locus in vitro (gel: left two lanes). Upon transplantation, the same clones induced myeloid leukemia that retained the initial rearrangement in vivo (gel: right two lanes) as assessed by nested PCR. GL = germline. (bottom panels) Methylcellulose-based replating of Myc-Bcl2+ GMPs generated myeloid clones that exclusively displayed the TCRβ locus in germline configuration. Cumulative survival of mice that received 106 cells of TCRβ-rearranged myeloid DN2-derived clones leading to death 33 ± 4 days after transplantation. Two DN2 clones were transplanted into five recipient mice. Download figure Download PowerPoint The myeloid potential of DN2 cells was stable across the leukemia transforming event, as transduction with the MLL-AF9 [t(9;11)] oncogenic fusion also generated mixed myeloid/T-cell clones at high frequency (Appendix Fig S2A and B). This suggests that the potential to produce myeloid blasts represents a feature that is endogenous to transformed DN2 cells. DN2 cells with T-cell receptor rearrangement can generate myeloid leukemic progeny The defining step in T-cell development is rearrangement of the T-cell receptor (TCR) locus. Thus, to underscore that T-cell progenitors can transform into myeloid blasts, we first identified rearrangement of the TCR beta locus in Myc/Bcl2+ DN2- and DN1-derived myeloid clones (Appendix Fig S2C) and subsequently transplanted one of each into mice. In both cases, the donor cells rapidly produced lethal myeloid leukemias, in which the blasts carried the same rearrangements as the initial clones (Fig 1E and F and data not shown). This experiment provides genetic evidence that myeloid leukemia can originate from T-cell progenitors. Transformed DN2 cells give rise to leukemias with clonal myeloid and T-lymphoid populations in vivo To directly assess the leukemic lineage potential in vivo, we first transduced DN2 cells with Myc/Bcl2 and then immediately transplanted them intravenously into sublethally irradiated congenic mice. In line with the in vitro results, Myc/Bcl2+ DN2 cells produced leukemias consisting of myeloid, T-cell, and biphenotypic fractions in the same recipients (Fig 2A). We also did this for Myc/Bcl2+ LSK cells, which also produced mixed-lineage leukemia, but with reduced T-lymphoid and biphenotypic fractions. The Myc/Bcl2+ GMPs exclusively produced myeloid leukemia. We also generated Myc/Bcl2+ DN3 cells, representing a more differentiated T-cell progenitor, which produced exclusively T-lymphoid leukemia, in part with dim co-expression of myeloid markers, but never myeloid leukemia (Appendix Fig S3A). Figure 2. Myc/Bcl2+ DN2 cells give rise to clonal myeloid and T-lymphoid leukemia in vivo Representative FACS plots of spleens from leukemic recipient mice that had received LSK, GMP, or DN2 cell grafts immediately after retroviral Myc/Bcl2 transduction. Numbers indicate frequencies (%) of CD45.2 donor cells within the indicated gates summarizing the mean ± SEM of at least eight recipients per group from a total of 10 independent experiments. Cumulative survival of mice that received 2 × 104 to 2 × 105 freshly Myc/Bcl2-transduced LSK, GMP, or DN2 cells. Graph shows n = 11 Myc-Bcl2+ LSK-, n = 14 DN2-, and n = 8 GMP-transplanted animals. Histological spleen sections of Myc-Bcl2+ LSK-, GMP-, or DN2-transplanted mice displaying myeloperoxidase staining. Leukemic cell engraftment in spleens ranged from 75.1% to 95.5%. Multiplex quantitative RT–PCR demonstrating strong expression of PU.1 in the myeloid (M3L: CD11b+Gr1+/CD3−CD4−CD8−) and biphenotypic T/myeloid (BL: CD11b+Gr1+/CD3+CD4+CD8+) fractions but not in the T-lymphoid (TL: CD11b−Gr1−/CD3+CD4+CD8+) fraction that were flow-sorted from diseased Myc-Bcl2+ DN2-transplanted mice. Myeloid progeny of Myc/Bcl2+ GMP-transplanted or T-lymphoid progeny of Myc/Bcl2+ DN3-transplanted animals as well as healthy thymocytes (Thy) and BM cells from WT mice served as controls. Results are shown as fold over actin transcripts. Error bars define standard deviation (SD), number of replicates n = 2. D-J rearrangement PCR of flow-sorted T-lymphoid (TL), biphenotypic (BL) and myeloid (ML) fractions of donor splenocytes of a diseased Myc/Bcl2+ DN2-transplanted mouse. All three fractions revealed a stable genomic reassembly of the Dβ1 TCR locus (Dβ1Jβ1.5) demonstrating clonality in DN2 leukemia. thy = thymus. FACS plot of diseased intrathymic grafts (CD45.2-gated), demonstrating that Myc/Bcl2+ DN2 cells could produce multi-lineage leukemia within a T-cell-supporting environment; Myc/Bcl2+ GMPs remained myeloid-restricted under these conditions. Numbers indicate cell frequencies within the gates (mean ± SEM) representing six independent experiments. Intrathymic transplantation of mock-infected non-leukemic DN2 cells produced predominantly T-lymphoid but also myeloid progeny 23 days after injection. This differentiation outcome resembles that of freshly isolated DN2 cells (Bell & Bhandoola, 2008; Richie Ehrlich et al, 2011) and indicates that the short-term culture necessary for retroviral transduction did not interfere with the physiological lineage potential of DN2 cells. Download figure Download PowerPoint The mean survival of Myc/Bcl2+ DN2, LSK and GMP recipients was comparably short (56 ± 5, 39 ± 5 and 49 ± 4 days, respectively) (Fig 2B). The animals showed massive infiltration of the BM with myeloid blasts, leukocytosis, infiltration of lymphatic tissues (splenomegaly, lymphadenopathy), and indications of extralympathic infiltration (hind limb paralysis). In many mice, we observed elevated white blood cell counts (> 50 × 103/μl as compared to 9.2 ± 2.4 × 103/μl in healthy animals; data not shown). A detailed flow-cytometric characterization of Myc/Bcl2+ DN2-leukemia is given in Appendix Fig S3B and C. This revealed heterogeneous marker expression in the biphenotypic population. The myeloid population expressed both Gr1 and/or CD11b, as well as myeloperoxidase and PU.1, a key myeloid identity transcription factor that is not expressed in T cells (Rothenberg et al, 2008; Fig 2C and D, and Appendix Fig S3B–F). Notably, TCR rearrangement analyses demonstrated clonal relationships between the T-lymphoid, biphenotypic, and myeloid fractions in leukemic Myc/Bcl2+ DN2-recipient mice (Fig 2E), suggesting that single DN2 cells have multi-lineage potential in vivo. To exclude the possibility that the short-term cultivation required for retroviral transduction imposed a myeloid lineage bias on DN2 cells, we showed that the in vivo differentiation capacity of mock virus-transduced cells was similar to that of DN2 cells freshly isolated from the thymus (Fig 2G; Richie Ehrlich et al, 2011). Moreover, to more closely model the physiological transformation process in vivo, we also transplanted Myc/Bcl2+ DN2 cells directly into the thymus, and confirmed their myeloid leukemic potential (Fig 2F). Intrathymic Myc/Bcl2+ DN2- or GMP-grafts developed full-blown leukemia after 36 ± 2 and 46 ± 5 days, respectively (Appendix Fig S3G). When comparing cell engraftment in BM, spleen, and thymus, we found that DN2- and GMP-derived blasts disseminated differently. Myc/Bcl2+ cells from GMPs preferentially infiltrated the organ(s) associated with their site of transplantation. Intravenously transplanted GMP-derived blasts primarily infiltrated the BM and spleen rather than the thymus (Appendix Fig S3H), while intrathymically transplanted GMP-derived blasts mainly stayed within the thymus. In contrast, Myc/Bcl2+ DN2-derived blasts infiltrated equally into all organs, suggesting an enhanced dissemination capacity. DN2-myeloid blasts have DC-like identity and retain a cell-of-origin memory To determine the molecular mechanisms that drive development of myeloid blasts from DN2 cells, we performed gene expression profiling of flow-sorted myeloid fractions of Myc/Bcl2+ DN2-, LSK- and GMP-derived leukemias from sick mice, along with normal DN2 und GMP cells from healthy wild-type (WT) mice (Dataset EV1). To better disclose the lineage identity of the diseased cells, we used gene set enrichment analysis (GSEA) to compare transcriptomes of the flow-sorted myeloid fractions to gene signatures defining different myeloid cell populations (Schonheit et al, 2013). Whereas the GMP- and LSK-derived myeloid blasts mainly demonstrated a neutrophil identity, DN2-derived myeloid blasts exhibited a pan-dendritic cell (DC) signature (Fig 3A), a finding confirmed with another DC signature (Chan et al, 2006; data not shown). This was in line with residual DC potential that has been assigned to early T-cell progenitors (Moore et al, 2012). Figure 3. DN2-myeloid leukemia retains a T-cell memory GSEA demonstrates that Myc/Bcl2+ DN2-derived myeloid blasts were significantly (FDR ≥ 0.05) enriched for expression of a pan-dendritic cell gene set. Gene sets were published in Schonheit et al (2013). Heat map of the top 100 differentially expressed genes (adjusted P-value of ≤ 0.05) in each myeloid fraction (ML: CD11b+Gr1+/CD3−CD4−CD8−) from GMP-, LSK-, and DN2-derived leukemias. Dendrogram revealing transcriptional relationships between Myc/Bcl2+ DN2-, LSK-, and GMP-derived myeloid blasts based on unsupervised hierarchical clustering of all probes on the microarray. GSEA demonstrates significant enrichment of a normal DN2-specific gene set (see 4 section) in the Myc/Bcl2+ DN2-derived myeloid blasts. Download figure Download PowerPoint Through a heat map comparison of the top 100 genes most differentially expressed (P ≤ 0.05) in each of the analyzed cell populations, we revealed that DN2- and GMP-derived myeloid blasts clearly differed from each other, but LSK-derived blasts had an intermediate phenotype (Fig 3B and Dataset EV2). Yet, unsupervised hierarchical clustering across the entire transcriptomes revealed that LSK- and GMP-derived cells are more closely related to each other than they are to DN2-derived cells (Fig 3C). Moreover, GSEA demonstrated that a signature of genes highly expressed in WT DN2 cells was significantly enriched in the DN2- but not in GMP- or LSK-derived myeloid blasts (Fig 3D and Dataset EV3). Collectively, gene expression profiling revealed that DN2-leukemia adopts a unique myeloid lineage identity, and still retains expression of a memory of its pre-malignant T-cell origin. The T-cell memory is required for DN2-leukemia growth We next assessed if the T-cell memory was involved in driving Myc/Bcl2+ DN2-leukemia. Because cell-specific gene programs are mainly regulated by transcription factors (Rosenbauer & Tenen, 2007; Rothenberg et al, 2008), we searched for genes in the thymic memory that encode transcription factors and identified Bcl11b and Gata3 as two promising candidates" @default.
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• W2513915644 title "Myeloid leukemia with transdifferentiation plasticity developing from T‐cell progenitors" @default.
• W2513915644 cites W1515023627 @default.
• W2513915644 cites W1520966126 @default.
• W2513915644 cites W1776790137 @default.
• W2513915644 cites W1965006511 @default.
• W2513915644 cites W1965584669 @default.
• W2513915644 cites W1969984078 @default.
• W2513915644 cites W1971186299 @default.
• W2513915644 cites W1971463847 @default.
• W2513915644 cites W1979521298 @default.
• W2513915644 cites W1980860344 @default.
• W2513915644 cites W1981082529 @default.
• W2513915644 cites W1982275421 @default.
• W2513915644 cites W1994041743 @default.
• W2513915644 cites W2009062738 @default.
• W2513915644 cites W2009114635 @default.
• W2513915644 cites W2010422802 @default.
• W2513915644 cites W2012831940 @default.
• W2513915644 cites W2020713861 @default.
• W2513915644 cites W2023497353 @default.
• W2513915644 cites W2024488862 @default.
• W2513915644 cites W2031527230 @default.
• W2513915644 cites W2034876218 @default.
• W2513915644 cites W2045556155 @default.
• W2513915644 cites W2045662960 @default.
• W2513915644 cites W2047137615 @default.
• W2513915644 cites W2053468615 @default.
• W2513915644 cites W2055160887 @default.
• W2513915644 cites W2055350603 @default.
• W2513915644 cites W2056777053 @default.
• W2513915644 cites W2060605982 @default.
• W2513915644 cites W2060618743 @default.
• W2513915644 cites W2060710804 @default.
• W2513915644 cites W2060935041 @default.
• W2513915644 cites W2062712297 @default.
• W2513915644 cites W2068981404 @default.
• W2513915644 cites W2077546113 @default.
• W2513915644 cites W2079853611 @default.
• W2513915644 cites W2081141657 @default.
• W2513915644 cites W2081988140 @default.
• W2513915644 cites W2094457561 @default.
• W2513915644 cites W2095588623 @default.
• W2513915644 cites W2096885745 @default.
• W2513915644 cites W2100126911 @default.
• W2513915644 cites W2115351357 @default.
• W2513915644 cites W2116634113 @default.
• W2513915644 cites W2116858301 @default.
• W2513915644 cites W2117466789 @default.
• W2513915644 cites W2119709204 @default.
• W2513915644 cites W2120757045 @default.
• W2513915644 cites W2132364074 @default.
• W2513915644 cites W2134237864 @default.
• W2513915644 cites W2142226489 @default.
• W2513915644 cites W2143320043 @default.
• W2513915644 cites W2145034432 @default.
• W2513915644 cites W2147373929 @default.
• W2513915644 cites W2148101855 @default.
• W2513915644 cites W2150690129 @default.
• W2513915644 cites W2157884213 @default.
• W2513915644 cites W2160026729 @default.
• W2513915644 cites W2161577298 @default.
• W2513915644 cites W2166306614 @default.
• W2513915644 cites W2167869412 @default.
• W2513915644 cites W2169405856 @default.
• W2513915644 cites W2170846829 @default.
• W2513915644 cites W2172033467 @default.
• W2513915644 cites W2343863476 @default.
• W2513915644 cites W2412936649 @default.
• W2513915644 doi "https://doi.org/10.15252/embj.201693927" @default.
• W2513915644 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/5109237" @default.
• W2513915644 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/27572462" @default.
• W2513915644 hasPublicationYear "2016" @default.

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