High Expression of Decoy Receptor TRAIL-R3 on AML Blasts Is Associated to Shortened Overall Survival. TRAIL-R3 Induced Apoptosis Resistance Can Be Overcome by Specifically Targeting TRAIL-R2 in Vitro

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2946-2946
Author(s):  
Martine Chamuleau ◽  
Linda van Dreunen ◽  
S.M.G. Jirka ◽  
A. Zevenbergen ◽  
G. Ossenkoppele ◽  
...  
Keyword(s):  
Overall Survival ◽  
Cell Lines ◽  
Leukemic Cell ◽  
Receptor Expression ◽  
Death Domain ◽  
Apoptosis Induction ◽  
Leukemic Cell Lines ◽  
Decoy Receptors ◽  
Myeloid Leukemic Cell

Abstract In vivo, effector T cells and NK cells do express membrane-bound TRAIL (TNF-related apoptosis-inducing ligand). T cells also secrete a soluble form of TRAIL (sTRAIL). Binding of TRAIL to one of the receptors on target cells can induce apoptosis. In human, 4 membranebound receptors for TRAIL have been identified: two of them (TRAIL-R1 and TRAIL-R2) contain a functional death domain, while the two other (TRAIL-R3 and TRAIL-R4) lack a functional death domain and function as decoy receptors. Most normal cells express TRAIL-R3 and TRAIL-R4, where many tumor cells express TRAIL-R1 and TRAIL-R2, making activation of the TRAIL mediated apoptotic pathway a possible new model for targeted therapy. We investigated bone-marrow samples from 119 patients with de novo AML and from 11 healthy donors for TRAIL receptor expression on CD45dim/SSClow blasts by flow-cytometry. Expression profiles of patients were correlated to survival data. Functional data were obtained by incubation of 6 myeloid leukemic cell lines and fresh AML samples with different concentrations of sTRAIL/Apo2L (n=10) and specific antibodies directly targeting R1 and R2 (HGS-ETR1 and HGS-ETR2) (n=18). Apoptosis was assessed by flow–cytometry after staining with 7AAD and annexinV. R3 expression on the cell surface of one cell line (MM6) was down-modulated with PI-PLC (Phosphatidyl-inositol phospholipase C ).When comparing AML and normal blasts, significantly more pro-apoptotic TRAIL-R1 and TRAIL-R2 receptor expression on myeloid leukemic blasts (p<0.0005) was measured however without association with outcome. About 30% of AML patients had a high anti-apoptotic TRAIL-R3 expression, which was strongly correlated to a shortened overall survival (log rank, p=0.0051). In multivariate analysis, R3 expression remained a significant prognostic factor next to cytogenetics (p=0.03 and p= 0.015 respectively). In vitro, studies on 6 myeloid leukemic cell lines confirmed sTRAIL sensitivity in cell lines that expressed presumably TRAIL-R1 and TRAIL-R2. Moreover, down-modulation of TRAIL-R3 by treatment with PI-PLC (TRAIL-R3 is the only GPI-linked TRAIL receptor) on cell lines improved sTRAIL sensitivity. Fresh myeloid leukemic samples were treated with sTRAIL. Mean apoptosis induction was 14% (0–54%). sTRAIL can bind to both the pro-apoptotic as the decoy receptors. Bypassing of the decoy R3 expression by treatment with the antibody directly targeting TRAIL-R2 (anti-TRAIL-R2) resulted in much higher rates of apoptosis (mean 30%, 0–80%). Apoptosis induction via direct targeting of TRAIL-R2 was, as expected, independent of TRAIL-R3 expression. All leukemic blasts that showed apoptosis with anti-TRAIL-R2 expressed the R2 receptor, but not all blasts that expressed TRAIL-R2 were sensitive for anti-TRAIL-R2, which could be due to inhibitory factors downstream the apoptotic cascade. In conclusion, AML blasts do express pro-apoptotic TRAIL receptors. However, co-expression with the decoy receptor TRAIL-R3 results in significant shortened overall survival. AML blasts are sensitive to targeting the pro-apoptotic TRAIL-R2 receptor, yielding a new therapeutic option for AML patients.

Repositioning of Difluorinated Propanediones as Inhibitors of Histone Methyltransferases and their Biological Evaluation in Human Leukemic Cell Lines

2019 ◽  
Vol 18 (13) ◽  
pp. 1892-1899 ◽  
Author(s):  
Tanushree Pal ◽  
Asmita Sharda ◽  
Bharat Khade ◽  
C. Sinha Ramaa ◽  
Sanjay Gupta
Keyword(s):  
Cell Lines ◽  
Physiological Role ◽  
Leukemic Cell ◽  
Docking Studies ◽  
Biological Evaluation ◽  
Leukemic Cell Lines ◽  
Histone Lysine ◽  
Histone Lysine Methyltransferase ◽  
Subsequent Decrease

Background: At present, ‘pharmaco-epigenomics’ constitutes the hope in cancer treatment owing to epigenetic deregulation- a reversible process and playing a role in malignancy. Objective: Chemotherapy has many limitations like host-tissue toxicity, drug resistance. Hence, it is imperative to unearth targets to better treat cancer. Here, we intend to repurpose a set of our previously synthesized difluorinated Propanediones (PR) as Histone lysine Methyltransferase inhibitors (HMTi). Methods: The cell lines of leukemic origin viz. histiocytic lymphoma (U937) and acute T-cell leukemia (JURKAT) were treated with PR-1 to 7 after docking studies with active pocket of HMT. The cell cycle analysis, in vitro methylation and cell proliferation assays were carried out to delineate their physiological role. Results: A small molecule PR-4, at 1 and 10µM, has shown to alter the methylation of histone H3 and H4 in both cell lines. Also, treatment shows an increase in G2/M population and a subsequent decrease in the G0/G1 population in U937. In JURKAT, an increase in both G2/M and S phase population was observed. The sub-G1 population showed a steady rise with increase in dose and prolonged time intervals in U937 and JURKAT cell lines. In SRB assay, the PR showed a cell growth of 42.6 and 53.4% comparable to adriamycin; 44.5 and 53.2% in U937 and JURKAT, respectively. The study suggests that PR-4 could emerge as a potential HMT inhibitor. Conclusion: The molecule PR-4 could be a lead in developing more histone lysine methyltransferases inhibitors with potential to be pro-apoptotic agents.

Differential expression of a novel proline-rich homeobox gene (Prh) in human hematolymphopoietic cells

Blood ◽  
1995 ◽  
Vol 85 (5) ◽  
pp. 1237-1245 ◽  
Author(s):  
G Manfioletti ◽  
V Gattei ◽  
E Buratti ◽  
A Rustighi ◽  
A De Iuliis ◽  
...  
Keyword(s):  
Cell Lines ◽  
Constitutive Expression ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Maturation Stage ◽  
Lymphoid Tissues ◽  
Specific Cell ◽  
B Cell Differentiation ◽  
Leukemic Cell Lines

Proline-rich homeobox (Prh) is a novel human homeobox-containing gene recently isolated from the CD34+ cell line KG-1A, and whose expression appears mainly restricted to hematopoietic tissues. To define the pattern of Prh expression within the human hematopoietic system, we have analyzed its constitutive expression in purified cells obtained from normal hematopoietic tissues, its levels of transcription in a number of leukemia/lymphoma cell lines representing different lineages and stages of hematolymphopoietic differentiation, and its regulation during in vitro maturation of human leukemic cell lines. Prh transcripts were not detected in leukemic cells of T-lymphoid lineage, irrespective of their maturation stage, and in resting or activated normal T cells from peripheral blood and lymphoid tissues. In contrast, high levels of Prh expression were shown in cells representing early stages of B lymphoid maturation, being maintained up to the level of circulating and tissue mature B cells. Terminal B-cell differentiation appeared to be conversely associated with the deactivation of the gene, since preplasmacytic and plasmocytoma cell lines were found not to express Prh mRNA. Prh transcripts were also shown in human cell lines of early myelomonocytic, erythromegakaryocytic, and preosteoclast phenotypes. Prh expression was lost upon in vitro differentiation of leukemic cell lines into mature monocyte-macrophages and megakaryocytes, whereas it was maintained or upregulated after induction of maturation to granulocytes and osteoclasts. Accordingly, circulating normal monocytes did not display Prh mRNA, which was conversely detected at high levels in purified normal granulocytes. Our data, which show that the acquisition of the differentiated phenotype is associated to Prh downregulation in certain hematopoietic cells but not in others, also suggest that a dysregulated expression of this gene might contribute to the process of leukemogenesis within specific cell lineages.

scholarly journals Akacid Medical Formulation Induces Apoptosis in Myeloid and Lymphatic Leukemic Cell Lines In Vitro and In Vivo

PLoS ONE ◽  
2015 ◽  
Vol 10 (2) ◽  
pp. e0117806 ◽  
Author(s):  
Hannes Neuwirt ◽  
Elisabeth Wabnig ◽  
Clemens Feistritzer ◽  
Iris E. Eder ◽  
Christina Salvador ◽  
...  
Keyword(s):  
Cell Lines ◽  
Leukemic Cell ◽  
Leukemic Cell Lines

Bioproduct obtained from probiotic microorganisms consortia-studies regarding the effects generated in vitro on two types of leukemic cell lines

2017 ◽  
Vol 655 (1) ◽  
pp. 275-286
Author(s):  
Nicoleta Radu ◽  
Viviana Roman ◽  
Marinela Bostan ◽  
Mariana Voicescu ◽  
Ciprian Tanasescu
Keyword(s):  
Cell Lines ◽  
Leukemic Cell ◽  
Leukemic Cell Lines

scholarly journals Bcr-abl translocation can occur during the induction of multidrug resistance and confers apoptosis resistance on myeloid leukemic cell lines

1997 ◽  
Vol 4 (8) ◽  
pp. 806-814 ◽  
Author(s):  
Francis Belloc ◽  
Sophie Cotteret ◽  
Gilles Labroille ◽  
Valérie Schmit ◽  
Claudine Jaloustre ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Cell Lines ◽  
Leukemic Cell ◽  
Apoptosis Resistance ◽  
Leukemic Cell Lines ◽  
Myeloid Leukemic Cell

scholarly journals Synthesis of Two Novel Homologous Polyphosphoesters Containing Aminophosphonate Units and Cytotoxicity of Some Low-Molecular and Polymeric Aminophosphonate Derivatives

2018 ◽  
Vol 2018 ◽  
pp. 1-8
Author(s):  
Ivanka Kraicheva ◽  
Georgi Momekov ◽  
Rositsa Mihaylova ◽  
Margarita Topashka-Ancheva ◽  
Ivelina Tsacheva ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Lines ◽  
Leukemic Cell ◽  
The Novel ◽  
Histological Subtype ◽  
New Materials ◽  
Antitumor Effects ◽  
Cytotoxicity Assays ◽  
Leukemic Cell Lines

Two novel polyphosphoesters containing anthracene- and furan-derived aminophosphonate moieties, namely, poly[oxyethylene(aminophosphonate-co-H-phosphonate)]s P-12 and P-13, were synthesized through an addition of poly(oxyethylene H-phosphonate) to 9-anthrylidene-furfurylamine and characterized. The novel polyphosphoester P-12 and a series of previously described anthracene-derived compounds including Schiff bases S-1 and S-2, α-aminophosphonates A-3–A-6, bis-aminophosphonate B-6, two enantiomers A-5a and A-5b, and polyphosphoesters P-8–P-11 containing aminophosphonate units were screened for antitumor activity against a panel of human leukemic cell lines, using cisplatin as a reference cytotoxic agent. As concluded from the cytotoxicity assays, both precursors S-1 and S-2 presented similar cytotoxicity profiles that are cisplatin-like only in the REH cell line. Leader compound of the α-aminophosphonates is A-4 with cell death-inducing properties fully equaling those of the referent drug in all of the screened leukemic cell lines with the only exception being the AML histological subtype HL-60. Some of the polymeric analogues elicited moderate (P-10 and P-12) to low (P-11) cytotoxic activity, whereas the polyphosphoesters P-8 and P-9 produced in vitro antitumor effects largely surpassing cisplatin’s. The compounds P-8, P-9, and A-4 could be potential new materials for anticancer therapeutic purposed.

Pioglitazone Inhibits the Growth of Human Leukemic Cell Lines and Primary Leukemic Cells in Vitro.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4493-4493 ◽  
Author(s):  
Yoshihiro Hatta ◽  
Minoru Saiki ◽  
Yuko Enomoto ◽  
Shin Aizawa ◽  
Umihiko Sawada ◽  
...  
Keyword(s):  
Cell Lines ◽  
Colony Formation ◽  
Mononuclear Cells ◽  
Hematopoietic Cells ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Peroxisome Proliferator ◽  
Leukemic Cell Lines ◽  
Ppar Γ

Abstract Troglitazone and pioglitazone are one of thiazolidinediones that are high affinity ligand for the nuclear receptor called peroxisome proliferator-activated receptor gamma (PPAR-γ). Troglitazone is a potent inhibitor of clonogenic growth of acute myeloid leukemia cells when combined with a retinoid. However, the effect of pioglitazone to neoplastic cells and normal hematopoietic cells has not been studied yet. Adult T-cell leukemia (ATL), prevalent in western Japan, is a highly aggressive malignancy of mature T lymphocyte. Therefore, we studied antitumor effect of pioglitazone against leukemic cells including ATL as well as normal hematopoietic cells. With 300 μM of pioglitazone, colony formation of ATL cell lines (MT1, MT2, F6T, OKM3T, and Su9T01) was completely inhibited. Colony formation of HUT102, another ATL cell line, was 12 % compared to untreated control. Clonogenic cells of other leukemic cell lines (K562, HL60, U937, HEL, CEM, and NALM1) was also inhibited to 0–30% of control. Colony formation of primary leukemic cells from 5 AML patients was decreased to 15 %. However, normal hematopoietic cells were weakly inhibited with 300 μM pioglitazone; 77 % of CFU-GM, 70 % of CFU-E, and 33 % of BFU-E survived. Cell cycle analysis showed that pioglitazone decreased the ratio of G2/M phase in HL60 cells, suggesting the inhibition of cell division. By Western blotting, PPAR-γ protein level was similar in all leukemic cells and normal bone marrow mononuclear cells. Taken together, pioglitazone effectively eliminate leukemic cells and could be used as an antitumor agent in vivo.

ARRY-429520, a Novel KSP Inhibitor with Potent Anti-Proliferative Activity in Leukemic Cell Lines.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4401-4401 ◽  
Author(s):  
Christine Lemieux ◽  
Walt DeWolf ◽  
Walt Voegtli ◽  
Eli Wallace ◽  
Rich Woessner ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Tumor ◽  
Leukemic Cell ◽  
Proliferating Cells ◽  
Single Digit ◽  
Leukemic Cell Lines ◽  
Microtubule Disruption ◽  
In Vitro Characterization ◽  
Ksp Inhibitor

Abstract Kinesins are eukaryotic microtubule-associated motor proteins. There are over 40 known kinesins, approximately 15 of which are closely associated with mitosis. Kinesin Spindle Protein (KSP), also known as Eg5, is a mitotic kinesin that is a required enzyme in mitosis (prophase / prometaphase). This protein plays a key role in the formation of the bipolar spindle, particularly related to its role in centrosome maturation/separation. Because KSP is expressed predominately in proliferating cells and is absent from postmitotic neurons, its inhibition should not produce the peripheral neuropathy associated with traditional microtubule disruption agents (taxanes and vinca alkaloids). We report here the in vitro characterization of a potent KSP inhibitor, ARRY-429520, a member of a series of KSP inhibitors discovered and optimized by structure-based design. ARRY-429520 inhibits human KSP, with an IC50 of 6 nM, by a mechanism which was demonstrated to be uncompetitive with respect to ATP and noncompetitive with respect to tubulin. It was shown to arrest cells in mitosis as measured by FACs analysis as well as the accumulation of phospho- histone H3, with an EC50 of 1.5 nM. Furthermore, this compound was demonstrated to be antiproliferative, with EC50s between 0.3 nM and 6.5 nM against a panel of human tumor cell lines, including various leukemia lines (K-562, KU-812, HL-60, KG-1, MOLT3, MOLT4). In addition, ARRY-429520’s potency in MDR-overexpressing cell lines was minimally impacted as compared to paclitaxel. Cellular imaging studies demonstrate that the normal mitotic spindle configuration was disrupted, with the formation of monopolar spindles, a hallmark feature of KSP inhibition, at single digit nanomolar concentrations of ARRY-429520. Markers of mitotic arrest and apoptosis were demonstrated in tumor xenografts from animals treated with ARRY-429520.

TG101348, An Orally Bioavailable JAK2-Selective Inhibitor, Potently Inhibits KITD816V and FIP1L1-PDGFRA in Vitro

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2807-2807 ◽  
Author(s):  
Jaya Kittur ◽  
Terra L Lasho ◽  
Joseph H Butterfield ◽  
Jolene Shorr ◽  
Ayalew Tefferi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Leukemic Cell ◽  
Colony Growth ◽  
Selective Inhibitor ◽  
Jak Inhibitor ◽  
Leukemic Cell Lines ◽  
Drug Concentrations ◽  
Orally Bioavailable ◽  
Eosinophil Colonies

Abstract Background: Systemic mastocytosis (SM) and hypereosinophilic syndrome (HES) are JAK2V617F-negative orphan diseases without effective long-term therapeutic options. Both of these diseases share clinical and laboratory features with FIP1L1-PDGFRA-positive chronic eosinophilic leukemia (CEL). SM is associated with KITD816V or other KIT mutations. HES is currently not molecularly characterized. The exquisite sensitivity of FIP1L1-PDGFRA to inhibition by imatinib mesylate (IM) underlies the efficacy of this agent in treating FIP1L1-PDGFRA-positive CEL. In contrast, KITD816V is resistant to inhibition by IM, and is only moderately sensitive to dasatinib (cellular IC50=micromolar concentrations), which may explain the relatively disappointing clinical responses in SM with these agents. TG101348 is an orally bioavailable JAK2-selective inhibitor that is currently being tested in a Phase 1 clinical trial for the treatment of myelofibrosis. The aim of the current study was to evaluate TG101348 (relative to the tyrosine kinase inhibitors IM, dasatinib, and sorafenib) for its ability to: inhibit growth of leukemic cell lines that carry KITD816V and FIP1L1-PDGFRA; and inhibit the in vitro growth of eosinophil colonies derived from progenitor cells from HES patients. Methods: The following cell lines were used for in vitro experiments: HMC-1, a mast cell leukemia line (KITD816V-positive); EOL-1, a CEL derived line (FIP1L1-PDFRA-positive), Ba/F3 T674I, a Ba/F3 line that express the IM-resistant FIP1L1-PDFRAT674I mutation, and HEL, a human erythroleukemia line (JAK2V617F-positive). Cell proliferation assays were performed in triplicate using the XTT assay (Leukemia. 2007 21:1658). Drug concentrations in these experiments ranged from 2.9 × 10−12 M to 10−4 M. JAK Inhibitor I (Calbiochem) is a non-selective JAK inhibitor tool compound. Eosinophil colonies were obtained by plating PBMCs from healthy controls or HES patients in methylcellulose, in the presence of IL-3, IL-5, and GM-CSF. Effects of IM and TG101348 on eosinophil colony growth were studied in parallel experiments. Results: Consistent with prior observations, the HMC-1 cell line was resistant to inhibition by IM (IC50>10 mM), but was moderately sensitive to dasatinib (Table). In contrast, TG101348 inhibited HMC-1 growth at nanomolar concentrations, similar to its effect on HEL cells that harbor the JAK2V617F mutation. TG101348 inhibited HMC-1 growth 53-fold and 4-fold more potently than IM and dasatinib, respectively (Figure). Sorafenib, a multikinase inhibitor that reportedly has activity against KIT kinase (enzyme IC50=68 nM), had only a limited effect on HMC-1 growth (IC50=4000 nM). In contrast to HMC-1, EOL-1 growth was inhibited by IM, dasatinib, sorafenib, and TG101348 at picomolar concentrations (Table). The T674I mutation (in the context of FIP1L1-PDGFRA) corresponds to the T315I mutation in BCR-ABL – it occurs infrequently in CEL and is usually found in the context of transformation to acute leukemia. Growth of Ba/F3 cells harboring FIP1L1-PDFGRA-T674I has been shown to be inhibited by sorafenib (confirmed in this report; Table) and nilotinib, at nanomolar concentrations. TG101348 is less potent at inhibiting growth of these cells, with cellular IC50 of ~2 mM – pharmacokinetic data from the ongoing Phase I study of TG101348 shows that this concentration is achievable in plasma with once daily dosing. Drug Cellular IC50 HMC-1 (nM) EOL-1 (pM) Ba/F3 T674I (nM) HEL (nM) Imatinib 18800 4 10700 - Dasatinib 1300 <1 30900 - Sorafenib 4000 2 16 - Jak Inhibitor I 926 30 4300 ? TG101348 355 <1 1950 300 Figure Figure Conclusions: The JAK2-selective inhibitor TG101348 is a potent inhibitor of KITD816V and FIP1L1-PDGFRA, warranting clinical trials using this drug in SM and FIP1L1- PDGFRA-positive CEL. Data regarding TG101348 effects on signaling intermediates in leukemic cell lines and on eosinophil colony growth will be presented at the meeting – data from the latter experiments may indicate a therapeutic role for TG10134 in the treatment of patients with HES.

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