Types Of Hematological Malignancies Research Articles

High Serum Levels of Extracellular Vesicles which Express Specific Markers of Malignancy Are Released in Patients with Various Types of Hematological Neoplastic Disorders

▪INTRODUCTION: Extracellular vesicles (EVs), including exosomes, shedding vesicles (microvesicles: MVs) and apoptotic bodies, are membrane vesicles of 40-3,000 nm that are released from many cell types, such as red blood cells, platelets, lymphocytes, dendritic cells, endothelial cells and tumour cells. They have a key role not only in the regulation of normal physiological processes, but also in the pathology underlying several diseases. Recently, it was discovered that EVs circulate in bodily fluids of cancer patients and could serve as potential diagnostic biomarkers. However, the presence and role of peripheral circulating EVs in hematological malignancies (HMs) remain unclear. The aim of this study was to investigate EVs in peripheral blood of HM patients, defining the EVs count, size and phenotype.METHODS: Peripheral blood samples were obtained prior to treatment from 87 subjects: 6 Chronic Lymphocytic Leukemia (CLL), 10 Non-Hodgkin’s Lymphoma (NHL; 5 follicular, 5 diffuse large B-cell), 6 Waldenstrom’s Macroglobulinemia (WM), 6 Hodgkin’s Lymphoma (HL), 6 Multiple Myeloma (MM), 5 Acute Myeloid Lukemia (AML), 19 Myeloproliferative Neoplasms [(MPNs; 5 Essential Thrombocythemia (ET), 5 Polycythemia Vera (PV), 5 Primary Myelofibrosis (PMF), 4 Chronic Myelogenous Leukemia (CML)], 5 Myelodysplastic Syndrome (MDS) and 28 healthy controls. EVs were isolated from serum of peripheral blood by ultracentrifugation steps. For calculation of counts, TruCOUNT beads were added immediately prior to analysis by flow cytometry (FACS). For size assessment, Megamix beads with specific size (0.3,0.9, 3.0 micrometer) were used. For phenotype characterization, a panel of specific antibodies (anti CD59, CD61, CD62E, CD19, CD20, CD38, CD138, CD13, CD30) were employed in a FACS analysis.RESULTS: MVs count was significantly higher in patients with CLL, WM, HL, MM, AML and CML (median values 890, 3227, 1218, 914, 656 and 1275 MVs/microliter of serum, respectively) compared to healthy controls (median value 270 MVs/ microliter of serum, ***P<0.001). MV levels in the other HMs, although sligthly higher when compared to healthy controls, did not mantain statistical significance when p value was corrected for the number of malignancies analysed. (Figure)In CLL and MDS, through the number of cases was small, MVs counts directly correlated with Rai stage and R-IPSS risk, respectively.As regards vesicles volume, all HMs generated a distinct population of MVs with smaller size when compared with controls. As expected, MVs were consistently Annexin V positive and expressed a common membrane protein, CD59, an ubiquitous complement regulator. Part of MVs originated from platelets (median value 400 CD61 positive/ microliter of serum) and endothelial cells (median value 60 CD62E positive/ microliter of serum).Interestingly, MV phenotype was disease specific: a relevant amount of MVs expressed surface specific proteins related to malignancy. In particular, we found MVs positive for CD19 and/or CD20 in CLL, WM and NHL, CD38 and/or CD138 in MM, CD13 in AML, MPNs and MDS, CD30 in HL. These markers were not significantly expressed on the surface of healthy subject MVs.CONCLUSIONS: In this study, for the first time, serum MVs in a panel of HMs were analysed. We found that patients with various types of HMs release elevated peripheral blood MV levels selectively displaying markers of the underlying malignancy. MVs could be therefore useful as potential novel biomarkers in HMs. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

Metabolic syndrome in hematologic malignancies survivors: a meta-analysis.

The results from the published studies on whether the risk of metabolic syndrome is high among hematologic malignancies survivors are conflicting. The aims of this study are to conduct a systematic search and assess the literature to determine the available evidence on the association between metabolic syndrome and hematologic malignancies. Two reviewers independently conducted a systemic search to identify the available evidence from PubMed, EMBASE, and Cochrane database from database inception to June 4, 2014. The articles were limited to human studies and published in English. A combined analysis was performed, followed by a subgroup analyses stratified by the type of hematologic malignancies, hematologic malignancies treatment modality, criteria of metabolic syndrome, and individual components of metabolic syndrome. Publication bias was assessed using the Begg and Egger's tests and visual inspection of funnel plot. All analysis was performed using Stata SE version 10.0 software package. Seven cross-sectional studies with a total of 3,550 participants were included in the analysis. Compared with the healthy control groups, the hematologic malignancies survivors were at an increased risk of metabolic syndrome, in the random-effect meta-analysis of all seven cross-sectional studies (OR 2.03; 95% confidence interval (CI) 1.23-3.35; I (2)=43.9%). In the subgroup meta-analysis by cancer treatment modality, a significant positive association was observed for hematopoietic stem cell transplantation (OR 2.40; 95% CI 1.71-3.35). However, no significant positive association was observed for other major cancer treatment modality, including chemotherapy and radiation therapy. Further subgroup analysis by individual metabolic syndrome components showed that the risk of high blood pressure among hematologic malignancies survivors (OR 1.67; 95% CI 1.10-2.53) was found to be higher compared with central obesity (OR 1.43; 95% CI 1.02-2.01), whereas no significant association between hematologic malignancies survivors and other individual components including high glucose levels, high triglyceride levels, and low HDL cholesterol levels was observed. Funnel plot inspection and Begg and Egger's tests showed no evidence of publication bias for combined analysis. Our meta-analysis of studies showed that hematologic malignancies survivors were at an increased risk of metabolic syndrome.

Sensitivity of leukemic T-cell lines to arsenic trioxide cytotoxicity is dependent on the induction of phosphatase B220/CD45R expression at the cell surface.

BackgroundArsenic trioxide (As2O3) is highly effective in treating acute promyelocytic leukemia (APL), but shows more variable therapeutic efficacy for other types of hematological malignancies. Previously, we reported that As2O3 selectively eliminates pathogenic B220-expressing T cells in autoimmune MRL/lpr mice. We investigated herein the relationship between As2O3 sensitivity of leukemic T-cell lines and the expression levels of the B220 isoform of transmembrane tyrosine phosphatase CD45.MethodsGSH content, O2- production, and B220, HSP70, Fas and FasL membrane expression was measured by flow cytometry. Subcellular localization of B220 was determined by imaging flow cytometry. Cell death was analyzed by morphological changes, annexin V and propidium iodide staining, and caspase 8 and 9 activation. B220 mRNA expression was analyzed by RT-PCR. Activated NF-κB p50 was quantified by a DNA binding ELISA.ResultsWe selected human (Jurkat, Jurkat variant J45.01, HPB-ALL) and mouse (EL-4, BW5147, L1210) T-cell lines for their marked differences in As2O3 sensitivity over a large range of doses (1 to 20 μM). Differences in redox status cannot explain the dramatic differences in As2O3 sensitivity observed among the T-cell lines. Unexpectedly, we found that B220 is differentially induced on As2O3-treated T-cell lines. As2O3 treatment for 24 h induced low (HPB-ALL), intermediate (Jurkat) and high (EL-4, BW5147) levels of B220 membrane expression, membrane-bound HSP70 and cell death, but inhibited NF-κB p50 nuclear translocation. When high levels of B220 expression were achieved with low doses of As2O3, the T-cell lines died by apoptosis only. When high doses of As2O3 were required to induce B220 expression, leukemic T cells died by both apoptosis and necrosis.ConclusionsCellular redox status is not essential for As2O3 sensitivity of leukemic T cells, suggesting the existence of additional factors determining their sensitivity to As2O3 cytotoxicity. Phosphatase B220 could be such a factor of sensitivity. As2O3 treatment inhibits NF-κB p50 nuclear translocation, and induces B220 expression and cell death in a dose and time dependent manner. The levels of B220 induction on leukemic T cells strictly correlate with both the extent and form of cell death, B220 might therefore play a checkpoint role in death pathways.Electronic supplementary materialThe online version of this article (doi:10.1186/1476-4598-13-251) contains supplementary material, which is available to authorized users.

Altered expression of immune-related genes in children with Down syndrome.

Individuals with Down syndrome (DS) have a high incidence of immunological alterations with increased susceptibility to bacterial and viral infections and high frequency of different types of hematologic malignancies and autoimmune disorders. In the current study, we profiled the expression pattern of 92 immune-related genes in peripheral blood mononuclear cells (PBMCs) of two different groups, children with DS and control children, to identify differentially expressed genes that might be of pathogenetic importance for the development and phenotype of the immunological alterations observed in individuals with DS. PBMCs samples were obtained from six DS individuals with karyotypically confirmed full trisomy 21 and six healthy control individuals (ages 2–6 years). Gene expression was profiled in duplicate according to the manufacturer's instructions provided by commercially available TaqMan Human Immune Array representing 92 immune function genes and four reference genes on a 96-plex gene card. A set of 17 differentially expressed genes, not located on chromosome 21 (HSA21), involved in immune and inflammatory pathways was identified including 13 genes (BCL2, CCL3, CCR7, CD19, CD28, CD40, CD40LG, CD80, EDN1, IKBKB, IL6, NOS2 and SKI) significantly down-regulated and four genes (BCL2L1, CCR2, CCR5 and IL10) significantly up-regulated in children with DS. These findings highlight a list of candidate genes for further investigation into the molecular mechanism underlying DS pathology and reinforce the secondary effects of the presence of a third copy of HSA21.

Exploring therapeutic potentials of baicalin and its aglycone baicalein for hematological malignancies

Despite tremendous advances in the targeted therapy for various types of hematological malignancies with successful improvements in the survival rates, emerging resistance issues are startlingly high and novel therapeutic strategies are urgently needed. In addition, chemoprevention is currently becoming an elusive goal. Plant-derived natural products have garnered considerable attention in recent years due to the potential dual functions as chemotherapeutics and dietary chemoprevention. One of the particularly ubiquitous families is the polyphenolic flavonoids. Among them, baicalin and its aglycone baicalein have been widely investigated in hematological malignancies because both of them exhibit remarkable pharmacological properties. This review focuses on the recent achievements in drug discovery research associated with baicalin and baicalein for hematological malignancy therapies. The promising anticancer activities of these two flavonoids targeting diverse signaling pathways and their potential biological mechanisms in different types of hematological malignancies, as well as the combination strategy with baicalin or baicalein as chemotherapeutic adjuvants for recent therapies in these intractable diseases are discussed. Meanwhile, the biotransformation of baicalin and baicalein and the relevant approaches to improve their bioavailability are also summarized.

Could valproic acid be an effective anticancer agent? The evidence so far

Valproic acid is an inhibitor of class I histone deacetylases. Epigenetic therapies in cancer have been focus of a keen interest and histone deacetylase inhibitors, in particular, have been approved for certain types of hematologic malignancies. Valproic acid is an attractive candidate for cancer therapy due to its mechanism of action, its low cost and generally good clinical tolerability. In the following editorial, we will review its role as monotherapy for cancer, its place in combination epigenetic therapy, and its role as chemosensitizer, and cancer preventative agent.

Simultaneous occurrence of two B-cell malignancies: A case report.

The present study describes the case of a 75-year-old male with coexisting multiple myeloma and diffuse large B-cell lymphoma. Although the two malignancies are mature B-cell neoplasms, their clinical manifestations are very different and the clinical approaches used to treat these two types of tumor vary. The patient in the present case was diagnosed with the simultaneous existence of two different B-cell tumors and was successfully treated using the DCEP regimen. The simultaneous presentation of two mature B-cell neoplasms, types of hematological malignancy, is very rare, thus the present case is considered to be interesting.

Tetraspanin CD9 modulates human lymphoma cellular proliferation via histone deacetylase activity

Non-Hodgkin Lymphoma (NHL) is a type of hematological malignancy that affects two percent of the overall population in the United States. Tetraspanin CD9 is a cell surface protein that has been thoroughly demonstrated to be a molecular facilitator of cellular phenotype. CD9 expression varies in two human lymphoma cell lines, Raji and BJAB. In this report, we investigated the functional relationship between CD9 and cell proliferation regulated by histone deacetylase (HDAC) activity in these two cell lines. Introduction of CD9 expression in Raji cells resulted in significantly increased cell proliferation and HDAC activity compared to Mock transfected Raji cells. The increase in CD9–Raji cell proliferation was significantly inhibited by HDAC inhibitor (HDACi) treatment. Pretreatment of BJAB cells with HDAC inhibitors resulted in a significant decrease in endogenous CD9 mRNA and cell surface expression. BJAB cells also displayed decreased cell proliferation after HDACi treatment. These results suggest a significant relationship between CD9 expression and cell proliferation in human lymphoma cells that may be modulated by HDAC activity.

Towards optimal clinical and epidemiological registration of haematological malignancies: Guidelines for recording progressions, transformations and multiple diagnoses

Haematological malignancies (HM) represent over 6% of the total cancer incidence in Europe and affect all ages, ranging between 45% of all cancers in children and 7% in the elderly. Thirty per cent of childhood cancer deaths are due to HM, 8% in the elderly. Their registration presents specific challenges, mainly because HM may transform or progress in the course of the disease into other types of HM. In the context of cancer registration decisions have to be made about classifying subsequent notifications on the same patient as the same tumour (progression), a transformation or a new tumour registration. Allocation of incidence date and method of diagnosis must also be standardised.We developed European Network of Cancer Registries (ENCR) recommendations providing specific advice for cancer registries to use haematology and molecular laboratories as data sources, conserve the original date of incidence in case of change of diagnosis, make provision for recording both the original as well as transformed tumour and to apply precise rules for recording and counting multiple diagnoses. A reference table advising on codes which reflect a potential transformation or a new tumour is included.This work will help to improve comparability of data produced by population-based cancer registries, which are indispensable for aetiological research, health care planning and clinical research, an increasing important area with the application of targeted therapies.

Splicing factor mutations and cancer

Recent advances in high-throughput sequencing technologies have unexpectedly revealed that somatic mutations of splicing factor genes frequently occurred in several types of hematological malignancies, including myelodysplastic syndromes, other myeloid neoplasms, and chronic lymphocytic leukemia. Splicing factor mutations have also been reported in solid cancers such as breast and pancreatic cancers, uveal melanomas, and lung adenocarcinomas. These mutations were heterozygous and mainly affected U2AF1 (U2AF35), SRSF2 (SC35), SF3B1 (SF3B155 or SAP155), and ZRSR2 (URP), which are engaged in the initial steps of RNA splicing, including 3' splice-site recognition, and occur in a large mutually exclusive pattern, suggesting a common impact of these mutations on RNA splicing. In this study, splicing factor mutations in various types of cancers, their functional/biological effects, and their potential as therapeutic targets have been reviewed.

Pattern of bloodstream infections in patients with hematological malignancies in a tertiary care centre.

The purpose of our study was to evaluate the clinical characteristics, to understand the pattern of bloodstream infections (BSIs), and to determine the risk factors contributing to high-risk febrile neutropenia in patients with hematological malignancy. A comprehensive review of retrospective data was done from 2004 till 2012 from a single center. There were total 171 consecutive febrile episodes with 103 acute lymphoblastic leukemia (ALL) patients and 63 acute myeloid leukemia (AML) patients. The highest number of febrile neutropenia episodes occurred during ALL and AML induction followed by consolidation treatment with high-dose cytarabine. In our study population, the most common organisms isolated were Gram-positive (20%) followed by Gram-negative (6.4%) organisms. The incidence of fungal sepsis was only 3%. In our study, it was seen that the recovery from febrile neutropenia depends upon the disease, ALL recovered rapidly compared to AML (P < 0.001) and also the on the phase of treatment, i.e consolidation recovered earlier than induction (P < 0.001). There was no death recorded in this study population during febrile neutropenia. The incidence of febrile neutropenia depends upon the type of haematological malignancy and the aggressiveness of therapy required treating the disease especially during induction. The improvement in antimicrobial coverage and its prompt use leads to the selective growth of Gram-positive organisms.

Association of an LMP2 Polymorphism with Acute Myeloid Leukemia and Multiple Myeloma

Hematological malignancies (HM) are a group of neoplasms derived from the cells of the bone marrow and lymphatic system. Genetic factors leading to susceptibility to HM have been investigated for years but little is known yet. Low molecular weight polypeptide (LMP) 2 and LMP7 genes are important subunits of the immunoproteasome and play significant role in antigen presentation. The polymorphisms of LMP genes have been reported to be risk factors for various types of diseases. The aim of this study was to investigate the association of LMP2 and LMP7 polymorphisms with the occurrence of particular types of HM. A total of 132 patients with HM and 130 control subjects were investigated. No significant difference was obtained in the distribution of genotype and allele frequencies of LMP7 gene in HM patients and the control group. On the other hand, the prevalence of LMP2-AA genotype was found to be higher in acute myeolid leukemia (AML) patients while it was significantly lower in multiple myeloma (MM) cases than in the control subjects. Our results suggested that LMP7 could not be a risk factor for susceptibility to HM, whereas LMP2 polymorphisms could play a role in the development of AML and MM.

Compare The Outcome Of Patients With Hematological Malignancies Treated With Allogeneic Stem Cell Transplant In Patients Age Over 50 Conditioned With Reduce Intensity Regimens With Those Under 50 Conditioned With Standard Intensity Regimen In a Rural University Hospital – A Single Institution Experience

BackgroundMajority of hematological malignancies occurs in older individuals and most can only be cured with high dose chemotherapy followed by stem cell transplantation. Unfortunately these patients who need transplantation the most cannot tolerate chemotherapy well. Development of reduced-intensity transplant regimens has allowed older patients be treated with allogeneic transplantation. AimsTo compare the outcome of allogeneic hematopoietic stem cell transplantation in patients age over 50 conditioned with reduce dose intensity regimens with those under 50 conditioned with standard dose intensity regimen MethodsWe analyzed retrospectively the data in a cohort of 50 consecutive patients with hematological malignancies treated with allogeneic stem cell transplant. We divided the patients into two groups, those under 50 years and those over 50 years. Those under 50 years received standard dose conditioning regimen, while those over 50 years received reduced dose regimen except for one patient. Cyclosporine and standard dose methotrexate was used as GVHD prophylaxis in patients under 50yrs while Cyclosporine and low dose methotrexate was used in patients over 50. Type of hematological malignancies, disease condition at transplant, type of transplant, conditioning regimen, dose of CD34+ stem cell and nucleated cell infused, day to myeloid and platelet engraftment and day 90 mortality is shown in the table. Patients who received unrelated donor transplant received ATG. Five high risk patients received peri-transplant Keratinocyte Growth Factor. All patients received G-CSF beginning day-0.Patients < 50 years oldPatients > 50 years oldNumber of Patients3020Age18 to 46 years (Median 31 yrs)50 to 69 years (Median 57.5 yrs)Indication for transplantAcute Leukemia & MDS (18), Aplastic Anemia (3), Chronic Leukemia (4), Lymphoma (5)Acute Leukemia & MDS (14), Myelofibrosis (1), Chronic Leukemia (1), Lymphoma (4)Type of transplanta) Sibling - 13b) MUD – 13 (43%)c) Cord Blood/Double Cord- 2/2a) Sibling - 13b) MUD – 6 (30%)c) Double Cord blood - 1Disease Status at Transplanta) Remission - 14b) Active – 15 (50%)c) Unknown - 1a) Remission - 5b) Active – 14 (70%)c) Unknown - 1Conditioning RegimenBu/CTX (11), Bu/Flu (9), CTX/TBI (2), ATG/CTX (2) Melphalan Containing Regimen (6).Mel/Flu (9), Bu/Flu (8), CTX/TBI (1), Bu/CTX (I), Mel/Thio (1).Dose of CD34+ hematopoietic stem cell infusedRange 0.12 x106 to 15.83 x 106 cells/kg (median 3.15 x 106 cells/kg).Range 1.51 x106 to 9.08 x 106 cells/kg (median 4.21 x 106 cells/kg).Dose of Nucleated cell infusedRange 0.22 x 108to 41.46 x 108 cells/kg (median 9.37 x 108nucleated cells/kg).Range 5.19 x 108to 18.32 x 108 cells/kg (median 9.08 108nucleated cells/kg).Day of Myeloid EngraftmentRange 10-30 days. Median 14 daysRange 9 to 21 days. Median 14 daysDay of Platelet Engraftment Range (Median day)Range 10 -42 days. (Median 16 days)Range 10-68 days. (Median 18 days)Engraftment Failure2 Patients (both received cord blood)0 PatientsEarly mortality (day 90)9/30 Patients (30%)9/20 Patients (45%) Results43% of patient < 50 years of age underwent match unrelated donor transplant vs. 30% in those over 50. Four underwent cord blood transplant in patient < 50 yrs. old vs. one in those over 50. 50% of patients had active disease in < 50 years group vs. 70% in those over 50. All but two patients (underwent cord blood transplantation <50 years of age) engrafted. Four patients died prior to engraftment in each group. Myeloid engraftment occurred at a median day 14 in each group. Platelet engraftment occurred at median day 16 in patients age under 50 years vs. day 18 in patients over 50. One patient with double cord blood transplant had platelet engraftment on day +68 (age >50). Two patients who received cord blood under 50 years group had engraftment failure. Ninety day mortality was 30% in those under 50 vs. 45% in those over 50. ConclusionsReduced intensity allogeneic stem cell transplantation is feasible and well tolerated in older patients with hematological malignancies with good engraftment however with high early (day 90) mortality. 70% of the patients had active disease at the time of transplant which could account for their high mortality. To prevent early peri-transplant mortality patients with active disease should not go to transplant or should be treated with newer chemotherapy or targeted agents to bring them into remission prior to transplantation Disclosures:No relevant conflicts of interest to declare.

Population Pharmacokinetics and Exposure-Safety Of Lenalidomide In Patients With Multiple Myeloma, Myelodysplastic Syndromes and Mantle Cell Lymphoma

IntroductionLenalidomide (LEN) has been approved for the treatment of multiple myeloma (MM: dose 25mg/day, in combination with dexamethasone), myelodysplastic syndromes (MDS: 10 mg/day), and mantle cell lymphoma (MCL: 25 mg/day). In patients with these 3 types of hematologic malignancies, grade 3/4 neutropenia and thrombocytopenia are the most common dose-limiting adverse events (AEs) associated with LEN treatment. Renal function has been used to guide the starting dose of LEN in these patients as renal excretion is the primary route for LEN clearance. However, questions remain whether other intrinsic factors (e.g., race, age, body weight, gender, mild hepatic impairment) should also be considered in determining LEN dose, and how hematologic AEs are related to LEN exposure. The aim of this study was to quantitatively assess the effect of various intrinsic factors on LEN clearance and to explore the exposure-response relationship for severe neutropenia and thrombocytopenia in patients with MM, MDS, and MCL. MethodsSingle- and multiple-dosing concentration data of LEN were pooled from 7 clinical studies yielding a total of 147 patients: 68 with MM, 25 with MDS, 24 with MCL, and 30 with renal impairment and without a malignant condition. In the entire analysis population, 39% of the patients had normal renal function (creatinine clearance [CrCl] ≥ 90 mL/min), 31% had mild renal impairment (RI) (CrCl = 60-89 mL/min, 26% had moderate to severe RI (CrCl = 15-59mL/min), and 4% of the patients were on hemodialysis. The LEN dose level studied was 5 mg [N = 6], 10 mg [N = 41], 25 mg [N = 80], or 50 mg [N = 20], respectively. Non-linear, mixed effects' modeling was used to develop a population pharmacokinetic (PK) model for LEN. Subsequently, Bayesian post-hoc population PK model parameters were used to generate LEN steady state area under the plasma concentration-time curve (AUC) and was analyzed via logistic regression to determine the probability of experiencing grade 3/4 neutropenia or thrombocytopenia during the treatment (N = 116). ResultsPlasma LEN concentrations were adequately described by a 2-compartment population PK model with first order absorption and elimination. LEN exhibited linear and time invariant PKs with moderate variability. The baseline CrCl was predictive of the apparent LEN clearance (CL/F). Inclusion of renal function (CrCl and hemodialysis) into the PK model explained 24% of the inter-individual variability in CL/F. The typical value of LEN CL/F at a CrCl level of 80 mL/min was 10.26 L/h, and each 10-mL/min decrease in CrCl resulted in approximately a 10% decrease in CL/F. Age (39-85 years), body weight (33-135 kg), gender, race (white [N=110]; Asian [N=27]; and other [N=10]), and mild hepatic impairment (total bilirubin > 1 to ≤ 1.5× upper level of normal [ULN] or alanine aspartate transaminase >ULN, N = 16) had no effect on LEN CL/F. All LEN PK parameters were comparable among MM, MDS, and MCL patients. Including all treatment cycles up to one year, and after adjusting for disease and baseline neutrophil or platelet counts, LEN AUC was a significant predictor of the probability experiencing Grade 3/4 thrombocytopenia (odds ratio [OR] = 3.337, 95%CI = 1.183 to 9.415) and it was also associated with an increased probability experiencing Grade 3/4 neutropenia (OR = 1.978, 95%CI = 0.999 to 3.917). However, these relationships were not apparent during the first treatment cycle. ConclusionCreatinine clearance is the only significant and clinically important predictor of LEN CL/F; race (White vs Asian), age, body weight, gender, and mild hepatic impairment had no effect. Also no difference in LEN disposition was observed between MM, MDS, and MCL patients. LEN AUC was a significant predictor of the probability of experiencing grade 3/4 thrombocytopenia and neutropenia. Disclosures:Chen:Celgene Corporation: Employment, Equity Ownership. Ette:Anoxis Corporation: Consultancy. Zhou:Celgene Corporation: Employment, Equity Ownership. Weiss:Celegene Corporation: Employment, Equity Ownership. Palmisano:Celgene Corporation: Employment, Equity Ownership.

Controlled Epstein-Barr Virus Reactivation After Allogenic Transplantation Improves Survival and Is Associated To An Expansion Of Natural Killer Cells (NK)

Background B-lymphocytes infected by the Epstein-Barr virus (EBV) are strongly controlled by the immune system, with the persistence of low counts of EBV-infected B lymphocytes in 98% of healthy individuals. Both T- and Natural Killer (NK)-lymphocytes play a main role in immune surveillance of EBV infection. EBV-reactivation (R) frequently occurs in patients having allogeneic transplantation (AT). Aims We evaluated the impact of controlled EBV-R on survival of allografted adult patients with haematological malignancies and analyzed the different circulating lymphocyte populations. Methods 190 consecutive adult patients were included in this study. Patients received fludarabine, busulfan, ATG for reduced intensity conditioning regimen or cyclophosphamide plus TBI for myeloablative conditioning regimen. Prophylaxis of acute GVHD was conventional. EBV viral load (EBV-VL) was monitored in PB, for all the patients after AT, weekly for the first 6 months, then monthly using standard PCR technique. EBV-R was defined by an EBV-VL ³500 copies/ml, increasing 1 week later. This cut-off level of EBV-VL was used to decide therapy, first reducing immunosuppression followed by rituximab if no EBV-VL decrease (375mg/m2, 4 injections weekly). Lymphocyte immunophenotyping (CD3,4,8,3+56+,3-56+,19,25,DR) was performed weekly for 3 months, then monthly using standard technique. In order to show interdependency between the parameters, a multivariate analysis was performed for OS and PFS including parameters like EBV-R, ATG usage. All analysis was performed with SAS 9.3. Results There were 105 acute leukaemias (26 ALL, 71 AML) and myelodysplastic syndromes (8), 71 lymphoproliferative disorders (17 lymphomas, 8 CLL and 46 MM), 5 aplastic anaemia and 9 CML. There were 76 females and 114 males (median age: 51y, range 18-69). 86 (45%) patients presented EBV-R, with 74 patients having EBV-VL&gt;1000 copies/mL. There was no statistical difference between the 2 groups of patients (EBV-R and no EBV-R), for age, sex, cancer type, conditioning regimen, particularly fludarabine, type of graft, and number of infused CD34+ hematopoietic progenitors. Patients with EBV-R received more frequently ATG (P =0.004). The median time of EBV-R detection was 73 days (IQR: 40-210) after transplantation. In case of EBV-R, the dose of immunosuppressive drugs was reduced before stopping. 69/86 patients received rituximab. Patients with no EBV-R did not receive rituximab. Within the 3 months after rituximab, GVHD intensity was decreased, particularly for acute GVHD, which was less frequent. The median follow up was 36.6 months (95%IC 31.5-45.7). Patients with EBV-R had a longer PFS than those with no EBV-R (at 2 years 51% vs. 69%, at 5 years 47% vs 38%) (P&lt;.04). Similarly, OS was longer in patients with EBV-R than that observed for patients with no EBV-R (at 2 years 76% vs. 64% and at 5 years 63 % vs. 47%, P&lt;.001). OS and PFS were not different between patients according to the type of hematological malignancy (respectively, P=.46 and P=.49). The use of ATG was not a significant factor on OS (univariate (p=0.93) and multivariate analysis (p=0.81)). Similarly, it was not a significant factor on PFS (univariate (p=0.47) or multivariate analysis (p=0.68)). Among the patients with EBV-R, OS and PFS were not significantly different for patients receiving or not rituximab (respectively p=0.67 and p=0.88). The median duration of lymphocyte measurement was 94 days (IQR 59-248) after AT. B-lymphocytes and particularly T-lymphocytes, including CD4+ and CD8+ lymphocytes were not significantly different between the 2 groups, respectively P=.29 and P=.48. Only circulating NK cells were significantly different, with a median increased by 22% (P=.03) in patients with EBV-R. CMV co-infection did not impact NK cell counts (P=.058). NK cell count was not different according the use of rituximab (P=.22). The use of rituximab did not modify NK cell counts at 3 and 6 months after stopping treatment (both P=.99). Conclusions Controlled EBV-R in allografted cancer patients is associated with better OS and PFS, in association with a significant increase in circulating NK cells. A prospective study is ongoing for analyzing different subpopulations of NK cells. Disclosures: No relevant conflicts of interest to declare.

Opticin, a small leucine-rich proteoglycan, is uniquely expressed and translocated to the nucleus of chronic lymphocytic leukemia cells

BackgroundOpticin (OPTC) is a member of the small leucine-rich proteoglycan (SLRP) family and is localized particularly in certain extracellular matrices. We have previously reported the unique expression of another SLRP, fibromodulin (FMOD) in the leukemic cells of patients with chronic lymphocytic leukemia (CLL). OPTC is located in the same region as FMOD on chromosome 1 (1q32.1). Cluster up-regulation of genes may be observed in malignancies and the aim of the present study was to analyze the expression of OPTC in CLL cells.MethodsThe expression of OPTC was tested by RT-PCR and realtime qPCR in PBMC from CLL patients, other hematological malignancies and healthy controls. The presence of OPTC protein, and its subcellular localization, was investigated using fractionation methods where the obtained lysate fractions were analyzed by Western blotting. Deglycosylation experiments were performed to investigate the glycosylation status of the CLL OPTC.ResultsOPTC was expressed at the gene level in all patients with CLL (n = 90) and in 2/8 patients with mantle cell lymphoma (MCL) but not in blood mononuclear cells of healthy control donors (n = 20) or in tumor samples from nine other types of hematological malignancies. OPTC was detected by Western blot in all CLL samples analyzed (n = 30) but not in normal leukocytes (n = 10). Further analysis revealed a CLL-unique unglycosylated 37 kDa core protein that was found to be located preferentially in the cell nucleus and endoplasmic reticulum (ER) of the CLL cells.ConclusionsA 37 kDa unglycosylated OPTC protein was detected in ER and in the nucleus of CLL cells and not in healthy control donors. The function of this OPTC core protein remains unclear but its CLL-specific expression and subcellular localization warrants further investigations in the pathobiology of CLL.

Low-dose cytosine arabinoside-induced syptomatic bradycardia in a patient with acute myeloid leukemia

Cytosine arabinoside (Ara-C) is one of the critical agents for the treatment of acute myeloid leukemia (AML). The toxicity profile of Ara-C is highly dependent on the dose and schedule of administration. Cardiologic complications associated with Ara-C are rare. These side effects were reported with high doses of cytarabine (1–3g/m2, 6–12 doses) in the literature. Herein, we report a patient who developed symptomatic sinus bradycardia while receiving low-dose Ara-C therapy for AML. A 45-year-old female patient diagnosed with AML was treated with standard remission induction chemotherapy protocol that includes 3 days of anthracycline and 7 days of low-dose (100–200mg/m2 2–1) Ara-C. The same chemotherapy regimen was applied again on the 15th day of admission. During the second chemotherapy cycle, the patient developed symptomatic sinus bradycardia. All causes except Ara-C were excluded after required investigational procedures. Ara-C infusion was discontinued for a while and after her symptoms passed chemotherapy was completed with atropine support. Cardiac toxicity is scarce with Ara-C. We want to remind that clinicians should be aware of this potential toxic manifestation even in low doses of the medication, especially as Ara-C is widely used in the treatment of leukemia.<Learning objective: Cytosine arabinoside is one of the historic chemotherapeutics in hematology. It was widely used to treat many types of hematological malignancies in high doses and is commonly available in worldwide markets. Symptomatic sinus bradycardia is a rare type of cardiologic side effect, but it should be kept in mind during chemotherapy.>

Targeting p53 by small molecules in hematological malignancies

p53 is a powerful tumor suppressor and is an attractive cancer therapeutic target. A breakthrough in cancer research came from the discovery of the drugs which are capable of reactivating p53 function. Most anti-cancer agents, from traditional chemo- and radiation therapies to more recently developed non-peptide small molecules exert their effects by enhancing the anti-proliferative activities of p53. Small molecules such as nutlin, RITA, and PRIMA-1 that can activate p53 have shown their anti-tumor effects in different types of hematological malignancies. Importantly, nutlin and PRIMA-1 have successfully reached the stage of phase I/II clinical trials in at least one type of hematological cancer. Thus, the pharmacological activation of p53 by these small molecules has a major clinical impact on prognostic use and targeted drug design. In the current review, we present the recent achievements in p53 research using small molecules in hematological malignancies. Anticancer activity of different classes of compounds targeting the p53 signaling pathway and their mechanism of action are discussed. In addition, we discuss how p53 tumor suppressor protein holds promise as a drug target for recent and future novel therapies in these diseases.

A case of bilateral pleural effusion as the first sign of multiple myeloma

Multiple myeloma (MM) is a type of hematological malignancy that can affect all types of tissues in human. However, it is extremely rare that pleural effusion presents as the first sign in MM patients. A 54-year-old male patient attended our department of respiratory medicine complaining of shortness of breath for the past 3 months. A chest computer tomography (CT) radiograph revealed a bilateral pleural effusion, which was further assessed as exudative type. Sinus spiral CT scan demonstrated diffuse bone destruction of craniofacial bone. A broad reduction of the lumbar bone signal was confirmed by MRI. Furthermore, pleural biopsy showed abnormal proliferation of plasmocytes whereas bone marrow biopsy showed active hyperplasia of plasmacytoid cells. Interestingly, Bence-Jones protein in urine and serum protein electrophoresis was negative. The patient was diagnosed as non-secretory MM. He then underwent chemotherapy with vincristine, adriamycin and dexamethasone. Partial regression of the pleural effusion was achieved after two rounds of chemotherapy, and the patient has been followed for more than one year.

High expression of the Ets-related gene (ERG) is an independent prognostic marker for relapse-free survival in patients with acute promyelocytic leukemia

In acute promyelocytic leukemia (APL), relapse occurs in about 15 % of cases and is a major cause for death. Molecular markers identifying patients at high risk for relapse are not well established. High expression of the transcription factor Ets-related gene (ERG) is associated with inferior overall survival (OS) and disease-free survival in different types of hematologic malignancies. There are no data available about the impact of ERG expression in APL. ERG expression levels were analyzed in bone marrow samples of 86 APL patients at initial diagnosis. High ERG expression was significantly associated with an inferior OS in patients who had reached first complete remission. It was also significantly correlated with inferior relapse-free survival (RFS) and time to relapse (i.e., relapse-free interval, RFI). In multivariate analysis, high ERG expression had an independent negative impact on RFS and RFI. High ERG expression was significantly associated with inferior OS, RFS, and RFI. Moreover, in multivariate analysis, it maintained its value as an independent negative prognostic factor with regard to RFS and RFI. Therefore, ERG expression might serve as a molecular marker for risk stratification in APL and might identify patients who could benefit from intensified treatment regimens.