45 results on '"Minden MD"'
Search Results
2. Alterations of p53 and c-myc in the clonal evolution of malignant lymphoma
- Author
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Chang, H, primary, Benchimol, S, additional, Minden, MD, additional, and Messner, HA, additional
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- 1994
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3. Expression of the Kit and KitA receptor isoforms in human acute myelogenous leukemia
- Author
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Piao, X, primary, Curtis, JE, additional, Minkin, S, additional, Minden, MD, additional, and Bernstein, A, additional
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- 1994
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4. Role of interleukin-6 in the proliferation of human multiple myeloma cell lines OCI-My 1 to 7 established from patients with advanced stage of the disease
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Hitzler, JK, primary, Martinez-Valdez, H, additional, Bergsagel, DB, additional, Minden, MD, additional, and Messner, HA, additional
- Published
- 1991
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5. Mutation of the p53 gene in human acute myelogenous leukemia
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Slingerland, JM, primary, Minden, MD, additional, and Benchimol, S, additional
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- 1991
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6. Analysis of molecular events in leukemic cells arrested at an early stage of T-cell differentiation
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Yumura-Yagi, K, Hara, J, Terada, N, Ishihara, S, Tawa, A, Takihara, Y, Champagne, E, Minden, MD, Mak, TW, and Kawa-Ha, K
- Abstract
We analyzed the rearrangement and expression of T-cell receptor (TCR) genes, including the recently identified TCR delta gene, in 21 patients with T-lineage leukemia/lymphoma. Among 8 patients with CD3-, CD4-, and CD8- (group I), 2 patients showed germline configuration of the TCR delta, gamma, beta, and alpha genes and 1 patient demonstrated only TCR delta gene rearrangement. All nine patients with CD3-, CD4+, and/or CD8+ (group II) showed concomitant rearrangements of the TCR delta, gamma, and beta genes. TCR alpha gene rearrangement was also observed in two patients. Three of four patients with CD3+ (group III) showed rearrangement of the TCR alpha gene with deletion of both alleles or of a single allele of the TCR delta gene. With Northern blot analysis, full-length transcripts of the TCR delta gene were detected in 3 of 15 examined patients. All were restricted to group I or group II. In contrast, full-length transcripts of TCR beta and alpha were observed mainly in samples from groups II and III. Based on these findings, rearrangement of the TCR delta gene may be the earliest event in T-cell differentiation, preceding rearrangements of the other TCR genes.
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- 1989
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7. The T-cell receptor delta chain locus is disrupted in the T-ALL associated t(11;14)(p13;q11) translocation
- Author
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Champagne, E, Takihara, Y, Sagman, U, de Sousa, J, Burrow, S, Lewis, WH, Mak, TW, and Minden, MD
- Abstract
Two T-ALL patients carrying a t(11;14)(p13;q11) translocation were analyzed. Southern blotting experiments demonstrated that both patients had rearranged their J delta genes and that the translocation involved the delta locus in both cases. In one patient, cloning, restriction mapping, and sequencing showed that the translocation occurred on a D delta 1-D delta 2-J delta 2 rearranged gene. In addition, the rearrangement on chromosome 11 occurred in both patients within a segment of less than or equal to 2 kb showing the presence in this region of a point of recurrent recombination.
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- 1989
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8. Clonogenic hemopoietic precursors in bone marrow transplantation
- Author
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Messner, HA, Curtis, JE, Minden, MD, Tritchler, D, Lockwood, G, Takahashi, T, Lepine, J, Jamal, N, Tweeddale, M, and Wandl, U
- Abstract
Multilineage and single-lineage hemopoietic precursors were studied in 102 bone marrow transplant recipients and their respective donors to determine their contribution to clinical outcome as measured by time to engraftment and survival. The patient population was heterogenous with respect to diagnosis and disease status. They included individuals with acute myeloid leukemia (AML), chronic myeloid leukemia (CML), acute lymphoblastic leukemia (ALL), aplastic anemia, and a few other hematopoietic malignancies. The frequency of various clonogenic precursors in the normal donor population varied considerably. The data yielded a symmetrical distribution. In contrast, most bone marrow transplant recipients presented with significantly reduced numbers of clonogenic cells before transplantation, resulting in skewed distribution profiles. Serial studies of recipients demonstrated a significantly lower than normal level of clonogenic precursors even 3 and 4 years after transplantation. The median values and distribution profiles approximated those observed before transplantation but did not return to measurements obtained for normal donors. Patients with ALL deviated from this pattern. The median values and distribution profiles of clonogenic precursors before transplantation approximated the pattern of normal donors. The frequency of clonogenic progenitors after transplantation, however, remained significantly lower than that of their respective donor or pretransplant values. Cell cycle studies performed after normalization of peripheral blood hematopoietic parameters demonstrated for most recipients that a higher than normal proportion of multipotent cells was in S-phase (P = .011). By univariate and multivariate approaches, clonogenic precursors and clinical parameters were assessed for their contributions to clinical outcome as measured by time to engraftment and survival time. The number of nucleated cells in the transplant inoculum contributed to survival independent of other risk factors. Patients with a higher cell load had a higher probability of surviving than did patients with a lower cell concentration in the transplant inoculum (P = .042). The frequency of clonogenic precursors in the transplant inoculum altered neither survival nor time to engraftment. The time to engraftment was significantly influenced by the frequency of clonogenic megakaryocyte precursors (CFU-M) observed in recipients prior to transplantation (P = .003). Patients with high values engrafted faster than did patients with a low frequency of CFU-M. This was independent of both diagnosis and disease status of the patients at time of transplantation.
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- 1987
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9. Chromosome-mediated transfer of the malignant phenotype by human acute myelogenous leukemic cells
- Author
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Minden, MD, Gusella, JF, and Housman, D
- Abstract
Acute myelogenous leukemia (AML) is a malignancy of the myeloid cells of the bone marrow. Recently, a number of groups have demonstrated that it is possible to study the malignant phenotype at the level of DNA through gene transfer experiments. We have used such an approach to determine whether it is possible to transfer the malignant phenotype of anchorage independence from human AML cells to anchorage-dependent rodent cells, using chromosomes as the source of genetic information. We found that chromosomes isolated from leukemic cell lines were capable of transferring the malignant phenotype of anchorage independence, whereas chromosomes derived from the lymphocytes of normal individuals were not active in this assay. Using Southern blot analysis of the DNA from transferants, we were able to show that the transfer of anchorage independence correlated with the presence of human DNA in the transferants. The pattern of human DNA in the transferants derived from different transfection experiments is compared.
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- 1984
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10. The structure of the T cell antigen receptor genes in normal and malignant T cells
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Minden, MD and Mak, TW
- Abstract
In this review the genomic structure and the RNA transcripts of the alpha and beta chain of the T cell antigen receptor have been discussed. Studies of the structure of TcR beta in hematologic malignancies have revealed rearrangement in almost all of the T cell malignancies and a small proportion of non-T cell malignancies. In addition, clonal involvement of T cells in diseases such as Hodgkin's disease, angioimmunoblastic lymphadenopathy, and chronic T cell lymphocytosis have been observed. The study of the structure of the TcR beta gene is thus a useful tool for identifying clonal expansions of cells and in conjunction with studies of the immunoglobulin gene structure, and cell surface markers a useful tool for identifying cell lineage. At the present time the evaluation of the structure of the alpha chain genes has not been as fruitful. However, chromosome translocations involving the TcR alpha chain genes have been recognized and, in one case, this rearrangement has been in association with a known oncogene. With the isolation of more probes to the alpha chain region it should be possible to test its utility in identifying clonal populations and cell lineage. The recent isolation of the gamma gene of the T cell will also permit such studies. Preliminary results of studies carried out with a probe to the gamma chain gene of the T cell have paralleled results obtained with the TcR beta probe (unpublished observation).
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- 1986
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11. Effects of recombinant GM-CSF on the blast cells of acute myeloblastic leukemia
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Hoang, T, Nara, N, Wong, G, Clark, S, Minden, MD, and McCulloch, EA
- Abstract
The effects of recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) were compared to those of media conditioned by the continuous bladder carcinoma line, HTB9 (HTB9-CM), using three criteria. First, both GM-CSF and HTB9-CM stimulated blast colony formation in methylcellulose cultures, patient-to-patient variations were seen in the dose-response curves, and GM-CSF was effective, but less so that HTB9-CM. Second, GM-CSF also enhanced growth of blast progenitors in suspension culture, indicating its capacity to support self-renewal. GM-CSF was as effective as HTB9-CM in the production of adherent cells during the growth of blast cells in suspension, a finding that is interpreted to mean that GM-CSF also supports postdeterministic events in blast differentiation. Finally, colonies growing in the presence of GM-CSF were not phenotypically different than those stimulated by HTB9-CM.
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- 1986
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12. Production of growth factors by malignant lymphoma cell lines
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Tweeddale, M, Jamal, N, Nguyen, A, Wang, XH, Minden, MD, and Messner, HA
- Abstract
Fourteen Epstein-Barr virus (EBV)-negative cell lines were raised from bone marrow (BM), peripheral blood (PB), or lymph node samples of patients with intermediate- or high-grade malignant lymphoma. The cell lines were propagated in liquid suspension culture. They contain clonogenic progenitors capable of forming lymphoma colonies in semi- solid culture medium. Cells of these lines were used to examine the growth factor requirements of their clonogenic progenitors and to assess their ability to produce their own growth factors. Two of the cell lines (OCI-Ly9 and OCI-Ly13.1) required addition of exogenous factors for colony growth. These factors were routinely provided by media conditioned by phytohemagglutinin-stimulated leukocytes (PHA- LCM). Three lines formed some and nine lines gave rise to optimal numbers of colonies without addition of growth factors. Eight of these factor-independent lines were able to function as feeder cells and promoted colony formation by both factor-dependent lines. Cell lines that displayed feeder cell function released activities into supernatants able to replace their cellular source. Some of these endogenously produced growth-promoting activities could be replaced by known hematopoietic growth factors. Both factor-dependent cell lines were cultured with recombinant IL-1 alpha, IL-2, IL-3, IL-6, and GM colony-stimulating factor (CSF) and semipurified B-cell growth factor (BCGF) interleukin-4 (IL-4). A heterogeneous response pattern was observed. Both lines formed colonies with IL-4. The colonies were comparable in frequency and size with colonies observed with (PHA-LCM). OCI-Ly9 responded to IL-6 but showed no growth with IL-2. In contrast, the TAC-positive line OCI-Ly13.1 gave rise to colonies with IL-2 while remaining unresponsive to IL-6. A moderate number of colonies was observed when cells of this line were cultured with GM-CSF. Colony formation of both lines was uninfluenced by IL1 alpha or IL-3.
- Published
- 1989
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13. A possible autocrine role for interleukin-6 in two lymphoma cell lines
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Yee, C, Biondi, A, Wang, XH, Iscove, NN, de Sousa, J, Aarden, LA, Wong, GG, Clark, SC, Messner, HA, and Minden, MD
- Abstract
Interleukin-6 (IL-6) is a growth factor with diverse biologic activity. Originally described as a T-cell product that enhances immunoglobulin (Ig) secretion in antigen-stimulated B cells, it also affects the growth of T cells, plasmacytomas, hybridomas, and hematopoietic stem cells. We report the expression and secretion of IL-6 by two lymphoma cell lines, OCI-LY3 and OCI-LY12. Addition of recombinant IL-6 stimulated their growth, whereas addition of polyclonal anti- recombinant IL-6 (anti-rIL-6) had a marked inhibitory effect on proliferation. These results suggest an autocrine role for IL-6 in the growth of these lymphoma cells in culture.
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- 1989
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14. T-cell receptor delta gene rearrangement in childhood T-cell acute lymphoblastic leukemia
- Author
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Biondi, A, Champagne, E, Rossi, V, Giudici, G, Cantu-Rajnoldi, A, Masera, G, Mantovani, A, Mak, TW, and Minden, MD
- Abstract
During the development of functional T lymphocytes, a variety of genes involved in antigen recognition undergo somatic rearrangement. These include the alpha, beta, and gamma chain genes. Recently a fourth rearranging gene, the delta chain gene, embedded in the alpha chain locus, has been described. We have determined the structure of the beta, gamma, and delta chain genes in 15 cases of T-cell acute lymphoblastic leukemia (T-ALL) representing stage I (CD7+, CD1-, CD3-) and stage II (CD7+, CD1+, CD3-) of intrathymic T-cell development. The alpha-delta locus was rearranged in 14 of the 15 cases. In three cases the delta constant region was deleted on both chromosomes, suggesting biallelic V-J alpha rearrangement. A limited pattern of rearrangement of the delta locus was observed in the remaining 11 cases. When the alpha-delta region was rearranged, there was rearrangement of the beta and gamma TcR in all cases except two; in these cases the beta chain was in the germline configuration. These findings support the hypothesis that delta chain gene rearrangement is an early event in T- cell development, possibly contemporary to gamma gene rearrangement, and that the delta locus has a limited repertoire.
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- 1989
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15. Structure and expression of genes of GM-CSF and G-CSF in blast cells from patients with acute myeloblastic leukemia
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Cheng, GY, Kelleher, CA, Miyauchi, J, Wang, C, Wong, G, Clark, SC, McCulloch, EA, and Minden, MD
- Abstract
The hematopoietic growth factors granulocyte/macrophage colony- stimulating factor (GM-CSF) and G-CSF, available as recombinant products, stimulate the growth in culture of blasts from patients with acute myeloblastic leukemia (AML). We used cDNA probes for each gene to study the genomic organization in blast cells of 22 patients and expression in the blast cells of 18 patients. Alteration in the structure of G-CSF (two instances) and GM-CSF (two instances) was found. In two patients in whom it was possible to study DNA from bone marrow obtained at remission, the new bands detected in the leukemic cells were not found. Fifteen of 18 patients showed no RNA expression of either growth factor. Both patients with GM-CSF abnormalities as seen by Southern analysis expressed an abnormally large GM-CSF message but no G-CSF messages. One patient with an abnormal Southern pattern with G-CSF expressed normal-sized G-CSF and GM-CSF messages. The biologic significance of these findings remains to be determined. Nonetheless, the abnormal Southern patterns may prove to be useful clonal markers in the study of AML.
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- 1988
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16. The effects of three recombinant growth factors, IL-3, GM-CSF, and G- CSF, on the blast cells of acute myeloblastic leukemia maintained in short-term suspension culture
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Miyauchi, J, Kelleher, CA, Yang, YC, Wong, GG, Clark, SC, Minden, MD, Minkin, S, and McCulloch, EA
- Abstract
The blast stem cells of acute myeloblastic leukemia (AML) respond in cell culture to growth factors by both self-renewal and terminal divisions. Both of these functions have been shown to be stimulated by the recombinant growth factors granulocyte-macrophage colony- stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF). In this paper, recombinant gibbon interleukin-3 (IL-3), homologous to human IL-3, was tested on blast cells and compared with the effects of GM-CSF, G-CSF, and medium conditioned by the bladder cell line 5637 (5637-CM). We found that IL-3 was an effective stimulator of blast renewal and terminal divisions. However, great patient-to-patient variation was found. A graphic method of presenting complex comparisons between growth factors is also included.
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- 1987
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17. The presence of clonogenic cells in high-grade malignant lymphoma: a prognostic factor
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Tweeddale, ME, Lim, B, Jamal, N, Robinson, J, Zalcberg, J, Lockwood, G, Minden, MD, and Messner, HA
- Abstract
A culture system has been developed that promotes growth of clonogenic lymphoma cells of some patients with intermediate and high-grade malignant lymphoma. The formation of colonies in bone marrow, lymph nodes, and peripheral blood samples is best supported by human plasma. Colony formation of some patients was dependent upon growth factors, which in this study were added in the form of medium conditioned by phytohemagglutinin (PHA)-stimulated leukocytes (PHA-LCM). Some gave rise to lymphoma colonies without PHA-LCM but improved their frequency with PHA-LCM; others were completely independent of PHA-LCM. Colonies grown in primary cultures were routinely recloned and propagated as Epstein-Barr virus (EBV)-negative cell lines with stable B cell phenotype. The cell lines showed the same immunoglobulin rearrangement pattern as that observed in the primary lymphoma sample. In addition, a significant clinical correlation was observed between culture data and clinical outcome. Survival of patients who formed lymphoma colonies at any time during their clinical course was significantly shorter than survival of patients who did not give rise to colonies (P = 0.0009). The same observation was made when the survival assessment was performed for the subset of patients studied at diagnosis (P = 0.0014).
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- 1987
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18. Synergism between recombinant growth factors, GM-CSF and G-CSF, acting on the blast cells of acute myeloblastic leukemia [published erratum appears in Blood 1987 Jul;70(1):339]
- Author
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Kelleher, C, Miyauchi, J, Wong, G, Clark, S, Minden, MD, and McCulloch, EA
- Abstract
The genes for the hemopoietic growth factors, GM colony-stimulating factor (CSF) and G-CSF have been cloned, and recombinant material is available for both. We tested these recombinant factors for their effects on the blast cells of acute myeloblastic leukemia (AML). Culture methods are available that support both colony formation by AML blasts and the growth of blast stem cells in suspension. Recombinant GM- CSF is active in both culture systems, although to a varying degree. We found that recombinant G-CSF was also effective; however, the two recombinant factors showed striking synergism for the stimulation of blast growth of cells from five of eight AML patients. In these cases, the combination was equivalent to the stimulating activity of supernatants from the continuous cell line 5637. This conditioned medium (HTB9-CM) is considered the standard for blast growth. Blasts from one of the patients grew without added factor. In another instance, recombinant GM-CSF alone was almost as effective as HTB9-CM. In the third case, both recombinant factors were active, but synergism was not observed and their combined effect was not equivalent to that of HTB9-CM. Both GM-CSF and G-CSF were active on normal bone marrow granulopoietic progenitors, but synergism was not observed. We conclude that the marked heterogeneity observed when AML blasts are examined by other criteria is also observed when their response to growth factors is evaluated.
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- 1987
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19. Proliferative state of blast cell progenitors in acute myeloblastic leukemia (AML)
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Minden, MD, Till, JE, and McCulloch, EA
- Abstract
Peripheral blood from patients with acute myeloblastic leukemia (AML) contains cells capable of giving rise to colonies in culture when stimulated by media conditioned by leukocytes (LCM) in the presence of phytohemagglutinin (PHA). Two types of colonies are recognized with high frequency: The first grows in the presence of low concentrations of PHA LCM, have a blast-like morphology, and are numerically correlated with morphologically identified blast cells. The second requires either high PHA LCM concentrations or PHA alone with or without 2-mercaptoethanol and consists of cells capable of forming rossettes with sheep erythrocytes and resembles. T-lymphocyte colonies from normal blood. Precursors of blast cell colonies from 15 leukemic patients were tested for cycle state, using either the 3H-thymidine or hydroxyurea techniques. All were found to have a high proportion of cells in the S phase of the cycle. In contrast, T lymphocyte precursors from three normal individual were quiescent. The data are consistent with the maintenance of the leukemic blast cell populations by the proliferative activity of a small subpopulation of blasts.
- Published
- 1978
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20. Self-renewal in culture of proliferative blast progenitor cells in acute myeloblastic leukemia
- Author
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Buick, RN, Minden, MD, and McCulloch, EA
- Abstract
We have proposed that colonies of cells with blastlike morphology growing in culture are derived from a blast subpopulation with high proliferative potential. To test whether or not these blast progenitors have the capacity for self-renewal, blast colonies grown from the peripheral blood of the 21 patients with acute myeloblastic leukemia were replated; secondary colonies were observed in 17 instances, and these were similar to primary colonies in size, morphology, and culture requirements. Great patient-to-patient variation was observed in the frequency of secondary colonies, but low secondary plating efficiency was significantly correlated with successful remission induction. We conclude that the blast progenitors detected in the assay have at least limited self-renewal capacity and that this capacity may, along with other risk factors, contribute to clinical outcome.
- Published
- 1979
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21. Separation of blast cell and T-lymphocyte progenitors in the blood of patients with acute myeloblastic leukemia
- Author
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Minden, MD, Buick, RN, and McCulloch, EA
- Abstract
The peripheral blood of acute myeloblastic leukemia (AML) patients often contains large numbers of two distinct cell populations, both capable of forming colonies in culture under similar conditions. The first population consists of the precursors of blast cells and has specificity for AML; the second population consists of T-lymphocyte precursors, also found in normal blood. The two progenitor populations can be separated by exploiting the capacity of T-lymphocyte (but not blasts) progenitors to form rosettes with sheep erythrocytes (E rosettes). After E-rosette formation, T-lymphocyte precursors can be removed by centrifugation on Ficoll-Hypaque. Such separation has a number of consequences: (1) Blast progenitors can be detected where unseparated mononuclear preparations have yielded either no colonies or only T-lymphocyte colonies (20 of 21 patients). (2) The stimulator requirements of the blast progenitors change, indicating that cell-cell interactions may take place between blast and T-lymphocyte progenitors. (3) It is feasible to characterize blast and T-lymphocyte precursors independently, even though they may coexist in peripheral blood. This may be important if progenitor properties are attributes contributing to the variance in outcome in AML.
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- 1979
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22. The structure of the T cell gamma chain gene in lymphoproliferative disorders and lymphoma cell lines [published erratum appears in Blood 1987 Jan;69(1):368]
- Author
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Griesser, H, primary, Feller, A, additional, Lennert, K, additional, Tweedale, M, additional, Messner, HA, additional, Zalcberg, J, additional, Minden, MD, additional, and Mak, TW, additional
- Published
- 1986
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23. Phase 3 trial of gilteritinib plus azacitidine vs azacitidine for newly diagnosed FLT3mut+ AML ineligible for intensive chemotherapy.
- Author
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Wang ES, Montesinos P, Minden MD, Lee JH, Heuser M, Naoe T, Chou WC, Laribi K, Esteve J, Altman JK, Havelange V, Watson AM, Gambacorti-Passerini C, Patkowska E, Liu S, Wu R, Philipose N, Hill JE, Gill SC, Rich ES, and Tiu RV
- Subjects
- Adult, Humans, Aged, Antineoplastic Combined Chemotherapy Protocols adverse effects, Pyrazines adverse effects, Azacitidine adverse effects, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute diagnosis
- Abstract
Treatment results for patients with newly diagnosed FMS-like tyrosine kinase 3 (FLT3)-mutated (FLT3mut+) acute myeloid leukemia (AML) ineligible for intensive chemotherapy are disappointing. This multicenter, open-label, phase 3 trial randomized (2:1) untreated adults with FLT3mut+ AML ineligible for intensive induction chemotherapy to receive gilteritinib (120 mg/d orally) and azacitidine (GIL + AZA) or azacitidine (AZA) alone. The primary end point was overall survival (OS). At the interim analysis (August 26, 2020), a total of 123 patients were randomized to treatment (GIL + AZA, n = 74; AZA, n = 49). Subsequent AML therapy, including FLT3 inhibitors, was received by 20.3% (GIL + AZA) and 44.9% (AZA) of patients. Median OS was 9.82 (GIL + AZA) and 8.87 (AZA) months (hazard ratio, 0.916; 95% CI, 0.529-1.585; P = .753). The study was closed based on the protocol-specified boundary for futility. Median event-free survival was 0.03 month in both arms. Event-free survival defined by using composite complete remission (CRc) was 4.53 months for GIL + AZA and 0.03 month for AZA (hazard ratio, 0.686; 95% CI, 0.433-1.087; P = .156). CRc rates were 58.1% (GIL + AZA) and 26.5% (AZA) (difference, 31.4%; 95% CI, 13.1-49.7; P < .001). Adverse event (AE) rates were similar for GIL + AZA (100%) and AZA (95.7%); grade ≥3 AEs were 95.9% and 89.4%, respectively. Common AEs with GIL + AZA included pyrexia (47.9%) and diarrhea (38.4%). Gilteritinib steady-state trough concentrations did not differ between GIL + AZA and gilteritinib. GIL + AZA resulted in significantly higher CRc rates, although similar OS compared with AZA. Results support the safety/tolerability and clinical activity of upfront therapy with GIL + AZA in older/unfit patients with FLT3mut+ AML. This trial was registered at www.clinicaltrials.gov as #NCT02752035., (© 2022 by The American Society of Hematology. Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution. All other rights reserved.)
- Published
- 2022
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24. Venetoclax enhances T cell-mediated antileukemic activity by increasing ROS production.
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Lee JB, Khan DH, Hurren R, Xu M, Na Y, Kang H, Mirali S, Wang X, Gronda M, Jitkova Y, MacLean N, Arruda A, Alaniz Z, Konopleva MY, Andreeff M, Minden MD, Zhang L, and Schimmer AD
- Subjects
- Adult, Antineoplastic Agents therapeutic use, Bridged Bicyclo Compounds, Heterocyclic therapeutic use, Cells, Cultured, Humans, Immunity, Cellular drug effects, Leukemia, Myeloid, Acute immunology, Reactive Oxygen Species immunology, Sulfonamides therapeutic use, T-Lymphocytes immunology, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Bridged Bicyclo Compounds, Heterocyclic pharmacology, Leukemia, Myeloid, Acute drug therapy, Sulfonamides pharmacology, T-Lymphocytes drug effects
- Abstract
Venetoclax, a Bcl-2 inhibitor, in combination with the hypomethylating agent azacytidine, achieves complete remission with or without count recovery in ∼70% of treatment-naive elderly patients unfit for conventional intensive chemotherapy. However, the mechanism of action of this drug combination is not fully understood. We discovered that venetoclax directly activated T cells to increase their cytotoxicity against acute myeloid leukemia (AML) in vitro and in vivo. Venetoclax enhanced T-cell effector function by increasing reactive oxygen species generation through inhibition of respiratory chain supercomplexes formation. In addition, azacytidine induced a viral mimicry response in AML cells by activating the STING/cGAS pathway, thereby rendering the AML cells more susceptible to T cell-mediated cytotoxicity. Similar findings were seen in patients treated with venetoclax, as this treatment increased reactive oxygen species generation and activated T cells. Collectively, this study presents a new immune-mediated mechanism of action for venetoclax and azacytidine in the treatment of AML and highlights a potential combination of venetoclax and adoptive cell therapy for patients with AML., (© 2021 by The American Society of Hematology.)
- Published
- 2021
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25. Very long chain fatty acid metabolism is required in acute myeloid leukemia.
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Tcheng M, Roma A, Ahmed N, Smith RW, Jayanth P, Minden MD, Schimmer AD, Hess DA, Hope K, Rea KA, Akhtar TA, Bohrnsen E, D'Alessandro A, Mohsen AW, Vockley J, and Spagnuolo PA
- Subjects
- Acyl-CoA Dehydrogenase, Long-Chain genetics, Acyl-CoA Dehydrogenase, Long-Chain metabolism, Cell Line, Tumor, Citric Acid Cycle, Fatty Acids genetics, Glycolysis, Humans, Ketone Oxidoreductases metabolism, Leukemia, Myeloid, Acute genetics, Neoplasm Proteins genetics, Neoplasm Proteins metabolism, Fatty Acids metabolism, Leukemia, Myeloid, Acute metabolism
- Abstract
Acute myeloid leukemia (AML) cells have an atypical metabolic phenotype characterized by increased mitochondrial mass, as well as a greater reliance on oxidative phosphorylation and fatty acid oxidation (FAO) for survival. To exploit this altered metabolism, we assessed publicly available databases to identify FAO enzyme overexpression. Very long chain acyl-CoA dehydrogenase (VLCAD; ACADVL) was found to be overexpressed and critical to leukemia cell mitochondrial metabolism. Genetic attenuation or pharmacological inhibition of VLCAD hindered mitochondrial respiration and FAO contribution to the tricarboxylic acid cycle, resulting in decreased viability, proliferation, clonogenic growth, and AML cell engraftment. Suppression of FAO at VLCAD triggered an increase in pyruvate dehydrogenase activity that was insufficient to increase glycolysis but resulted in adenosine triphosphate depletion and AML cell death, with no effect on normal hematopoietic cells. Together, these results demonstrate the importance of VLCAD in AML cell biology and highlight a novel metabolic vulnerability for this devastating disease., (© 2021 by The American Society of Hematology.)
- Published
- 2021
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26. CRISPR screen identifies genes that sensitize AML cells to double-negative T-cell therapy.
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Soares F, Chen B, Lee JB, Ahmed M, Ly D, Tin E, Kang H, Zeng Y, Akhtar N, Minden MD, He HH, and Zhang L
- Subjects
- Adoptive Transfer, Animals, CRISPR-Cas Systems, Cells, Cultured, Female, Gene Expression Regulation, Leukemic, Humans, Mice, Inbred NOD, Receptors, IgG genetics, Mice, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute therapy, T-Lymphocytes transplantation
- Abstract
Acute myeloid leukemia (AML) remains a devastating disease in need of new therapies to improve patient survival. Targeted adoptive T-cell therapies have achieved impressive clinical outcomes in some B-cell leukemias and lymphomas but not in AML. Double-negative T cells (DNTs) effectively kill blast cells from the majority of AML patients and are now being tested in clinical trials. However, AML blasts obtained from ∼30% of patients show resistance to DNT-mediated cytotoxicity; the markers or mechanisms underlying this resistance have not been elucidated. Here, we used a targeted clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) screen to identify genes that cause susceptibility of AML cells to DNT therapy. Inactivation of the Spt-Ada-Gcn5-acetyltransferase (SAGA) deubiquitinating complex components sensitized AML cells to DNT-mediated cytotoxicity. In contrast, CD64 inactivation resulted in resistance to DNT-mediated cytotoxicity. Importantly, the level of CD64 expression correlated strongly with the sensitivity of AML cells to DNT treatment. Furthermore, the ectopic expression of CD64 overcame AML resistance to DNTs in vitro and in vivo. Altogether, our data demonstrate the utility of CRISPR/Cas9 screens to uncover mechanisms underlying the sensitivity to DNT therapy and suggest CD64 as a predictive marker for response in AML patients., (© 2021 by The American Society of Hematology.)
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- 2021
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27. CC-90009, a novel cereblon E3 ligase modulator, targets acute myeloid leukemia blasts and leukemia stem cells.
- Author
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Surka C, Jin L, Mbong N, Lu CC, Jang IS, Rychak E, Mendy D, Clayton T, Tindall E, Hsu C, Fontanillo C, Tran E, Contreras A, Ng SWK, Matyskiela M, Wang K, Chamberlain P, Cathers B, Carmichael J, Hansen J, Wang JCY, Minden MD, Fan J, Pierce DW, Pourdehnad M, Rolfe M, Lopez-Girona A, Dick JE, and Lu G
- Subjects
- Acetamides therapeutic use, Animals, CRISPR-Cas Systems, Cell Line, Tumor, Humans, Isoindoles therapeutic use, Mice, Mice, Inbred NOD, Mice, SCID, Models, Molecular, Neoplastic Stem Cells enzymology, Nuclear Factor 45 Protein physiology, Nuclear Factor 90 Proteins physiology, Peptide Termination Factors metabolism, Piperidones therapeutic use, Proteasome Endopeptidase Complex metabolism, Protein Conformation, Protein Processing, Post-Translational drug effects, Proteolysis, Small Molecule Libraries, Stress, Physiological, TOR Serine-Threonine Kinases physiology, U937 Cells, Ubiquitination drug effects, Xenograft Model Antitumor Assays, Acetamides pharmacology, Adaptor Proteins, Signal Transducing antagonists & inhibitors, Isoindoles pharmacology, Leukemia, Myeloid, Acute pathology, Molecular Targeted Therapy, Neoplasm Proteins antagonists & inhibitors, Neoplastic Stem Cells drug effects, Piperidones pharmacology, Ubiquitin-Protein Ligases antagonists & inhibitors
- Abstract
A number of clinically validated drugs have been developed by repurposing the CUL4-DDB1-CRBN-RBX1 (CRL4CRBN) E3 ubiquitin ligase complex with molecular glue degraders to eliminate disease-driving proteins. Here, we present the identification of a first-in-class GSPT1-selective cereblon E3 ligase modulator, CC-90009. Biochemical, structural, and molecular characterization demonstrates that CC-90009 coopts the CRL4CRBN to selectively target GSPT1 for ubiquitination and proteasomal degradation. Depletion of GSPT1 by CC-90009 rapidly induces acute myeloid leukemia (AML) apoptosis, reducing leukemia engraftment and leukemia stem cells (LSCs) in large-scale primary patient xenografting of 35 independent AML samples, including those with adverse risk features. Using a genome-wide CRISPR-Cas9 screen for effectors of CC-90009 response, we uncovered the ILF2 and ILF3 heterodimeric complex as a novel regulator of cereblon expression. Knockout of ILF2/ILF3 decreases the production of full-length cereblon protein via modulating CRBN messenger RNA alternative splicing, leading to diminished response to CC-90009. The screen also revealed that the mTOR signaling and the integrated stress response specifically regulate the response to CC-90009 in contrast to other cereblon modulators. Hyperactivation of the mTOR pathway by inactivation of TSC1 and TSC2 protected against the growth inhibitory effect of CC-90009 by reducing CC-90009-induced binding of GSPT1 to cereblon and subsequent GSPT1 degradation. On the other hand, GSPT1 degradation promoted the activation of the GCN1/GCN2/ATF4 pathway and subsequent apoptosis in AML cells. Collectively, CC-90009 activity is mediated by multiple layers of signaling networks and pathways within AML blasts and LSCs, whose elucidation gives insight into further assessment of CC-90009s clinical utility. These trials were registered at www.clinicaltrials.gov as #NCT02848001 and #NCT04336982)., (© 2021 by The American Society of Hematology.)
- Published
- 2021
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28. Mitochondrial carrier homolog 2 is necessary for AML survival.
- Author
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Khan DH, Mullokandov M, Wu Y, Voisin V, Gronda M, Hurren R, Wang X, MacLean N, Jeyaraju DV, Jitkova Y, Xu GW, Laister R, Seneviratne A, Blatman ZM, Ketela T, Bader GD, Marhon SA, De Carvalho DD, Minden MD, Gross A, and Schimmer AD
- Subjects
- Acetylation, Animals, CRISPR-Cas Systems, Cell Differentiation, Cell Line, Tumor, Cell Nucleus metabolism, Fetal Blood cytology, Gene Expression Regulation, Leukemic genetics, Gene Knockdown Techniques, Histones metabolism, Humans, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute pathology, Mice, Mice, Inbred C57BL, Myeloid-Lymphoid Leukemia Protein physiology, Oncogene Proteins, Fusion physiology, Protein Processing, Post-Translational, Pyruvic Acid metabolism, RNA Interference, RNA, Small Interfering genetics, RNA, Small Interfering pharmacology, Leukemia, Myeloid, Acute metabolism, Mitochondria metabolism, Mitochondrial Membrane Transport Proteins physiology, Neoplasm Proteins physiology
- Abstract
Through a clustered regularly insterspaced short palindromic repeats (CRISPR) screen to identify mitochondrial genes necessary for the growth of acute myeloid leukemia (AML) cells, we identified the mitochondrial outer membrane protein mitochondrial carrier homolog 2 (MTCH2). In AML, knockdown of MTCH2 decreased growth, reduced engraftment potential of stem cells, and induced differentiation. Inhibiting MTCH2 in AML cells increased nuclear pyruvate and pyruvate dehydrogenase (PDH), which induced histone acetylation and subsequently promoted the differentiation of AML cells. Thus, we have defined a new mechanism by which mitochondria and metabolism regulate AML stem cells and gene expression., (© 2020 by The American Society of Hematology.)
- Published
- 2020
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29. A stemness screen reveals C3orf54/INKA1 as a promoter of human leukemia stem cell latency.
- Author
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Kaufmann KB, Garcia-Prat L, Liu Q, Ng SWK, Takayanagi SI, Mitchell A, Wienholds E, van Galen P, Cumbaa CA, Tsay MJ, Pastrello C, Wagenblast E, Krivdova G, Minden MD, Lechman ER, Zandi S, Jurisica I, Wang JCY, Xie SZ, and Dick JE
- Subjects
- Animals, Cell Cycle Checkpoints, Cell Line, Tumor, Female, Humans, Leukemia, Myeloid, Acute pathology, Male, Mice, Inbred NOD, Neoplastic Stem Cells cytology, Neoplastic Stem Cells pathology, Up-Regulation, p21-Activated Kinases analysis, Gene Expression Regulation, Leukemic, Intracellular Signaling Peptides and Proteins genetics, Leukemia, Myeloid, Acute genetics, Neoplastic Stem Cells metabolism
- Abstract
There is a growing body of evidence that the molecular properties of leukemia stem cells (LSCs) are associated with clinical outcomes in acute myeloid leukemia (AML), and LSCs have been linked to therapy failure and relapse. Thus, a better understanding of the molecular mechanisms that contribute to the persistence and regenerative potential of LSCs is expected to result in the development of more effective therapies. We therefore interrogated functionally validated data sets of LSC-specific genes together with their known protein interactors and selected 64 candidates for a competitive in vivo gain-of-function screen to identify genes that enhanced stemness in human cord blood hematopoietic stem and progenitor cells. A consistent effect observed for the top hits was the ability to restrain early repopulation kinetics while preserving regenerative potential. Overexpression (OE) of the most promising candidate, the orphan gene C3orf54/INKA1 , in a patient-derived AML model (8227) promoted the retention of LSCs in a primitive state manifested by relative expansion of CD34
+ cells, accumulation of cells in G0 , and reduced output of differentiated progeny. Despite delayed early repopulation, at later times, INKA1 -OE resulted in the expansion of self-renewing LSCs. In contrast, INKA1 silencing in primary AML reduced regenerative potential. Mechanistically, our multidimensional confocal analysis found that INKA1 regulates G0 exit by interfering with nuclear localization of its target PAK4, with concomitant reduction of global H4K16ac levels. These data identify INKA1 as a novel regulator of LSC latency and reveal a link between the regulation of stem cell kinetics and pool size during regeneration., (© 2019 by The American Society of Hematology.)- Published
- 2019
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30. Distinct patterns of clonal evolution in patients with concurrent myelo- and lymphoproliferative neoplasms.
- Author
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Kennedy JA, Medeiros JJF, Dobson SM, Arruda A, Sukhai MA, Stockley T, Tierens A, Minden MD, Kamel-Reid S, Dick JE, and Gupta V
- Subjects
- Aged, Aged, 80 and over, Female, Humans, Karyotype, Lymphoproliferative Disorders complications, Lymphoproliferative Disorders pathology, Male, Middle Aged, Myeloproliferative Disorders complications, Myeloproliferative Disorders pathology, Polymorphism, Single Nucleotide, Primary Myelofibrosis complications, Primary Myelofibrosis genetics, Primary Myelofibrosis pathology, Clonal Evolution, Lymphoproliferative Disorders genetics, Mutation, Myeloproliferative Disorders genetics
- Published
- 2018
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31. Leveraging increased cytoplasmic nucleoside kinase activity to target mtDNA and oxidative phosphorylation in AML.
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Liyanage SU, Hurren R, Voisin V, Bridon G, Wang X, Xu C, MacLean N, Siriwardena TP, Gronda M, Yehudai D, Sriskanthadevan S, Avizonis D, Shamas-Din A, Minden MD, Bader GD, Laposa R, and Schimmer AD
- Subjects
- Animals, Cell Line, Tumor, Cells, Cultured, DNA Replication, Humans, Mice, SCID, NM23 Nucleoside Diphosphate Kinases metabolism, Nucleoside-Phosphate Kinase metabolism, Signal Transduction, Tumor Cells, Cultured, Zalcitabine metabolism, DNA, Mitochondrial genetics, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute metabolism, Oxidative Phosphorylation, Phosphotransferases metabolism
- Abstract
Mitochondrial DNA (mtDNA) biosynthesis requires replication factors and adequate nucleotide pools from the mitochondria and cytoplasm. We performed gene expression profiling analysis of 542 human acute myeloid leukemia (AML) samples and identified 55% with upregulated mtDNA biosynthesis pathway expression compared with normal hematopoietic cells. Genes that support mitochondrial nucleotide pools, including mitochondrial nucleotide transporters and a subset of cytoplasmic nucleoside kinases, were also increased in AML compared with normal hematopoietic samples. Knockdown of cytoplasmic nucleoside kinases reduced mtDNA levels in AML cells, demonstrating their contribution in maintaining mtDNA. To assess cytoplasmic nucleoside kinase pathway activity, we used a nucleoside analog 2'3'-dideoxycytidine (ddC), which is phosphorylated to the activated antimetabolite, 2'3'-dideoxycytidine triphosphate by cytoplasmic nucleoside kinases. ddC is a selective inhibitor of the mitochondrial DNA polymerase γ. ddC was preferentially activated in AML cells compared with normal hematopoietic progenitor cells. ddC treatment inhibited mtDNA replication, oxidative phosphorylation, and induced cytotoxicity in a panel of AML cell lines. Furthermore, ddC preferentially inhibited mtDNA replication in a subset of primary human leukemia cells and selectively targeted leukemia cells while sparing normal progenitor cells. In animal models of human AML, treatment with ddC decreased mtDNA, electron transport chain proteins, and induced tumor regression without toxicity. ddC also targeted leukemic stem cells in secondary AML xenotransplantation assays. Thus, AML cells have increased cytidine nucleoside kinase activity that regulates mtDNA biogenesis and can be leveraged to selectively target oxidative phosphorylation in AML., (© 2017 by The American Society of Hematology.)
- Published
- 2017
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32. International phase 3 study of azacitidine vs conventional care regimens in older patients with newly diagnosed AML with >30% blasts.
- Author
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Dombret H, Seymour JF, Butrym A, Wierzbowska A, Selleslag D, Jang JH, Kumar R, Cavenagh J, Schuh AC, Candoni A, Récher C, Sandhu I, Bernal del Castillo T, Al-Ali HK, Martinelli G, Falantes J, Noppeney R, Stone RM, Minden MD, McIntyre H, Songer S, Lucy LM, Beach CL, and Döhner H
- Subjects
- Aged, Aged, 80 and over, Blast Crisis pathology, Female, Follow-Up Studies, Humans, International Agencies, Leukemia, Myeloid, Acute pathology, Male, Neoplasm Staging, Prognosis, Prospective Studies, Survival Rate, Antimetabolites, Antineoplastic therapeutic use, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Azacitidine therapeutic use, Blast Crisis drug therapy, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myeloid, Acute mortality
- Abstract
This multicenter, randomized, open-label, phase 3 trial evaluated azacitidine efficacy and safety vs conventional care regimens (CCRs) in 488 patients age ≥65 years with newly diagnosed acute myeloid leukemia (AML) with >30% bone marrow blasts. Before randomization, a CCR (standard induction chemotherapy, low-dose ara-c, or supportive care only) was preselected for each patient. Patients then were assigned 1:1 to azacitidine (n = 241) or CCR (n = 247). Patients assigned to CCR received their preselected treatment. Median overall survival (OS) was increased with azacitidine vs CCR: 10.4 months (95% confidence interval [CI], 8.0-12.7 months) vs 6.5 months (95% CI, 5.0-8.6 months), respectively (hazard ratio [HR] was 0.85; 95% CI, 0.69-1.03; stratified log-rank P = .1009). One-year survival rates with azacitidine and CCR were 46.5% and 34.2%, respectively (difference, 12.3%; 95% CI, 3.5%-21.0%). A prespecified analysis censoring patients who received AML treatment after discontinuing study drug showed median OS with azacitidine vs CCR was 12.1 months (95% CI, 9.2-14.2 months) vs 6.9 months (95% CI, 5.1-9.6 months; HR, 0.76; 95% CI, 0.60-0.96; stratified log-rank P = .0190). Univariate analysis showed favorable trends for azacitidine compared with CCR across all subgroups defined by baseline demographic and disease features. Adverse events were consistent with the well-established safety profile of azacitidine. Azacitidine may be an important treatment option for this difficult-to-treat AML population. This trial was registered at www.clinicaltrials.gov as #NCT01074047., (© 2015 by The American Society of Hematology.)
- Published
- 2015
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33. AML cells have low spare reserve capacity in their respiratory chain that renders them susceptible to oxidative metabolic stress.
- Author
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Sriskanthadevan S, Jeyaraju DV, Chung TE, Prabha S, Xu W, Skrtic M, Jhas B, Hurren R, Gronda M, Wang X, Jitkova Y, Sukhai MA, Lin FH, Maclean N, Laister R, Goard CA, Mullen PJ, Xie S, Penn LZ, Rogers IM, Dick JE, Minden MD, and Schimmer AD
- Subjects
- Cell Death, Cell Respiration, Electron Transport, Humans, Mitochondrial Size, Reactive Oxygen Species metabolism, Tumor Cells, Cultured, Leukemia, Myeloid, Acute metabolism, Leukemia, Myeloid, Acute pathology, Oxidative Stress physiology, Oxygen Consumption physiology
- Abstract
Mitochondrial respiration is a crucial component of cellular metabolism that can become dysregulated in cancer. Compared with normal hematopoietic cells, acute myeloid leukemia (AML) cells and patient samples have higher mitochondrial mass, without a concomitant increase in respiratory chain complex activity. Hence these cells have a lower spare reserve capacity in the respiratory chain and are more susceptible to oxidative stress. We therefore tested the effects of increasing the electron flux through the respiratory chain as a strategy to induce oxidative stress and cell death preferentially in AML cells. Treatment with the fatty acid palmitate induced oxidative stress and cell death in AML cells, and it suppressed tumor burden in leukemic cell lines and primary patient sample xenografts in the absence of overt toxicity to normal cells and organs. These data highlight a unique metabolic vulnerability in AML, and identify a new therapeutic strategy that targets abnormal oxidative metabolism in this malignancy., (© 2015 by The American Society of Hematology.)
- Published
- 2015
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34. Treatment outcomes following leukemic transformation in Philadelphia-negative myeloproliferative neoplasms.
- Author
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Kennedy JA, Atenafu EG, Messner HA, Craddock KJ, Brandwein JM, Lipton JH, Minden MD, Schimmer AD, Schuh AC, Yee KW, and Gupta V
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Antimetabolites, Antineoplastic therapeutic use, Azacitidine analogs & derivatives, Azacitidine therapeutic use, Combined Modality Therapy, Cytarabine therapeutic use, Decitabine, Female, Humans, Male, Middle Aged, Myeloproliferative Disorders genetics, Myeloproliferative Disorders mortality, Registries, Survival Analysis, Transplantation, Homologous, Treatment Outcome, Young Adult, Cell Transformation, Neoplastic pathology, Hematopoietic Stem Cell Transplantation, Myeloproliferative Disorders pathology, Myeloproliferative Disorders therapy, Philadelphia Chromosome
- Abstract
Leukemic transformation (LT) is a rare but fatal complication of Philadelphia-negative myeloproliferative neoplasms (MPNs) for which optimal treatment strategies are not known. At our center, we have adopted a treatment approach for LT where patients within the transplant age group who have a reasonable fitness level are treated with curative intent and offered induction chemotherapy. Subsequently, those who respond and have a suitable donor are considered for allogeneic hematopoietic cell transplantation (HCT). In this study, we evaluated the clinical outcomes of this treatment approach in 75 patients with LT. The 2-year overall survival (OS) from the time of LT was 15%. A total of 39 patients (52%) were treated with curative intent (induction ± HCT) and had a 2-y OS of 26% compared with 3% in those noncuratively treated (P < .0001). In the curative intent group, 18 individuals (46%) achieved complete remission (CR) or CR with incomplete recovery and 12 (31%) reverted to a chronic MPN phase, with 17 patients undergoing HCT. Survival of patients posttransplant was significantly improved compared with those who responded to induction but were not transplanted (2-y OS of 47% vs 15%; P = .03). Thus, induction chemotherapy followed by HCT has the potential for long-term disease control in select patients with LT preceded by a MPN.
- Published
- 2013
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35. A small molecule screening strategy with validation on human leukemia stem cells uncovers the therapeutic efficacy of kinetin riboside.
- Author
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McDermott SP, Eppert K, Notta F, Isaac M, Datti A, Al-Awar R, Wrana J, Minden MD, and Dick JE
- Subjects
- Adenosine analysis, Adenosine isolation & purification, Adenosine pharmacology, Animals, Antineoplastic Agents analysis, Antineoplastic Agents isolation & purification, Antineoplastic Agents pharmacology, Antineoplastic Agents therapeutic use, Humans, Kinetin analysis, Kinetin isolation & purification, Kinetin pharmacology, Mice, Mice, Inbred NOD, Mice, Nude, Mice, SCID, Neoplastic Stem Cells pathology, Treatment Outcome, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, Adenosine therapeutic use, High-Throughput Screening Assays methods, Kinetin therapeutic use, Leukemia drug therapy, Leukemia pathology, Neoplastic Stem Cells drug effects, Small Molecule Libraries analysis
- Abstract
Gene regulatory networks that govern hematopoietic stem cells (HSCs) and leukemia-initiating cells (L-ICs) are deeply entangled. Thus, the discovery of compounds that target L-ICs while sparing HSC is an attractive but difficult endeavor. Presently, most screening approaches fail to counter-screen compounds against normal hematopoietic stem/progenitor cells (HSPCs). Here, we present a multistep in vitro and in vivo approach to identify compounds that can target L-ICs in acute myeloid leukemia (AML). A high-throughput screen of 4000 compounds on novel leukemia cell lines derived from human experimental leukemogenesis models yielded 80 hits, of which 10 were less toxic to HSPC. We characterized a single compound, kinetin riboside (KR), on AML L-ICs and HSPCs. KR demonstrated comparable efficacy to standard therapies against blast cells in 63 primary leukemias. In vitro, KR targeted the L-IC-enriched CD34(+)CD38(-) AML fraction, while sparing HSPC-enriched fractions, although these effects were mitigated on HSC assayed in vivo. KR eliminated L-ICs in 2 of 4 primary AML samples when assayed in vivo and highlights the importance of in vivo L-IC and HSC assays to measure function. Overall, we provide a novel approach to screen large drug libraries for the discovery of anti-L-IC compounds for human leukemias.
- Published
- 2012
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36. The antiparasitic agent ivermectin induces chloride-dependent membrane hyperpolarization and cell death in leukemia cells.
- Author
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Sharmeen S, Skrtic M, Sukhai MA, Hurren R, Gronda M, Wang X, Fonseca SB, Sun H, Wood TE, Ward R, Minden MD, Batey RA, Datti A, Wrana J, Kelley SO, and Schimmer AD
- Subjects
- Animals, Antineoplastic Agents pharmacology, Antiparasitic Agents pharmacology, Calcium metabolism, Cell Death drug effects, Cell Line, Tumor, Cell Membrane drug effects, Cell Size drug effects, Chlorides metabolism, Cytarabine pharmacology, Daunorubicin pharmacology, Drug Synergism, Gene Expression Regulation, Leukemic drug effects, Humans, Ivermectin pharmacology, Mice, Mice, SCID, Reactive Oxygen Species metabolism, Tumor Cells, Cultured, Antineoplastic Agents therapeutic use, Antiparasitic Agents therapeutic use, Cell Survival drug effects, Ivermectin therapeutic use, Leukemia drug therapy
- Abstract
To identify known drugs with previously unrecognized anticancer activity, we compiled and screened a library of such compounds to identify agents cytotoxic to leukemia cells. From these screens, we identified ivermectin, a derivative of avermectin B1 that is licensed for the treatment of the parasitic infections, strongyloidiasis and onchocerciasis, but is also effective against other worm infestations. As a potential antileukemic agent, ivermectin induced cell death at low micromolar concentrations in acute myeloid leukemia cell lines and primary patient samples preferentially over normal hematopoietic cells. Ivermectin also delayed tumor growth in 3 independent mouse models of leukemia at concentrations that appear pharmacologically achievable. As an antiparasitic, ivermectin binds and activates chloride ion channels in nematodes, so we tested the effects of ivermectin on chloride flux in leukemia cells. Ivermectin increased intracellular chloride ion concentrations and cell size in leukemia cells. Chloride influx was accompanied by plasma membrane hyperpolarization, but did not change mitochondrial membrane potential. Ivermectin also increased reactive oxygen species generation that was functionally important for ivermectin-induced cell death. Finally, ivermectin synergized with cytarabine and daunorubicin that also increase reactive oxygen species production. Thus, given its known toxicology and pharmacology, ivermectin could be rapidly advanced into clinical trial for leukemia.
- Published
- 2010
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37. An upstream insulator regulates DLK1 imprinting in AML.
- Author
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Khoury H, Suarez-Saiz F, Wu S, and Minden MD
- Subjects
- Alleles, Base Sequence, Calcium-Binding Proteins, Cell Line, Tumor, DNA Methylation genetics, DNA Mutational Analysis, Gene Expression Regulation, Leukemic, Humans, Molecular Sequence Data, Genomic Imprinting genetics, Insulator Elements genetics, Intercellular Signaling Peptides and Proteins genetics, Leukemia, Myeloid, Acute genetics, Membrane Proteins genetics
- Abstract
DLK1 is an imprinted gene on chromosome 14. Using informative coding single nucleotide polymorphisms, we found DLK1 expression to be monoallelic in normal bone marrow, whereas it was biallelic in 76% of acute myeloid leukemia (AML) overexpressing DLK1 (61% of all AML). Quantitative methylation analysis of 7 cytosine-phosphate-guanosine-rich areas (3 upstream of or within DLK1, the putative intergenic-differentially methylated region and 3 upstream of or within MEG3) revealed a strong association between biallelic DLK1 expression and hypermethylation of a cytosine-phosphate-guanosine-rich region 18 kb upstream of DLK1. Allele-specific methylation analysis of this region revealed the alleles to be differentially methylated in normal bone marrow and monoallelic DLK1 AML, whereas there was increased methylation of both alleles in AML with biallelic expression. Moreover, chromatin immunoprecipitation analysis revealed that CCTC-binding factor binds to this region in monoallelic but not biallelic expression samples. Taken together, our data indicate that an insulator located 18 kb upstream of DLK1 plays an important role in regulating DLK1 imprinting.
- Published
- 2010
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38. Small-molecule XIAP inhibitors derepress downstream effector caspases and induce apoptosis of acute myeloid leukemia cells.
- Author
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Carter BZ, Gronda M, Wang Z, Welsh K, Pinilla C, Andreeff M, Schober WD, Nefzi A, Pond GR, Mawji IA, Houghten RA, Ostresh J, Brandwein J, Minden MD, Schuh AC, Wells RA, Messner H, Chun K, Reed JC, and Schimmer AD
- Subjects
- Cytarabine pharmacology, Female, Humans, Male, Middle Aged, Proteins metabolism, Proto-Oncogene Proteins c-bcl-2 metabolism, Tumor Cells, Cultured, X-Linked Inhibitor of Apoptosis Protein, Aniline Compounds pharmacology, Apoptosis drug effects, Caspases metabolism, Leukemia, Myeloid, Acute metabolism, Leukemia, Myeloid, Acute pathology, Phenylurea Compounds pharmacology, Proteins antagonists & inhibitors
- Abstract
We tested the effects of small-molecule XIAP antagonists based on a polyphenylurea pharmacophore on cultured acute myelogenous leukemia (AML) cell lines and primary patient samples. X-linked inhibitor of apoptosis protein (XIAP) antagonist N-[(5R)-6-[(anilinocarbonyl)amino]-5-((anilinocarbonyl){[(2R)-1-(4-cyclohexylbutyl)pyrrolidin-2-yl]methyl}amino)hexyl]-N-methyl-N'-phenylurea (1396-12), but not a structurally related control compound, induced apoptosis of primary leukemia samples with a lethal dose (LD50) of less than 10 microM in 16 of 27 (60%) samples. In contrast, XIAP antagonist 1396-12 was not lethal to the normal hematopoietic cells in short-term cytotoxicity assays. Response of primary AML specimens to XIAP inhibitor correlated with XIAP protein levels, with higher levels of XIAP associated with sensitivity. The XIAP antagonist 1396-12 induced activation of downstream caspases 3 and 7 prior to the activation of upstream caspase 8 and caspase 9. Apoptosis induction was also independent of B-cell lymphoma protein-2 (Bcl-2) or caspase 8, indicative of a downstream effect on apoptotic pathways. Thus, polyphenylurea-based XIAP antagonsists directly induce apoptosis of leukemia cells and AML patient samples at low micromolar concentrations through a mechanism of action distinct from conventional chemotherapeutic agents.
- Published
- 2005
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39. Heterozygous PU.1 mutations are associated with acute myeloid leukemia.
- Author
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Mueller BU, Pabst T, Osato M, Asou N, Johansen LM, Minden MD, Behre G, Hiddemann W, Ito Y, and Tenen DG
- Subjects
- Acute Disease, DNA Mutational Analysis, DNA, Neoplasm genetics, Ethnicity genetics, Heterozygote, Humans, Leukemia, Myeloid ethnology, Mutation, Polymerase Chain Reaction, Sequence Analysis, DNA, Leukemia, Myeloid genetics, Neoplasm Proteins genetics, Proto-Oncogene Proteins genetics, Trans-Activators genetics
- Published
- 2003
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- View/download PDF
40. Heterozygous PU.1 mutations are associated with acute myeloid leukemia.
- Author
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Mueller BU, Pabst T, Osato M, Asou N, Johansen LM, Minden MD, Behre G, Hiddemann W, Ito Y, and Tenen DG
- Subjects
- Acute Disease, Binding Sites genetics, Cell Differentiation genetics, DNA Mutational Analysis, Gene Expression Regulation drug effects, Gene Expression Regulation genetics, Granulocytes cytology, Heterozygote, Humans, Leukemia, Myeloid etiology, Leukemia, Myeloid pathology, Leukocytes, Mononuclear pathology, Protein Binding genetics, Proto-Oncogene Proteins pharmacology, Proto-Oncogene Proteins physiology, Receptor, Macrophage Colony-Stimulating Factor genetics, Receptor, Macrophage Colony-Stimulating Factor metabolism, Trans-Activators pharmacology, Trans-Activators physiology, Leukemia, Myeloid genetics, Mutation, Proto-Oncogene Proteins genetics, Trans-Activators genetics
- Abstract
The transcription factor PU.1 is required for normal blood cell development. PU.1 regulates the expression of a number of crucial myeloid genes, such as the macrophage colony-stimulating factor (M-CSF) receptor, the granulocyte colony-stimulating factor (G-CSF) receptor, and the granulocyte-macrophage colony-stimulating factor (GM-CSF) receptor. Myeloid cells derived from PU.1(-/-) mice are blocked at the earliest stage of myeloid differentiation, similar to the blast cells that are the hallmark of human acute myeloid leukemia (AML). These facts led us to hypothesize that molecular abnormalities involving the PU.1 gene could contribute to the development of AML. We identified 10 mutant alleles of the PU.1 gene in 9 of 126 AML patients. The PU.1 mutations comprised 5 deletions affecting the DNA-binding domain, and 5 point mutations in 1) the DNA-binding domain (2 patients), 2) the PEST domain (2 patients), and 3) the transactivation domain (one patient). DNA binding to and transactivation of the M-CSF receptor promoter, a direct PU.1 target gene, were deficient in the 7 PU.1 mutants that affected the DNA-binding domain. In addition, these mutations decreased the ability of PU.1 to synergize with PU.1-interacting proteins such as AML1 or c-Jun in the activation of PU.1 target genes. This is the first report of mutations in the PU.1 gene in human neoplasia and suggests that disruption of PU.1 function contributes to the block in differentiation found in AML patients.
- Published
- 2002
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- View/download PDF
41. Receptor- and mitochondrial-mediated apoptosis in acute leukemia: a translational view.
- Author
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Schimmer AD, Hedley DW, Penn LZ, and Minden MD
- Subjects
- Acute Disease, Adult, Amino Acid Sequence, Cytochrome c Group physiology, Humans, Leukemia drug therapy, Leukemia genetics, Mitochondria pathology, Molecular Sequence Data, Proto-Oncogene Proteins c-bcl-2 chemistry, Proto-Oncogene Proteins c-bcl-2 physiology, Translocation, Genetic, Apoptosis genetics, Leukemia pathology, Mitochondria physiology, Receptors, Tumor Necrosis Factor physiology
- Published
- 2001
- Full Text
- View/download PDF
42. Regulation of drug sensitivity by ribosomal protein S3a.
- Author
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Hu ZB, Minden MD, McCulloch EA, and Stahl J
- Subjects
- Animals, Antibiotics, Antineoplastic pharmacology, Antimetabolites, Antineoplastic pharmacology, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Apoptosis, Cell Line, Cryopreservation, Cytarabine pharmacology, DNA Replication, DNA, Neoplasm biosynthesis, Doxorubicin pharmacology, Gene Targeting, Humans, Leukemia, Myeloid, Acute drug therapy, Leukemia, Myelomonocytic, Chronic drug therapy, Neoplasm Proteins deficiency, Neoplasm Proteins genetics, Neoplastic Stem Cells metabolism, Paclitaxel pharmacology, Phosphorylation, Phosphoserine analysis, Protein Processing, Post-Translational, Rats, Ribosomal Proteins deficiency, Ribosomal Proteins genetics, Tissue Preservation, Treatment Outcome, Antineoplastic Agents pharmacology, Drug Resistance, Neoplasm physiology, Leukemia, Myeloid, Acute pathology, Leukemia, Myelomonocytic, Chronic pathology, Neoplasm Proteins physiology, Neoplastic Stem Cells drug effects, Ribosomal Proteins physiology, Tretinoin pharmacology
- Abstract
When bcl-2 is immunoprecipitated from (32)P-labeled cell extracts of all-trans retinoic acid (ATRA)-treated acute myeloblastic leukemia (AML) blasts, a phosphorylated protein of approximately 30 kd is coprecipitated. This protein has been identified as ribosomal protein S3a. The biologic effects of S3a include favoring apoptosis and enhancing the malignant phenotype. We sought to determine whether S3a, like bcl-2, influenced the response of cells to chemotherapeutic drugs and ATRA. Cell lines were studied in which S3a was genetically increased or disrupted; increased S3a was regularly associated with increased plating efficiency and increased sensitivity to either cytosine arabinoside (ara-C) or doxorubicin (DNR). S3a did not affect the sensitivity of cells to paclitaxel. Pulse exposures to either (3)HTdR or ara-C showed a greater percentage of clonogenic cells in the S phase of the cell cycle in cells with increased S3a than in controls. Cells with increased S3a responded to ATRA by increased ara-C or DNR sensitivity, whereas cells with reduced S3a protein were either protected by ATRA or not affected. We studied cryopreserved blast cells from patients with AML or chronic myelomonocytic leukemia (CMML). S3a protein levels were heterogeneous in these populations. In 32 cryopreserved blast populations, S3a levels were significantly correlated with both bcl-2 and with cell growth in culture. As in cell lines, high S3a in cryopreserved blasts was associated with ATRA-induced sensitization to ara-C. No significant association was seen between S3a levels and response to treatment.
- Published
- 2000
43. Increased sensitivity of acute myeloid leukemias to lovastatin-induced apoptosis: A potential therapeutic approach.
- Author
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Dimitroulakos J, Nohynek D, Backway KL, Hedley DW, Yeger H, Freedman MH, Minden MD, and Penn LZ
- Subjects
- Acute Disease, Adult, Aged, Aged, 80 and over, Antineoplastic Agents therapeutic use, Female, Flow Cytometry, Humans, Hydroxymethylglutaryl-CoA Reductase Inhibitors therapeutic use, Leukemia, Myeloid enzymology, Lovastatin therapeutic use, Male, Microscopy, Electron, Middle Aged, Tumor Cells, Cultured, Antineoplastic Agents pharmacology, Apoptosis drug effects, Hydroxymethylglutaryl-CoA Reductase Inhibitors pharmacology, Leukemia, Myeloid drug therapy, Leukemia, Myeloid pathology, Lovastatin pharmacology
- Abstract
We recently demonstrated that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme of de novo cholesterol synthesis, was a potential mediator of the biological effects of retinoic acid on human neuroblastoma cells. The HMG-CoA reductase inhibitor, lovastatin, which is used extensively in the treatment of hypercholesterolemia, induced a potent apoptotic response in human neuroblastoma cells. This apoptotic response was triggered at lower concentrations and occurred more rapidly than had been previously reported in other tumor-derived cell lines, including breast and prostate carcinomas. Because of the increased sensitivity of neuroblastoma cells to lovastatin-induced apoptosis, we examined the effect of this agent on a variety of tumor cells, including leukemic cell lines and primary patient samples. Based on a variety of cytotoxicity and apoptosis assays, the 6 acute lymphocytic leukemia cell lines tested displayed a weak apoptotic response to lovastatin. In contrast, the majority of the acute myeloid leukemic cell lines (6/7) and primary cell cultures (13/22) showed significant sensitivity to lovastatin-induced apoptosis, similar to the neuroblastoma cell response. Of significance, in the acute myeloid leukemia, but not the acute lymphocytic leukemia cell lines, lovastatin-induced cytotoxicity was pronounced even at the physiological relevant concentrations of this agent. Therefore, our study suggests the evaluation of HMG-CoA reductase inhibitors as a therapeutic approach in the treatment of acute myeloid leukemia.
- Published
- 1999
44. Phosphorylation of BCL-2 after exposure of human leukemic cells to retinoic acid.
- Author
-
Hu ZB, Minden MD, and McCulloch EA
- Subjects
- Dimerization, Electrophoresis, Gel, Two-Dimensional, Humans, Immunosorbent Techniques, Paclitaxel pharmacology, Phosphoamino Acids analysis, Phosphoric Monoester Hydrolases metabolism, Phosphorus Radioisotopes, Phosphorylation, Phosphoserine metabolism, Recombinant Proteins, Tumor Cells, Cultured, Leukemia, Myeloid, Acute metabolism, Proto-Oncogene Proteins c-bcl-2 metabolism, Tretinoin pharmacology
- Abstract
Serine phosphorylation of bcl-2 has been reported after treatment of cells with protein kinase C, okadaic acid, taxol, and other chemotherapeutic agents that attack microtubules. We report here that bcl-2 is phosphorylated on serine in acute myeloblastic leukemia (AML) blasts exposed to all trans retinoic acid (ATRA). Two-dimension gels (isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis [SDS-PAGE]) disclosed a novel acidic isoform of bcl-2 in ATRA-treated blast cells from a continuous line and from two AML patients; when the cell lysates were digested with lambda-phosphatase, bcl-2 reverted to the control position, indicating that it was phosphorylated. Metabolic labeling experiments using 32Pi showed that, while control bcl-2 was labeled, incorporation was greatly increased when cells were treated with ATRA. A comparison of bcl-2 from blasts treated with ATRA or taxol showed that bcl-2 was phosphorylated on serine in cells treated with either agent; however, both qualitative and quantitative differences were seen. Qualitatively, the phosphorylated isoform from taxol-treated cells was slightly larger than the native isoform and could be distinguished on 10% to 20% SDS-polyacrylamide gradient gels, while the phosphorylated bcl-2 after ATRA ran as a single band on gradient gels at the same position as control bcl-2. Quantitatively, all bcl-2 from ATRA-treated cells was in the phosphorylated isoform, while after taxol, both phosphorylated and native bcl-2 was present; incorporation of 32Pi into bcl-2 was stimulated to greater extent in ATRA-treated compared with taxol-treated cells. We used immunoprecipitation experiments to ask if bcl-2 phosphorylated after ATRA or taxol had altered capacity to dimerize with bax. No change in dimerization was demonstrated. We conclude that: bcl-2 is phosphorylated on serine after treatment of AML blasts with ATRA; bcl-2 phosphorylation after ATRA is different from that seen after taxol; bcl-2 phosphorylated after either agent retains capacity to dimerize with bax. The ATRA or taxol-induced phosphorylation of bcl-2 can also be seen in blast cells obtained from AML patients., (Copyright 1998 by The American Society of Hematology.)
- Published
- 1998
45. Defects of the mismatch repair gene MSH2 are implicated in the development of murine and human lymphoblastic lymphomas and are associated with the aberrant expression of rhombotin-2 (Lmo-2) and Tal-1 (SCL).
- Author
-
Lowsky R, DeCoteau JF, Reitmair AH, Ichinohasama R, Dong WF, Xu Y, Mak TW, Kadin ME, and Minden MD
- Subjects
- Adaptor Proteins, Signal Transducing, Animals, Basic Helix-Loop-Helix Transcription Factors, DNA-Binding Proteins genetics, Exons genetics, Homeodomain Proteins biosynthesis, Homeodomain Proteins genetics, Humans, LIM Domain Proteins, Metalloproteins genetics, Mice, Mice, Knockout, Molecular Sequence Data, MutS Homolog 2 Protein, Oncogene Proteins biosynthesis, Oncogene Proteins genetics, Precursor Cell Lymphoblastic Leukemia-Lymphoma pathology, Proto-Oncogenes, T-Cell Acute Lymphocytic Leukemia Protein 1, DNA Repair genetics, DNA-Binding Proteins biosynthesis, DNA-Binding Proteins physiology, Fungal Proteins, Gene Expression Regulation, Neoplastic, Metalloproteins biosynthesis, Precursor Cell Lymphoblastic Leukemia-Lymphoma genetics, Proto-Oncogene Proteins, Transcription Factors
- Abstract
Mutations in the DNA mismatch repair (MMR) gene hMSH2 underlie a novel pathway of tumorigenesis for some cancers of epithelial origin. Mice deficient in MSH2 are susceptible to lymphomas but defects in this gene have not been identified in human lymphoid tumors. To determine if the lymphomas these mice develop are related to a particular subtype of human lymphoma we evaluated 20 clinically ill homozygous MSH2-/- mice ranging in age from 2 to 13 months. The murine tumors comprised a single histopathologic entity representing the malignant counterpart of precursor thymic T cells and closely resembled human precursor T-cell lymphoblastic lymphoma (LBL). Evaluation of the expression of three T-cell malignancy associated genes showed that Rhombotin-2 (RBTN-2 also known as Lmo-2), TAL-1 (also known as SCL), and HOX-11 were expressed in 100%, 40%, and 0% of the murine tumors, respectively. The MSH2-/- murine model of precursor T-cell LBL was substantiated by the finding of a nearly identical expression profile of RBTN-2, TAL-1, and HOX-11 in 10 well-characterized cases of human LBL. Direct evidence for MSH2 abnormalities in human LBL was established by sequence analysis of exon 13 of hMSH2, which revealed coding region mutations in 2 of 10 cases. Our findings implicate defects in the MMR system with the aberrant expression of T-cell specific proto-oncogenes and define a new pathway of human lymphomagenesis.
- Published
- 1997
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