bluuebunny/crossref_metadata_2025 huggingface dataset:
CD38 Expression in B-Cell Chronic Lymphocytic Leukemia: Association with Clinical Presentation and Laboratory Features in 64 Newly Diagnosed Patients.
Authors: Matias Kleber , Freire Ana Dulce , Matias Carolina , Neves Maria Amélia , Machado Cintia
Published: October 2019

Background: B-cell chronic lymphocytic leukemia (B-CLL) is characterized by the progressive accumulation of a neoplastic clone consisting of CD5/CD19/CD23/surface immunoglobulin cells. Despite this homogeneity in phenotype, B-CLL can follow either an indolent or a progressive course. CD38 is a transmembrane glycoprotein expressed on the surface of leukemic cells in a significant percentage of patients with B-CLL. Recent studies suggest that CD38 expression in CLL is a reliable prognostic marker, leading to an unfavorable clinical course with a more advanced stage of disease, poor responsiveness to chemotherapy and a shorter survival rate, along with others markers like ZAP-70 that reflect more accurately the mutagenic status of VH gene region, unfortunately not yet available in clinical practice routinely.

Objectives: To assess the association of CD38+ expression with clinical and laboratory parameters at diagnosis in patients with B-CLL.

Patients and Methods: CD38 expression was analyzed in 64 unselected newly diagnosed B-CLL patients from January 2003 to May 2005 seen at the Hematology Center of Pernambuco-Brazil (HEMOPE). According to Damle et al. patients with 30% or more B cells expressing CD38 were considered positive(CD38+), and those with less than 30% were considered negative(CD38−). Various patient characteristics were studied including age, sex, Binet stage, hemoglobin, ß2 microglobulin and lactate dehydrogenase levels in the serum. Statistical differences between each group (CD38+ vs CD38−) were analyzed using χ2 tests for categorical variables and Student’s t-tests for continuous variables.

Results: Thirty-six patients (56.25%) were CD38+ and 28 (43.75%) were CD38-. Their median age at diagnosis was 64 years (range, 39–83) and the male to female ratio was 1.46:1. There were no differences between age, sex, hemoglobin and ß2 microglobulin levels in the CD38+ and CD38- groups. ß2 microglobulin level, however, was not available for assessment in all patients. Lactate dehydrogenase level was significantly higher in the CD38+ group (p=0.007). It was observed a trend toward a higher Binet stage (B or C) in the CD38+ compared with CD38- group (63.4 vs. 36.6% respectively), although not statistically significant (p=0.09) possibly because of the small number of patients studied.

Conclusion: CD38 expression was associated with a higher lactate dehydrogenase level and a tendency for more aggressive clinical stage. CD38 evaluation is a measurable biological parameter that is not as subjective as the evaluation of some clinical parameters and should be considered a stantard clinical test for newly diagnosed B-CLL patients.
Cytogenetic Classification Systems and Overall Survival Following Bone Marrow Transplant (BMT) for Acute Myelogenous Leukemia (AML).
Authors: Kalaycio Matt , Sekeres Mikkael , Sobecks Ronald , Rybicki Lisa , Pohlman Brad , Advani Anjali , Kuczkowski Elizabeth , Bolwell Brian
Published: October 2019

Risk-adapted therapy for AML in first complete remission generally calls for allogeneic BMT for patients with poor risk cytogenetics. However, poor risk cytogenetics is defined differently in commonly-applied classification schemes. We hypothesized that differences in cytogenetic classification might result in differences in survival after BMT. From September 1991 to December 2003, we treated 47 patients with AML in first complete remission with high-dose busulfan-containing preparative regimens and an HLA-matched sibling BMT. The median age was 42 years (range 18 to 60). At the time of diagnosis, 35 patients had either a normal karyotype or at least one clonal abnormality. Cytogenetic analysis was unavailable for the other 12 patients. The 47 patients were then classified according to the SWOG/ECOG (Blood96:4075, 2000), MRC (Blood92:2322, 1998), or CALGB (Blood100:4325, 2002) cytogenetic classification systems and analyzed for overall survival. With a median follow-up of 4.9 years, 3 year overall survival for various cytogenetic risk groups are displayed in the table:

SWOG/ECOG CALGB MRC Intermediate risk 69% 63% 74% Poor risk 43% 42% 18% Unknown risk 45% 52% 45%

Concordance indices were 0.58 for CALGB, 0.60 for SWOG, and 0.66 for MRC, demonstrating a slight superiority of the MRC system. Consistent with reports of large, prospective randomized trials of BMT for AML in first remission, patients with poor risk cytogenetics have worse overall survival compared to patients with intermediate risk cytogenetics. Overall survival appears to be different for patients classified as poor risk by MRC criteria. Larger, prospective studies are needed to confirm this observation, but our results suggest that standardized, international cytogenetic risk criteria are needed to develop risk-adapted strategies for the treatment of AML.
Simplifying Complex Microarray Data To Derive Gene Expression Profiles Which Identify Childhood Acute Lymphoblastic Leukaemia Patients at Risk of Relapse.
Authors: Guo Dachuan , Fong Alex , Lail Andy , O’Sullivan Maree , Stone Glenn , Kiiveri Harri , Henry Michael , Stephan Dietrich , Dalla-Pozza Luce , Catchpoole Daniel R.
Published: October 2019

The optimal treatment of patients with childhood acute lymphoblastic leukaemia (ALL) depends on establishing accurate diagnosis. Our investigations seek to strategically develop the application of microarray gene expression profiling to identify ALL patients with clinically homogenous presentations but which may respond differently to established treatment regimens. We have determined the gene expression profiles of ALL bone marrow (BM) samples taken from patients at diagnosis. Data analysis has focussed on the use of a novel and innovative statistical technology, Gene-RaVE. This series of patent protected algorithms builds a multinomial regression model using Bayesian variable selection. Gene-RaVE leads to the generation of a parsimonious and simple diagnostic gene expression signature, but which provides increased predictive ability over current analysis approaches. We describe our analysis of both Affymetrix (HU133A) and cDNA (10.5K) microarray gene expression profiles generated from diagnostic BM from >100 ALL patients covering the broad ALL subtypes including T and B lineage as well as T cell lymphoma leukaemia. Comparison of gene expression data failed to identify clearly distinguishing profiles between patients identified as ‘standard risk’ from ‘medium risk’ according to BFM95 clinical criteria. Gene expression profiles from a cohort of ALL patients, identified as ‘standard risk’ at diagnosis, were compared on the basis of their overall clinical outcome: relapse within 2 yrs vs non-relapse. Using a range of analyses including t-test, Gene-RaVE, discriminant analysis approaches and principle component analysis, we have discovered that small subsets of genes (<10), all of which included Nedd4BP3 and Ribosomal Protein L38 (RPL38), can be used to distinguish the two outcome groups. Subsequent validation using real time PCR supports the increase in Nedd4BP3 expression in standard risk patients which do not respond well to established treatment regimens. The Gene-RaVE algorithm also provides a generic framework for survival analysis. This approach indicates that the expression of these Nedd4BP3, RPL38 and inositol 1, 4, 5-triphosphate receptor, type 2 can be used to build a survival ‘index’ which correlates with the time to a relapse event in standard risk childhood ALL patients. Our results are suggestive of a way forward in the development of an informative, yet efficient diagnostic tool for this childhood malignancy using microarray gene expression analysis technology.
A New Predictive and Prognostic Marker for De Novo AML: Peripheral Blood CD34 (pCD34) Count at Recovery Following Remission Induction (RI) Therapy (Supp. by Ankara University-2003-0809114).
Authors: Arat Mutlu , Topcuoglu Pervin , Yuksel Meltem K. , Dalva Klara , Gultekin Gulhis , Gurman Gunhan , Arslan Onder , Ozcan Muhit , Beksac Meral , Konuk Nahide , Uysal Akin , Koc Haluk , Ilhan Osman
Published: October 2019

In EORTC-LCG AML-10 study patients with low CD34 mobilization profile showed a better outcome (Leukemia,2003,17:60-7). We hypothesized that patients with low pCD34 at the recovery period after RI therapy with de novo AML should have a better clinical outcome or vice versa and launched this prospective study. Between Jan 2002 and Oct 2004, 40 patients with AML (median age: 40 yrs, 17–60; 22M/18F) were enrolled to this study. Using flow cytometrical method CD34-expressing cells were measured in peripheral venous blood samples once WBC was between 1x10e9/L to 3x10e9/L at the recovery period after first RI therapy.

Results: The median time for estimation of pCD34 was 23 days (7–35) after RI. The pCD34 cell count was lower in patients responding to first RI therapy (12x10e6/L vs 68x10e6/L, p=0.011). We observed a weak, but statistically significant positive correlation between CD34 expression of the blasts at diagnosis and pCD34 at recovery period (p=0.048, r2:0.322). We could not show any significant negative impact of CD34 expression of the blasts at diagnosis and response to first RI (p=0.059). Only two of 8 patients not responding to the 1st RI achieved a complete remission after re-induction. The two-year probability of OS, LFS, relapse incidence and mortality rates were 47.1%, 37.2%, 55% and 42.5%, respectively (fig 1,2). If we set up a cut-off value of 18x10e6/L for pCD34, we did not observe any impact of pCD34 on relapse incidence but the mortality rate was significantly increased in patients with high CD34+ cell count (p=0.025).

Conclusion: We were able to show a positive impact of pCD34 estimations after first RI in AML patients on both remission and mortality rate. AML Patients with lower pCD34 after 1st RI tends to have a higher hematological remission rate and lower mortality. This impact of pCD34 as a predictive and prognostic marker in AML should be verified in large cohorts using multivariate analysis.

Table 1: Distribution of risk factors at diagnosis according RI response Variables 1stCRafter1stRI (n=32 ) NoCR after 1stRI (n=8 ) P *p<0.05; Abbr: NS: Non-Significant; ND: Not-Done pCD34 (x10e6/L) 12.4 (0–268) 68.6 (4.4–3820) 0.011* CD34 expr at dx 12% (0%–94%) 44% (9–89) 0.059 Age, years 38.5 (17–57) 43 (20–60) NS WBC at dx (x10e9/L) 11.85 (0.7–175.0) 5.10 (2.7–114.7) 0.934 EM involvement (Y/N) 7/25 0/8 NS Table 2: The outcome of AML patients according to median pCD34 at recovery Variables pCD34<Med 18x10e6/kg (n=20 ) pCD34 ≥ Med 18x10.6/kg (n=20) p CR1 (Y/N) after 1st RI 19/1 13/7 0.044* Relapse incidence 44% 54.5% NS Mortality rate 25% 60% 0.025* DFS 46.19%±12.6% 29.2%±10.4 NS OS 64.95%±13.2% 35.6%±11.3 NS

Figure Figure
Complementary Diagnostic and Prognostic Value of Parallel Karyotyping and Molecular Diagnostics in Patients with Acute Leukemias.
Authors: Ejduk Anna , Majewski Miroslaw , Solarska Iwona , Makuch-Lasica Hanna , Kania Iwona , Nowak Grazyna , Seferynska Ilona , Konopka Lech , Warzocha Krzysztof
Published: October 2019

Chromosomal aberrations analyzed by cytogenetic and molecular methods are major prognostic factors determining treatment options in patients with acute leukemias. The aim of this study was to compare cytogenetic and molecular methods as diagnostic and prognostic tools in patients with acute leukemias. Sixty one previously untreated patients with acute leukemias (AML - 43, ALL - 18) were studied for the presence of chromosomal translocations and corresponding fusion genes [AML1-ETO t(8;21), CBFB-MYH11 inv(16), PML-RARA t(15;17), BCR-ABL p190, p210 t(9;22), MLL-AF4 t(4;11), TEL-AML1 t(12;21), E2A-PBX1 t(1;19), SIL-TAL1 del(1)] using both molecular (RT-PCR, nested PCR) and cytogenetic (GTG) methods. Molecular diagnostics was performed from RNA isolated from bone marrow samples according to BIOMED-1 protocol. Cytogenetic studies were carried out with classical GTG and FISH methods. G-banded mitoses of bone marrow specimen were analysed according to ISCN. Chromosomal aberrations were found in 32.8% patients using GTG method while the parallel molecular tests revealed related fusion genes in 50.8% patients. In 8.5% patients cytogenetic analysis was not performed because of lack of metaphases in cultured cells. All cytogenetic aberrations found in GTG were also confirmed in RT-PCR. Stratification into cytogenetic risk groups was performed after applying combined analysis of karyotyping and molecular tests. Low cytogenetic risk group consisted of 32.8% patients including 11.5% diagnosed with GTG and additionally 21.3% patients after applying molecular tests. The intermediate and high cytogenetic risk group consisted of 32.8% and 34.4% respectively using combined cytogenetic and molecular diagnostics. In low cytogenetic risk group, 85% of patients achieved complete remission (CR), early deaths were found in 15% and none of the patients presented primary chemotherapy resistance. In intermediate risk group CR were obtained in 80%, chemoresistance in 10% and early deaths were observed in 10% of patients. In high cytogenetic risk group, CR were achieved in only 23.8% and chemoresistance occured in 76.2% of the patients. In conclusion we suggest that molecular and cytogenetic tests are complementary methods and should be used in parallel in the initial diagnosis of patients with acute leukemias. This seems to be critical for obtaining the accurate diagnosis, cytogenetic risk assessment and choosing an optimal treatment options.
Laser Tweezers Raman Spectroscopy Detects Differences between Normal Human Lymphocytes, Activated Lymphocytes and Leukemia.
Authors: Taylor Douglas Scott , Chan James W. , Zwerdling Theodore , Lane Stephen M. , Tuscano Joeseph , Huser Thomas
Published: October 2019

Raman spectroscopy is a laser-based analytical technique that enables chemical characterization of molecules in a sample. It is based on the inelastic scattering of photons by molecular bond vibrations. A Raman spectrum obtained from single cells is an intrinsic molecular fingerprint of the sample, revealing detailed information about DNA, RNA, protein, lipid, and carbohydrate content as well as macromolecular conformations. This technique is rapid, non-invasive and non-destructive. We have demonstrated that single-cell laser tweezers Raman spectroscopy (LTRS) reliably discriminates between normal human T- and B-lymphocytes and their transformed, Jurkat-T and Raji-B, counterparts. Herein the same technique is used to discern normal human T- and B-cells from T- and preB-leukemia cells. Also, demonstrated are Raman spectra changes that result from the activation of normal T-cells with a combination of TPA and ionomycin, and normal B-cells by cross linkage of surface immunoglobulin. T- and B-cells were isolated from healthy volunteers. T- and preB-leukemia cells were isolated from patient samples. Spectra are both compared directly and analyzed by principal component analysis. Significant Raman spectra differences in characteristic DNA and protein signals discriminate between normal cells and their leukemia counterparts. Human leukemia cells were indistinguishable from Jurkat-T and Raji-B cells. Changes in spectra that result from cell activation and proliferation only partially account for the spectra changes of leukemia cells. These data demonstrate the utility of LTRS to discern normal lymphocytes from both transformed cells and leukemia cells.
Laser Tweezers Raman Spectroscopy Detects Individual Neoplastic and Normal Hemotopoietic Cells.
Authors: Taylor Douglas Scott , Chan James W. , Zerdling Theodore , Lane Stephen M. , Ihara Ko , Huser Thomas
Published: October 2019

Current methods for identifying neoplastic cells and discerning them from normal counterparts are often non-specific, slow, biologically perturbing, or a combination, thereof. Here, we show that single-cell, laser-tweezers Raman spectroscopy (LTRS) averts these shortcomings and also permits mechanical manipulation of the single cell under investigation. LTRS is used to characterize the biomolecular Raman signature of both normal human lymphocytes and transformed Jurkat and Raji lymphocyte cell lines from single, unfixed cells in suspension. We demonstrate that single-cell Raman spectra provide a highly reproducible biomolecular fingerprint. Characteristic peaks, mostly due to different DNA and protein concentrations, allow for discerning normal lymphocytes from transformed lymphocytes with high confidence (p << 0.05). Spectra are also compared and analyzed by principal component analysis (PCA) to demonstrate that normal and transformed cells form distinct clusters that can be defined using just two principal components. The method is shown to have a sensitivity of 98.3% for cancer detection, with 97.2% of the cells being correctly classified as being normal or transformed. These results demonstrate the potential application of LTRS as a clinical tool, research instrument, and for cell sorting based on its intrinsic biomolecular signature, therefore eliminating the need for exogenous fluorescent labeling.
Endogenous TNFα Mediates Cell Survival and Chemotherapy Resistance by Activating the PI3K/Akt Pathway in Childhood Acute Lymphoblastic Leukemia.
Authors: Gu Lubing , Findley Harry W. , Zhu Ningxi , Zhou Muxiang
Published: October 2019

Expression of endogenous TNFα (enTNF) by tumor cells has been shown to confer a survival advantage and resistance to chemotherapy. Here we report that constitutive activation of the PI3K/Akt pathway is an important mechanism for cell survival and chemotherapy resistance in enTNF+ acute lymphoblastic leukemia (ALL). We analyzed two pediatric ALL cell lines, EU-1 (enTNF+) and EU-3 (enTNF-) for the regulation of Akt by enTNF in association with cell growth and sensitivity to doxorubicin. EU-1 cells constitutively expressed activated/phosphorylated Akt (p-Akt) and were resistant to doxorubicin, whereas EU-3 cells were negative for p-Akt and sensitive to this drug. Treatment of EU-3 cells with supernatant from EU-1 cultures rapidly induced expression of p-Akt. Conversely, treatment with either anti-TNFα or anti-TNF-receptor 2 (TNFR2) antibodies rapidly downregulated constitutive p-Akt in EU-1 cells, and blocked p-Akt induction in EU-3 cells cultured with EU-1 supernatant. Inhibition of p-Akt either by a PI3K inhibitor Ly294002 or by TNF siRNA suppressed EU-1 growth and sensitized these cells to doxorubicin. These results suggest that enTNF produced by EU-1 activates Akt in both EU-1 and supernatant-treated EU-3 cells by binding to cell-surface TNFR2, and that enTNF-activation of the PI3K/Akt pathway may be associated with drug-resistance in pediatric ALL.
The Methylation Status of p21 Is Not Relevant in Adult Acute Lymphoblastic Leukemia Patients.
Authors: Gregorj Chiara , De Cave Fabiana , Ricciardi Maria R. , Scerpa Maria C. , Precupanu Cristina M. , Gervasoni Jacopo , Iannitto Maria L. , Fazi Paola , Gubbiotti Stefania , Specchia Giorgina , Cupri Alessandra , Ferrara Felicetto , Mondello Francesca , Rinaldi Ciro , Fioritoni Giuseppe , Cascavilla Nicola , Miraglia Eustachio , Annino Luciana , Audisio Ernesta , Mazza Patrizio , Peta Antonio , D’Arco Alfonso M. , Bigatti Catia , Tedeschi Alessandra , Foà Robin , Tafuri Agostino , Petrucci Maria Teresa
Published: October 2019

Methylation of CpG islands in the 5′ gene region is associated with transcriptional silencing of gene expression. The hypermethylation of tumor suppressor genes has been described in various tumor tissues, as in gastric and pancreatic cancer, as well as in acute myeloid leukemia, suggesting its potential role in tumorigenesis. Among the three members of the Kip/Cip family of cyclin dependent kinase inhibitors (CKI) p21, p27 and p57, little is known of their methylation status in hematological malignancies and contrasting studies have been reported on the role of p21 hypermethylation in the pathogenesis of acute lymphoblastic leukemia (ALL). The aim of our study was to analyze in primary blasts from adult ALL enrolled in the GIMEMA protocols 0496 and LAL2000 the methylation status of p21, defining in addition its protein expression by Western blot using the monoclonal antibody p21-WAF1 (Santa Cruz, CA). Primary samples from 81 untreated ALL patients were processed using a widely accepted method based on bisulfite modification of DNA, followed by the use of methylation-specific PCR assay (MSP). The human lymphoblastic cell lines (Jurkat, RPMI8866 and CEM), the myeloid cell line OCI-AML3 and normal peripheral blood lymphocytes (PBL) from 10 healthy donors were characterized by a consistent p21 promoter unmethylation (negative controls). In contrast, it was weakly methylated in the Raji cell line and strongly methylated in the Rael (Burkitt’s lymphoma) cell line (positive controls). This assay was further validate in vitro by SsI methylase. In the present study we analyzed 54 B-lineage ALLs, 25 T-ALLs and 2 biphenothypic leukemias; the mean WBC value at diagnosis was 125.6x109/L and 20 samples were Philadelphia chromosome positive. 71/81 of patients studied for p21 methylation were evaluated for response: 53 (74.6%) achieved complete remission (CR) after induction therapy, 8 (11.3%) patients were resistance and 10 (14.1%) died during induction therapy. DNA methylation was not observed in any of the adult ALL patients. p21 protein expression was found in OCI-AML3, Raji and RPMI8866 cell lines, while resulted negative in the Jurkat cell line and in normal PBL. Preliminary results obtained in the ALL samples showed that this protein was expressed in 8/29 (27.6%) cases. In summary, we demonstrated in a large number of primary ALL cases studied at presentation that the p21 gene is not methylated in this population and therefore that the status of p21 methylation does not play a role in the pathogenesis of adult ALL.
Marrow Plasma Levels of Stromal Cell-Derived Factor-1α in Patients with Hematopoietic Malignancies Might Be Associated with the Migration of Leukemic Cells.
Authors: Qian Jun , Lin Jiang , Chen Zi-xing , Chen Yong-jun , Cen Jiang-nong , Zhang Yong-ning , Ji Yong-hui , Qian Zhen
Published: October 2019

The chemokine, stromal cell-derived factor-1 (SDF-1), which is mostly produced by marrow stromal cells, has various effects on hematopoietic cell functions. Its crucial role in the survival, trafficking, and homing of hematopoietic stem cells and progenitor cells is currently established. Here we described the potential significance of bone marrow plasma levels of SDF-1α in patients with hematopoietic malignancies. The bone marrow plasma was obtained from 74 aspirate samples from 38 patients with acute myeloid leukemia (AML), 12 acute lymphoblastic leukemia (ALL) and 24 myelodysplastic syndrome (MDS). The concentration of SDF-1α was determined by a sandwich enzyme-linked immunosorbent assay (ELISA). The results revealed a significant negative correlation of marrow plasma levels of SDF-1 with peripheral white blood cells counts (r=−0.304, P<0.05). The patients with low or normal WBC count (<=10×109/L) had significantly higher SDF-1 levels than those with high WBC count (>10×109/L) (358.88±251.64 pg/mL vs 149.33±129.16 pg/mL, P<0.001). The correlation of SDF-1α levels with platelet counts was also observed (r=0.279, P=0.002). No statistically significant differences were observed in the marrow plasma levels of SDF-1α between normal and abnormal karyotypic groups. There was no significant difference between patients with AML and ALL (279.24±318.50 pg/mL vs 209.78±120.96 pg/mL, P>0.05), however, MDS patients had significantly higher SDF-1α levels (528.65±351.57 pg/mL) than AML and ALL patients (P<0.001). In the subtypes of AMLs, granulocytic leukemia has higher marrow plasma levels of SDF-1α (337.81±350.92 pg/mL, n=28) than mononuclear leukemia (104.94±96.12 pg/mL, n=7). Our results suggest that the migration of malignant leukemic hematopoietic cells, especially of mononuclear leukemic cells, might be associated with the marrow SDF-1α level.
Serum Dependency of t(8;21) AML Cell Line Is Associated with VEGF/VEGFR Pathway and Early Phosphorylation of Akt.
Authors: Imai Norikazu , Shikami Masato , Miwa Hiroshi , Hattori Akiko , Hiramatsu Akihito , Watarai Masaya , Satoh Atsushi , Itoh Masato , Imamura Akira , Mihara Hidetsugu , Nitta Masakazu
Published: October 2019

Most human leukemia cell lines are dependent on serum supplementation (usually fetal calf serum (FCS)), although the extent of serum dependency differs among each cell line. Kasumi-1, a t(8;21) AML cell line is one of the most serum-dependent cell lines. Since growth and survival of many leukemia cell lines are associated with phosphorylation of Akt, we examined the Akt phosphorylation by FCS treatment. In Kasumi-1, Akt was phosphorylated by culture with FCS in a dose-dependent manner, although no such Akt phosphorylation was observed in NB-4, a t(15;17) cell line. By FCS stimulation, Akt (Thr308, Ser473) was phosphorylated from 0.5 hr and the phosphorylation sustained until 48 hours in Kasumi-1. Then, we tested the effect of VEGF/VEGFR signaling in phosphorylation of Akt by FCS. The addition of VEGFR1/Fc and VEGFR2/Fc (which bind external VEGF and abrogate its function) inhibited the Akt phosphorylation from 2 hours until 10 hours, although the growth of Kasumi-1 was not inhibited. The addition of VEGFR2 kinase inhibitor (which inhibits internal VEGF signal) inhibited the Akt phosphorylation from 0.5 hr until 2 hours, and the growth of Kasumi-1 was greatly inhibited.

Taken together, it is suggested that serum dependency of Kasumi-1 is at least in part attributed to VEGF/VEGFR pathway. Then, both external and internal VEGF/VEGFR pathways work in Kasumi-1, which in turn phosphorylate Akt. However, blockade of only internal VEGF signal (by VEGFR2 kinase inhibitor) inhibit the early Akt phosphorylation (0.5 hr), which resulted in growth inhibition, indicating the importance of early Akt phosphorylation.
Results of Acute Myeloid Leukemia Treated with Triple-Drug Regimen Based on HA as Induction Chemotherapy and Its Relationship with Karyotype.
Authors: Wang Jianxiang , Mi Yingchang , Xue Yangping , Yu Wenjuan , Bian Shougeng
Published: October 2019

243 untreated de novo acute myeloid leukemia (AML) patients were treated with (homoharringtonine+AraC, HA) based induction therapy which composing of three chemotherapeutic drugs (HAD, HAM, HAA, HAE) in our hospital for recent 12 years. Complete remission (CR) rate, disease free survival (DFS) and overall survival (OS) of the patients were calculated. 184 patients who had karyotype results were divided into four groups according to SWOG (Southwestern Oncology Group) criteria. Differences of CR rate, DFS and OS of different groups were evaluated. The CR rate of all 243 cases was 77.4%, and 94.6% out of them were for 1~2 courses. Median DFS of the 188 CR patients was 28.5 (range from 1.0 to 153) months. DFS rates at 3 years and 5 years were 45.4%, 40.2% respectively. The median OS of 243 patients was 18.4 (range from 0.5 to 154) months. OS rates at 3 years and 5 years were 36.9%, 31.4% respectively. CR rate, DFS and OS of the different cytogenetic risk groups were also be analyzed. According to SWOG criteria, patients were classified into favorable, intermediate, adverse and unknown groups. CR rate, median DFS and OS were 97.8%, 87.4 months and 89.0 months in favorable group; 81.9%, 17.6 months and 22.3 months in intermediate group; 61.5%, 9 months and 11.5 months in the adverse group; 79.3%, 29.0 months, 19.9 months in the unknown group, respectively. The differences among the four groups were statistically significant. Multivariates analyses confirmed contribution of cytogenetics and courses of post-remission chemotherapy to DFS and OS. If we incorporated unknown group into the intermediate group, the CR rate is 80.6%. DFS rates at 3 years and 5 years were 40.5%, 38.1% respectively. OS rates at 3 years and 5 years were 40.3%, 37.4% respectively. We conclude triple-drugs induction regimens based on HA are highly effective in adult AML in China. Cytogenetics is the important prognostic factor. SWOG karyotype subtyping criteria is appropriate to our patients and unknown group can be incorporated into the intermediate group.
Possible Eradication of Minimal Residual Disease in Acute Leukemia.
Authors: Hijikata Yasuki , Maeda Yasuhiro , Yamaguchi Terufumi , Tanaka Miyako , Hirase Chikara , Shunsuke Takai , Morita Yasuyoshi , Matsuda Mitsuhiro , Kanamaru Akihisa
Published: October 2019

The complete remission (CR) rate in acute leukemia (AL) has been improved due to progressive chemotherapy. However, relapse frequently occurs as minimal residual disease (MRD). To eradicate MRD, immunotherapy using autologous cytotoxic T-lymphocytes has been available. Leukemia cells (LC) of the patients were stored frozen before chemotherapy. Furthermore, after achieving CR, an EBV-transformed lymphoblastoid B-cell line (LCL) was established in each case. To induce Cytotoxic T-lymphocytes (CTL), co-culture system was carried out in two types. One is co-culture of peripheral blood mononuclear cells (PBMCs) and original leukemia cells (LC), the other is co-culture of PBMCs and the patient’s own LCL. After co-culture for 5 days, cytotoxic activity was examined by FACS with PKH-26 staining. Interestingly, cytotoxicity against not only autologous LC but the patient’s own LCL was observed in almost cases. This phenomenon is called cross-killing. Moreover, regulatory T-cells (Treg) were detected in the effecter phase determined with FACS (CD4+ CD25+) and FOXP3 mRNA by RT-PCR. Enhanced cytotoxicity was observed in inverse relation to a low proportion of Treg. These results suggested that more cytotoxicity may be induced by deletion of the Treg population. In conclusion, cell therapy as an immunotherapy should be useful tool to improve the prognosis, and if stocked LC is not available, CTL activity must be induced by co-culture with the patient’s own LCL.
Acute Myeloid Leukemia (AML) in the Elderly. To Treat or Not To Treat: A Decision Analysis.
Authors: Kusminsky Gustavo D. , Rivas Maria M. , O’Flaherty Martin , Maggini Norberto J. , Pose Cabarcos Julio M. , Pinto Miguel Tezanos
Published: October 2019

AML is a disease frequently observed in elderly patients (pts) Treating this population with intensive chemotherapy or palliative treatment is a controversial decision. We analysed the available evidence with a decision model tree considering prognosis and comorbidities.

Method: A decision tree was designed to model significant events and prognostic variables that have an impact in survival at 5 years. The probabilities of this items were obtained from a literature review and for those items without literature support, assumptions were made by an expert opinion based on investigator’s consensus. Utility of each strategy was defined in an ordinal scale and the preference for each was calculated as the expected utility (utility x probability of the result)

Results: Probabilities were: unfavourable prognosis 95%, incidence of comorbidities 48%, death in induction without comorbidities 25%, response to induction in unfavourable prognosis 42%, response to induction in favourable prognosis 63%, adequate performance status after induction without comorbidities 43%, probability of finishing consolidation 64%, survival at 5 years in pts with unfavourable AML 11%, survival at 5 ys in pts with favourable AML 27%. The effect of comorbidities on survival to induction and adequate PS post induction was estimated as 50% higher than in pts without comorbidities. The survival at 5 ys in pts with no response or those who did not complete induction was considered equal to palliative treatment, and S at 5 ys in pts with comorbidities who finished consolidation 30% less than in those with no comorbidities. Utility for “not to treat” was 11, and for “to treat” was 6. The preference for “not to treat” was maintained for any modification of the survival to the induction chemotherapy independently of comorbidities. In sensitivity analysis, the only modification of the observed preference was that “To treat” strategy would be preferable only if the good PS after induction chemotherapy would be observed in 65% of pts or more. In conclusion, the preferred strategy as suggested by the model is “not to treat”, however many relevant questions are still unanswered, reinforcing the need to include these pts in clinical trials for better evidence to base the decisions.
Prospective, Randomized, Multicenter Phase III, Polish Adult Leukemia Group (PALG) Study Comparing DAF (DNR+AraC+Fludarabine), DAC (DNR+AraC+Cladribine), and Standard DA Regimen in Induction of Untreated Acute Myeloid Leukemia Patients - First Report.
Authors: Holowiecki Jerzy , Grosicki Sebastian , Kyrcz-Krzemien Slawomira , Kloczko Janusz , Sulek Kazimierz , Hellmann Andrzej , Skotnicki Aleksander , Kuliczkowski Kazimierz , Zawilska Krystyna , Robak Tadeusz , Dmoszynska Anna , Warzocha Krzysztof
Published: October 2019

The goal of this study is to compare the efficacy and safety of an original DAF regimen: daunorubicine (DNR) 60 mg/m2/d iv, days 1–3; cytarabine (AraC) 200 mg/m2/d ci, d 1–7, and fludarabine 25 mg/m2 2h inf. iv d 1–5 versus previously studied DAC regimen* (DNR, AraC, Cladribine), and versus standard DA in de novo acute myeloid leukemia (AML) patients below 60. Primary objective is complete remission (CR) rate after single course of induction and overall survival, secondary objectives - overall CR rate, toxicity, leukemia-free survival rate, assessment of lymphocyte subpopulations levels and survival in patients submitted to bone marrow allotransplantation (alloBMT) immediately after CR assessment.

Patients achieving CR and did not submit to alloBMT received two courses of subsequent intensive consolidation: 1) HAM (HD AraC, mitoxantrone) 2) HD AraC. In case of partial remission (PR) after the first induction course the same regimen was repeated, Patients with no remission (NR) or PR/NR after 2 induction courses were withdrawn with the study. We are planning to enrol to the study 600 patients in 3 years. Between 09.2004 and 07.2005, 147 adult AML patients, aged 48 (19–60)y, sex: male 57, female 90, treated in 16 co-operating Polish Adult leukemia Group (PALG) centres were randomised to either DAF (n=44), DAC (n=49) or DA (n=54) arm. PML/RAR alfa positive - FAB M3 cases were excluded. Both study groups were well balanced in respect of age, sex, FAB subtype, and WBC.

The final CR rate and CR rate after the first induction course equalled: for DAF 65% and 60%, for DAC 70% and 60%, and for DA 55% and 47%, respectively (p=NS).

The median times to ANC recovery > 0.5 G/L, and PLT >50 G/L in each arms were similar (22–26 d.) (p=NS). All patients developed WHO grade IV thrombocytopenia and agranulocytosis. The frequency and severity of infections, mucositis, vomiting, diarrhea, alopecia, polyneuropathy as well as of cardiac, liver or kidney failure were comparable in each treatment arms. Early death was noted in 6% (n=2) in DAF, in 9% (n=3) in DAC, and in 13% (n=5) in DA group, because of bacterial sepsis in every cases.

In conclusion, this original study proves that the addition of fludarabine to the standard DNR+AraC regimen (DAF) comparing to DAC and DA regimen is a potent antileukemic treatment without increased toxicity.
Clinical Investigation of G-CSF-Priming Regimen in Treatment of Relapsing, Refractory and Senile Acute Myeloid Leukemia.
Authors: Cai Zhen , Han Xiaoyan , Huang He , Xie Wanzhuo , Zheng Weiyan , Lin Maofang
Published: October 2019

Objective:To evaluate the clinical outcome of granulocyte colony-stimulating factor (G-CSF) priming regimen in treatment of refractory, relapsing and senile aute myeloid leukemia.

Materials and Methods:16 patients, 10 men and 6 women ranged in age from 15 to 78 years (median 48), with refractory, relapsing or senile acute myeloid leukemia were treated in our hospital between March 2002 and January 2004. All patients were diagnosed with MIC (morphologic, immunologic, cytogenetic) classification. The control group consists of 18 patients (12 men and 6 women, median age 48) with refractory, relapsing or senile acute myeloid leukemia treated in our hospital between 1999 and 2002. All of them were also diagnosed with MIC. All patients enrolled were treated with G-CSF-priming regimen consisting of low-dose cytosine arabinoside (Ara-C 10mg.m−2.12h−1, subcutaneously,day1–14), aclarubicin (Acla5-7mg.m−2.d−1, continuous intravenous infusion, day 1–8; or 10–14mg.m−2.d−1, continuous intravenous infusion, day1–4) or homoharringtonine (HHT 1mg.m−2.d−1, continuous intravenous infusion, day1–14), and concurrent use of G-CSF (100ug.m−2.12h−1, subcutaneously, day 1–14). In the course of chemotherapy, blood routine examinations were made every other day, and G-CSF was suspended while WBC>20x109/L. Chemotherapy intermission was 14–21 days. If two courses were of no effect, then the investigation was terminated. Patients in the control group received intensive chemotherapy with medium /high dosed Ara-C (1-3g.m−2.12h−1× 3–5d), combined with mitoxantrone or daunorubicin (DNR) or etoposide (vp-16).Statistics:Enumeration data was analyzed by Chi-square test.

Results: All patients accomplished the chemotherapy, with no treatment related death. 9 of the 16 patients achieved complete remission. The CR rate was 56.25%, compared with 22.2% of the control group (Χ2=4.15, P<0.05). However, the total effective rate (68.75%) was not statistically higher than the control group (44.4%) (Χ2=2.03, P>0.05). Therapy related mortality (0.0%) was apparently decreased in priming group than in control group (38.9%) (Χ2=7.84, P<0.01).

Conclusions:Our study shows that the priming regimen is effective and safe to refractory, relapsing, secondary or senile acute myeloid leukemia. Further studies are necessary to evaluate the long-term effects of this new therapeutic regimen in AML.
Pilot Study of Low Dose Melphalan and Bortezomib for Treatment of Acute Myelogenous Leukemia and High-Risk Myelodysplastic Syndromes.
Authors: Bubis Jeffrey A. , Ely Pamela , Meehan Kenneth R. , Stearns Diane , Schaal Anna D. , Kimtis Elizabeth A. , Lowrey Christopher , Bengtson Elizabeth M. , Hill John , Gautier Marc
Published: October 2019

Elderly patients with AML (>60) have a poor prognosis due to high risk cytogenetics and a high incidence of secondary AML. Comorbid conditions make induction therapy more risky and less effective. There is a need for less toxic and more effective treatment for these patients. The combination of bortezomib (B) and melphalan (M) demonstrate synergistic cytotoxicity in vitro against AML cell lines providing a rationale for this combination in patients with AML. We report the initial results of a pilot study of low-dose M and B for the treatment of patients with AML and high-risk myelodysplastic syndromes. Eligibility requirements included a diagnosis of AML or high-risk MDS and subjects were not candidates for standard induction therapy or had failed therapy. Strata 1 included patients who had received no previous therapy and strata 2 patients with refractory or relapsed disease. Treatment consisted of M (2mg) oral daily dose and B (1.0mg/M2) on day 1, 4, 8 &11 of a 28 day cycle. Ten patients have been enrolled. Median age was 69 (range 54–79), 8 patients w/ AML and 2 w/ refractory MDS. Five patients were previously treated (strata 2); regimens included induction (7(3), auto SCT (1) and azacytadine (2). Cytogenetic abnormalities were present in 70% of patient specimens. To date 25 cycles of treatment have been given (range 1–6). Six patients are evaluable for response. Two patients (both with MDS) were withdrawn after 1 cycle for cytopenias. Two patients with AML achieved CR by bone marrow morphology. One patient, a 78 year old man with multiple cytogenetic abnormalities and no previous treatment achieved remission with normalization of cytogentetics after 4 cycles. A 74 year old man who had failed previous therapy for his AML achieved a morphologic remission with persistent cytogenetic abnormalities after 6 cycles. Three patients required admission for management of febrile episodes in association with neutropenia that were disease related. There were no treatment related admissions or grade 3/4 non hematologic toxicity events. The 74 year old man who achieved a CR with no “in hospital” days while on study had previously failed to respond to two induction attempts which required a 44 day hospitalization. Accrual continues. In summary, the combination of low dose oral melphalan with low dose bortezomib appears to be a novel, active and well tolerated outpatient treatment for elderly patients with high risk MDS and AML.
A Phase I Study of the Histone Deacetylase Inhibitor MGCD0103 (MG-0103) Given as a Three-Times Weekly Oral Dose in Patients with Leukemia or Myelodysplastic Syndromes (MDS).
Authors: Garcia-Manero Guillermo , Minden Mark , Estrov Zeev , Verstovsek Srdan , Fiorentino Jackie , Kantarjian Hagop , Reid Gregory , Martell Robert E.
Published: October 2019

MG-0103 is a novel non-hydroxamic acid inhibitor of human histone deacetylases (HDACs), with selectivity for the cancer-associated isoforms of class I HDACs. Deacetylation of histones by HDACs is postulated to inactivate tumour suppressor genes leading to neoplastic transformation, and therefore inhibition of this enzyme may result in antineoplastic activity. To study the safety and activity of MG-0103, we have developed a phase I open-label dose escalation study of MG-0103 administered orally, three-times weekly in patients with leukemia or MDS, with the primary endpoints being the determination of the maximum tolerated dose (MTD) and the pharmacokinetic (PK) and pharmacodynamic (PD) characteristics of MG-0103. Eligibility criteria included appropriate performance status and renal and hepatic functions. Patients with relapsed/refractory leukemia or MDS and older patients with untreated AML/MDS were eligible. Eight patients have been enrolled at two dose levels (20 &40 mg/m2) so far. Their characteristics are: median age 70 years (range 33–79); all patients so far treated have AML; median number of prior therapies is 1 (range 0–3). All patients had complex cytogenetics including one elderly patient with t(8;21) that had not achieved CR with high dose ara-C-based therapy. MG-0103 has been well tolerated with no dose-limiting toxicities. No drug-related adverse events (AEs) >= Grade (Gr) 3 have been observed. The only Gr 2 drug-related AE has been heartburn (1 patient). All other drug-related AEs were grade 1. No cardiac abnormalities have been observed so far. Two of 3 patients at the first dose level have been treated for 3 or more cycles. PK evaluations are shown in the table below. Analysis of peripheral blood cell HDAC activity indicates that HDAC inhibition occurs in a dose-dependent manner. Enrolment is currently proceeding at a dose of 80 mg/m2. In summary, MG-0103 is a well-tolerated HDAC inhibitor in patients with AML at doses and exposures that result in target inhibition in peripheral blood.

Dose 20 mg/m2 40 mg/m2 Pharmacokinectic characteristics AUC (0-∞) ng.hr/ml 260 +/− 10.8 528 +/−275 T 1/2 h 8.09 +/− 2.98 6.24 +/− 0.373
Acute Myeloid Leukemia in the Elderly, Intensive or Maintenance Therapy? Our Experience in Patients over 65 Years.
Authors: Mettivier Vincenzo , Pezzullo Luca , Rocco Stefano , Finizio Olimpia , Bene Lucia , De Rosa Carlo , Nunziata Giuseppe Rodolfo , Liberati Marina
Published: October 2019

The treatment of acute myeloid leukemia in elderly with age > 65 years is still debated. Various reported studies have valued the feasibility of intensive chemotherapy in these setting of patients. The aim of the our study is to value the difference in DFS and OS among 2 groups of elderly patients with AML treated with intensive chemotherapy (IC) or maintenance (M). From June 2001 to May 2005 we have accepted in our Division 45 patients affected by AML aged up to 65 years. 25 were male and 20 were female, with a median age of 75 years (range 67–86 years). 22 patients (12 M and 10 F, median age: 71 years) received intensive chemotherapy (I.C. Flag and MICE) and 23 (13 M and 10 F, median age: 78 years) received or maintenance therapy (low dose cytarabine) or only support. In IC group 11 patients (50%) have obtained to complete remission (CR) with a DSF and OS media of 7,5 and 6,3 months respectively, and a TRM rate of 27%. In the M group the CR has been documented in 6 patients (35%) treated with low dose of cytarabine, with a DFS and OS media of 6,9 and 4,4 months respectively (graph 1–2). This results have shown, as expected, a better CR rate in the IC group, but without any advantage in terms of OS and DFS when compared with the M group. Difference is not statistically significant between the two groups (p: 0.5). Despite a good CR rate, Intensive chemotherapy do not improves survival in elderly patients. This is probably due to the combination of either a high TRM in patients treated with intensive chemotherapy or a high relapse rate. New therapeutics strategies are needed to improve DFS and OS in these patients. Interesting is the use of specific monoclonal antibodies (anti CD33) in this poor risk disease especially in maintenance after a CR obtainable with a low intensive or low dose chemotherapy.

Figure Figure
Treatment of Children with Relapsed Acute Myeloid Leukemia with Gemtuzumab in Combination with Mitoxantrone and Cytarabine.
Authors: Aplenc Richard , Alonzo Todd A. , Gerbing Robert , Smith Franklin O. , Eric Sievers , Arceci Robert J.
Published: October 2019

Despite improvements in the treatment of children with de novo acute myeloid leukemia (AML), a substantial fraction of patients experience relapse. The Children’s Oncology Group Phase I trial, AAML00P2, sought to test the safety and feasibility of combining Gemtuzumab (GMTZ) with a standard AML reinduction regimen of mitoxantrone and cytarabine in a dose de-escalation design. Patients received cytarabine 1000 mg/m2/dose every 12 hours on days 1–4 with mitoxantrone 12 mg/m2/dose daily on days 3–6. The starting dose of GMTZ for dose finding was 3 mg/m2 once on day 7. Doses for patients under 3 years of age were as follows: cytarabine 33 mg/kg/dose, mitoxantrone 0.4 mg/kg/dose, and GMTZ 0.1 mg/kg/dose. Three patients were enrolled in the dose finding strata and an additional 18 patients were subsequently enrolled to acquire further toxicity data. Of these, 15 were male and 6 female. The median patient age was 11.3 years, range 1.2 to 19.2 years. No patients in the initial dose finding cohort experienced a dose limiting toxicity. Toxicities observed in the entire cohort of 21 patients included hematologic and infectious complications typical of AML therapy. One patient had grade III bilirubin elevation. No patients had grade III or IV hepatic toxicity or grade III or IV veno-occlusive disease (VOD). Data on toxicity in stem cell transplant post GMTZ exposure is limited. However, no episodes of VOD have been reported. In summary, GMTZ combined with cytarabine and mitoxantrone is reasonably well tolerated in pediatric AML patients.