scholarly journals Cell cycle phase-dependent effect of retinoic acid on the induction of granulocytic differentiation in HL-60 promyelocytic leukemia cells

FEBS Letters ◽  
1993 ◽  
Vol 318 (2) ◽  
pp. 193-199 ◽  
Author(s):  
Eric Ka-Wai Hui ◽  
Benjamin Yat-Ming Yung
Keyword(s):  
Cell Cycle ◽  
Retinoic Acid ◽  
Promyelocytic Leukemia ◽  
Cell Cycle Phase ◽  
Leukemia Cells ◽  
Cycle Phase ◽  
Granulocytic Differentiation ◽  
Dependent Effect

scholarly journals Vinblastine sensitizes leukemia cells to cyclin-dependent kinase inhibitors, inducing acute cell cycle phase-independent apoptosis

2011 ◽  
Vol 12 (4) ◽  
pp. 314-325 ◽  
Author(s):  
Darcy J.P. Bates ◽  
Bethany L. Salerni ◽  
Christopher H. Lowrey ◽  
Alan Eastman
Keyword(s):  
Cell Cycle ◽  
Kinase Inhibitors ◽  
Cell Cycle Phase ◽  
Leukemia Cells ◽  
Cycle Phase ◽  
Cyclin Dependent Kinase

APX3330 inhibition of the redox function of ape-1/ref-1 (Ref-1) in promyelocytic leukemia cells enhances retinoic acid (ATRA) induced myeloid differentiation and limits cell proliferation as an approach to the prevention of the retinoic acid syndrome (RAS)

2009 ◽  
Vol 27 (15_suppl) ◽  
pp. e14613-e14613
Author(s):  
K. A. Robertson ◽  
E. S. Colvin ◽  
M. R. Kelley ◽  
M. L. Fishel
Keyword(s):  
Flow Cytometry ◽  
Retinoic Acid ◽  
Cell Growth ◽  
Real Time ◽  
Real Time Pcr ◽  
Mapk Pathway ◽  
Promyelocytic Leukemia ◽  
Leukemia Cells ◽  
Myeloid Differentiation ◽  
Granulocytic Differentiation

e14613 Background: ATRA + chemotherapy has improved the treatment of promyelocytic leukemia(APL). However, 25% of ATRA treated APL patients experience toxicities that comprise the RAS (life-threatening respiratory distress, edema, renal failure, hypotension, coagulopathy and rising blast count). One approach to prevent RAS is to limit blast proliferation and enhance myeloid differentiation. Ref-1 is a DNA repair protein that functions in redox regulation of cellular proteins, such as Fos, Jun, p53, and NFkB. HL60 myeloid leukemia cells are promyeloblasts that respond to ATRA with granulocytic differentiation/growth arrest. Prior studies suggest Ref-1 redox control is integral to ATRA-induced differentiation. To define the role of the redox function of Ref-1, we used the Ref-1 specific drug, APX3330, to block Ref-1 redox function and examined the response of HL60 cells to ATRA. Methods: Cell growth assessed using trypan blue. Differentiation was evaluated by morphology and expression of CD11b by flow cytometry. Apoptosis was assayed by annexin-PI staining on flow cytometry and cell cycle analysis assayed with propidium iodide flow cytometry. To assess activation of the MAPK pathway, BLR-1 expression was determined by real time PCR. Results: 1) APX3330 blockade of Ref-1 redox function resulted in limited cell growth yet a profound increase in differentiation and a moderate increase in apoptosis. 2) dose dependent studies with ATRA showed a similar degree of differentiation in cells treated with 10 μM ATRA to cells treated with APX3330 + 0.01 μM ATRA; allowing HL60 cells + APX3330 to give a similar response to a 1000 fold lower dose of ATRA. APX3330 alone did not induce differentiation and induced only minimal apoptosis but in combination with ATRA, increased the number of cells in G1/G0 phase significantly. 3) APX3330 + ATRA increased BLR-1 expression significantly by real time PCR suggesting enhanced activation of the MAPK pathway. Conclusions: APX3330 + ATRA limits HL60 growth and dramatically enhances terminal granulocytic differentiation. These finding may provide a therapeutic approach for prevention of the RAS. No significant financial relationships to disclose.

Combination of retinoic acid and tumor necrosis factor overcomes the maturation block in a variety of retinoic acid-resistant acute promyelocytic leukemia cells

Blood ◽  
2004 ◽  
Vol 104 (10) ◽  
pp. 3335-3342 ◽  
Author(s):  
Michael Witcher ◽  
Hoi Ying Shiu ◽  
Qi Guo ◽  
Wilson H. Miller
Keyword(s):  
Retinoic Acid ◽  
Tumor Necrosis Factor ◽  
Cell Line ◽  
Acute Promyelocytic Leukemia ◽  
Tumor Necrosis ◽  
Promyelocytic Leukemia ◽  
Leukemia Cells ◽  
Monocytic Differentiation ◽  
Granulocytic Differentiation ◽  
Necrosis Factor

Abstract Retinoic acid (RA) overcomes the maturation block in t(15:17) acute promyelocytic leukemia (APL), leading to granulocytic differentiation. Patients receiving RA alone invariably develop RA resistance. RA-resistant cells can serve as useful models for the development of treatments for both APL and other leukemias. Previously, we showed that RA and tumor necrosis factor (TNF) promote monocytic differentiation of the APL cell line NB4 and U937 monoblastic cells. Here, we report that combining TNF with RA leads to maturation of several RA-resistant APL cells along a monocytic pathway, whereas UF-1, a patient-derived RA-resistant cell line, showed characteristics of granulocytic differentiation. We found distinct differences in gene regulation between UF-1 cells and cells showing monocytic differentiation. Although IRF-7 was up-regulated by TNF and RA in all cells tested, expression of c-jun and PU.1 correlated with monocytic differentiation. Furthermore, synergistic induction of PU.1 DNA binding and macrophage colony-stimulating factor receptor (m-CSF-1R) mRNA was observed only in cells differentiating into monocytes. Using neutralizing antibodies against m-CSF-1R or its ligand, we found that inhibiting this pathway strongly reduced CD14 expression in response to RA and TNF, suggesting that this pathway is essential for their synergy in RA-resistant leukemia cells. (Blood. 2004;104:3335-3342)

Impact of Protein Phosphatase PP2A on ATRA-Induced Activation of FoxO3a In Acute Promyelocytic Leukemia Cells

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2492-2492
Author(s):  
Yasuhiko Sakoe ◽  
Kumi Sakoe ◽  
Haruo Shimazaki ◽  
Keita Kirito ◽  
Norio Komatsu
Keyword(s):  
Retinoic Acid ◽  
Cell Growth ◽  
Fusion Protein ◽  
Acute Promyelocytic Leukemia ◽  
Cellular Responses ◽  
Promyelocytic Leukemia ◽  
Leukemia Cell Line ◽  
Leukemia Cells ◽  
Atra Treatment ◽  
Granulocytic Differentiation

Abstract Abstract 2492 Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia caused by reciprocal translocations of the long arms of chromosomes 15 and 17, which prevent cellular differentiation into mature neutrophils. The translocation of the promyelocytic leukemia (PML) gene on chromosome 15 and a retinoic acid receptor α (RARα) gene on chromosome 17 generates a PML-RARα fusion protein that inhibits PML-dependent apoptotic pathways in a dominant negative fashion. This fusion protein also blocks granulocytic differentiation by direct transcriptional inhibition of retinoic acid target genes. All-trans retinoic acid (ATRA) arrests cell growth, granulocytic differentiation, and apoptosis in APL cells via proteasome-dependent degradation of PML-RARα fusion protein and subsequent PML-nuclear body (NB) formation. Although PML is the essential component of PML-NBs and functions as a tumor suppressor, disruption of PML-NBs by the PML-RARα fusion protein inhibits endogenous PML tumor-suppressive functions in APL cells. Therefore, degradation of PML-RARα fusion protein and reorganization of PML-NBs during ATRA treatment are regarded as critical cellular responses, similar to the cell growth arrest and apoptosis of leukemia cells. Recently we demonstrated that FoxO3a (also named FKHRL1), a member of the Forkhead family of transcription factors, is a key molecule for the ATRA-induced cellular responses in APL cells (Blood 2010; 115: 3787–3795). In this study, we investigated the mechanism by which FoxO3a is activated by ATRA treatment in a human promyelocytic leukemia cell line NB4. Okadaic acid, a potent PP2A inhibitor, cancelled ATRA-induced dephosphorylation of AKT and its downstream molecule FoxO3a in NB4 cells. Knockdown of endogenous PP2A by siRNA significantly enhanced phosphorylation of both AKT and FoxO3a. These results suggested that PP2A is involved in ATRA-induced dephosphorylation of AKT and FoxO3a. Concomitantly, PP2AC, a catalytic subunit of PP2A, was dephoshorylated at tyrosine 307, and phosphatase activity of PP2A increased after ATRA treatment. Co-immunoprecipitation assay revealed that PP2A constitutively and directly binds to FoxO3a. Using artificial oligopeptides, we demonstrated that enhanced PP2A activity by ATRA directly dephosphorylates phosphothreonine 32 on FoxO3a. In addition, we found that 14-3-3 epsilon binded to phosphorylated FoxO3a in the cytoplasm in the absence of ATRA. After ATRA treatment, however, dephosphorylated FoxO3a dissociated from 14-3-3 epsilon and moved into the nucleus. Confocal microscopic analysis revealed that PP2A-FoxO3a complex partially co-localized with PML-NBs in the nucleus after ATRA treatment. Together, PML orchestrates nuclear networking with PP2A and FoxO3a for ATRA-induced granulocytic differentiation and apoptosis of APL cells. Disclosures: No relevant conflicts of interest to declare.

Oncogene-Induced DNA Repair Defects Promote PARP1-Mediated “Dual Synthetic Lethality” To Eradicate Quiescent and Proliferating Leukemia Stem and Progenitor Cells

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 810-810 ◽  
Author(s):  
Margaret Nieborowska-Skorska ◽  
Artur Slupianek ◽  
Grazyna Hoser ◽  
Elisabeth Bolton-Gillespie ◽  
Alexei Tulin ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Synthetic Lethality ◽  
Cell Cycle Phase ◽  
Leukemia Cells ◽  
Cycle Phase ◽  
Dsb Repair ◽  
Normal Cells ◽  
Quiescent Cells ◽  
Repair Pathways ◽  
Parp1 Inhibitors

Abstract Leukemia stem cells (LSCs), including quiescent cells, are disease initiating and therapy-refractory cells. Therefore, even if a treatment clears a disease burden consisting mostly of leukemia progenitor cells (LPCs), it usually fails to eradicate LSCs and residual LPCs which developed therapy-resistance. Leukemia cells expressing BCR-ABL1 (CML, Ph+ALL), FLT3(ITD) (AML), and AML1-ETO (AML) accumulate high numbers of spontaneous DNA double strand breaks (DSBs). Leukemia cells can survive numerous DSB, the most lethal DNA lesions, due to enhanced DSB repair activity by homologous recombination (HR; active in S and G2 cell cycle phase) and/or non-homologous end-joining (NHEJ; active in G0/G1 and S cell cycle phase). Altogether, leukemia cells may be “addicted” to DSB repair pathways. Normal cells usually employ BRCA1/2-RAD51 -mediated HRR and Ku70/86-DNA-PKcs -dependent NHEJ. BCR-ABL1 causes BRCA1 and DNA-PKcs deficiency, FLT3(ITD) inhibits Ku86, and AML1-ETO downregulates DNA-PKcs and RAD51. Accordingly, leukemia cells expressing these oncogenes are forced to employ alternative DSB repair pathways, such as PARP-LigIII –mediated NHEJ. Since NHEJ plays a predominant role in quiescent cells and also supports HR in proliferating cells, we postulated that targeting PARP should exert “dual synthetic lethality” to eradicate quiescent LSCs and proliferating LSCs/LPCs, with negligible effect on normal cells. We showed that PARP inhibitor olaparib, which is in clinical trials for the treatment of solid tumors displaying BRCA1/2 mutations, abrogated NHEJ activity in BCR-ABL1 cells. Olaparib reduced the number of imatinib-naïve and imatinib-treated Lin-CD34+CD38-CTVmax quiescent CML-CP LSCs in hypoxia and normoxia mimicking bone marrow niche and arterial peripheral blood, respectively. In addition, olaparib enhanced the anti-proliferative effect of imatinib in Lin-CD34+CML-CP and CML-AP LPCs. These effects are probably due to imatinib-mediated inhibition of BCR-ABL1 kinase-dependent anti-apoptotic activity and olaparib-induced increase of the number of lethal DSBs, resulting in accumulation of annexin V-positive apoptotic quiescent LSCs and hyper-activation of caspase-3 in imatinib+olaparib treated LPCs. Inhibition of PARP almost completely abrogates DSB repair in DNA-PKcs-deficient quiescent LSCs and diminishes resolution of ROS-induced stalled replication forks in BRCA1-deficient proliferating LSCs/LPCs. The combination of imatinib+olaparib did not affect normal quiescent hematopoietic stem cells, but exerted modest inhibitory effect on normal proliferating progenitors. Since olaparib does not discriminate neither between PARP family members nor other enzymatic pathways involving NAD+, its long-term application may generate side-effects. Our genetic studies involving Parp1-/- mice and PARP1(E988K) catalytic-deficient mutant identified PARP1 as major player in NHEJ in BCR-ABL1 cells. Using high throughput screening we identified 5F2, a small molecule which abrogated histone 4-dependent PARP1 activation and exerted synthetic lethality in BRCA1-deficient, but not BRCA1-proficient carcinoma cells. 5F2, similarly to olaparib, reduced the number of imatinib-naïve and imatinib-treated Lin-CD34+CD38-CTVmax quiescent LSCs and inhibited colony formation by Lin-CD34+LPCs. However 5F2, in contrast to olaparib, did not affect normal cells. In addition, olaparib and/or 5F2 reduced the number of imatinib-naïve and imatinib-treated Ph+ALL cells harvested from patients at diagnosis. Moreover, PARP1 inhibitors exerted anti-leukemia effect against ponatinib-naïve and ponatinib-treated Ph+ALL cells carrying BCR-ABL1 T315I mutation, and against lestaurtinib/quizartinib-naïve and lestaurtinib/quizartinib-treated FLT3(ITD)-positive AML cells. PARP1 inhibitors also abrogated the growth of leukemia cells expressing AML1-ETO (AML), but not of these expressing PML-RAR (APL) or overexpressing HOXA9+MEIS1 (AML). In conclusion, targeting PARP1 resulted in the induction of “dual synthetic lethality” and eradication of quiescent and proliferating CML cells displaying specific defects in DSB repair pathways. Similar effect is induced in other leukemias carrying specific, oncogene-induced DSB repair deficiencies. PARP1 inhibitors are currently tested in vivo using primary leukemia xenografts. Disclosures: Valent: Novartis: Consultancy, Honoraria, Research Funding.

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