Daunorubicin Group Research Articles

Effect of TRIM59 Expression Interference on Daunorubicin Chemosensitivity of Chronic Myeloid Leukemia K562 Cells and Its Mechanism

To investigate the effect of tripipartite motif 59 (TRIM59) expression interference on the chemosensitivity of daunorubicin (DNR) in chronic myeloid leukemia (CML) K562 cells and the related molecular mechanism. The expressions of TRIM59 mRNA in bone marrow tissues of patients with CML and K562 cells were detected by RT-qPCR. Liposome-based transfection technology was used to transfect TRIM59-specific siRNA (si-TRIM59) into K562 cells which then were treated with DNR. The proliferation and apoptosis of cells were detected by CCK-8 assay and flow cytometry, respectively, and the expressions of apoptosis-related protein and Wnt/β-catenin signaling pathway-related protein were detected by Western blot. Compared with the bone marrow tissue of CML patients at the time of initial treatment, the expression of TRIM59 mRNA in bone marrow tissue of CML patients at the time of chemotherapy resistance was significantly increased (P <0.05). Compared with control group, the cell proliferation inhibition rate and apoptosis rate in si-TRIM59 group and DNR group were significantly increased (P <0.05), the expression of Bax, Caspase3 and Cleaved-Caspase3 protein were significantly increased (P <0.05), while the expressions of Bcl-2, Wnt3α, GSK-3β protein and the ratio of p-β-catenin/β-catenin were significantly decreased (P <0.05). Compared with si-TRIM59 group and DNR group, the proliferation inhibition rate and apoptosis rate of si-TRIM59+DNR group were significantly increased (P <0.05), the expression of Bax, Caspase3 and Cleaved-Caspase3 protein were significantly increased, while the expression of Bcl-2, Wnt3α, GSK-3β protein and the ratio of p-β-catenin/β-catenin were significantly decreased (P <0.05). TRIM59 expression interference may enhance the chemosensitivity of K562 cells to DNR, and its mechanism may be related to the regulation of Wnt/β-catenin signaling pathway.

Study on the Effect and Mechanism of Tetrandrine on Bone Marrow Mesenchymal Stem Cell-Mediated Drug Resistance in Leukemia

To explore the role of bone marrow mesenchymal stem cells (BMSC)，an essential element of the bone marrow microenvironment, in multidrug resistance(MDR) of K562 cells, as well as the reversal effect of tetrandrine (TET) on BMSC-mediated MDR and its potential mechanism. A mixed co-culture system and a transwell co-culture system for BMSC and K562 cells were established, and the cells were divided into different groups and treated with daunorubicin (DNR) alone or combined with TET and DNR. The CCK-8 assay was used to detect the proliferation of K562 cells in each group, and the cell inhibition rate was calculated. Cytometric bead array (CBA) was used to detect the expression levels of IFN, IL-2, IL-6 and IL-10 in the supernatant of different groups. RT-qPCR and Western blot were used to detected the expression of STAT3 at mRNA and protein levels, respectively. Compared with K562+DNR group, the inhibition rate of DNR on K562 cell proliferation in K562+BMSC+DNR group was significantly decreased (P < 0.05), while the levels of IL-6 in the culture supernatant and phosphorylated STAT3 in K562 cells were significantly increased (P < 0.05). Compared with K562+BMSC+DNR group, the inhibition rate of DNR on K562 cell proliferation in K562+BMSC+DNR+TET group was significantly increased (P < 0.05), while the level of IL-6 and phosphorylated STAT3 was significantly decreased (P < 0.05). BMSC can promote the drug resistance of leukemia cells, and TET may reverse the BMSC-mediated drug resistance via inhibiting IL-6/STAT3 signaling pathway.

Basic Research on Therapy of Leukemia with Nanoparticles Drug Delivery System Modified by Transferrin Receptor Monoclonal Antibody

To investigate the therapeutic effect of targeted drug-loaded nanoparticles modified by transferrin receptor monoclonal antibody (TfR mAb) on acute leukemia and its potential anti-tumor mechanism. Nanoparticles drug delivery system, which was composed of poly (lactic-co-glycolic acid), poly-l-lysine, polyethylene glycol, TfR mAb (TfR mAb-PLGA-PLL-PEG)-daunorubicin (DNR), was first synthesized. After drug intervention, the intracellular accumulation in leukemia HL60 cells was observed under a fluorescent microscope and concentration of DNR was determined by flow cytometry (FCM). Meanwhile, cell apoptosis rate was measured by FCM and the expression levels of apoptosis related protein Cleaved-caspase 3 was determined by Western blot. Under an inverted fluorescent microscope, intracellular accumulation of DNR autofluorescence in HL60 cells was observed in both TfR mAb-PLGA-PLL-PEG-DNR group and DNR group. FCM analysis showed that the intracellular concentration of DNR in TfR mAb-PLGA-PLL-PEG-DNR group was higher than that in DNR group（P<0.05）. The apoptotic rate of HL60 cells in TfR mAb-PLGA-PLL-PEG-DNR group was higher than that of DNR group（P<0.05）. Moreover, the expression levels of apoptosis-related protein Cleaved-caspase 3 in TfR mAb-PLGA-PLL-PEG-DNR group was significantly higher than that in DNR group（P<0.05）. TfR mAb-PLGA-PLL-PEG nanoparticle drug delivery system can target chemotherapy drugs to leukemia cells and enhance anticancer ability through apoptotic pathway.

Effects and Mechanisms of Homoharringtonine on Expression of CEBPA Protein

To investigate the effect of homoharringtonine (HHT) on CEBPA protein and explore the mechanism of HHT in the treatment of acute myeloid leukemia (AML) with double CEBPA mutations. The K562 cell line expressing CEBPA p30 (K562 CEBPA p30) was established. Western blot was used to determine the changes of the expression of CEBPA protein in K562 CEBPA p30, U937 and MOLM-13 cell lines before and after treatments with HHT, daunorubicin (DNR) or cytarabine (Ara-C). The effects of protease inhibitors and protein synthesis inhibitors on the expression of CEBPA protein were also determined. RNA-seq was used to analyze the difference of gene expressions and pathway enrichments between HHT group and DNR group. Both the endogenous CEBPA protein in U937 and MOLM-13 cell lines and the exogenous CEBPA protein in K562 CEBPA p30 were decreased by HHT (P<0.05) while were not by DNR or Ara-C. Proteasome inhibitors can increase the expression of CEBPA protein (P<0.05) while protein synthesis inhibitors can decrease the expression of CEBPA protein (P<0.05). The ribosome biogenesis related pathways in K562 CEBPA p30 were upregulated in HHT group while were not in DNR group. HHT can inhibit the synthesis of CEBPA and reduce the expression of CEBPA protein and this may be the mechanism of HHT in the treatment of CEBPA-double-mutant AML.

The Mechanism of Artesunate Combined with Cytarabine and/or Daunorubicin on the Apoptosis of MV4-11 MLL-rearranged Acute Myeloid Leukemia Cell Line

To investigate the effect and mechanism of artesunate (ARTS) combined with cytarabine(Ara-C) and/or daunorubicin (DNR) on the proliferation and apoptosis of MV4-11 human mixed-lineage leukemia rearranged（MLL-r） acute myeloid leukemia (AML) cell line. CCK-8 assay was used to detect the proliferation effect of individual or in combination of ARTS, DNR, Ara-C on MV4-11 cells. The IC50 of ARTS, DNR and Ara-C was calculated separately. The cell apoptosis and expression of receptors DR4 and DR5 were detected by flow cytometry. Western blot was used to detect the expression of Caspase-3 and Caspase-9 in each groups. The inhibition effect of ARTS, Ara-C and DNR on the proliferation of MV4-11 were all dose-dependently (r=0.99, 0.90 and 0.97, respectively). The IC50 of ARTS, Ara-C and DNR on MV4-11 for 48 hours were 0.31 μg/ml, 1.43 μmol/L and 22.47 nmol/L, respectively. At the dose of ARTS 0.3 μg/ml， Ara-C 1.0 μmol/L and DNR 15 nmol/L, the proliferation rate for 48 hours of the tri-combination treatment was significantly lower than that of the bi-combination treatment, while both were significantly lower than that of the individual treatment (all P＜0.05). In terms of bi-combination treatment, the cells proliferation rate for 48 hours of the ARTS+Ara-C group was significantly lower than that of the ARTS+DNR group, while both were significantly lower than that of the Ara-C+DNR group (all P＜0.05). The cooperativity index (CI) of bi- and tri-combination treatment were all less than 1. After 48 hours of drug action, the cell apoptosis rate of the ARTS+DNR+Ara-C group was significantly higher than that of the Ara-C+DNR group, while both were significantly higher than that of the ARTS+DNR group (all P＜0.05). Meanwhile, the was no statistical difference between the cells apoptotic rate of the ARTS+DNR+Ara-C group and the ARTS+Ara-C group (P>0.05). The expression of DR4 and DR5 also showed no difference between control group and drug group. Compared with the DNR+Ara-C group, the expressions of Caspase-3 were significantly down-regulated in both the ARTS+DNR+Ara-C group and the ARTS+Ara-C group (all P＜0.05). The down-regulation of Caspase-3 expression was the most significantly in the combination group of three drugs, while the Caspase-9 expressions in different groups showed no apparent change. The in vitro study showed that tri-combination of ARTS+Ara-C+DNR and bi-combination of ARTS+Ara-C could inhibit the proliferation and promote apoptosis of MV4-11 cell line. The inhibition effect of these two combinations were significantly superior to that of the traditional Ara-C+DNR treatment. The mechanism underlying this finding may be identified by the down regulation of Caspase-3, while no altered expression was observed of Caspase-9, DR4 and DR5.

Impact of the Type of Anthracycline and of Allogeneic Stem Cell Transplantation in the Treatment of Younger Acute Myeloid Leukemia Patients: Long-Term Follow-up of the EORTC-Gimema AML-10 Trial

FB, FE and SS contributed equally to the work.Background. The best post-remission treatment for younger acute myeloid leukemia (AML) patients remains controversial (Cornelissen JJ & Blaise D, Blood 2016, 127, 62-70). Most prospective studies comparing autologous (auto-) to allogeneic (allo-) hematopoietic stem cell transplantation (HSCT) have been published with a relatively short follow-up.Aim. The main objective of this study was to provide a long-term follow-up evaluation of patients previously enrolled in the EORTC/GIMEMA AML-10 study.Methods. In the EORTC/GIMEMA AML-10 prospective trial, 2157 patients aged 15-60 years were randomized to receive either daunorubicin (DNR) (50 mg/m2/d), mitoxantrone (MXR) (12 mg/m2/d), or idarubicin (IDA) (10 mg/m2/d) on days 1, 3 and 5 in addition to standard-dose cytarabine and etoposide for induction chemotherapy. A second cycle of induction was administered in patients who achieved a partial response while patients who achieved a CR or a CR with incomplete counts recovery (CRi) after 1 or 2 courses of induction chemotherapy received consolidation chemotherapy with the same anthracycline as in the induction course plus intermediate dose cytarabine. Young (≤ 45 years old) patients were then scheduled to receive an allo-HCT in first CR/CRi if they had an HLA-identical sibling donor or an auto-HCT otherwise. Median follow-up at the time of the initial report was 5.6 years (Mandelli et al., J Clin Oncol 2009, 27, 5397-5403). For the current analyses, cytogenetics were centrally reviewed and re-classified using the UK Medical Research Council (MRC) classification.Results. In the current report, median follow-up was 11 (95% CI, 9-15) years. The 5-, 10-, 15- and 20-year overall survival (OS) rates were 33.2%, 30.1%, 28.0% and 26.1% respectively. No significant difference between the 3 randomized groups regarding OS was observed (Table 1). In young patients, the OS was comparable in the 3 treatment groups, whereas, in older patients, who had generally a worse outcome, OS was prolonged in the MXT and IDA groups as compared to the DNR group (Table 1). As expected, the initial MRC cytogenetic risk group had an important impact on OS. Within each subgroup, treatment outcomes were quite homogeneous (Table 1).The impact of auto- versus allo-HCT was assessed in a cohort of 839 patients ≤ 45 years old who achieved a CR/CRi after induction chemotherapy, who generally received consolidation chemotherapy, and who were HLA-typed. A total of 330 patients had an HLA-identical sibling (donor group) while the remaining 509 patients had not (Table 2). The 10- and 15-year DFS and OS rates from CR were approximately 10% higher in patients with a donor as compared to those without (Table 2). Assessing the impact of donor availability on the DFS according to cytogenetic risk group, we observed that patients with a donor had a higher 10- and 15- yr DFS rates in all but those in the favorable MRC cytogenetic subgroup (table 2).Among patients aged 46-60 years, 635 reached CR/CRi. The outcomes (DFS and OS) of CR/CRi patients with a donor were only marginally prolonged as compared to those without a donor, which can be explained by the positive effect of decreased relapse rate being neutralized by an increased risk of death in CR/CRi. The 10- and 15-yr DFS rates were increased by 6-11% in the MXR and IDA groups as compared to the DNR (overall logrank P=0.12); the HR (MXR vs DNR) was 0.88 (99% CI, 0.66-1.17), and the HR (IDA vs DNR) was 0.79 (99% CI, 0.58-1.06). The 10- and 15-yr OS rates were also increased, by 7-14% in the MXR and IDA groups as compared to the DNR (overall logrank P=0.04); the HR (MXR vs DNR) was 0.86 (99% CI, 0.64-1.16) and the HR (IDA vs DNR) was 0.74 (99% CI, 0.54-1.10).Conclusions. Long-term follow-up of the EORTC/GIMEMA AML-10 trial confirmed that, overall, similar outcomes were obtained by using either DNR, MXR or IDA. In young patients without favorable cytogenetic features, who reached CR/CRi, the long-term outcome of those with a HLA-identical donor, who generally received AlloHSCT, was improved as compared to those without a donor. In older patients who reached CR/CRi, allo-HSCT and auto-HSCT yielded similar outcomes; however, MXR and IDA administered during induction and consolidation, provided better results than DNR. [Display omitted] DisclosuresEfficace:Incyte: Consultancy; Lundbeck: Research Funding; TEVA: Consultancy, Research Funding; Seattle Genetics: Consultancy; Amgen: Consultancy, Research Funding. Guimaraes:Janssen: Speakers Bureau; Pfizer: Honoraria; Servier: Honoraria; Roche: Honoraria; Incyte: Speakers Bureau.

Acute Promyelocytic Leukemia: A Long-Term Retrospective Study in Mexico

BackgroundThe present retrospective study reviewed acute promyelocytic leukemia (APL) cases recorded in Mexico between January 2007 and January 2017. The primary objective of the study was to evaluate overall survival (OS) in Mexican patients with APL. Secondary objective was to evaluate the impact of induction treatment with different anthracyclines on OS, event-free survival (EFS) and complications in this patient population.MethodsThe medical charts of patients referred to medical institutions in Mexico from January 2007 through January 2017 for the treatment of suspected APL were reviewed retrospectively. Patients aged 15 - 75 years, in whom the diagnosis of APL was confirmed, who had an Eastern Cooperative Group performance status of 0 - 2, and who were eligible for combined treatment with intensive chemotherapy and all-trans retinoic acid (ATRA), were included in the study. Study participants received induction and consolidation treatment with ATRA plus either daunorubicin or idarubicin, followed by 2 years of single-agent ATRA as maintenance therapy. Patients who were unable to pay for ATRA treatment received anthracycline-based induction and consolidation, with methotrexate plus mercaptopurine as maintenance therapy.ResultsA total of 360 patients from 21 public and private hospitals were included in the study. The median age of the population was 37 years, and 51% were male. Of the 360 patients, 205 (57%) vs. 155 (43%) received daunorubicin vs. idarubicin as induction treatment for APL. ATRA was administered to 201 (98%) patients in the daunorubicin group vs. 138 (89%) in the idarubicin group (P = 0.001), and was initiated at diagnosis in 92% vs. 73% of recipients, respectively (P = 0.0001). At 150 months, OS and EFS for the entire population were 84% and 79%, respectively. Both OS (90% vs. 76%, P = 0.003) and EFS (85% vs. 72%, P = 0.001) were significantly prolonged in daunorubicin vs. idarubicin recipients. Rates of complications were similar in the two groups.ConclusionsAs arsenic trioxide (ATO) is not currently available in Mexico, anthracycline plus ATRA is the mainstay of treatment for APL here. Our results confirm the efficacy of this strategy, with high OS and EFS rates being observed 12.5 years after diagnosis.

The effectiveness of three different 7 + 3 induction regimes in China: A retrospective analysis in adult patients with acute myeloid leukemia.

Objectives:In China, for economic reasons, induction regimes for acute myeloid leukemia (AML) often involve domestically produced idarubicin (IDA) rather than imported IDA. Our objective was to compare the effectiveness of induction regimens in combination with cytarabine; involving imported or domestic IDA, or daunorubicin (DNR).Methods:The study was conducted from 1st July 2012 to 30th November 2015 at Baoding No.1 Central Hospital. This was a retrospective cohort study of patients with newly diagnosed AML admitted to Baoding First Central Hospital, China. Patients were divided into three groups according to their treatment regimen: the IA-imported group, the IA-domestic group, and the DNR group. Clinical data, complete remission (CR), partial remission (PR), non-remission (NR) rates, and side effects were compared.Results:Total 282 patients were enrolled, including 123 patients in the IA-imported group, 98 in the IA-domestic group and 61 in the DNR group. The IA-domestic and IA-imported groups’ remission rates were similar (P=0.123) but significantly different from the DNR group (both P<0.05). Side effects were similar in all three groups and no severe side effects were reported.Conclusion:Cytarabine induction regimens showed similar remission rates in combination with IDA produced in China compared to imported IDA and were more effective than DNR.

LY294002 Enhaces Chemosensitivity of K562 Cells to Daunorubicin

To investigate the proliferation inhibition and pro-apoptosis effect of LY294002 (PI3K/AKT inhibtor) combined with daunorubicin (DNR) on the chronic myeloid leurenia cell line K562 and its possible mechanisms. The effect of LY294002 and DNR on the proliferation of K562 cells in different treating time and concentration were measured by MTT assay. The cell cycle was determined by flow cytometry, the mRNA and protein expression of SKP2 , P27, BCL-2 and BAX were determined by RT-PCR and Western blot. LY294002 and DNR were able to inhibit the growth of K562 cells and promote apoptosis in time- and concentration-dependent manner (P＜0.05), both the cell proliferation-inhibiting rate and apoptosis rate in combination therapy group were higher than that in DNR-monotherapy group (P＜0.05). After K562 cells treated by LY294002 combined DNR for 36 h, the cells were statistically significantly reduced in G2/M phase (P＜0.05), as compared with control group and DNR group. Compared with DNR group, the cell level of G0/G1 phase rased (P＜0.05) and cell level of S phase decreased (P＞0.05). Compared with DNR group, the expresson of SKP2 and BCL-2 mRNA decreased, and the expression of P27 mRNA increased in the combination therapy group (P＜0.05). The expression of BAX mRNA was not significantly different between different groups. The same result was found in the protein expression. LY294002 has the sensibilizative effect on DNR chemotherapy, which may be relative with blocking the cell cycle and inducing cell apoptosis.

LY294002 Enhaces Chemosensitivity of K562 Cells to Daunorubicin

To investigate the proliferation inhibition and pro-apoptosis effect of LY294002 (PI3K/AKT inhibtor) combined with daunorubicin (DNR) on the chronic myeloid leurenia cell line K562 and its possible mechanisms. The effect of LY294002 and DNR on the proliferation of K562 cells in different treating time and concentration were measured by MTT assay. The cell cycle was determined by flow cytometry, the mRNA and protein expression of SKP2 , P27, BCL-2 and BAX were determined by RT-PCR and Western blot. LY294002 and DNR were able to inhibit the growth of K562 cells and promote apoptosis in time- and concentration-dependent manner (P＜0.05), both the cell proliferation-inhibiting rate and apoptosis rate in combination therapy group were higher than that in DNR-monotherapy group (P＜0.05). After K562 cells treated by LY294002 combined DNR for 36 h, the cells were statistically significantly reduced in G2/M phase (P＜0.05), as compared with control group and DNR group. Compared with DNR group, the cell level of G0/G1 phase rased (P＜0.05) and cell level of S phase decreased (P＞0.05). Compared with DNR group, the expresson of SKP2 and BCL-2 mRNA decreased, and the expression of P27 mRNA increased in the combination therapy group (P＜0.05). The expression of BAX mRNA was not significantly different between different groups. The same result was found in the protein expression. LY294002 has the sensibilizative effect on DNR chemotherapy, which may be relative with blocking the cell cycle and inducing cell apoptosis.

Applications of daunorubicin-loaded PLGA-PLL-PEG-Tf nanoparticles in hematologic malignancies: an in vitro and in vivo evaluation.

BackgroundWith the development of drug delivery, novel tools and technological approaches have captured the attention of researchers in recent years. Several target drug delivery systems (DDSs) including nanoparticles (NPs) have been developed as an important strategy to deliver classical medicine.ObjectiveThe objective of this study was to evaluate the application of novel daunorubicin (DNR)-loaded poly(lactic-co-glycolic acid)-poly-l-lysine-polyethylene glycol-transferrin (Tf) nanoparticles (DNR-loaded NPs) in hematologic malignancies in vitro and in vivo.Materials and methodsDNR-loaded NPs were prepared by the modified double-emulsion solvent evaporation/diffusion method, and its microscopic form was observed under scanning electron microscope. Intracellular distribution of DNR was directly detected by fluorescence microscopy. After establishment of a tumor xenograft model by injecting K562 cells into the left leg of nude mice, the therapeutic effect of the DNR-loaded NPs on the growth of tumors was measured by calculating the tumor size, and the relative expression of Caspase-3 protein was detected by immunohistochemical staining. Furthermore, intracellular concentration of DNR and the extent of cell apoptosis in primary leukemia cells were quantified by flow cytometry.ResultsDNR-loaded NPs had a spherical shape of about 180 nm in diameter. DNR-loaded NP group showed a significant enhancement of cellular uptake in K562 cells compared with DNR group. Tumor inhibition rate was higher in DNR-loaded NP group in comparison with DNR group, and the relative expression of Caspase-3 protein was upregulated in DNR-loaded NP group compared with DNR group. Furthermore, DNR-loaded NPs obviously increased intracellular concentration of DNR in primary leukemia cells compared with DNR group, but there was no significant difference in primary cell apoptosis between the two groups. These findings suggest that the novel NP DDS can enhance the performance of conventional antitumor drugs and may be suitable for further application in the treatment of hematologic malignancies.

Addition of Cladribine: Improving Standard of Care in Newly Diagnosed Acute Myeloid Leukemia

Acute myeloid leukemia (AML) continues to be one of the most common myeloid malignancies in the United States. Achieving a complete response (CR) after induction with standard daunorubicin and cytarabine (7+3) remains suboptimal, occurring in 55-65% of patients. No significant advances have improved these outcomes in the last decade. A recent phase III randomized clinical trial by the Polish Adult Leukemia Group (PALG), showed that adding cladribine 5mg/m2 to standard 7+3 (Daunorubicin 60mg/m2, Ara-C 200mg/m2) improved CR rates to 68% vs 56% with standard 7+3 alone. Achieving a higher CR is important in the outcome of high-risk patients who require allogeneic stem cell transplantation (Allo-SCT) as a curative measure. At our center, we conducted a retrospective analysis evaluating the addition of cladribine to the current standard of care treatment in newly diagnosed AML. We hypothesized an improvement in CR rates in all-risk patients will allow more patients to proceed to Allo-SCT.Adult patients 18 years of age or older with newly diagnosed AML who received remission induction chemotherapy between May 2012 and May 2018 were included in the study. A total of 118 AML patients were screened and those who did not complete induction or did not have bone marrow biopsy at end of induction chemotherapy were excluded. A total of 100 patients were evaluable of which 26 (26%) received DAC and 74 (74%) received 7+3. Within the 7+3 group, 29 (39%) received a Daunorubicin dose of 60mg/m2 (D60) and 45 (61%) received a dose of 90mg/m2 (D90). A higher absolute CR rate was noted after one induction in the DAC group as compared to the combined 7+3 group (21 [80.7%] vs. 49 [66.2%], respectively). The improvement in CR between DAC and D60 reached statistical significance (p=0.043) but there was no statistically significance difference in CR rates between DAC vs D90 (p=0.48). It is interesting to note that more patients in D60 (39%) and D90 (26%) group required re-induction as compared with DAC (12%) (p=0.067). Median overall survival (OS) for the DAC group was not reached, while median OS for D60 was 10 (DAC vs D60 p=0.076) and for D90 was 28 months (DAC vs D90 p=0.97). In the subgroup of patients who underwent Allo-SCT, median OS in the D90 group was 30 months (DAC vs D90 p=0.43) while not reached in the DAC and D60 group (DAC vs D60 p=0.84). For all patients, the median relapse-free survival (RFS) for the DAC cohort was not reached. Median RFS was 12 months for D60 (DAC vs D60 p=0.047) and 25 months for D90 (DAC vs D90 p=0.166) groups. There was no difference in OS and RFS between all three groups who were transplanted in first CR (CR1).In conclusion CR rates following induction chemotherapy were higher in DAC vs Daunorubicin groups, however only statistically significant with respect to D60. Additionally a statistically significant improvement in RFS and a trend towards improved OS was noted in favour of DAC vs D60. Regardless of the induction chemotherapy, there was no difference in OS and RFS of patients who were transplanted in CR1. These results have encouraged us to perform a prospective comparison of standard 7+3 with Dauno 90mg/m2 and DAC. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Synergistic antitumoral efficacy of a novel replicative adenovirus SG611-PDCD5 and daunorubicin in human leukemic cells.

BackgroundDaunorubicin is a traditional chemotherapeutic agent that plays a pivotal role in leukemia therapy. However, the dose-related toxicity remains a considerable challenge. The apoptosis-regulating gene, PDCD5, is downregulated in various tumors, including leukemias, and may provide a potential target for the diagnosis and treatment of leukemia. The purpose of this study was to construct a triple-regulated oncolytic adenovirus carrying a PDCD5 gene expression cassette (SG611-PDCD5) and explore the combined antitumor efficacy of SG611-PDCD5 in combination with low dose daunorubicin on leukemic cells.Materials and methodsA variety of leukemic cell lines, including K562, MEG-01, KG-1a, HL-60, SUP-B15, and BV-173, were cultured according to the providers’ instructions. The insertion and orientation of all recombined plasmids were confirmed by restriction enzyme digestion and PCR. The tumor-selective replication of the constructed conditionally replicating SG611-PDCD5 and its antitumor efficacy in combination with daunorubicin were characterized in leukemic cell lines in vitro and in a nude mouse xenograft model. Cell viability was detected using cell-counting kit-8. Apoptosis was detected in whole living cells using flow cytometry and in paraffin-embedded tumor tissues using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay.ResultsThe triple-regulated CRAd carrying SG611-PDCD5 and nude mouse xenograft models of K562 cells were successfully constructed. In vitro treatment with SG611-PDCD5 in combination with low-dose daunorubicin elicited more potent anti-proliferative and proapoptotic effects in leukemic cells in a dose-dependent manner. The Chou-Talalay analysis revealed synergistic anti-proliferative effects in all of the above cell lines. In the nude mice xenograft model, the tumor size in the control, daunorubicin, SG611-PDCD5, and combined treatment groups on day 10 were 170.1±47.8, 111.9±81.1, 60.7±12.3, and 33.2±17.5 mm3, respectively (all P<0.05). The results of the TUNEL assay showed significantly more apoptotic cells in the SG611-PDCD5 plus daunorubicin group than in the SG611-PDCD5 or daunorubicin groups alone (25±0.82, 12.5±2.27, and 7.8±2.67 apoptotic cells/field, respectively) (P<0.05).ConclusionThe findings suggest that combined treatment with SG611-PDCD5 and daunorubicin may be a promising strategy for enhancing chemosensitivity and thus lowering the dose-related toxicity of daunorubicin in leukemia therapy.

Abstract 1313: Targeting micelles eradicate acute myeloid leukemia stem cells in a patient-derived leukemia xenograft model

Abstract Patients with acute myeloid leukemia (AML) have a very poor prognosis related to a high rate of recurrence rate drug-related toxicity. The current standard “7+3” regimen, with cytarabine and an anthracycline such as daunorubicin (DNR), can achieve complete remission in up to 70-80% of patients, but the majority of patients ultimately suffer from relapse and as many as 30% of patients die from drug-related toxicity during induction therapy. The ability of leukemia stem cells (LSC) to survive chemotherapy is primarily responsible for relapse, and eliminating LSC is ultimately essential for cure. The goal of this project is to develop novel LSC-targeting nanometer-scale micelles, called disulfide-crosslinked CLL1-targeting micelles (DC-CTM), which can deliver high concentrations of DNR into both leukemia and leukemia stem cells and eradicate AML from the very root. In this project, we successfully synthesized DC-CTM-DNR. The nanoparticles were 17+/- 6 nm in size and had a payload of 2mg/ml DNR with 100% loading efficiency. DC-CTM were very stable, while an intracellular reductive agent, Glutathione, could reduce the disulfide bonds resulting in micelle dissociation after uptake. Furthermore, using an AML patient-derived xenograft (PDX) model, we showed that 1 mg/kg DC-CTM-DNR-treated mice had significantly less leukemia burden in the bone marrow at week 2 and 4 than both DNR- and PBS-treated mice. Although all mice developed end-stage leukemia by week 8, the DC-CTM-DNR-treated group had significantly smaller leukemia-infiltrated spleens than the PBS group. We further evaluated the effect of treatment on LSC in vivo by secondary transplantation of total bone marrow cells from DNR- and DC-CTM-DNR-treated AML PDX bearing mice into naïve NSG mice. At 8 weeks post secondary transplantation, 100% (7/7) and 37.5% (3/8) of mice developed leukemia in the bone marrow from the 1 mg/kg DNR- and DC-CTM-DNR-treated groups, respectively. Spleens from secondary transplant mice in the DC-CTM-DNR group were significantly smaller than the DNR group indicating less leukemia infiltration. Collectively, DC-CTM-DNR exhibited superior anti-leukemia efficacy and could eradicate LSC more effectively than free DNR. Lastly, pharmacokinetic studies were performed with free DNR and DC-CTM-DNR in rats. Consistent with previous results, DC-CTM-DNR had a longer T1/2a, T1/2b, and accumulated under curve area, than free DNR. Toxicology studies also confirmed that DC-CTM-DNR exhibited a superior toxicity profile, with significantly less cardiotoxicity. In conclusion, we have developed a novel LSC-targeting micelle delivery system that can effectively target AML LSC and bulk leukemia cells with decreased systemic toxicity. Ultimately, DC-CTM-DNR has the potential to significantly enhance the complete remission rate and disease-free survival time and decrease the relapse rate and treatment-related mortality for patients with AML. Citation Format: Tzu-Yin Lin, Yuanpei Li, Yanjun Zhu, Hongyong Zhang, Rick Harse, Kit Lam, Brian Jonas, Chong-xian Pan. Targeting micelles eradicate acute myeloid leukemia stem cells in a patient-derived leukemia xenograft model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1313.

Homoharringtonine in newly diagnosed acute promyelocytic leukemia treatment: a prospective, randomized controlled trial

目的比较高三尖杉酯碱（HHT）±柔红霉素（DNR）联合全反式维甲酸（ATRA）和以DNR为基础的方案治疗初诊中低危急性早幼粒细胞白血病（APL）患者的疗效及耐受性。方法采用前瞻性随机对照研究方法，将96例初诊中低危APL患者随机分配至HHT组、DNR组和HHT+DNR组，比较三组患者的完全缓解（CR）率、总生存（OS）及无事件生存（EFS）情况。结果HHT组31例，DNR组33例，HHT+DNR组32例，三组患者一般资料具有可比性。三组治疗方案诱导治疗CR率均为100%，无早期死亡患者。三组诱导治疗期间WBC达峰值的中位时间分别为9(1~27）、7(1~27）和4(1~23）d，HHT+DNR组短于HHT组（P=0.008）和DNR组（P=0.240）。三组治疗方案在维甲酸综合征的发生率，达CR的中位时间，WBC峰值及血小板、红细胞、血浆输注量方面的差异均无统计学意义（P值均>0.05）。所有患者均在巩固治疗阶段达分子生物学完全缓解（CMR），三组患者达CMR的时间差异无统计学意义（P>0.05）。HHT组、DNR组及HHT+DNR组的3年OS率分别为95.0%、100.0%、91.0%；3年EFS率分别为93.0%、90.0%、85.0%，差异均无统计学意义（P值分别为0.595和0.382）。三组患者不良事件发生率比较差异无统计学意义（P>0.05）。结论以HHT联合ATRA为基础的诱导、巩固治疗方案可以有效治疗初诊APL，疗效与以蒽环类药物为基础的方案类似，可以作为APL的治疗选择之一。

Proapoptotic effects of heme oxygenase-1 inhibitor on Kasumi-1 cells via the ATF4/CHOP/Ire-1α pathway.

We evaluated the effects of down-regulated heme oxygenase (HO)-1 expression on the proliferation of the acute myelocytic leukemia Kasumi-1 cell line by using the HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX) in combination with daunorubicin (DNR), and evaluated the mechanism. The proliferation rates of cells treated with 10 mg/mL DNR and 10 mM ZnPPIX individually or in combination for different time periods were detected using the MTT assay. The apoptotic outcomes of the blank control, ZnPPIX, DNR, and ZnPPIX groups in combination with the DNR group were detected by flow cytometry. The expression of HO-1, activating transcription factor 4, CCAAT-enhancer-binding protein homologous protein, and inositol-requiring enzyme-α mRNA and proteins were detected by fluorescent quantitative real-time polymerase chain reaction and western blotting, respectively. Combined administration inhibited the cells most potently and time-dependently, decreased the expression of HO-1, and significantly increased the expression of activating transcription factor 4, CCAAT-enhancer-binding protein homologous protein, and inositol-requiring enzyme-α expression levels. The cell apoptotic rates in the blank control, DNR, ZnPPIX, and combined administration groups were 8.32 ± 0.53, 39.16 ± 1.46, 10.46 ± 0.88, and 56.26 ± 2.24%, respectively. Inhibiting HO-1 expression can enhance the damaging effects of DNR on Kasumi-1 cells, providing experimental evidence for the improvement of therapeutic effects on acute myelocytic leukemia in clinical practice.

GW24-e2277 Cardiac Diastolic Dysfunction may Hold Promise as Markers of Cardiotoxicity during Daunorubicin Treatment in Acute Nonlymphocytic Leukemia Patients

ObjectivesDaunorubicin causes cardiac injury in acute nonlymphocytic leukemia. Early identifying caridotoxicity induced by daunorubicin might help individualise cardiac protect treatment. Dexrazoxane can reduce cardiac damage. In this study, we assess...

GW24-e1850 Dexrazoxane prevents daunorubicin induced cardiotoxicity in acute nonlymphocytic leukemia patients

ObjectivesThe usefulness of daunorubicin in acute nonlymphocytic leukemia patients is limited by their cardiotoxicity. Current prophylaxis relies upon early detection of systolic and/or diastolic dysfunction. Dexrazoxane reduces cardiac damage during...

Telmisartan inhibits the progression of cardiomyopathy in daunorubicin treated rats: the role of advanced glycation end products

Background: Anthracyclines have been reported to induce cardiotoxicity through mechanisms involving formation of advanced glycation end-products (AGEs), including pentosidine and Nє”-(carboxymethyl) lysine (CML). We investigated the potential utility of telmisartan (TML), an angiotensin II receptor antagonists (ARB) on anthracycline-induced cardiotoxicity. Methods: Three groups of Sprague-Dawley rats were treated as follows: The first group received daunorubicin (DNR) 3 mg/kgBW every alternating day to reach a cumulative dose of 9 mg/kg DNR . The second group received DNR plus TLM at a dose10 mg/kgBW, by oral gavage for 6 weeks, and the third group served as control group (CTL) which only received vehicle of DNR. Mean blood pressure (MBP) peak left ventricular pressure (LVP), LV end-diastolic pressure (LVEDP), and intra-ventricular contractility (±dP/dt) were recorded by using Powerlab instrumentation. Ejection fraction (EF), and fractional shortening (FS) were measured by echocardiography. Expression of receptor of AGE (RAGE), pentosidine and CML were measured by immunohistochemistry and Western blot in LV tissue. Results: DNR treatment was associated with significant weakening of some hemodynamic parameters which could be reversed by TML (LVP: 124.3 ± 6.0; 111 ± 7; and 115.1 ± 5.4 mmHg, respectively in CTL, DNR and DNR-TLM groups; LVEDP: 7.5 ± 0.9; 10.7 ± 0.3; 8.7 ± 0.4 mmHg, respectively; +dP/dt: 6813 ± 541; 4800 ± 345; 5950 ± 398 mmHg/s, respectively). The same phenomenons were also observed on echocardiographic parameters (EF: 78.9 ± 1.8; 59.6 ± 1.4; 76.2 ± 2.75 %, resepectively; FS: 42.8 ± 1.7; 29.1 ± 1.3; 41 ± 2.7 %) respectively. Expression of RAGE as well as pentosidine and CML were increased in DNR-rats. TML treatment ameliorated these changes. Conclusion: These results suggested the role of AGE formation in DNR-induced cardiotoxicity and telmisartan could inhibit the progression of cardiac toxicity at least in part by reduction RAGE expressiom. (Med J Indones 2011; 20:255-62)

Analysis of efficacy and cost-effectiveness of high-dose arabinoside versus daunorubicin chemotherapy in older adult patients with acute myeloid leukemia by cytogenetic risk profile: retrospective review from China

High-dose arabinoside (HiDAC) and daunorubicin (DNR)-based chemotherapy are the primary consolidation treatment options for older adults (50-60years old) with acute myeloid leukemia in China. We analyzed the event-free survival (EFS) and hospital treatment charges of older adult patients with different cytogenetic risk profiles. In patients with a better/intermediate risk profile, the average total treatment cost of HiDAC was similar to that of DNR (P=0.11). A 5-year follow-up of patients with better/intermediate cytogenetic risk profiles revealed that the median EFS of patients who received HiDAC was significantly longer than for patients who received the DNR-based regimen (27 vs. 20months, P=0.03). Average cost per year of life saved was 18,746.84 USD for HiDAC, compared to 32,733.37 USD for DNR. In contrast, for patients with a poor cytogenetic risk profile, the average total treatment cost for HiDAC was higher than for DNR (P<0.005). In addition, the median EFS in the HiDAC protocol group was significantly lower than in the DNR group (11 vs. 20months, P=0.003). Meanwhile, in this risk group, the average cost per year of life saved was 103,237.70 USD compared to 32,277.93 USD, respectively, in the HiDAC and DNR regimens. We conclude that HiDAC is a more efficacious and cost-effective consolidation treatment regimen in the better/intermediate risk group, while the DNR-based regimen is more cost-effective in the poor risk group.