35 results on '"Roger, T"'
Search Results
2. MLL fusion proteins preferentially regulate a subset of wild-type MLL target genes in the leukemic genome
- Author
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Wang, Qian-fei, Wu, George, Mi, Shuangli, He, Fuhong, Wu, Jun, Dong, Jingfang, Luo, Roger T., Mattison, Ryan, Kaberlein, Joseph J., Prabhakar, Shyam, Ji, Hongkai, and Thirman, Michael J.
- Published
- 2011
- Full Text
- View/download PDF
3. The full transforming capacity of MLL-Af4 is interlinked with lymphoid lineage commitment
- Author
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James C. Mulloy, Michael J. Thirman, Shan Lin, Mahesh Shrestha, and Roger T. Luo
- Subjects
0301 basic medicine ,Myeloid ,Oncogene Proteins, Fusion ,Immunology ,Biology ,Biochemistry ,Leukemogenic ,Mice ,03 medical and health sciences ,hemic and lymphatic diseases ,medicine ,Animals ,Humans ,Cell Lineage ,Lymphocytes ,Cell Self Renewal ,Progenitor cell ,neoplasms ,Lymphoid Neoplasia ,Myeloid leukemia ,Cell Biology ,Hematology ,medicine.disease ,Fusion protein ,Leukemia ,Haematopoiesis ,Cell Transformation, Neoplastic ,030104 developmental biology ,medicine.anatomical_structure ,Cellular Microenvironment ,Cancer research ,Stem cell ,Myeloid-Lymphoid Leukemia Protein - Abstract
Chromosome rearrangements involving the mixed-lineage leukemia gene (MLL) create MLL-fusion proteins, which could drive both acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). The lineage decision of MLL-fusion leukemia is influenced by the fusion partner and microenvironment. To investigate the interplay of fusion proteins and microenvironment in lineage choice, we transplanted human hematopoietic stem and progenitor cells (HSPCs) expressing MLL-AF9 or MLL-Af4 into immunodeficient NSGS mice, which strongly promote myeloid development. Cells expressing MLL-AF9 efficiently developed AML in NSGS mice. In contrast, MLL-Af4 cells, which were fully oncogenic under lymphoid conditions present in NSG mice, displayed compromised transformation capacity in a myeloid microenvironment. MLL-Af4 activated a self-renewal program in a lineage-dependent manner, showing the leukemogenic activity of MLL-Af4 was interlinked with lymphoid lineage commitment. The C-terminal homology domain (CHD) of Af4 was sufficient to confer this linkage. Although the MLL-CHD fusion protein failed to immortalize HSPCs in myeloid conditions in vitro, it could successfully induce ALL in NSG mice. Our data suggest that defective self-renewal ability and leukemogenesis of MLL-Af4 myeloid cells could contribute to the strong B-cell ALL association of MLL-AF4 leukemia observed in the clinic.
- Published
- 2017
4. ELL-associated factor 2 (EAF2), a functional homolog of EAF1 with alternative ELL binding properties
- Author
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Simone, Federico, Luo, Roger T., Polak, Paul E., Kaberlein, Joseph J., and Thirman, Michael J.
- Published
- 2003
- Full Text
- View/download PDF
5. The full transforming capacity of MLL-Af4 is interlinked with lymphoid lineage commitment
- Author
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Lin, Shan, Luo, Roger T., Shrestha, Mahesh, Thirman, Michael J., and Mulloy, James C.
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- 2017
- Full Text
- View/download PDF
6. EAF1, a novel ELL-associated factor that is delocalized by expression of the MLL-ELL fusion protein
- Author
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Simone, Federico, Polak, Paul E., Kaberlein, Joseph J., Luo, Roger T., Levitan, Denise A., and Thirman, Michael J.
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- 2001
- Full Text
- View/download PDF
7. BCL2/BCL-XL inhibition induces apoptosis, disrupts cellular calcium homeostasis, and prevents platelet activation
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David Dinsdale, Gerald M. Cohen, Hassan A. Hamali, Roger T. Snowden, Edward T. W. Bampton, Meike Vogler, Xiao-Ming Sun, Alison H. Goodall, and Martin J. S. Dyer
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Blood Platelets ,medicine.medical_specialty ,Programmed cell death ,Immunology ,bcl-X Protein ,Gene Expression ,chemistry.chemical_element ,Antineoplastic Agents ,Apoptosis ,Biology ,Calcium ,Biochemistry ,Piperazines ,Calcium in biology ,Thrombopoiesis ,Nitrophenols ,hemic and lymphatic diseases ,Internal medicine ,medicine ,Homeostasis ,Humans ,Platelet ,Calcium Signaling ,Lymphocytes ,Molecular Targeted Therapy ,Platelet activation ,Calcium signaling ,Sulfonamides ,Aniline Compounds ,Biphenyl Compounds ,Cell Biology ,Hematology ,Platelet Activation ,Leukemia, Lymphocytic, Chronic, B-Cell ,Thrombocytopenia ,Kinetics ,Endocrinology ,Proto-Oncogene Proteins c-bcl-2 ,chemistry ,Cancer research ,Platelet aggregation inhibitor ,Apoptosis Regulatory Proteins ,Platelet Aggregation Inhibitors - Abstract
Apoptosis in megakaryocytes results in the formation of platelets. The role of apoptotic pathways in platelet turnover and in the apoptotic-like changes seen after platelet activation is poorly understood. ABT-263 (Navitoclax), a specific inhibitor of antiapoptotic BCL2 proteins, which is currently being evaluated in clinical trials for the treatment of leukemia and other malignancies, induces a dose-limiting thrombocytopenia. In this study, the relationship between BCL2/BCL-XL inhibition, apoptosis, and platelet activation was investigated. Exposure to ABT-263 induced apoptosis but repressed platelet activation by physiologic agonists. Notably, ABT-263 induced an immediate calcium response in platelets and the depletion of intracellular calcium stores, indicating that on BCL2/BCL-XL inhibition platelet activation is abrogated because of a diminished calcium signaling. By comparing the effects of ABT-263 and its analog ABT-737 on platelets and leukemia cells from the same donor, we show, for the first time, that these BCL2/BCL-XL inhibitors do not offer any selective toxicity but induce apoptosis at similar concentrations in leukemia cells and platelets. However, reticulated platelets are less sensitive to apoptosis, supporting the hypothesis that treatment with ABT-263 induces a selective loss of older platelets and providing an explanation for the transient thrombocytopenia observed on ABT-263 treatment.
- Published
- 2011
8. The Transcriptome Heterogeneity of MLL-Fusion ALL Is Driven By Fusion Partners Via Distinct Chromatin Binding
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Lin, Shan, primary, Ptasinska, Anetta, additional, Assi, Salam Adli, additional, Kerry, Jon, additional, Meetei, Ruhikanta A., additional, Luo, Roger T., additional, Thirman, Michael J., additional, Milne, Thomas, additional, Bonifer, Constanze, additional, and Mulloy, James C., additional
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- 2016
- Full Text
- View/download PDF
9. ELL-associated factor 2 (EAF2), a functional homolog of EAF1 with alternative ELL binding properties
- Author
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Federico Simone, Joseph J. Kaberlein, Roger T. Luo, Michael J. Thirman, and Paul E. Polak
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Oncogene Proteins, Fusion ,Recombinant Fusion Proteins ,Molecular Sequence Data ,Immunology ,Glutamic Acid ,Sequence Homology ,RNA polymerase II ,Transfection ,Biochemistry ,Conserved sequence ,Mice ,Bone Marrow ,Proto-Oncogenes ,Serine ,Animals ,Humans ,Amino Acid Sequence ,Binding site ,Peptide sequence ,Conserved Sequence ,Immunosorbent Techniques ,Cell Nucleus ,chemistry.chemical_classification ,Aspartic Acid ,Binding Sites ,biology ,Histone-Lysine N-Methyltransferase ,Cell Biology ,Hematology ,Blotting, Northern ,Hematopoietic Stem Cells ,Fusion protein ,Molecular biology ,Peptide Fragments ,Amino acid ,DNA-Binding Proteins ,Elongation factor ,Leukemia, Myeloid, Acute ,Cell Transformation, Neoplastic ,Retroviridae ,chemistry ,biology.protein ,Myeloid-Lymphoid Leukemia Protein ,Transcriptional Elongation Factors ,HeLa Cells ,Transcription Factors - Abstract
The (11;19)(q23;p13.1) translocation in acute leukemia results in the formation of an MLL-ELL fusion protein. ELL is an RNA polymerase II elongation factor that interacts with the recently identified EAF1 protein. To characterize the normal functions of ELL and its aberrant activities when fused to MLL, we isolated a second protein that interacts with ELL named EAF2 for ELLAssociated Factor 2. EAF2 is highly homologous to EAF1, with 58% identity and 74% amino acid conservation. Using specific antibodies generated to EAF2, we coimmunoprecipitated ELL and EAF2 from multiple cell lines. Confocal microscopy revealed that endogenous EAF2 and ELL colocalized in a nuclear speckled pattern. Database comparisons with EAF2 identified a region with a high content of serine, aspartic acid, and glutamic acid residues that is conserved with EAF1 and exhibited amino acid similarity with several translocation partner proteins of MLL, including AF4 and ENL. We found that EAF2 and EAF1 both contain transcriptional activation domains within this region. Using retroviral bone marrow transduction, we observed that a heterologous fusion of EAF2 to MLL immortalized hematopoietic progenitor cells. In contrast to EAF1, EAF2 does not bind to the carboxy-terminus of ELL. We identified a protein-protein interaction domain within the amino-terminus of ELL that binds to both EAF1 and EAF2. This amino-terminal interaction domain is disrupted in the formation of the MLL-ELL fusion protein. Thus, MLL-ELL retains an interaction domain for EAF1 but not for EAF2. Taken together, these data suggest that MLL-ELL may disrupt the normal protein-protein interactions of ELL.
- Published
- 2003
10. The Transcriptome Heterogeneity of MLL-Fusion ALL Is Driven By Fusion Partners Via Distinct Chromatin Binding
- Author
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Michael J. Thirman, Ruhikanta A. Meetei, Anetta Ptasinska, Roger T. Luo, Salam A. Assi, James C. Mulloy, Jon Kerry, Thomas A. Milne, Constanze Bonifer, and Shan Lin
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Regulation of gene expression ,Chromatin binding ,Immunology ,Cell Biology ,Hematology ,Computational biology ,Gene signature ,Biology ,Biochemistry ,Fusion protein ,Chromatin ,Transcriptome ,hemic and lymphatic diseases ,Histone methyltransferase ,neoplasms ,Gene - Abstract
Chromosome rearrangements involving the Mixed Lineage Leukemia (MLL) gene on chromosome 11q23 account for 15-20% of acute lymphoid leukemia (ALL) and confer poor prognosis. Such rearrangements generate the MLL-fusion proteins, in which the N-terminus of the MLL protein mediating chromatin interactions is fused with one of more than 70 different partner proteins. Proteins that are frequently involved in MLL translocations, including AF4, ENL, AF9 and AF10, were identified as components of the super elongation complex (SEC) or DOT1L complex (an H3K79 histone methyltransferase). Based on these observations, a consensus model of MLL-fusion leukemogenesis has been proposed, which suggests that all fusion proteins bind to the targets of wildtype MLL and lead to the aberrant transcriptional elongation and H3K79 methylation via the recruitment of SEC or DOT1L complex and thus the uncontrolled activation of the target genes. Therefore, regardless of the nature of fusion partners, all MLL-fusion proteins work in a similar fashion by dysregulating the same pathways. Our group has successfully established xenograft models of MLL-AF4 and MLL-AF9 B-ALL using human CD34+ hematopoietic stem and progenitor cells transduced with FLAG-tagged MLL-Af4 or MLL-AF9, which faithfully recapitulate the clinical features of the disease. We generated MLL-Af4 and -AF9 ALL using matched units of human CD34+ cells to directly test the consensus model. Interestingly, although having the same genetic background, the immunophenotype of the two ALL are unique, with CD34 expressed only in MLL-Af4 but not -AF9 cells. The transcriptomes of the two ALL were analyzed by RNAseq and dysregulated genes were defined by comparison with the transcriptome of normal pro-B cells (p≤0.05, fold-change≥2). Strikingly, only 40% of MLL-Af4-regulated genes overlap with those of MLL-AF9. This transcriptome heterogeneity is mirrored in clinical samples, where the gene signature generated from our model leukemia can be utilized to accurately classify patient samples in unsupervised hierarchical clustering analysis, with MLL-AF4 patient samples readily distinguishable from MLL-AF9 samples. To identify the mechanisms accounting for this heterogeneity, we performed ChIP-seq analysis using anti-FLAG antibody to compare the chromatin occupancy of MLL-Af4 and MLL-AF9 in our model ALL cells. The MLL-Af4 ChIP-seq signal displayed a clear correlation with those of published MLL-AF4 ChIP-seq datasets from patient-derived cell lines, in the range of 70-90%, highlighting the faithfulness of our model. Surprisingly, MLL-Af4 shows a distinct genome-wide distribution compared to MLL-AF9, with only 20% of MLL-Af4 peaks and 35% of MLL-AF9 peaks overlapping. In contrast to MLL-Af4 which predominantly binds to promoter regions, MLL-AF9 has a relatively greater enrichment at intra- and inter-genic regions. Intriguingly, MLL-AF9 tends to bind at repetitive sequences in introns, suggesting these repetitive sequences may serve as regulatory elements for gene expression. Integration with RNAseq data reveals a significant association between differentially-expressed MLL-Af4 and -AF9 targets and specific chromatin binding of different MLL-fusion proteins. This data demonstrates that chromatin binding is not solely controlled by the MLL portion of the fusion protein and that differential target recognition of different fusion proteins is one molecular mechanism driving gene expression heterogeneity. To test whether distinct co-factor recruitment by MLL-fusions adds another layer regulating gene expression heterogeneity beyond DNA binding, we purified the core complexes of MLL-Af4 and -AF9 from ALL cells by anti-FLAG immunoprecipitation and analyzed by mass spectrometry. These experiments identified both common and fusion-specific interacting proteins. MLL-Af4 showed a higher affinity with SEC component EAF2 but a lower affinity with DOT1L compared to MLL-AF9, suggesting that MLL-fusions have distinct associations with complex components which may achieve differential gene regulation. In summary, our data question the consensus model of MLL-fusion leukemia and emphasize that MLL-fusion ALL is a heterogeneous disease. These findings have important implications for therapy development as each MLL-fusion leukemia could have its own Achilles' heel and customized therapy may need to be introduced for each type of disease. Disclosures No relevant conflicts of interest to declare.
- Published
- 2016
11. Lymphoid Lineage Preference of MLL-AF4 Is Revealed in a Species-Specific Model
- Author
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Lin, Shan, primary, Luo, Roger T., additional, Wunderlich, Mark, additional, Kaberlein, Joseph J., additional, Rayes, Ahmad, additional, Anastasi, John, additional, O'Brien, Maureen M, additional, Mulloy, James C., additional, and Thirman, Michael J., additional
- Published
- 2015
- Full Text
- View/download PDF
12. Eotaxin Induces a Rapid Release of Eosinophils and Their Progenitors From the Bone Marrow
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Paul D. Collins, Timothy J. Williams, Sara M. Rankin, and Roger T. Palframan
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Eotaxin ,Chemokine ,biology ,Immunology ,Cell Biology ,Hematology ,respiratory system ,Granulocyte ,Eosinophil ,Biochemistry ,Allergic inflammation ,medicine.anatomical_structure ,biology.protein ,medicine ,Eosinophilia ,Bone marrow ,medicine.symptom ,Interleukin 5 - Abstract
The CC-chemokine eotaxin is a potent eosinophil chemoattractant that stimulates recruitment of eosinophils from the blood to sites of allergic inflammation. Mobilization from the bone marrow is an important early step in eosinophil trafficking during the allergic inflammatory response. In this paper we examine the potential of eotaxin to mobilize eosinophils and their progenitors from bone marrow. Eotaxin stimulated selective, dose-dependent chemotaxis of guinea pig bone marrow eosinophils in vitro. Intravenous injection of eotaxin (1 nmol/kg) into guinea pigs in vivo stimulated a rapid blood eosinophilia (from 3.9 ± 1.2 to 28 ± 9.9 × 104eosinophils/mL at 30 minutes) and a corresponding decrease in the number of eosinophils retained in the femoral marrow (from 9.0 ± 0.8 to 4.8 ± 0.8 × 106 eosinophils per femur). To show a direct release of eosinophils from the bone marrow an in situ perfusion system of the guinea pig femoral bone marrow was developed. Infusion of eotaxin into the arterial supply of the perfused femoral marrow stimulated a rapid and selective release of eosinophils into the draining vein. In addition, eotaxin stimulated the release of colony-forming progenitor cells. The cytokine interleukin-5 was chemokinetic for bone marrow eosinophils and exhibited a marked synergism with eotaxin with respect to mobilization of mature eosinophils from the femoral marrow. Thus, eotaxin may be involved in both the mobilization of eosinophils and their progenitors from the bone marrow into the blood and in their subsequent recruitment into sites of allergic inflammation.
- Published
- 1998
13. Lymphoid Lineage Preference of MLL-AF4 Is Revealed in a Species-Specific Model
- Author
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Mark Wunderlich, Shan Lin, James C. Mulloy, Ahmad Rayes, John Anastasi, Maureen M. O'Brien, Michael J. Thirman, Joseph J. Kaberlein, and Roger T. Luo
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Myeloid ,Immunology ,CD33 ,CD34 ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,CD19 ,Leukemia ,Haematopoiesis ,Immunophenotyping ,medicine.anatomical_structure ,hemic and lymphatic diseases ,medicine ,Cancer research ,biology.protein ,neoplasms ,Interleukin 3 - Abstract
The Mixed Lineage Leukemia (MLL) gene on chromosome 11q23 is fused by reciprocal translocation to a diverse group of partner genes that drive both acute myeloid and acute lymphoid leukemia (AML and ALL). As a result of the t(4;11)(q21;q23), MLL fuses to AF4 (also referred to as AFF1), one of the most common MLL fusion partner proteins. Unlike several other MLL fusions that are frequently identified in AML, for example MLL-AF9 caused by t(9;11)(p22;q23), MLL-AF4 is almost exclusively associated with B-cell ALL with a pro-B immunophenotype. It is the most frequent MLL fusion in ALL and accounts for 10-15% of ALL cases. Patients with t(4;11) have a poor prognosis compared with other cytogenetically defined subsets. Although many MLL-fusion leukemia models have successfully been established, it has not been possible to generate a t(4;11) pro-B leukemia model that accurately recapitulates the human disease, hampering research into the molecular mechanisms that underlie the development of this subtype of leukemia. Here we present a faithful human cell based model of t(4;11) pro-B-ALL that fully recapitulates the immunophenotypic and molecular aspects of the human disease. Transduced with a modified MLL-AF4 fusion gene, human hematopoietic CD34+ cells successfully initiate ALL in a xenograft system with high penetrance. The leukemia cells have a CD19+CD34+ pro-B immunophenotype and are CD10(-), a common feature in MLL-AF4 patients. The effect of the oncogene is species-specific, as retroviral transduction and transplantation of murine hematopoietic cells with MLL-AF4 results in only AML using either lymphoid or myeloid conditions. An MLL-AF4 specific gene signature derived from patients is significantly enriched in our model cells, as shown by RNAseq, and the model samples group tightly with MLL-AF4 patients, even when compared with other MLL-fusions in unsupervised hierarchical clustering analysis. Interestingly, using gene profiles of normal pro-B and pre-B cells as reference, our MLL-AF4 leukemia cells show strong enrichment for pro-B genes, while instead, pre-B but not pro-B genes are overrepresented in our MLL-AF9 B-ALL leukemia cells. This differential developmental stage blockage of MLL-fusions is also reflected in patient samples. More strikingly, in accordance with the distinct lineage bias of MLL-fusions observed in the clinic, human cells expressing MLL-AF4 have a strong predilection for the lymphoid lineage and a demonstrated resistance to reprogramming in response to myeloid signals compared to human cells expressing MLL-AF9. This difference in lineage predisposition of MLL-AF4 compared to MLL-AF9 can be attributed to differential effects on lineage-specific gene expression. Under myeloid-priming conditions, phenotypically (CD33+CD19-) and morphologically myeloid MLL-AF4 cells are still able to initiate pro-B ALL in immunodeficient mice, while only AML is generated by MLL-AF9 myeloid cells. Accordingly, an active lymphoid molecular program with lower expression of critical myeloid genes is observed in MLL-AF4 myeloid cells compared to MLL-AF9 myeloid cells. Interestingly, we find that the polycomb gene BMI1, which was reported to be critical to prevent lymphoid priming in normal hematopoiesis, is expressed at significantly lower levels in MLL-AF4 than in MLL-AF9 myeloid cells. Strikingly, this decreased BMI1 expression is evident in primary B-ALL patient samples as well, with MLL-AF9 B-ALL samples demonstrating increased BMI1 relative to MLL-AF4. Reintroduction of BMI1 into MLL-AF4 cells enables AML generation with variable penetrance, while control vector transduced cells always result in B-ALL. Our results demonstrate that lineage fate in response to MLL-fusion protein expression involves a complex interplay of oncogene, intra- and extra-cellular microenvironmental cues. In addition, our data clearly demonstrate the species specificity associated with the t(4;11) oncogene and highlight the limitations of using murine cells in human disease modeling. The model provides a valuable tool to unravel the pathogenesis of MLL-AF4 leukemogenesis. Disclosures Thirman: AbbVie: Research Funding; Pharmacyclics: Research Funding; Gilead: Research Funding.
- Published
- 2015
14. MLL fusion proteins preferentially regulate a subset of wild-type MLL target genes in the leukemic genome
- Author
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Shuangli Mi, Jingfang Dong, Joseph J. Kaberlein, Jun Wu, George Wu, Roger T. Luo, Ryan J. Mattison, Shyam Prabhakar, Michael J. Thirman, Hongkai Ji, Fuhong He, and Qianfei Wang
- Subjects
Oncogene Proteins, Fusion ,Immunology ,Biology ,Biochemistry ,Methylation ,Histones ,Mice ,hemic and lymphatic diseases ,Cell Line, Tumor ,Gene expression ,Tumor Cells, Cultured ,Animals ,Humans ,Transcription factor ,Gene ,neoplasms ,Genetics ,Regulation of gene expression ,Homeodomain Proteins ,Leukemia ,Myeloid Neoplasia ,Gene Expression Regulation, Leukemic ,Intracellular Signaling Peptides and Proteins ,Nuclear Proteins ,Cell Biology ,Hematology ,Histone-Lysine N-Methyltransferase ,Fusion protein ,Up-Regulation ,Histone ,Genetic Loci ,Histone methyltransferase ,biology.protein ,Myeloid-Lymphoid Leukemia Protein ,Protein Tyrosine Phosphatases ,Protein Binding - Abstract
MLL encodes a histone methyltransferase that is critical in maintaining gene expression during embryonic development and hematopoiesis. 11q23 translocations result in the formation of chimeric MLL fusion proteins that act as potent drivers of acute leukemia. However, it remains unclear what portion of the leukemic genome is under the direct control of MLL fusions. By comparing patient-derived leukemic cell lines, we find that MLL fusion-bound genes are a small subset of that recognized by wild-type MLL. In an inducible MLL-ENL model, MLL fusion protein binding and changes in H3K79 methylation are limited to a specific portion of the genome, whereas wild-type MLL distributes to a much larger set of gene loci. Surprisingly, among 223 MLL-ENL–bound genes, only 12 demonstrate a significant increase in mRNA expression on induction of the fusion protein. In addition to Hoxa9 and Meis1, this includes Eya1 and Six1, which comprise a heterodimeric transcription factor important in several developmental pathways. We show that Eya1 has the capacity to immortalize hematopoietic progenitor cells in vitro and collaborates with Six1 in hematopoietic transformation assays. Altogether, our data suggest that MLL fusions contribute to the development of acute leukemia through direct activation of a small set of target genes.
- Published
- 2011
15. Dasatinib (BMS-354825) inhibits KITD816V, an imatinib-resistant activating mutation that triggers neoplastic growth in most patients with systemic mastocytosis
- Author
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Cem Akin, Francis Y. Lee, Marjolein L. Donker, Roger T. Luo, Yibin Jiang, Neil P. Shah, and Faculteit Medische Wetenschappen/UMCG
- Subjects
Models, Molecular ,Protein Conformation ,Dasatinib ,TYROSINE KINASE ,Ligands ,Biochemistry ,Piperazines ,Mice ,hemic and lymphatic diseases ,MAST-CELL LEUKEMIA ,Systemic mastocytosis ,ABL ,GASTROINTESTINAL STROMAL TUMORS ,Myeloid leukemia ,Hematology ,PROTOONCOGENE C-KIT ,Protein-Tyrosine Kinases ,Mast cell leukemia ,Proto-Oncogene Proteins c-kit ,Benzamides ,Imatinib Mesylate ,Tyrosine kinase ,medicine.drug ,Immunology ,Biology ,Structure-Activity Relationship ,Mastocytosis, Systemic ,Cell Line, Tumor ,medicine ,Animals ,Humans ,Computer Simulation ,WILD-TYPE ,CHRONIC MYELOID-LEUKEMIA ,Protein Kinase Inhibitors ,ABL GENE ,ACUTE LYMPHOBLASTIC-LEUKEMIA ,Cell Proliferation ,Binding Sites ,Imatinib ,Cell Biology ,medicine.disease ,Protein Structure, Tertiary ,KINASE DOMAIN MUTATIONS ,Thiazoles ,Imatinib mesylate ,Pyrimidines ,Drug Resistance, Neoplasm ,Case-Control Studies ,Mutation ,Cancer research ,ACQUIRED-RESISTANCE - Abstract
Mastocytosis is associated with an activating mutation in the KIT oncoprotein (KITD816V) that results in autophosphorylation of the KIT receptor in a ligand-independent manner. This mutation is inherently resistant to imatinib and, to date, there remains no effective curative therapy for systemic mastocytosis associated with KITD816V. Dasatinib (BMS-354825) is a novel orally bioavailable SRC/ABL inhibitor that has activity against multiple imatinib-resistant BCR-ABL isoforms in vitro that is presently showing considerable promise in early-phase clinical trials of chronic myeloid leukemia (CML). Pharmacokinetic analysis suggests that high nanomolar concentrations of dasatinib can be achieved safely in humans. In this study, we demonstrate significant inhibitory activity of dasatinib against both wild-type KIT and the KITD816V mutation in the nanomolar range in in vitro and cell-based kinase assays. Additionally, dasatinib leads to growth inhibition of a KITD816V-harboring human masto-cytosis cell line. Significantly, dasatinib selectively kills primary neoplastic bone marrow mast cells from patients with systemic mastocytosis while sparing other hematopoietic cells. Computer modeling suggests that the KITD816V mutation destabilizes the inactive conformation of the KIT activation loop to which imatinib binds, but it is not predicted to impair binding of KIT by dasatinib. Based upon our results, further evaluation of dasatinib for the treatment of systemic masto-cytosis in clinical trials is warranted. Moreover, dasatinib may be of clinical utility in other disease settings driven by activating KIT mutations. (Blood. 2006;108:286-291)
- Published
- 2006
16. The full transforming capacity of MLL-Af4 is interlinked with lymphoid lineage commitment.
- Author
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Shan Lin, Luo, Roger T., Shrestha, Mahesh, Thirman, Michael J., and Mulloy, James C.
- Subjects
- *
LYMPHOBLASTIC leukemia treatment , *LYMPHOBLASTIC leukemia , *CHROMOSOMAL rearrangement , *CHIMERIC proteins , *GENE expression , *PATIENTS - Abstract
Chromosome rearrangements involving the mixed-lineage leukemia gene (MLL) create MLL-fusion proteins, which could drive both acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). The lineage decision of MLL-fusion leukemia is influenced by the fusion partner and microenvironment. To investigate the interplay of fusion proteins and microenvironment in lineage choice, we transplanted human hematopoietic stem and progenitor cells (HSPCs) expressingMLL-AF9 or MLL-Af4 into immunodeficient NSGS mice, which strongly promote myeloid development. Cells expressing MLL-AF9 efficiently developed AML in NSGS mice. In contrast, MLL-Af4 cells, which were fully oncogenic under lymphoid conditions present in NSG mice, displayed compromised transformation capacity in a myeloid microenvironment. MLL-Af4 activated a self-renewal program in a lineage-dependent manner, showing the leukemogenic activity of MLL-Af4 was interlinked with lymphoid lineage commitment. The C-terminal homology domain (CHD) of Af4 was sufficient to confer this linkage. Although the MLL-CHD fusion protein failed to immortalize HSPCs in myeloid conditions in vitro, it could successfully induce ALL in NSG mice. Our data suggest that defective self-renewal ability and leukemogenesis of MLL-Af4 myeloid cells could contribute to the strong B-cell ALL association of MLL-AF4 leukemia observed in the clinic. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
17. Low-penetrance genetic susceptibility and resistance loci implicated in the relative risk for radiation-induced acute myeloid leukemia in mice
- Author
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Roger T. Snowden, A.L. Knight, Mark Plumb, Helen Cleary, Emma Boulton, and Clare Cole
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Risk ,Myeloid ,Genotype ,Genetic Linkage ,Immunology ,Locus (genetics) ,Penetrance ,Biology ,Biochemistry ,Mice ,Genetic linkage ,hemic and lymphatic diseases ,medicine ,Animals ,Genetic Predisposition to Disease ,Inbreeding ,Genetics ,Leukemia, Radiation-Induced ,Mice, Inbred BALB C ,Myeloid leukemia ,Cell Biology ,Hematology ,medicine.disease ,Molecular biology ,Mice, Inbred C57BL ,Leukemia ,Haematopoiesis ,Leukemia, Myeloid, Acute ,medicine.anatomical_structure ,Mice, Inbred DBA ,Mice, Inbred CBA ,Bone marrow ,Stem cell - Abstract
Inbred CBA/H mice are susceptible to radiation-induced acute myeloid leukemia (r-AML), and C57BL/6 mice are resistant. A genome-wide screen for linkage between genotype and phenotype (r-AML) of 67 affected (CBA/H × C57BL/6)F1 × CBA/H backcross mice has revealed at least 2 suggestive loci that contribute to the overall lifetime risk for r-AML. Neither is necessary or sufficient for r-AML, but relative risk is the net effect of susceptibility (distal chromosome 1) and resistance (chromosome 6) loci. An excess of chromosome 6 aberrations in mouse r-AML and bone marrow cells up to 6 months after irradiation in vivo suggests the locus confers a proliferative advantage during the leukemogenic process. The stem cell frequency regulator 1 (Scfr1) locus maps to distal chromosome 1 and determines the frequency of hemopoietic stem cells (HSCs) in inbred mice, suggesting that target size may be one factor in determining the relative susceptibility of inbred mice to r-AML.
- Published
- 2002
18. Mimetic Peptide to CDK Inhibitor p16 INK4a Induces Cell Death In Mantle Cell Lymphoma Cells: A New Strategy to Eradicate Minimal Residual Disease
- Author
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Greg Koval, Steven F. Dowdy, Amittha Wickrema, Greg Malnassy, Roger T. Luo, Michael J. Thirman, Hongtao Liu, Hui Liu, Pamela Ihonor, Wendy Stock, and Koen van Besien
- Subjects
biology ,Immunology ,Cell Biology ,Hematology ,Cell cycle ,medicine.disease ,Biochemistry ,Minimal residual disease ,Cyclin D1 ,Autologous stem-cell transplantation ,Cyclin-dependent kinase ,hemic and lymphatic diseases ,biology.protein ,medicine ,Cancer research ,Mantle cell lymphoma ,Stem cell ,Progenitor cell - Abstract
Abstract 3921 Autologous stem cell transplantation (ASCT) is an effective treatment strategy for Mantle Cell Lymphoma (MCL) that has been shown to improve disease free survival. However, data from recent trials suggest that the presence of minimal residual disease (MRD) contributes to relapse following current intensive treatment strategies, including ASCT. Thus, we sought to test a highly selective approach to eradication of MRD in stem cell collections from MCL patients using targeted small peptides that disrupt the aberrant cyclin/CDKs interactions that are involved in the pathogenesis of MCL. In the current study, the efficacy of a mimetic peptide to the CDK inhibitor p16INK4a to eliminate MRD was tested. Using (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) cell viability assays, we demonstrated that a transducible TAT-p16 mimetic peptide was able to induce cell death in the MCL cell line, Jeko-1, in a dose and time-dependent fashion (see Figure). A TAT-p16 mimetic was able to induce cell death in approximately 60% of Jeko-1 cells after 4 hours of incubation, and approximately 80% of Jeko-1 cells at 24 hours. In contrast, the same dose of TAT-p16 mimetic had no effect on cell survival when it was incubated with CD34+ rich apheresis samples from healthy donors. Furthermore, CFU-GM, BFU-E colony assays showed that there was no toxicity of the TAT-p16 mimetic when compared to untreated cells or cells treated with a control, scrambled TAT-p16 peptide. TAT-p16 mimetic did not impair erythroblast differentiation but induced mild apoptotic cell death in the erythroblast cells. The strategy was further tested using a CD34+ enriched apheresis sample collected from a patient with MCL with documented MRD who had been treated on a clinical trial of ASCT for MCL. Using Real-time quantitative PCR for IgH and BCL1 gene rearrangements, we demonstrated that the TAT-p16 mimetic was able to reduce MRD level by 40% compared with the scrambled TAT-p16 peptide. These data suggest that the use of a peptide-mimetic to p16 selectively and effectively can reduce MRD in MCL cells and in an apheresis (stem cell) sample from a patient with MCL. Treatment with the peptide allowed differentiation of CD34+ progenitor cells. A combinational approach using additional targeted cell cycle regulatory peptides that block p21CIP1/WAF1 and cyclin D2 is being explored to optimize the efficacy of this purging technique. Thus, this novel strategy may be an effective, selective and non-toxic method for eradication of MRD in MCL. Disclosures: No relevant conflicts of interest to declare.
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- 2010
19. Identification of Genes Deregulated in Both Human and Murine MLLRearrangement Leukemias.
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Li, Zejuan, primary, Luo, Roger T., additional, Mi, Shuangli, additional, Sun, Miao, additional, Chen, Ping, additional, Neilly, Mary Beth, additional, Zhang, Yanming, additional, Rowley, Janet D., additional, Thirman, Michael J., additional, and Chen, Jianjun, additional
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- 2008
- Full Text
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20. MicroRNA Expression Profiles in Acute Myeloid Leukemia with Common Translocations.
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Li, Zejuan, primary, Lu, Jun, additional, Sun, Miao, additional, Mi, Shuangli, additional, Zhang, Hao, additional, Luo, Roger T., additional, Qian, Zhijian, additional, Neilly, Mary Beth, additional, Wang, Yungui, additional, Jin, Jie, additional, Zhang, Yanming, additional, Bohlander, Stefan K., additional, Larson, Richard, additional, LeBeau, Michelle, additional, Thirman, Michael J., additional, Golub, Todd R., additional, Rowley, Janet D., additional, and Chen, Jianjun, additional
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- 2007
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21. Identification of Genes Deregulated in Both Human and Murine MLLRearrangement Leukemias
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Shuangli Mi, Jianjun Chen, Zejuan Li, Roger T. Luo, Michael J. Thirman, Janet D. Rowley, Mary Beth Neilly, Ping Chen, Yanming Zhang, and Miao Sun
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Genetics ,Candidate gene ,ABL ,Immunology ,Myeloid leukemia ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Molecular biology ,Leukemia ,hemic and lymphatic diseases ,Gene expression ,medicine ,Serial analysis of gene expression ,Hox gene ,Gene - Abstract
Chromosome translocations are among the most common genetic abnormalities in human leukemia. The mixed lineage leukemia (MLL) gene was identified as a common target of chromosomal translocations associated with human acute leukemias; it is located on human chromosome 11 band q23 and on mouse chromosome 9. More than 50 different loci are rearranged in11q23 leukemias involving MLL, resulting in either acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). In general, MLL rearrangements are associated with a poor prognosis. MLL-ELL and MLL-ENL resulting from t(11;19)(q23;p13.1) and t(11;19)(q23;p13.3) respectively are two common examples of these rearrangements. These two fusions are frequently involved in human AML, while MLL-ENL is also involved in human ALL. There is a common observation that important biological properties are often conserved across species. Cross-species sequence comparison has been widely used to infer gene function, but it is becoming apparent that sequence similarity is not always proportional to functional similarity. To determine the function of a gene precisely, therefore, we need to investigate not only its sequence characteristics but also its expression characteristics. Model organisms have contributed substantially to our understanding of the etiology of human disease and the development of new treatment methodologies. However, although genetically engineered mouse leukemia models have been established for many years, there are few systematic studies to identify and study the genes that exhibit similar abnormal expression patterns in both human leukemia and mouse leukemia model cells. To perform an interspecies gene expression comparative study in leukemia, we used the serial analysis of gene expression (SAGE) technique to examine gene expression profiles between MLL-ELL or MLL-ENL myeloid leukemia progenitor cells and normal myeloid progenitor cells in both humans and mice. We obtained 484,303 total SAGE tags for the nine samples and a total of 103,899 unique SAGE tags from five human and 60,993 from four mouse samples. We identified 88 genes that appeared to be significantly deregulated (32 up- and 56 down-regulated) in both human and murine MLL-ELL and/or MLL-ENL leukemia. Fifty-seven genes have not been reported previously. A large-scale quantitative real-time PCR (qPCR) assay was performed to validate the candidate genes, and 84% (36/43) of the tested SAGE candidate genes were confirmed. The most up-regulated genes include several HOX genes (e.g., HOX A5, HOXA9 and HOXA10) and a HOX cofactor MEIS1; their overexpression is a hallmark of MLL-rearrangement leukemia. The top down-regulated genes include LTF, LCN2, MMP9, S100A8, S100A9, PADI4, TGFBI and CYBB. Remarkably, up-regulated genes have a much higher percentage of enrichment in Gene Ontology (GO) terms related to gene expression and transcription, whereas down-regulated genes are more enriched in GO terms related to apoptosis, signal transduction and response. Thus, the up-regulation of genes responsible for gene expression and transcription but down-regulation of genes responsible for apoptosis, signal transduction and response, can promote cell proliferation and inhibit apoptosis, and thereby contribute to the development of leukemia. We showed that the CpG islands of several significantly down-regulated genes including LIF, TGFBI and G0S2 are hypermethylated. We also examined the expression of microRNAs from the mir-17–92 cluster, which are overexpressed in human MLL-rearrangement leukemias, and showed that seven individual microRNAs (i.e., miR-17-5p, miR-17-3p, miR-18a, miR-19a, miR-20a, miR-19b and miR-92) within this cluster are also overexpressed in mouse MLL-rearrangement leukemia cells. Nineteen putative targets (i.e., APP, RASSF2, SH3BP5, DBN1, ELK3, FLT1, GNAI1, HIF1A, ITGA6, MN1, POU4F1, RB1, RGL1, RNF167, SASH1, SLC24A3, TNFRSF21, WWP1 and YES1) of these microRNAs were reported and/or confirmed by our qPCR to be down-regulated in MLL-rearrangement leukemias. We further confirmed both APP and RASSF2 as direct targets of miR-17 through luciferase reporter and mutagenesis assay. The identification and validation of gene expression changes in MLL-rearrangement human and murine leukemia provides important insights into the genetic pathways that are important for MLL fusion-induced leukemogenesis.
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- 2008
22. Complex Interaction of the MoeA-N and MoeA-C Domains of Gephyrin Is Required for the Immortalization Activity of MLL-Gephyrin.
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Imamura, Toshihiko, primary, Luo, Roger T., additional, and Thirman, Michael J., additional
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- 2006
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23. Identification of Genes Abnormally Expressed in Human MLL-AF4 Leukemia.
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Mi, Shuangli, primary, Sun, Miao, primary, Li, Zejuan, primary, Luo, Roger T., primary, Jayathilaka, Nimanthi, primary, Neilly, Mary Beth, primary, Kocherginsky, Masha, primary, Zhang, Yanming, primary, Lee, Sanggyu, primary, Karrison, Theodore G., primary, Thirman, Michael J., primary, Wang, San Ming, primary, Rowley, Janet D., primary, and Chen, Jianjun, primary
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- 2006
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24. Identification of Genes Abnormally Expressed in Both Human and Murine MLL-ELL and/or MLL-ENL Leukemia.
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Li, Zejuan, primary, Sun, Miao, additional, Mi, Shuangli, additional, Luo, Roger T., additional, Bao, Jingyue, additional, Neilly, MeryBeth, additional, Jayathilaka, Nimanthi, additional, Zhang, Yanming, additional, Johnson, Deborah S., additional, Wang, Lili, additional, Lavau, Catherine, additional, Tseng, Charles, additional, Zhang, Xiuqing, additional, Wang, Jian, additional, Yu, Jun, additional, Yang, Huanming, additional, Thirman, Michael J., additional, Wang, San Ming, additional, Rowley, Janet D., additional, and Chen, Jianjun, additional
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- 2006
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25. Genes Similarly Abnormally Expressed in Both Human and Murine MLL-Associated Leukemia.
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Chen, Jianjun, primary, Sun, Miao, primary, Luo, Roger T., primary, Bao, Jingyue, primary, Kocherginsky, Masha, primary, Johnson, Deborah S., primary, Wang, Lili, primary, Lee, Sanggyu, primary, Lavau, Catherine, primary, Karrison, Theodore G., primary, Tseng, Charles, primary, Zhang, Xiuqing, primary, Wang, Jian, primary, Yu, Jun, primary, Yang, Huanming, primary, Thirman, Michael J., primary, Wang, San Ming, primary, and Rowley, Janet D., primary
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- 2005
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26. MicroRNA Expression Profiles in Acute Myeloid Leukemia with Common Translocations
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Jun Lu, Shuangli Mi, Hao Zhang, Richard A. Larson, Stefan K. Bohlander, Janet D. Rowley, Yungui Wang, Jie Jin, Jianjun Chen, Michael J. Thirman, Zhijian Qian, Yanming Zhang, Miao Sun, Mary Beth Neilly, Todd R. Golub, Michelle M. LeBeau, Roger T. Luo, and Zejuan Li
- Subjects
Genetics ,Acute leukemia ,Immunology ,Myeloid leukemia ,Chromosomal translocation ,Cell Biology ,Hematology ,Biology ,Biochemistry ,Gene expression profiling ,microRNA ,Cancer research ,Epigenetics ,DNA microarray ,Gene - Abstract
Acute myeloid leukemia (AML) is the most common type of acute leukemia in adults. It is estimated that 13,410 cases will be diagnosed and 8,990 will die of AML in the United States in 2007 (http://seer.cancer.gov). AML is a genetically diverse hematopoietic malignancy with variable response to treatment. Expression profiling of protein-coding genes using DNA microarray in AML has resulted in inconsistent data from different laboratories. Therefore, further validation of these observations in large cohorts and in independent studies is definitely required before clinical application becomes feasible. Recently, Golub and colleagues described a new, bead-based flow cytometric microRNA (miRNAs, miRs) expression profiling method that could successfully classify tumors. MiRNAs are endogenous ∼22 nucleotide non-coding RNAs, which can function as oncogenes and tumor suppressors. To provide new insights into the complex genetic alterations in leukemogenesis and to identify novel markers for diagnosis and treatment of AML, we performed a genome-wide analysis of miRNA expression profiles using the bead-based method on 54 AML samples with common translocations including t(15;17), t(8;21), inv(16), and 11q23 rearrangement, along with normal controls. In both unsupervised and supervised hierarchical cluster analyses, we observed that t(15;17) samples grouped together as one cluster, as do the 11q23 rearrangement samples. Interestingly, t(8;21) and inv(16), both CBF (core-binding factor) AMLs, grouped together as a unique cluster. Forty-one miRNAs exhibited significantly differential expression between different subtypes of AMLs, and/or between AMLs and normal controls. Notably, expression signature of a minimal number of two, three, and seven miRNAs could be used for class prediction of CBF, t(15;17), and 11q23 rearrangement AMLs, respectively, with an overall diagnostic accuracy of 94–96%. We further showed that overexpression of the two discriminatory miRNAs in CBF AML is associated with epigenetic regulation, rather than DNA copy number amplification. Moreover, several important target genes of these discriminatory miRNAs have also been validated. We are currently exploring the role of these discriminatory miRNAs and their critical target genes in the development of AML using in vitro and in vivo models. This work will enhance our understanding of the biological role of these miRNAs and their targets in leukemogenesis.
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- 2007
27. Complex Interaction of the MoeA-N and MoeA-C Domains of Gephyrin Is Required for the Immortalization Activity of MLL-Gephyrin
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Toshihiko Imamura, Michael J. Thirman, and Roger T. Luo
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chemistry.chemical_classification ,Gephyrin ,biology ,Immunology ,Cell Biology ,Hematology ,Subcellular localization ,Biochemistry ,Fusion protein ,Tubulin binding ,Amino acid ,Cell nucleus ,medicine.anatomical_structure ,chemistry ,Cytoplasm ,hemic and lymphatic diseases ,biology.protein ,medicine ,Glycine receptor - Abstract
More than 40 proteins have been identified as partners of MLL in acute leukemia. The partner proteins are categorized based on their subcellular localization into either the nucleus or cytoplasm. Recent studies have proposed that the mechanism of transformation of MLL-cytoplasmic protein fusion products is mediated by the oligomerization ability of partner proteins. Gephyrin, which is involved in synaptic anchoring of glycine receptor and certain GABAA receptor subtypes, is a rare partner of MLL in patients with AML. Using an in vivo cross linking assay, we confirmed the dimerization activity of a domain within the MoeA-N domain of Gephyrin, which is located at amino acids 456–476. However, we found that this domain was dispensable for immortalization in the methylcellulose colony-forming assay, indicating that the presence of a dimerization domain was not sufficient for transformation. We also observed that the Tubulin binding domain, which is located at amino acids 322–376, was also dispensable. To evaluate which domain of Gephyrin is critical for the immortalization activity of MLL-Gephyrin, we performed a colony-forming assay in methylcellulose and found that both the MoeA-N (amino acids 354–528) and the MoeA-C (amino acids 691–769) domains of Gephyrin are essential for immortalization. In addition, we observed that a MLL- Gephyrin (444–476) deletion mutant formed dimers using the in vivo cross linking assay. To explore the possibility that both MoeA-N and MoeA-C domains are associated with dimer formation of MLL- Gephyrin, we performed co-immunoprecipitation experiments with three fragments of Gephyrin, MoeA-N, MoCF (amino acids 532–690) and MoeA-C. Our data showed that both the MoeA-N and MoeA-C domains interacted with the MoCF domain of Gephyrin. Taken together, these findings show that Gephyrin has multiple dimerization domains, and that the interaction that links the MoeA-N, MoeA-C, and MoCF domains is critical for the immortalization activity of the MLL- Gephyrin fusion protein. Our data indicate that complex patterns of dimerization exist among MLL partners and suggest that specific types of dimerization domains might be critical for MLL-associated transforming activity.
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- 2006
28. Identification of Genes Abnormally Expressed in Human MLL-AF4 Leukemia
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Zejuan Li, Sanggyu Lee, Michael J. Thirman, Masha Kocherginsky, Mary Beth Neilly, Yanming Zhang, Janet D. Rowley, San Ming Wang, Shuangli Mi, Miao Sun, Jianjun Chen, Theodore Karrison, Nimanthi Jayathilaka, and Roger T. Luo
- Subjects
Candidate gene ,biology ,Immunology ,Salvia officinalis ,Syk ,Cell Biology ,Hematology ,medicine.disease ,Biochemistry ,Molecular biology ,CD19 ,food.food ,Cell biology ,Infant Acute Lymphoblastic Leukemia ,Leukemia ,food ,hemic and lymphatic diseases ,medicine ,biology.protein ,Bruton's tyrosine kinase ,Tyrosine kinase - Abstract
The t(4;11)(q21;q23) translocation is a hallmark of infant acute lymphoblastic leukemia (ALL), which results in the fusion of the MLL gene on chromosome 11 and the AF4 gene on chromosome 4. MLL-AF4 fusion is the most common consequence of chromosomal translocations in infant leukemia and is associated with a poor prognosis. To identify leukemia-related genes, we used the SAGE technique to compare gene expression profiles between two MLL-AF4 patient samples and one normal sample (CD19+ progenitor B cells; 216,464 tags in total). We identified 61 candidate genes that appear to be abnormally expressed in the leukemia samples (29 up- and 32 down-regulated). Remarkably, we found that many candidate genes appear to play important role in the development of B cells. In addition, many candidate genes can bind with and/or regulate other candidates in the candidate gene list. For example, SYK, BTK and BLNK can bind directly and regulate each other. SYK can also bind directly with TNFRSF1B. In addition, EBF may positively regulate BLK, while BLK can bind directly with BTK. All six of these genes are significantly down-regulated in MLL-AF4 leukemia samples. BTK, SYK and BLK are tyrosine kinases. BTK (B-cell progenitor tyrosine kinase) is a key regulator in B-lymphocyte differentiation and activation. BLK (B-lymphocyte-specific tyrosine kinase) is expressed only in B lymphocytes, and controls pre-B cell development. SYK (spleen tyrosine kinase) is widely expressed in hematopoietic cells, which can phosphorylate BLNK (B-cell linker protein). BLNK represents a central linker protein that bridges the B-cell receptor-associated kinases and may regulate B-cell function and development. EBF (early B-cell factor) is a tissue-specific and differentiation stage-specific DNA-binding protein, and mice lacking Ebf are unable develop B lymphoid cells. TNFRSF1B is strongly expressed on stimulated T and B lymphocytes. Moreover, previous studies indicate that BTK, BLK, SYK, BLNK and TNFRSF1B can positively regulate apoptosis, while BTK can also positively regulate differentiation. Thus, their down-regulation may inhibit apoptosis and differentiation, and thereby contribute to leukemogenesis. In contrast, GNA12, a transforming oncogene which can enhance proliferation and transformation and can bind directly with BTK, is significantly up-regulated in MLL-AF4 leukemia cells. Its up-regulation may also be important to leukemogenesis. Taken together, the deregulation of the important candidate genes may contribute to leukemogenesis through inhibiting apoptosis and differentiation while promoting proliferation of hematopoietic cells. We have validated the expression patterns of the candidate genes with real-time quantitative RT-PCR and are studying the functions and pathways of the validated candidate genes using RNAi and retrovirus transduction over-expression methods. In addition, we will also establish knock-in or knock-out mouse models for the most promising functional candidate genes to see the effect on the development of leukemia. Our studies will provide important insights into the complex functional pathways related to MLL rearrangements in the development of acute lymphoblastic leukemia, which may lead to more effective therapy for these leukemias.
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- 2006
29. Identification of Genes Abnormally Expressed in Both Human and Murine MLL-ELL and/or MLL-ENL Leukemia
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Jianjun Chen, Janet D. Rowley, Jian Wang, Shuangli Mi, Charles C. Tseng, Xiuqing Zhang, MeryBeth Neilly, Michael J. Thirman, Huanming Yang, Yanming Zhang, Zejuan Li, Jingyue Bao, Nimanthi Jayathilaka, Miao Sun, Roger T. Luo, Catherine Lavau, Jun Yu, Lili Wang, Deborah S. Johnson, and San Ming Wang
- Subjects
Candidate gene ,Acute leukemia ,Myeloid ,JUNB ,Immunology ,Myeloid leukemia ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Molecular biology ,Fusion gene ,Leukemia ,Haematopoiesis ,medicine.anatomical_structure ,hemic and lymphatic diseases ,medicine ,Cancer research - Abstract
Chromosome translocations are among the most common genetic abnormalities in human leukemia. Their abnormally expressed genes identify specific markers for their clinical diagnosis. Important biological properties are often conserved across species. However, although genetically engineered mouse leukemia models are well-established, few systematic studies have validated the genes that exhibit similar abnormal expression patterns in both human and mouse leukemia models. MLL-ELL and MLL-ENL fusion genes resulting from t(11;19)(q23;p13.1) and t(11;19)(q23;p13.3), respectively, are frequently involved in human acute leukemia, and in retrovirus-mediated mouse leukemia models. We used the SAGE technique to compare gene expression profiles between MLL-ELL or MLL-ENL myeloid leukemia progenitor cells and normal myeloid progenitor cells in both human and mouse. We analyzed four patient samples (two with each fusion) and two retrovirally-induced mouse leukemias containing either MLL-ELL or MLL-ENL fusions, and a leukemia cell line with an MLL-ELL fusion. 484,303 SAGE tags were identified from the nine samples, yielding 103,899 unique tags in human and 60,993 in mouse samples. We identified 40 candidate genes that appear to be abnormally expressed in both human and murine MLL-ELL leukemias (2 up- and 38 down-regulated), and 72 in both human and murine MLL-ENL leukemias (23 up and 49 down). 25 candidate genes are down-regulated in both types of leukemias, and many of them can bind with and/or regulate other candidate genes in the candidate list. For example, LCN2 can bind directly with and positively regulate MMP9; MMP9 and TMSB4X may positively regulate FOS; FOS and JUNB can bind directly and positively regulate each other. JUNB may inhibit proliferation and promote apoptosis, and it was reported that inactivation of JunB in LT-HSC leads to MPD while its inactivation in committed myeloid progenitors also predisposes to leukemia evolution. LCN2 may also positively regulate apoptosis. Meanwhile, some important candidate genes are observed only in one type of leukemia. For example, both PXN and ARHGEF1 are down-regulated only in MLL-ELL leukemias. PXN can bind directly with ARHGEF1, and the latter may inhibit proliferation. Similarly, MYB is significantly upregulated only in MLL-ENL leukemias, which was reported to play a role in MLL-ENL-mediated transformation. Taken together, some common pathways may exist in the development of both types of leukemias, whereas each may also have their own pathway. The deregulation of the important candidate genes may contribute to leukemogenesis through inhibiting apoptosis while promoting proliferation of hematopoietic cells. We have validated the expression patterns of the candidate genes, and are studying the functions and pathways of the validated candidate genes. Our studies will provide important insights into the complex functional pathways related to MLL rearrangements in the development of acute myeloid leukemia, which may lead to more effective therapy for these leukemias.
- Published
- 2006
30. Genes Similarly Abnormally Expressed in Both Human and Murine MLL-Associated Leukemia
- Author
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Jingyue Bao, Charles C. Tseng, San Ming Wang, Deborah S. Johnson, Roger T. Luo, Catherine Lavau, Jianjun Chen, Jun Yu, Janet D. Rowley, Jian Wang, Miao Sun, Sanggyu Lee, Michael J. Thirman, Huanming Yang, Xiuqing Zhang, Masha Kocherginsky, Lili Wang, and Theodore Karrison
- Subjects
Regulation of gene expression ,Genetics ,Candidate gene ,Immunology ,Myeloid leukemia ,Cell Biology ,Hematology ,Biology ,medicine.disease ,Biochemistry ,Molecular biology ,Gene expression profiling ,Leukemia ,hemic and lymphatic diseases ,Sense (molecular biology) ,medicine ,Serial analysis of gene expression ,Gene - Abstract
Although more than 50 different loci translocate to the MLL gene at chromosome band 11q23, resulting in either acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL), no unifying property is shared by all partner genes. The translocations result in a functional fusion of the N-terminal part of MLL gene and the C-terminal part of each partner gene, presumably leading to changes in the expression of the normal target genes, most of which have not been identified. Although genetically engineered mouse leukemia models have been widely used, few systematic studies have evaluated whether such models are valid equivalents of human leukemia. We used serial analysis of gene expression (SAGE) to obtain genome-wide gene expression profiles in normal myeloid progenitor cells from human CD15+ and mouse Gr-1+ cells. We also analyzed four patient samples (two with each fusion) and two retrovirally-induced mouse leukemias containing either MLL-ELL [t(11;19)(q23;p13.1)] or MLL-ENL [t(11;19)(q23;p13.3)] fusions, and a cell line from a leukemia mouse transduced with an MLL-ELL fusion. MLL-ELL and MLL-ENL fusions are frequently involved in human AML, while MLL-ENL is also seen in human ALL. 484,303 SAGE tags were identified from the nine samples (40,000 to 100,000 tags per sample), yielding 103,899 unique SAGE tags in the human and 60,993 in the mouse samples. Analysis of the SAGE data identified 43 candidate genes that appear to be abnormally expressed in both human and mouse myeloid leukemia progenitor cells with either MLL-ELL or MLL-ENL fusions (9 up-regulated and 34 down-regulated; Table 1). Increasing evidence suggests that endogenous antisense RNAs may play critical roles in gene regulation and cancer. Natural antisense RNAs include cis-encoded antisense RNAs transcribed from the opposite strand of the same genomic locus as the sense target genes, and trans-encoded antisense RNAs such as microRNAs (miRNAs) transcribed from a genomic locus different from the sense target genes. 26 of the 43 candidate genes have antisense partners (with a total of 7 cis-encoded antisense RNAs and 36 trans-encoded miRNAs) and thereby might be regulated by endogenous antisense RNAs. We are currently validating the expression pattern of the 43 candidate genes in at least 30 different human and mouse leukemia and normal control samples with quantitative RT-PCR, and measuring the level of expression of all known miRNAs via microarray in these samples. Our studies on the abnormally expressed genes and their potential antisense partners will provide important insights into the complex functional pathways related to MLL rearrangements in the development of acute leukemia, which may lead to more effective therapy for these leukemias. Table 1. Genes deregulated in both human and mouse leukemiasa Total number Up-regulated genes Down-regulated genes Genes with antisense partner(s) aThe genes have at least 3 fold difference in expression with a significance P < 0.05 between each leukemia sample and the normal control sample. In MLL-ELL fusions 21 1 20 12 In MLL-ENL fusions 33 8 25 21 In both types of fusions 11 0 11 7 Total unique genes 43 9 34 26
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- 2005
31. Eotaxin Induces a Rapid Release of Eosinophils and Their Progenitors From the Bone Marrow
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Palframan, Roger T., primary, Collins, Paul D., additional, Williams, Timothy J., additional, and Rankin, Sara M., additional
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- 1998
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32. BCL2/BCL-XLinhibition induces apoptosis, disrupts cellular calcium homeostasis, and prevents platelet activation
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Vogler, Meike, Hamali, Hassan A., Sun, Xiao-Ming, Bampton, Edward T.W., Dinsdale, David, Snowden, Roger T., Dyer, Martin J.S., Goodall, Alison H., and Cohen, Gerald M.
- Abstract
Apoptosis in megakaryocytes results in the formation of platelets. The role of apoptotic pathways in platelet turnover and in the apoptotic-like changes seen after platelet activation is poorly understood. ABT-263 (Navitoclax), a specific inhibitor of antiapoptotic BCL2 proteins, which is currently being evaluated in clinical trials for the treatment of leukemia and other malignancies, induces a dose-limiting thrombocytopenia. In this study, the relationship between BCL2/BCL-XLinhibition, apoptosis, and platelet activation was investigated. Exposure to ABT-263 induced apoptosis but repressed platelet activation by physiologic agonists. Notably, ABT-263 induced an immediate calcium response in platelets and the depletion of intracellular calcium stores, indicating that on BCL2/BCL-XLinhibition platelet activation is abrogated because of a diminished calcium signaling. By comparing the effects of ABT-263 and its analog ABT-737 on platelets and leukemia cells from the same donor, we show, for the first time, that these BCL2/BCL-XLinhibitors do not offer any selective toxicity but induce apoptosis at similar concentrations in leukemia cells and platelets. However, reticulated platelets are less sensitive to apoptosis, supporting the hypothesis that treatment with ABT-263 induces a selective loss of older platelets and providing an explanation for the transient thrombocytopenia observed on ABT-263 treatment.
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- 2011
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33. BCL2/BCL-XL inhibition induces apoptosis, disrupts cellular calcium homeostasis, and prevents platelet activation.
- Author
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Vogler, Meike, Hamali, Hassan A., Xiao-Ming Sun, Bampton, Edward T. W., Dinsdale, David, Snowden, Roger T., Dyer, Martin J. S., Goodall, Alison H., and Cohen, Gerald M.
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B cell lymphoma , *APOPTOSIS , *INTRACELLULAR calcium , *HOMEOSTASIS , *BLOOD platelet aggregation , *MEGAKARYOCYTES , *PREVENTION - Abstract
Apoptosis in megakaryocytes results in the formation of platelets. The role of apoptotic pathways in platelet turnover and in the apoptotic-like changes seen after platelet activation is poorly understood. ABT-263 (Navitoclax), a specific inhibitor of antiapoptotic BCL2 proteins, which is currently being evaluated in clinical trials for the treatment of leukemia and other malignancies, induces a dose-limiting thrombocytopenia. In this study, the relationship between BCL2/BCL-XL inhibition, apoptosis, and platelet activation was investigated. Exposure to ABT-263 induced apoptosis but repressed platelet activation by physiologic agonists. Notably, ABT-263 induced an immediate calcium response in platelets and the depletion of intracellular calcium stores, indicating that on BCL2/BCL-XL inhibition platelet activation is abrogated because of a diminished calcium signaling. By comparing the effects of ABT-263 and its analog ABT-737 on platelets and leukemia cells from the same donor, we show, for the first time, that these BCL2/BCL-XL inhibitors do not offer any selective toxicity but induce apoptosis at similar concentrations in leukemia cells and platelets. However, reticulated platelets are less sensitive to apoptosis, supporting the hypothesis that treatment with ABT-263 induces a selective loss of older platelets and providing an explanation for the transient thrombocytopenia observed on ABT-263 treatment. [ABSTRACT FROM AUTHOR]
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- 2011
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34. MLL fusion proteins preferentially regulate a subset of wild-type MLL target genes in the leukemic genome.
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Qian-fei Wang, Wu, George, Shuangli Mi, Fuhong He, Jun Wu, Jingfang Dong, Luo, Roger T., Mattison, Ryan, Kaberlein, Joseph J., Prabhakar, Shyam, Hongkai Ji, and Thirman, Michael J.
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METHYLTRANSFERASES , *HISTONES , *GENE expression , *HEMATOPOIESIS , *CELL lines , *GENOMES - Abstract
MLL encodes a histone methyltransferase that is critical in maintaining gene expression during embryonic development and hematopoiesis. 11q23 translocations result in the formation of chimeric MLL fusion proteins that act as potent drivers of acute leukemia. However, it remains unclear what portion of the leukemic genome is under the direct control of MLL fusions. By comparing patient-derived leukemic cell lines, we find that MLL fusion-bound genes are a small subset of that recognized by wild-type MLL. In an inducible MLL-ENL model, MLL fusion protein binding and changes in H3K79 methylation are limited to a specific portion of the genome, whereas wild-type MLL distributes to a much larger set of gene loci. Surprisingly, among 223 MLL-ENL-bound genes, only 12 demonstrate a significant increase in mRNA expression on induction of the fusion protein. In addition to Hoxa9 and Meis1, this includes Eya1 and Six1, which comprise a heterodimeric transcription factor important in several developmental pathways. We show that Eya1 has the capacity to immortalize hematopoietic progenitor cells in vitro and collaborates with Six1 in hematopoietic transformation assays. Altogether, our data suggest that MLL fusions contribute to the development of acute leukemia through direct activation of a small set of target genes. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
35. Histone deacetylase inhibitors impair innate immune responses to Toll-like receptor agonists and to infection.
- Author
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Roger T, Lugrin J, Le Roy D, Goy G, Mombelli M, Koessler T, Ding XC, Chanson AL, Reymond MK, Miconnet I, Schrenzel J, François P, and Calandra T
- Subjects
- Animals, Cells, Cultured, Disease Models, Animal, Down-Regulation drug effects, Down-Regulation genetics, Drug Evaluation, Preclinical, Female, Gene Expression Profiling, Gene Expression Regulation drug effects, Humans, Immunity, Innate genetics, Infections pathology, Macrophages drug effects, Macrophages immunology, Macrophages metabolism, Mice, Mice, Inbred BALB C, Microarray Analysis, Histone Deacetylase Inhibitors pharmacology, Immunity, Innate drug effects, Infections immunology, Toll-Like Receptors agonists
- Abstract
Regulated by histone acetyltransferases and deacetylases (HDACs), histone acetylation is a key epigenetic mechanism controlling chromatin structure, DNA accessibility, and gene expression. HDAC inhibitors induce growth arrest, differentiation, and apoptosis of tumor cells and are used as anticancer agents. Here we describe the effects of HDAC inhibitors on microbial sensing by macrophages and dendritic cells in vitro and host defenses against infection in vivo. HDAC inhibitors down-regulated the expression of numerous host defense genes, including pattern recognition receptors, kinases, transcription regulators, cytokines, chemokines, growth factors, and costimulatory molecules as assessed by genome-wide microarray analyses or innate immune responses of macrophages and dendritic cells stimulated with Toll-like receptor agonists. HDAC inhibitors induced the expression of Mi-2β and enhanced the DNA-binding activity of the Mi-2/NuRD complex that acts as a transcriptional repressor of macrophage cytokine production. In vivo, HDAC inhibitors increased the susceptibility to bacterial and fungal infections but conferred protection against toxic and septic shock. Thus, these data identify an essential role for HDAC inhibitors in the regulation of the expression of innate immune genes and host defenses against microbial pathogens.
- Published
- 2011
- Full Text
- View/download PDF
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