Vitamin E in Chronic Myeloid Leukemia (CML) Prevention

The resistance to inhibitors of tyrosine kinase necessitates novel approaches to the therapy of chronic myeloid leukemia (CML). The progression of CML to blast crisis is associated with down-regulation of C/EBP-alpha being involved in the differentiation block in leukemic blast cells. Moreover, lowered C/EBP-alpha expression correlates with resistance to imatinib in CML. We have demonstrated that vitamin E up-regulates expression of C/EBP-alpha and down-regulates expression of Snail transcription factor in K562 cells in vitro contributing to the putative recovery of myeloid differentiation potential. In parallel with increased CEBP alpha expression, Vitamin E treatment results in the decreasing expression of placental-like alkaline phosphatase and increasing expression of tissue non-specific alkaline phosphatase. We suggest that vitamin E could be used as the plausible biological modulator to prevent the progression to blast crisis and to overcome drug resistance of leukemic cells in CML.

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Synergic Effects of Proteasome Inhibitor PS-341 and Tyrosine Kinase Inhibitor STI571 on Chronic Myeloid Leukemia Cells and BCR-ABL Oncoprotein In Vitro and In Vivo.

Blood ◽

10.1182/blood.v108.11.4771.4771 ◽

2006 ◽

Vol 108 (11) ◽

pp. 4771-4771

Author(s):

Guangbiao Zhou ◽

Zheng Hu ◽

Dapeng Liu ◽

Fuqun Wu ◽

Jiang Zhu ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Tumor Growth ◽

Proteasome Inhibitor ◽

Myeloid Leukemia ◽

K562 Cells ◽

Leukemic Cells ◽

Great Promise ◽

Dependent Manner

Abstract STI571/Gleevec/imatinib, a rationally-designed agent that occupies the ATP-binding site of BCR-ABL and stabilizes the protein in its closed, inactive conformation, has been a remarkable success for the treatment of chronic myeloid leukemia (CML). However, a significant proportion of patients chronically treated with STI571 develop resistance because of the acquisition of mutations in the kinase domain of BCR-ABL. Furthermore, the effects of STI571 on CML patients in accelerated phase or blastic crisis are unsatisfactory since many patients relapse after transient remission. Hence, additional drugs or STI571-based combination regimens are desired to circumvent resistance and to improve response rates. Here we reported that PS-341, a proteasome inhibitor which offers great promise to patients with multiple myeloma (MM), significantly enhanced the antileukemia activity of STI571 in vitro and in vivo. We found a synergy exists between low concentrations of PS-341 (5–10 nM) and STI571 (0.1–0.2 μM) in inhibition of cell growth and induction of apoptosis in K562 cell line and CD34+ leukemic cells isolated from CML patients. In K562 cells, combined use of PS-341 and STI571 accelerated activation of caspase-3, 9, and facilitated cleavage of poly-(ADP-ribose) polymerase (PARP) as compared to those in cells treated with PS-341 or STI571 alone. Moreover, PS-341/STI571 combination resulted in potentiated degradation of BCR-ABL and downregulation of phosphorylated BCR-ABL as compared to those in mono treatment. In nude mice inoculated subcutaneously with K562 cells, treatment with PS-341 (injected intraperitoneally, ip) alone (at doses of 0.05, 0.5, 1 mg/kg/d, twice a week for 4 weeks, respectively) decreased tumor growth in a dose-dependent manner. STI571 (ip) at 10 mg/kg/d also inhibited tumor growth. Intriguingly, combinatory administration of low dose PS-341 (0.05 mg/kg/d, twice a week for 4 weeks) and STI571 (10 mg/kg/d) yielded a much more profound inhibition of tumor growth and even clearance of leukemic cells in mice compared to either monotherapy. Taken together, these results demonstrate synergic effects of PS-341 and STI571, and provide the rationale to evaluate PS-341/STI571 combination in treating CML aiming to further improve clinical outcome of patients.

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Complete Remission of Accelerated Phase Chronic Myeloid Leukemia by Treatment With Leukemia-Reactive Cytotoxic T Lymphocytes

Blood ◽

10.1182/blood.v94.4.1201.416k08_1201_1208 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1201-1208 ◽

Cited By ~ 1

Author(s):

J.H. Frederik Falkenburg ◽

Amon R. Wafelman ◽

Peter Joosten ◽

Willem M. Smit ◽

Cornelis A.M. van Bergen ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Progenitor Cells ◽

Myeloid Leukemia ◽

Cytotoxic T Lymphocyte ◽

Blast Crisis ◽

Chronic Phase ◽

Donor Lymphocyte Infusion ◽

Leukemic Cells

Relapse of chronic myeloid leukemia (CML) in chronic phase after allogeneic stem cell transplantation (SCT) can be successfully treated by donor lymphocyte infusion (DLI). However, relapse of accelerated phase CML, blast crisis, or acute leukemia after allogeneic SCT are resistant to DLI in the majority of cases. In vitro-selected and expanded leukemia-reactive T-cell lines may be more effective in inducing an antileukemic response in vivo. To treat a patient with accelerated phase CML after allogeneic SCT, leukemia-reactive cytotoxic T-lymphocyte (CTL) lines were generated from her HLA-identical donor. Using a modification of a limiting dilution assay, T cells were isolated from the donor, selected based on their ability to inhibit the in vitro growth of CML progenitor cells, and subsequently expanded in vitro to generate CTL lines. Three CTL lines were generated that lysed the leukemic cells from the patient and inhibited the growth of leukemic progenitor cells. The CTL did not react with lymphocytes from donor or recipient and did not affect donor hematopoietic progenitor cells. The 3 leukemia-reactive CTL lines were infused at 5-week intervals at a cumulative dose of 3.2 × 109 CTL. Shortly after the third infusion, complete eradication of the leukemic cells was observed, as shown by cytogenetic analysis, fluorescence in situ hybridization, molecular analysis of BCR/ABL-mRNA, and chimerism studies. These results show that in vitro cultured leukemia-reactive CTL lines selected on their ability to inhibit the proliferation of leukemic progenitor cells in vitro can be successfully applied to treat accelerated phase CML after allogeneic SCT.

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High Levels of Circulating HSP70 Levels in Patients with Chronic Myeloid Leukemia and Correlation with Resistance to Therapy with Imatinib.

Blood ◽

10.1182/blood.v110.11.2922.2922 ◽

2007 ◽

Vol 110 (11) ◽

pp. 2922-2922

Author(s):

Richard Tseng ◽

Chen-Hsiung Yeh ◽

Iman Jilani ◽

Zhong Zhang ◽

Hagop Kantarjian ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Myeloid Leukemia ◽

Blast Crisis ◽

Death Receptor ◽

Chronic Phase ◽

Leukemic Cells ◽

Hsp70 Protein ◽

N 93

Abstract Cellular increase in the heat shock protein 70 (HSP70) inhibits death receptor and mitochondria-initiated signaling for apoptosis. In Vitro and in vivo data from patients with chronic myeloid leukemia (CML) indicates that overexpression of HSP70 in leukemic cells is associated with resistance to imatinib. The HSP70 protein is detectable in the plasma and has been reported to represent a marker for cardiovascular stress as well as prostate cancer. We measured HSP70 in the plasma of 139 patients with CML using a sandwich assay based on meso scale technology and correlated levels of HSP70 protein in plasma with clinical behavior. All samples were collected prior to initiating imatinib therapy. The levels of HSP70 in CML patients in chronic phase (N=93) were not significantly (P=0.08) different from those in patients with CML in accelerated/blast crisis (N=46) (median 33.24 ng/ml, range: 3.892–128.172 ng/ml Vs median 26.57 ng/ml, range: 4.498–114.746 ng/ml, respectively). However, CML patients had significantly (P<0.0001) higher levels of HSP70 than normal control (N=95, median=4.17 ng/ml, range:1.746–24.684 ng/ml). There was significant correlation between plasma levels of HSP70 and platelets (r=0.54), WBC (r=0.32), and basophils (r=0.31) in patients in chronic phase and with platelets (r=0.72), blasts (r=0.39), and basophils (r=0.50) in patients in Acc/Bl phase. Patients in chronic phase and high levels above the median had significantly (P=0.02) higher rate of progression to ACC/Bl phase while on therapy with imatinib (figure). In addition these patients appear to have a tendency toward shorter survival (P=0.07). There was no significant correlation between plasma HSP70 levels and survival in patients with ACC/Bl phase. This data support the reported role of HSP70 in the resistance to imatinib in patients with CML and potentially plasma HSP70 levels may represent a marker for resistance in patients with chronic phase CML. Figure Figure

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Complete Remission of Accelerated Phase Chronic Myeloid Leukemia by Treatment With Leukemia-Reactive Cytotoxic T Lymphocytes

Blood ◽

10.1182/blood.v94.4.1201 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1201-1208 ◽

Cited By ~ 214

Author(s):

J.H. Frederik Falkenburg ◽

Amon R. Wafelman ◽

Peter Joosten ◽

Willem M. Smit ◽

Cornelis A.M. van Bergen ◽

...

Keyword(s):

Chronic Myeloid Leukemia ◽

Progenitor Cells ◽

Myeloid Leukemia ◽

Cytotoxic T Lymphocyte ◽

Blast Crisis ◽

Chronic Phase ◽

Donor Lymphocyte Infusion ◽

Leukemic Cells

Abstract Relapse of chronic myeloid leukemia (CML) in chronic phase after allogeneic stem cell transplantation (SCT) can be successfully treated by donor lymphocyte infusion (DLI). However, relapse of accelerated phase CML, blast crisis, or acute leukemia after allogeneic SCT are resistant to DLI in the majority of cases. In vitro-selected and expanded leukemia-reactive T-cell lines may be more effective in inducing an antileukemic response in vivo. To treat a patient with accelerated phase CML after allogeneic SCT, leukemia-reactive cytotoxic T-lymphocyte (CTL) lines were generated from her HLA-identical donor. Using a modification of a limiting dilution assay, T cells were isolated from the donor, selected based on their ability to inhibit the in vitro growth of CML progenitor cells, and subsequently expanded in vitro to generate CTL lines. Three CTL lines were generated that lysed the leukemic cells from the patient and inhibited the growth of leukemic progenitor cells. The CTL did not react with lymphocytes from donor or recipient and did not affect donor hematopoietic progenitor cells. The 3 leukemia-reactive CTL lines were infused at 5-week intervals at a cumulative dose of 3.2 × 109 CTL. Shortly after the third infusion, complete eradication of the leukemic cells was observed, as shown by cytogenetic analysis, fluorescence in situ hybridization, molecular analysis of BCR/ABL-mRNA, and chimerism studies. These results show that in vitro cultured leukemia-reactive CTL lines selected on their ability to inhibit the proliferation of leukemic progenitor cells in vitro can be successfully applied to treat accelerated phase CML after allogeneic SCT.

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Aberrant expression of placental-like alkaline phosphatase in chronic myeloid leukemia cells in vitro and its modulation by vitamin E

Experimental Oncology ◽

10.32471/exp-oncology.2312-8852.vol-42-no-1.14285 ◽

2020 ◽

Vol 42 (1) ◽

Author(s):

G.D. Telegeev ◽

◽

D.F. Gluzman ◽

M.P. Zavelevich ◽

L.P. Shvachko ◽

...

Keyword(s):

Alkaline Phosphatase ◽

Vitamin E ◽

Chronic Myeloid Leukemia ◽

Myeloid Leukemia ◽

Leukemia Cells ◽

Aberrant Expression

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Fucose binding lectin for characterizing acute myeloid leukemia progenitor cells

Blood ◽

10.1182/blood.v68.1.41.41 ◽

1986 ◽

Vol 68 (1) ◽

pp. 41-45 ◽

Cited By ~ 14

Author(s):

R Delwel ◽

I Touw ◽

F Bot ◽

B Lowenberg

Keyword(s):

Acute Myeloid Leukemia ◽

Myeloid Leukemia ◽

Fluorescence Analysis ◽

Leukemic Cells ◽

Ulex Europaeus ◽

Acute Myeloid Leukemia Cells ◽

Blast Cells ◽

Binding Lectin ◽

Acute Myeloid

Abstract The reactivity of acute myeloid leukemia cells (AML) was determined in 29 patients using the fucose binding lectin Ulex europaeus agglutinin (UEA) as surface marker. We show a marked heterogeneity in the UEA- binding abilities of the cells in these patients as determined by fluorescence analysis of the blasts labeled with the UEA coupled to the fluorescent molecule FITC. The results suggest a correlation between the capability of AML blast cells to bind UEA and cytologic maturation, because in 1 of 10 M1, 3 of 8 M2, 6 of 8 M4, and 1 of 3 M5 cytology types UEA binding to the leukemic cells was apparent. In 13 cases, the cells gave rise to colonies in vitro. The amount of UEA binding to AML colony-forming cells (AML-CFU) was determined by cell sorting and subsequent colony culture of UEA-negative, intermediately positive, and highly fluorescent cells. AML-CFU from none of the four patients with M1 cytology were UEA positive, whereas they showed intense reactivity with the lectin in 1 of 4 cases with M2 cytology and in all 4 cases of M4. In these five cases with strongly UEA positive AML-CFU, the fluorescence distribution of the colony formers differed from that of the total leukemia population, indicating that AML-CFU represent a subpopulation of AML cells with specific UEA-binding properties. Normal bone marrow myeloid and multipotential colony-forming cells (CFU-GM, CFU-GEMM) showed low or no binding of UEA. UEA-FITC appears a useful reagent for membrane analysis of AML-CFU. In certain cases, UEA-FITC labeling may be applied to discriminate AML-CFU from normal hematopoietic progenitors.

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Phenotypic heterogeneity of primitive leukemic hematopoietic cells in patients with chronic myeloid leukemia

Blood ◽

10.1182/blood.v80.10.2522.2522 ◽

1992 ◽

Vol 80 (10) ◽

pp. 2522-2530 ◽

Cited By ~ 38

Author(s):

C Udomsakdi ◽

CJ Eaves ◽

PM Lansdorp ◽

AC Eaves

Keyword(s):

Chronic Myeloid Leukemia ◽

Myeloid Leukemia ◽

Philadelphia Chromosome ◽

Significant Proportion ◽

Hematopoietic Cells ◽

Chronic Phase ◽

Leukemic Cells ◽

Clonogenic Cell ◽

Clonogenic Cells

Abstract The peripheral blood of chronic myeloid leukemia (CML) patients with chronic-phase disease and elevated white blood cell (WBC) counts typically contains markedly increased numbers of a variety of neoplastic pluripotent and lineage-restricted hematopoietic progenitors. These include cells detected in standard colony assays as well as their more primitive precursors. The latter are referred to as long-term culture-initiating cells (LTC-IC) because of their ability to generate clonogenic cell progeny detectable after a minimum of 5 weeks incubation on competent fibroblast feeder layers. In this study, we have investigated a number of the properties of the LTC-IC and clonogenic cells present in the blood of such CML patients with high WBC counts. This included an analysis of the light scattering properties of these progenitors, as well as their expression of CD34 and HLA-DR, Rhodamine-123 staining, and in vitro sensitivity to 4- hydroperoxycyclophosphamide. In the case of LTC-IC, the production of different types of lineage-restricted and multipotent progeny was also analyzed. Most of the circulating LTC-IC and clonogenic cells in the CML patients studied (on average approximately 70% and approximately 90%, respectively) showed features of proliferating or activated cells. This is in marked contrast to the majority of progenitors in the blood of normal individuals and most of the LTC-IC in normal marrow, all of which exhibit a phenotype expected of quiescent cells. Interestingly, a significant proportion of the circulating clonogenic cells and LTC-IC in the CML samples studied (on average approximately 10% and approximately 30%, respectively) appeared to be phenotypically similar to normal circulating progenitors, although their absolute numbers were indicative of a neoplastic origin. Both phenotypes of circulating CML clonogenic cells and LTC-IC could be obtained at approximately 10% to 20% purity by differential multiparameter sorting. These findings suggest that expansion of the Philadelphia chromosome-positive clone at the level of the earliest types of hematopoietic cells results from the activation of mechanisms that enable some, but not all, signals that block the cycling of normal stem cells to be bypassed or overcome. In addition, they provide strategies for purifying these primitive leukemic cells that should facilitate further analysis of the mechanisms underlying their abnormal proliferative behavior.

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DC-Based Vaccines Show in-Vitro Activity against Chronic Myeloid Leukemia Cells

Blood ◽

10.1182/blood.v112.11.4276.4276 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4276-4276

Author(s):

Yuen-Fen Tan ◽

Soon-Keng Cheong ◽

Chooi-Fun Leong ◽

SAW Fadhilah

Keyword(s):

T Cells ◽

Immune System ◽

Chronic Myeloid Leukemia ◽

Myeloid Leukemia ◽

Residual Disease ◽

Leukemic Cells ◽

Gm Csf ◽

Dc Vaccine ◽

Hla Dr

Abstract Chronic Myeloid leukemia is a common myeloproliferative disease. Despite recent advances in targeted therapy, only 7–12% of patients achieve molecular remission. Leukemic cells arrange multiple mechanisms to avoid recognition by the immune system. Dendritic cells (DC) are professional antigen presenting cells of the immune system playing a crucial role in the induction of anti-tumor responses. The use of DC is an attractive immunotherapeutic strategy against cancers, especially in minimal residual disease state. In this study, DC vaccine against chronic myeloid leukemia was generated and evaluated in-vitro. Monocytes were isolated and enriched from peripheral blood. These monocytes were subsequently cultured in RPMI medium supplemented with GM-CSF and IL-4 to induce them to become DC. These DC were then co-cultured with tumor lysates obtained from CML cell line in culture medium supplemented with GM-CSF, IL-4 and TNF alpha to become DC-based CML vaccine. The generated DC-based CML vaccines retained their DC morphology, showed strong expression of CD 86 and HLA-DR, and were negative for CD14. Mixed lymphocyte reaction indicated that the generated DC-based CML vaccines were capable of inducing proliferative responses to allogeneic lymphocytes. DC-based CML vaccines were shown to stimulate T cells to express DC-ligands, ie CD28 and CD154, as well as HLA-DR, CD71 and CD 25. In addition, the stimulated T cells were cytotoxic to CML cells used to prepare tumor lysates.

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Arsenic induces apoptosis of multidrug-resistant human myeloid leukemia cells that express Bcr-Abl or overexpress MDR, MRP, Bcl-2, or Bcl-xL

Blood ◽

10.1182/blood.v95.3.1014.003k04_1014_1022 ◽

2000 ◽

Vol 95 (3) ◽

pp. 1014-1022 ◽

Cited By ~ 162

Author(s):

Charles Perkins ◽

Caryn N. Kim ◽

Guofu Fang ◽

Kapil N. Bhalla

Keyword(s):

Myeloid Leukemia ◽

Blast Crisis ◽

K562 Cells ◽

Cell Types ◽

Multidrug Resistant ◽

Leukemia Cells ◽

Protein Levels ◽

Growth Inhibitory ◽

Induced Apoptosis

We investigated the in vitro growth inhibitory and apoptotic effects of clinically achievable concentrations of As2O3 (0.5 to 2.0 μmol/L) against human myeloid leukemia cells known to be resistant to a number of apoptotic stimuli. These included chronic myelocytic leukemia (CML) blast crisis K562 and HL-60/Bcr-Abl cells, which contain p210 and p185 Bcr-Abl, respectively, and HL-60 cell types that overexpress Bcl-2 (HL-60/Bcl-2), Bcl-xL(HL-60/Bcl-xL), MDR (HL-60/VCR), or MRP (HL-60/AR) protein. The growth-inhibitory IC50 values for As2O3 treatment for 7 days against all these cell types ranged from 0.8 to 1.5 μmol/L. Exposure to 2 μmol/L As2O3 for 7 days induced apoptosis of all cell types, including HL-60/Bcr-Abl and K562 cells. This was associated with the cytosolic accumulation of cyt c and preapoptotic mitochondrial events, such as the loss of inner membrane potential (▵Ψm) and the increase in reactive oxygen species (ROS). Treatment with As2O3 (2 μmol/L) generated the activities of caspases, which produced the cleavage of the BH3 domain containing proapoptotic Bid protein and poly (ADP-ribose) polymerase. Significantly, As2O3-induced apoptosis of HL-60/Bcr-Abl and K562 cells was associated with a decline in Bcr-Abl protein levels, without any significant alterations in the levels of Bcl-xL, Bax, Apaf-1, Fas, and FasL. Although As2O3 treatment caused a marked increase in the expression of the myeloid differentiation marker CD11b, it did not affect Hb levels in HL-60/Bcr-Abl, K562, or HL-60/neo cells. However, in these cells, As2O3 potently induced hyper-acetylation of the histones H3 and H4. These findings characterize As2O3 as a growth inhibiting and apoptosis-inducing agent against a variety of myeloid leukemia cells resistant to multiple apoptotic stimuli.

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