bluuebunny/crossref_metadata_2025 huggingface dataset:
Cost-Effectiveness of Targeted Cancer Therapies for Hematological Malignancies.
Authors: Topaloglu Ozlem , Stevens Chuck , Aggarwal Saurabh
Published: October 2019

Abstract 4547

Targeted cancer therapies for hematological malignancies have rapidly gained uptake in different types of indications, and some have become gold standard of treatment. However, the high cost of these therapies poses challenges for payers, patients and physicians.

Methods

To understand the cost-effectiveness of these new classes of drugs we reviewed the published cost-effectiveness studies and analyzed their cost/QALY, time horizon, comparators, indications and model approaches.

Results

Targeted therapies for hematological cancers have much lower and less variable cost/QALY than branded drugs indicated for solid tumors. For example, incremental cost effectiveness ratio (ICER) for imatinib ranges from $40,000 to $45,000/QALY, while ICER for bevacizumab ranges from $125,000 to $325,000/QALY. Secondly, the choice of comparator in some occasions may depend on the country of use, therefore models are required to be customized to local regimens. In one of the cost-effectiveness analysis for bortezomib for refractory multiple myeloma patients, NICE questioned the use of high dose dexamethasone (HDD) as comparator. Third, there is also some variability in methods used by different manufacturers, some used cost/QALY whereas others used cost/LYG. Conclusion: This analysis shows the range, variability and methods used for calculation of ICER values for these targeted cancer therapies and provides lessons for executives and policy makers.

Disclosures:

No relevant conflicts of interest to declare.
A New Model of Care: A Combined Pediatric/ Adult Thrombosis and Hemostasis (PATH) Clinic – One Center's Experience.
Authors: Richardson Matthew W , Steingart Richard H
Published: October 2019

Abstract 4553

Background

Patients of all ages with thrombosis are frequently tested for heritable thrombophilias (HT). Similarly, patients with bleeding or bruising may be tested for heritable bleeding (HB) conditions. We developed the Pediatric/Adult Thrombosis & Hemostasis (PATH) Clinic, a once-monthly clinic where children, adults and their families in whom the diagnosis of HT or HB was established or being considered could be assessed simultaneously by a pediatric and “adult” hematologist. Reasons for the clinic's creation included 1. HT/ HB conditions are lifelong. A combined clinic aids transition from a pediatric provider to an internist at the same institution. 2. Adolescence is when many HT/ HB disorders manifest themselves. This group has medical and developmental traits that cross pediatric and internal medicine approaches and expertise. 3. Adult experience. Much treatment for pediatric thromboses is extrapolated from adult data. An adult hematologist present at the evaluation of a pediatric patient with HT provides a valuable resource for the child and pediatric hematologist. 4. The finding of HT or HB in a patient has implications for family members regardless of age. 5. Convenience. All family members can be evaluated at a single visit where several members might need screening.

Objective

To review the experience of a combined pediatric/adult clinic created to evaluate potential HT/ HB in patients of all ages.

Method

IRB-approved chart review.

Results

92 patients have been evaluated (61 for HT, 31 HB). Mean age 18 yr., median 16 yr. Female 64%. The most common reason for referral was evaluation of HT in an asymptomatic patient (42%) because of a family member having had a thrombosis or recent diagnosis of HT. Those evaluated for potential HT were less likely to be symptomatic (i.e. had not experienced a thrombosis) than those with HB (31% vs. 65%, p < 0.01). Adolescents (13–21 yr old) comprised 52% of all patients (57% of all females). Adolescents were more likely to be referred for HT than other ages (75% vs. 56%, p < 0.01).

Conclusions

The model of the PATH clinic – simultaneous evaluation of patients of all ages with or suspicion for HT/ HB by a pediatric and “adult” hematologist – may be useful to other centers. Adolescents (in transition from the pediatric to adult age group) comprised the single largest cohort, emphasizing the potential importance of a combined approach. Adolescent women were especially represented, likely due to the new potential for bleeding (menarche) leading to a diagnosis of an HB and the emergence of acquired thrombophilic risk factors (estrogen contraception and pregnancy) in this group that have implications in families with HT. Physicians in such a clinic should have particular knowledge of bleeding and thrombophilic conditions in adolescent women and of counseling for asymptomatic HT.

Disclosures:

No relevant conflicts of interest to declare.
During Terminal Differentiation, HbF-Inducing Cytokines Cause Decreased Expression and Reduced Globin Locus Occupancy of BCL11A in Human Erythroblasts.
Authors: Sripichai Orapan , Kiefer Christine M. , Lee Y. Terry , Meier Emily Riehm , Byrnes Colleen , Dean Ann , Miller Jeffery L.
Published: October 2019

Abstract 457

Genetic association studies and gene regulation studies demonstrate that the transcription factor BCL11A is a regulator of fetal hemoglobin (HbF) expression in humans. Cytokine signal transduction also regulates fetal hemoglobin expression in cultured adult human erythroblasts. To further explore the potential for BCL11A in the cytokine-mediated induction of HbF during adult erythropoiesis, transcript and protein expression levels of BCL11A were measured during erythroblast differentiation. BCL11A expression was detected at all stages of erythroid differentiation with the highest level expression in proerythroblasts during the first week in culture under both low-HbF (%HbF ≤3) and high-HbF (%HbF ≥30) culture conditions. Despite a reduction in BCL11A mRNA expression, Western analyses failed to demonstrate reduced levels of BCL11A nuclear protein expression at the proerythroblast stage of differentiation. However, BCL11A protein expression in the high-HbF producing cells was reduced relative to the low-HbF cells during the later period of culture as the cells underwent terminal differentiation. During this later culture period, hemoglobinization occurred, and cells grown in the high-HbF condition revealed a pancellular distribution of HbF compared with a heterocellular distribution in the low-HbF culture condition. Chromatin immunoprecipitation further demonstrated that the addition of HbF-inducing cytokines caused a nearly complete loss of BCL11A chromatin occupancy within the beta-globin locus under the high-HbF culture condition. Specifically, the loss of chromatin occupancy was detected in a region approximately 3 kb downstream of the (A)gamma-globin gene. Further examination of this genomic region demonstrated several BCL11A binding domains located on a cluster of non-coding, intronless RNAs previously named “BGL3” that possess an expression pattern in vivo that is largely restricted to the fetal-liver. In addition to increased and pancellular expression of fetal hemoglobin in the high-HbF erythroblasts, the loss of BCL11A chromatin occupancy in that region of the beta-globin locus was associated with increased expression of BGL3 mRNA (GenBank: AY034471) measured by RT-PCR. These findings demonstrate that defined combinations of cytokines regulate the expression level and chromatin occupancy of BCL11A in adult human erythroblasts as they undergo terminal differentiation. In addition to inheritance and ontogeny, the data also support a role for BCL11A in the regulation of HbF by cytokine signal transduction.

Disclosures:

No relevant conflicts of interest to declare.
Biological Characteristics of Bone Marrow-Derived Mesenchymal Stem Cells From a Patient with β-Thalassemia.
Authors: Yoon Jinsun , Song Sung Heon , Kim Seoju , Kim Eun Shil , Kong Gu , Kim Byung-Kook , Lee Young
Published: October 2019

Abstract 4579

Thalassemia syndromes are considered as a heterogeneous group of inherited anemias leading to decrease or loss of synthesis of one or more globin chain subunits of the adult hemoglobin tetramer (Hb A). Beta-thalassemia results from a decrease in β-chain production of hemoglobin relative to alpha-chain production. Hematopoietic stem cell transplantation from HLA-identical siblings is curative for thalassemia syndrome. Bone marrow (BM)-derived mesenchymal stem cells (MSCs) are primitive, undifferentiated cells which are capable of self-renewal, and differentiating morphologically and functionally into different cell lineages including adipocytes, chondrocytes, myocytes, astrocytes, tenocytes, and hepatocytes. There have been contrasting data on whether BM derived MSCs are altered in various hematological disorders including leukemia, multiple myeloma, and myelodysplastic syndrome as well as autoimmune disease. To date, functional characterization of MSCs has never been performed in thalassemia syndrome. In the present study, we isolated BM-derivied MSCs from a patient with β-thalassemia in order to compare phenotypic and functional characteristics to those from normal healthy donor. No differences were observed between MSCs from β-thalassemia and those from normal healthy donor in terms of morphology, phenotype, karyotype, multi-differentiation capacity. In mitogen-stimulated T cell proliferation assay and mixed lymphocyte reaction, MSCs from β-thalassemia strongly inhibited the proliferation of allogeneic T cells in association with reduced proportion of CD3+, CD4+ and CD8+ cells. Furthermore, the fraction of CD4+CD25+Foxp3+ cells (Treg cells) was increased under the culture with MSCs from β-thalassemia, suggesting that MSCs from β-thalassemia exerts normal immunomodulatory function. In addition, β-thalassemia-derived MSCs expressed hematopoietic cytokines and supported hematopoiesis which were comparable to those from normal BM-dereived MSCs. In summary, β-thalassemia-derived MSCs exhibited normal phenotype, karyotype as well as normal immunomodulatory function, and autologous MSCs from patients with β-thalassemia may be an attractive source of stem cell in terms of hematopoietic support as well as immunomodulatory activity.

Disclosures:

No relevant conflicts of interest to declare.
Lentiviral Short Hairpin RNA Silencing of DNM3Decreases Megakaryoycte Progenitor Proliferation and Inhibits the Formation of the Demarcation Membrane System.
Authors: Reems Jo Anna , Wang Wenjing , Sijie Sun , Wu Xiaoping , Gilligan Diana M.
Published: October 2019

Abstract 4594

Dynamin 3 (DNM3) is a mechanochemical enzyme, which participates in membrane dynamics by hydrolyzing nucleotides to link cell membranes to the actin cytoskeleton. Recently, we identified DNM3 to be differentially up-regulated during megakaryocyte (MK) development and reported that DNM3 participates in the proliferation of MK progenitors. We report here that dynamin 3 not only participates in MK progenitor amplification, but is also involved in the formation of the demarcation membrane system (DMS). To explore the role of dynamin 3 during MK development, we used three different lentiviral DNM3 shRNA constructs to infect human umbilical cord blood (UCB) CD34+ cells that were then cultured to differentiate along the MK lineage. Relative to controls, CD34+ cells transfected with DNM3 shRNAs significantly decreased the total number of cells produced in culture as well as the number of colony forming unit-megakaryocytes (CFU-MKs). As a result, the total number of CD41+ and CD61+ progeny cells was also reduced. To investigate the role dynamin 3 plays during the maturation of MKs, a Src family kinase inhibitor, SU6656, was used to induce high polyploidization and the development of a complex DMS in the UT-7/mpl cell line. The styryl membrane dye di-8-ANEPPS was used to monitor the formation of the DMS. In the absence of SU6656 treatment, UT-7/mpl cells showed a peripheral staining pattern that was indicative of cells that did not have extensive plasma membrane invaginations. In contrast, the staining pattern of UT-7/mpl cells treated with SU6656 clearly showed a well-developed plasma membrane system with invaginations. When DNM3 was silenced in SU6656 treated UT-7/mpl cells the results showed a significant reduction in di-8-ANEPPS staining relative to empty vector and non-specific negative controls. These findings are consistent with the conclusion that dynamin 3 participates in the morphogenesis of the DMS.

Disclosures:

No relevant conflicts of interest to declare.
Demographic and Socio-Economic Features of a Cohort of Adult Sickle Cell Disease Patients.
Authors: Haider Syed N , Tang Jun , Osarogiagbon Raymond U.
Published: October 2019

Abstract 4616

Introduction

Despite improvements in diagnosis, preventive care and treatment, care of sickle cell disease (SCD) patients remains difficult. The majority are from lower socio-economic strata, poorly educated and from single parent households. The racial divide between patients (who tend to be almost all black) and their health care providers (who tend to be predominantly non black) can adversely affect the quality of care when stereotypes are assumed. We examined the demographic and socio-economic features of patients in our adult sickle cell disease program.

Patients and Methods

We retrospectively reviewed data on 118 patients that entered our program between February 2005 and October 2008. Our catchment area included parts of rural Mississippi, Tennessee and Arkansas as well as a major urban community. We reviewed the following demographic characteristics: age, gender, patient's marital status, parent's marital status, employment status, highest level of education, health insurance status and referral source. We then contrasted the characteristics of those with SC (which tends to be milder) with SS/Sβ0 sickle cell disease (which is phenotypically more severe). Continuous variables were evaluated using t-test. Chi-square test was used to assess categorical variables.

Results

The results are depicted in the table.

Conclusions

We describe the demographic and socio-economic features of a cohort of 118 patients seen in an academic center with a diverse catchment area. In spite of adverse family circumstances, relatively high proportion of the cohort was college educated (32%) or graduated from high school/GED (55%). 61% of the patients are either currently employed or current students. These results are quite contrasting to the general perception about the adult sickle cell disease population.

Disclosures:

No relevant conflicts of interest to declare.
Non Transferrin Bound Labile Plasma Iron and Iron Overload in Sickle Cell Disease: a Comparative Study Between Sickle Cell Disease and β Thalassemic Patients.
Authors: Koren Ariel , Fink Daniel , Admoni Osnat , Tennenbaum-Rakover Yardena , Levin Carina
Published: October 2019

Abstract 4625

BACKGROUND

Blood transfusions are the standard of care in β thalassemia and transfusions are also indicated in Sickle Cell Disease (SCD) patients with hypersplenism, recurrent vaso-occlusive crises and for stroke prevention. Iron overload caused by blood transfusions in thalassemia and in SCD may affect morbidity and mortality. Recent studies of iron overload in SCD suggest that the biologic features of SCD and the chronic inflammatory state may protect SCD patients from iron damage.

DESIGNS AND METHODS

In view of the controversy regarding the effect of iron overload in patients with SCD we studied the iron status, including non transferrin bound iron (NTBI) and labile plasma iron (LPI) levels in a cohort of thirty six SCD patients and compare the results with 43 thalassemia patients.

RESULTS

Our results indicate that none of the SCD patients had clinical symptoms of iron overload. Only two SCD patients had NTBI values in the grey zone (0.4 units) and none had positive values. By contrast, 14 patients with Thalassemia Major and 3 with Thalassemia Intermedia had NTBI values above 0.6, level that are in the positive pathological range. Similarly, four thalassemia patients, but only one SCD patient had positive LPI levels.

CONCLUSIONS

The parameters of iron status in SCD patients, even after frequent transfusions are different when compared to patients with thalassemia. The low NTBI and LPI levels found in patients with SCD are in keeping with the absence of clinical signs of iron overload in this disease.

Disclosures:

Koren: Novartis Oncology: Research Funding. Levin:Novartis Oncology: Research Funding.
Lack of Sustained Benefit of Red Cell Exchange in Patients with Sickling Hemoglobinopathies and Coexisting Warm Autoantibody Hemolysis.
Authors: Baron Beverly , Baron Joseph M.
Published: October 2019

Abstract 4626

Introduction

Red cell exchange (RCE) can be beneficial in the management of patients (pts) with sickling hemoglobinopathies by rapidly reducing hemoglobin (Hb) S levels and raising Hb A levels, thereby decreasing hemolysis and improving organ perfusion. We present 2 pts with sickle beta+ thalassemia (SBT) whose clinical courses were complicated by coexisting warm autoantibody hemolysis and prolonged refractory pain. They had less benefit from RCE than expected due to rapid rebound of Hb S levels and decline in Hb A levels within 48 to 72 hrs after RCE. Each subsequently improved with immunosuppressive therapy.

Case Reports

We report 2 African American female pts with SBT who presented with severe anemia, high absolute reticulocyte counts (arc), evident hemolysis, and lack of sustained response of their Hb levels to red cell transfusions. Each had multiple alloantibodies and warm autoantibodies. Patient 1 (see Fig. A), age 34 years, presented with anemia (initial Hb 7.5 g/dL), arc 267 K/μL, and total serum bilirubin (bili) 4.3 mg/dL. Between hospital days 5 and 8, she received 8 units of compatible red cells, but her Hb the next day was only 5.7 g/dL. She then underwent a 5 unit RCE (starting Hb S was 79.3%, Hb A was 14%). Hb S fell to 60.6% the next day; Hb A rose to 33%. Three days post RCE, arc was 854 K/μL, Hb S rose to 68.5%, and Hb A fell to 24.8%. Subsequently intravenous immune globulin (IVIG) was given (48 g per day for 3 days). Her Hb stabilized, began to rise, and she experienced clinical improvement. Patient 2 (see Fig. B), age 36 years, also presented with anemia (Hb 6.8 g/dL), arc 694 K/μL, and bili 2.7 mg/dL. Over the first 10 hospital days she received 3 units of red cells. She was on prednisone (5 mg daily) maintenance for warm autoantibody (IgG) hemolysis. On day 13 a 4 unit RCE was performed. Pre-RCE Hb S was 61.9%, Hb A was 26.9%. Post RCE Hb S was 40.5%, Hb A 50.9%, but 48 hrs post RCE, Hb S level had risen to 52.8% and Hb A fell to 38.8%. Arc was 843 K/μl; Hb was 6.3 g/dL. Prednisone dose was increased to 100 mg per day, and IVIG (2 gm total per kg body weight) was given. Her Hb stabilized and she improved clinically.

Conclusions

Lack of sustained benefit from RCE in these pts likely resulted from 2 factors: (1) high reticulocyte counts, leading to rapid replacement of Hb S cells; and (2) shortened survival of Hb A cells, secondary to their destruction by the warm autoantibodies. Immunosuppressive therapy targeted to the autoimmune component of the anemia provided substantial clinical benefit.

Disclosures:

No relevant conflicts of interest to declare.
Human Germinal Center-Associated Lymphoma (HGAL) Protein Expression Is Associated with Improved Failure-Free Survival in Brazilian Patients with Classical Hodgkin Lymphoma.
Authors: Azambuja Denize , Lossos Izidore , Biasoli Irene , Morais José Carlos , Britto Luciana , Scheliga Adriana , Pulcheri Wolmar , Natkunam Yaso , Spector Nelson
Published: October 2019

Abstract 4632

BACKGROUND

The HGAL gene has prognostic value in diffuse large B-cell lymphoma and expression of its cognate protein is germinal center–specific. A previous study had suggested that HGAL protein expression might also be related to outcome in patients with Hodgkin lymphoma.

PATIENTS AND METHODS

The aim of the current study was to confirm the prognostic impact of HGAL protein expression in an independent, well-characterized cohort of 232 classic Hodgkin lymphoma patients treated uniformly with the ABVD regimen from 1997 to 2004 in Brazil. The median follow-up was 6.2 years.

RESULTS

Tissue microarray analysis showed HGAL staining in 188 specimens (81%). The overall survival was better in patients with young age, early-stage, absence of B symptoms, low-risk international prognostic score (IPS) and good performance status. Failure-free survival was superior in patients with early-stage disease, low-risk IPS and HGAL-positive patients. The estimated 5-year failure-free survival for HGAL-positive and HGAL-negative patients was 82% and 67%, respectively (P=0.03). When stage, B symptoms and performance status were included along with HGAL in a multivariate analysis, advanced stage and absence of HGAL staining were independent predictors of a worse failure-free survival.

CONCLUSION

This study confirms and validates recent findings of a correlation between HGAL expression and outcome in classical Hodgkin lymphoma.

Disclosures:

No relevant conflicts of interest to declare.
BK-Viruria and Hemorrhagic Cystitis – a Retrospective Analysis of a Patient Cohort After Allogeneic Bone Marrow Transplantation for Hematological Malignancies.
Authors: Ostermann Helmut , Rieger Christina T , Peterson Lisa
Published: October 2019

Abstract 4657

Background

BK-virus-associated hemorrhagic cystitis is a major issue among patients with hematologic malignancies after allogeneic bone marrow transplantation and associated with significant morbidity and mortality. Risk factors for its development are still poorly understood and sufficient data lacking. This is a retrospective analysis of all patients with viruria after allogeneic BMT of the years 2007-2008 transplanted at the centre for BMT of the University of Munich.

Methods

All patients displaying at least one event of viruria in the period of January 2007 until December 2008 were included in this analysis. We recorded demographic factors, transplantation setting, clinical manifestation of HC, conditioning regimens, GvHD, immunosuppression and relevant co-infections.

Results

56 patients (39 ‰/ 17 š) with median age of 48,5 years were recorded. Underlying diseases were AML (31 pts. / 55,3%), ALL (8 pts./14,2%), CLL (2 pts./ 3,5%), malignant lymphoma (5 pts./ 8,9%), multiple myeloma (4 pts./ 7,1%) and other hematological malignancies (6 pts./ 10,7%). 18 patients were transplanted from a related, 38 from an unrelated donor, two of those from cord blood. 29 patients (51.8%) were registered alive at the time of data collection, 23 patients (41%) were recorded deceased, 4 patients (7,1%) could not be tracked.

The concomitant or preceeding incidence of GvHD received a special interest in our analysis. Notably, 47 pts. (84%) showed clinically apparent manifestation of GvHD, only 9 pts. (16%) displayed no sign of GvHD. Of these, 22 pts. (39%) developed an overall grade III (severe) and grade IV (life-threatening) GvHD. Furthermore, 27 pts. (48%) developed a grade III GvHD and 11 pts. (19,6%) a grade IV GvHD of a single organ, giving an overall of 67,6% of grade III or IV single organ-GvHD in this patient collective. The organ system most affected was the skin, 47pts. (84%), followed by the gut, 21 pts. (37,5%), the liver, 10 pts. (17,8%) and other organ systems 4 pts. (7,1%).

Conclusion

BK-viruria is a frequent event among patients with bone marrow transplantation for hematological malignancies. Among our patient collective, we observed a high incidence of high grade GvHD. Whether there is an independent risk factor for both or there is a causative link between these two events is not clear. However, our retrospective analysis represents one of the largest data collections regarding BK-virus-infections in allogeneic BMT patients so far and further studies are required.

Disclosures:

No relevant conflicts of interest to declare.
Detection of GATA1 Mutation in the Blood of 3 Mothers Giving Birth to Children with Trisomie 21 and TMD.
Authors: Kabisch Hartmut , Obser Tobias , Bartl Sonja , Kordes Uwe , Blohm Martin , Schneppenheim Reinhard
Published: October 2019

Abstract 4680

One of the most common human chromosomal abnormalities, Down syndrome, is caused by trisomy 21. Transient myeloproloferative disorders (TMD) occur in 10-20 % of newborns with Down syndrome. About 30 % of those TMD patients develop Acute Megakaryoblastic Leukemia (AMKL) within 4 years. Therefore both disorders have their origin in the fetus. There is emerging evidence that the protein product of GATA1 mutations, GATA1s, directly contributes to leukemogenesis. Although an in utero origin of GATA1 mutations and consecutively TMD is established it was never analyzed if GATA1 mutations could be detected in maternal PBL collected before and after the delivery of a child with Down syndrome and TMD. We therefore initiated a study to detect mutant GATA1 in three pairs of mother and child (A, B, C).

After the identification of the specific GATA1 mutations of the newborn with TMD, PBL samples of the mothers drawn 4 days after delivery (mother A), 7 days after delivery (mother B) and 4 week before as well as just before sectio cesarea ( mother C) were analyzed.

Results

All 3 patients had duplications in exon 2. Patient A had a duplication of 191 bp including the start codon,, patient B showed a duplication of 2 bp, and patient C had a duplication of 13 bp, respectively.

The three mothers harbored the TMD specific GATA1 mutations in PBL analyzed around the birth of their children.

The GATA1s signal in mother A and B had disappeared 1 month after birth. No analysis at this time point was done in mother C,whereas she was positive for the specific GATA1 mutation one month before delivery. At that time TMD was suggested by ultrasound which revealed clinical signs of hydrops in the fetus.

In mother B and C the particular GATA1 mutations could also be detected in their plasma, in the plasma sample of mother C even 4 weeks before delivery (not done in mother A).

Conclusion

In our study we could show for the first time that GATA1 mutations of leukemic cells could be detected in PBL of the mothers around birth. Cell free GATA1 DNA was also detectable. The prenatal finding of TMD specific mutations in the mothers' blood suggests that feto-maternal transfer of leukemic cells and DNA may already occur up to 4 weeks before delivery.

Disclosures:

No relevant conflicts of interest to declare.
P15 and P16 Promoters Methylation Status in APL Patients Treated by Arsenic Trioxide.
Authors: Yeganeh Omid , Alimoghaddam Kamran , Ghaffari Seyed Hamidolah , Rostami Shahrbano , Jalili Mahdi , Shamshiri Ahmad Reza , Ghavamzadeh Ardeshir
Published: October 2019

Abstract 4682

Introduction

Inactivation of tumor suppressor genes P15 & P16 due to Hypermethylation has a critical role in progression of cancer. We analyzed the meltylation status of promoter region of these two genes and their correlation with patients survival in APL patients treated with ATO without chemotherapy and /or ATRA.

Methods

Patients treated by Arsenic Trioxide without ATRA or chemotherapy and 45 newly diagnosed and 5 relapsed were enrolled in our study. Whole blood DNA was extracted and modified by sodium bisulphite reaction. Then methylation status was studied by MS-PCR and gel electrophoresis. Also for newly diagnosed patients, early mortality, disease-free and overall survival compared between methylated or non-methylated groups.

Results

Of 45 new cases, 31patients(69%) had methylated P15 but no patient had methylated P16. Eighty percent of relapsed patients had methylated P15 too, and no relapsed patient had methylated P16. DFS and OS correlated significantly with methylation status of P15 ( P=0.05 and 0.04) for patients who achieve to CR after treatment but it has no correlation with early motility during treatment course. Actually methyaltion of P15 improve chance of DFS and OS.

Conclusion

P15 but not P16 is frequently methylated in APL. Although in some studies, methyaltion of P15 negatively affected prognosis of APL patients treated by ATRA but it has no adverse effect when Arsenic Trioxide used in induction and cosolidation.

Disclosures:

No relevant conflicts of interest to declare.
Familial Platelet Disorder and Predisposition to Myeloid Leukemia (FPD/AML) in the Absence of RUNX1 Mutations- A Report of Three Families.
Authors: Berman Jason N. , Couban Stephen , Ebbin Allan , Frangoul Haydar , Quisumbing Maria B. , Aikie Thomas E , Riddell Christie , Ferguson Meghan , Ludman Mark D. , Fernandez Conrad V. , Cantor Alan B.
Published: October 2019

Abstract 4689

Familial Platelet Disorder with Predisposition to Acute Myeloid Leukemia (FPD/AML) has been attributed to abnormalities of the RUNX1 gene, located at human chromosome 21q22. The protein product of this gene forms a critical component of the core binding factor (CBF) complex, which is required for normal hematopoiesis. Truncating mutations and deletions of RUNX1 (particularly in exons 3, 4, and 5, encoding the DNA binding and CBF-beta interaction domains) that result in haploinsufficiency or dominant-negative activity have been identified as the genetic mechanisms underlying FPD/AML. Clinically, these patients present with mild to moderate thrombocytopenia, platelet dysfunction, bleeding, and about a 35% risk of developing AML. We report on three unrelated families who share histories of autosomal dominant thrombocytopenia over two to five generations, bleeding, platelet aggregation defects, and development of myelodysplastic syndrome, AML or intriguingly, chronic myeloid leukemia (CML). Detailed genetic interrogation of the RUNX1 locus in the proband from each family, including complete sequencing of all 8 exons, flanking 50 base pair regions, and P1 and P2 promoters, as well as gene dosage studies, failed to demonstrate a causative lesion in the RUNX1 gene. Our findings strongly suggest that genetic loci other than RUNX1 are involved in some cases of autosomal dominant thrombocytopenia with a predisposition to both AML and CML. Identification of the disease-causing genes in these families will allow for prospective testing of family members, appropriate surveillance and early intervention in affected individuals, and potentially new molecular insights into leukemogenesis. We have broadened the genotype-phenotype correlation in FPD/AML beyond the RUNX1 gene and suggest that this syndrome may be more genetically heterogeneous than initially described.

Disclosures:

No relevant conflicts of interest to declare.
Loss of Structural Maintenance of Chromosomes 1A Protein Expression Is Associated with a Poor Prognosis in Acute Myelogenous Leukemia.
Authors: Krug Utz O. , Hömme Claudia , Tidow Nicola , Buerger Horst , Koehler Gabriele , Heinecke Achim , Buchner Thomas , Berdel Wolfgang E , Koschmieder Steffen , Mueller-Tidow Carsten
Published: October 2019

Abstract 4693

Loss of Structural Maintenance of Chromosomes 1A Protein Expression is Associated with a Poor Prognosis in Acute Myeloid Leukemia

Utz Krug, Claudia Hömme, Nicola Tidow, Horst Bürger, Gabriele Köhler, Achim Heinecke, Thomas Büchner, Wolfgang E. Berdel, Steffen Koschmieder, Carsten Müller-Tidow

Introduction

Acute myelogenous leukemia is a genetically heterogenous disease with many risk factors for a poor prognosis. One of the most important independent risk factor in AML is the age at diagnosis. Older patients with AML generally have a poor prognosis which suggests that the biology of AML in elderly patients differs from AML in younger patients.

Methods

Gene expression profiling was carried out to identify age related changes in AML blasts of 67 AML patients of different age (range: 17 to 80 years). Among the genes that correlated with age, SMC1A was selected for protein expression studies. A tissue array was created containing bone marrow histology samples of 135 patients with newly diagnosed AML of different ages and probed with an antibody against SMC1A and protein expression was quantified by the DAKO score.

Results

131 genes showed a significant correlation between mRNA expression levels and patient age. Increasing age was associated with significantly decreased mRNA levels of SMC1A. 116 patient samples were evaluable for SMC1A protein expression and expression of SMC1A protein was low or absent in 74 out of 116 AML specimens. SMC1A protein expression did not show a correlation with patients' age at diagnosis. Both event free survival (2.6 months vs. 10.3 months, p=0.003, see figure) and overall survival (10.4 months vs. 22.6 months, p=0.015, see figure) were significantly worse in patients with low or absent SMC1A protein expression. In a multivariate analysis, SMC1A protein expression level remained a significant prognostic factor for event free survival (p=0.014) with a borderline significance for overall survival (p=0.066).

Conclusions

We identified 131 genes with putative age-dependent microarray mRNA expression and identified low levels of SMC1A protein expression as a marker for poor prognosis in patients with newly diagnosed acute myeloid leukemia.

Disclosures:

No relevant conflicts of interest to declare.
Detection of NFκB Activity in Primary CLL Cells Using a Sensitive Oligo-Based Chemiluminescent ELISA.
Authors: Mertens Daniel , Bhattacharya Nupur , Sarno Antonio , Idler Irina , Zenz Thorsten , Döhner Hartmut , Stilgenbauer Stephan
Published: October 2019

Abstract 4709

Contemporary research on cellular signaling in oncogenesishas undergone a shift of focus from qualitative measurements of single signaling pathways to high-throughput quantitation of comprehensive signaling networks. Notably, NFkB is a family of transcription factors involved in immune and inflammatory responses, developmental processes and especially in cellular growth and apoptosis, which are central in oncogenesis: NFkB signaling is deregulated in a number of hematologic malignancies like Hodgkin's lymphoma or chronic lymphocytic leukemia. We have established a chemiluminescent oligonucleotide-based ELISA (co-ELISA) that is simple and quantitative. In contrast to currently used assays, it allows quantitation of all NFkB components (i.e. RelA, p50, p52, RelB, and c-Rel). In addition, it can make use of whole extract and does not require cumbersome nuclear/cytosolic fractionation, saving time and resources. Co-ELISA has a 3.5- to 43-fold higher signal-over-noise ratio than currently available assays, while the standard error (SEM) is 3-to 6-fold lower. Co-ELISA is suited for the assessment of NFkB activity in primary tumor cells or in high-throughput experimentation i.e. functional evaluation of novel compounds. The method is faster than EMSA, not restricted to transfectable cells as is the case for luciferase reporter assays, making it applicable to primary tumor cells, and ten times more cost-efficient than commercially available ELISA assays. Co-ELISA is a quantitative, sensitive, fast, and cost-efficient measurement of the activity of all DNA-binding NFkB proteins, also in primary tumor cells.

Disclosures:

No relevant conflicts of interest to declare.
Expression of KDR Plus Detection of Flt3-ITD and NPM1 Mutation as a Prognostic Marker in AML Patients with Normal Karyotype.
Authors: Liang Aibin , Xiu Bing , Huang Binbin , Han Ying , Bo Lanjun
Published: October 2019

Abstract 4715

From July 2006 to February 2009, total of 146 patients with non-APL AML were admitted in our department. Cytogenetic analyses, detection of Flt3-ITD, NPM1 gene mutation and VEGF and its receptors (Flt-1, KDR) were performed on all patients. Forty-nine patients less than 60 years of old and with normal karyotype were selected for prognostic analyses. Flt3-ITD was positive detected in 15 cases (30.61%), NPM1 mutation in 18 cases (36.73%), VEGF in 46 cases (93.88%), Flt-1 in 41 patients (84.04%) and KDR in 38 cases (77.55%). After one or two courses of induction therapy with IDA regimen (Idarubicin + cytarabine: 3+7) in all 49 patients, total CR rate was 67.43% (33/49). One patient died because of severe invasive fungal infection. Among the remaining 15 non-CR patients, 10 were Flt3-ITD positive and NPM1 negative, and all with higher expression of VEGF and KDR. All the CR patients were treated with consolidation regimen with high dose cytarabine (3g/m2, q 12h iv for 3 consecutive days) for 6 courses. After follow-up at least for 6 months, only 12 patients are alive up to now. In these 12 patients, Flt3-ITD were all negative expressed, 6 patients were NPM1 positive, 2 patients VEGF negative, 3 patients both KDR and Flt-1 negative. There are no patients alive with positive expressed of Flt3-ITD, KDR and negative expressed of NPM1. Therefore, we supposed that negative expression of Flt3-ITD and KDR plus positive expression of NPM1 could be a favorable parameter for outcome prediction in AML patients with normal karyotype.

Disclosures:

No relevant conflicts of interest to declare.
Aven and Survivin Expression in Egyptian Acute Leukemia and Their Relation to Apoptosis.
Authors: Assem Magda , Hamid Thoraya Mohamed Abdel , Basset Gihan Abedel , Hilal Amany , Kamel Mahmoud , Khalil Iman , Alsharkaway Nahla , Metwally Aymen
Published: October 2019

Abstract 4717

Background

Several anti apoptotic signals have been recently identified. Aven and Survivin are broadly expressed and are conserved in mammalian species.

Patients and Methods

Sixty-four patients with acute leukemia [39 with acute myeloblastic leukemia (AML) and 25 with acute lymphoblastic leukemia (ALL)] were used as the study group. Reverse transcriptase (RT –PCR) was used, Aven and Survivin expression were detected at the messenger (mRNA) level .DNA fragmentation was carried out on daily basis before &24 hours after the therapeutic dose every day for 4 times during the initial induction of chemotherapy (everyday for ALL& every otherday for AML).

Results

Survivin expression was found in (66%) of acute leukemia, near significantly (p=0.06) more in AML (74%) than in ALL (52%). While, Aven expression was (40.6%) in acute leukemia equally expressed in AML (41%) and ALL (40%).Patients were categorized into 3 groups based on DNA fragmentation, whereby group I: DNA fragmentation found on day 1 or day 1+day 2, group II: DNA fragmentation found on days 1, 2 and 3, group III where fragmentation was found on days 1+2+3+4 or/and day5. Absence of Aven expression significantly (p<0.001) contributed to DNA fragmentation as 24 /35 (68.6%) of acute leukemia with Aven negative were in group III (the best group). Absence of Survivin did not contribute as much, Where 18 out of 31 (58%) of group III were Survivin positive (P= 0.1).None of the concordant both positive Survivin and Aven were in group III (the good 5 day fragmentation), this was statistically highly significant (P< 0.001) relation. Survivin was statistically related to CD7 expression (P<0.001) in AML only, while Aven was statistically related to CD34 expression (P<0.014) in the whole acute leukemia cases. Aven was statistically correlated to alkaline phosphatase (P< 0.036).There was a significant dissociation between Aven and Survivin in AML (p=0.03) and near significant dissociation in ALL (p=0.07).

Conclusion

Aven seems to be more important as an inhibitor of apoptosis than survivin in acute leukemia. The presence of both Aven & survivin confers a survival disadvantage & a significantly worse DNA fragmentation pattern, thus suggesting a synergistic inhibition of apoptosis when present together, although, they tend not to be expressed together. The highly significant relation between CD7 & surviving expression, might suggest their involvement in a common signal transduction pathway.

Key words: Survivin, Aven, AML, ALL and DNA fragmentation.

Disclosures:

No relevant conflicts of interest to declare.
Cytogenetic and Molecular Features of 1331 Chinese Patients with Primary Acute Lymphoblastic Leukemia (ALL).
Authors: Chen Bing , Wang Xiang , Yao Ying , Zhu Yong-Mei , He Hai-Yan , Jiang Xiao-Xing , Zhou Ji-Fang , Chen Hai-Min , Cao Qi , Yang Guang , Jiang Chun-Lei , Weng Xiang-Qin , Zhang Xu-Xi , Jiao Bo , Zhao Wei Li , Li Jun-Min , Chen Yu , Xiong Shu-Min , Shen Zhi-Xiang , Gu Long-Jun , Jiang Hui , Mi Jian-Qing , Chen Zhu , Chen Sai-Juan
Published: October 2019

Abstract 4718

Cytogenetic-molecular signature plays an important role in diagnosis, treatment and prognosis evaluation in acute lymphoblastic leukemia (ALL). With examination of karyotype and gene rearrangement, we investigated retrospectively the cytogenetic characteristics in 1331 adults and children with primary acute lymphoblastic leukemia (ALL) diagnosed from November 1989 to July 2007.

Cytogenetic and molecular genetic analysis was carried out successfully in 1030 (77.39%) of 1331 cases, abnormal clone was detected in 629 (61.07%) cases. The abnormalities of t(9;22)/BCR-ABL and/or t(4;11)/MLL-AF4 were presented more commonly in adults than in children (25.39% Vs 10.34%,P<0.001; 4.69% Vs 1.68%,P=0.0067). On the contrary, hyperdiploid(>50) was more frequently seen in children than in adults (13.18% Vs 4.30%,P<0.001). In addition, t(8;14)/IGH-MYC,t(12;21)/TEL-AML1 and t(11;19)/MLL-ENL were only observed in pediatric ALL. Compared with data of western countries, t(9;22)/BCR-ABL involvement in Chinese ALL patients, either in adults or in children, was more common. Simultaneously, a lower frequency of t(12;21)/TEL-AML1 was found in Chinese pediatric ALL patients.

Ik6, one of IKZF1 deletion isoforms, was detected in 72.29% of ALL patients with Ph chromosome using reverse transcription-PCR amplification.

In T-ALL, the frequency of the expression of SIL-TAL,CALM-AF10,HOX11,HOX11L2 gene and the mutation of NOTCH1 gene was 12.38%,4.69%,25.49%,24.51% and 32.99%, respectively.

The overall survival of the children was superior to that of the adults, which was the same as previous reports by many other groups. According to the prognosis of patients, these cytogenetic-molecular signatures could be further classified into three subgroups in adult and four subgroups in children. Patients with Ik6 deletion isoform of IKZF1 gene had a significantly worse prognosis than those with wild type isoform (P=0.009). In T-ALL, patients with NOTCH1 mutation were related with poor outcome (P=0.007).

Disclosures:

No relevant conflicts of interest to declare.
Investigation On Exosomes Carrying TSA Derived From Healthy Human White Buffy Coat with Positive HLA-A2 and PolyI:C On Subcellular Antitumor Vaccination.
Authors: Ren Weina , Chang Chunkang , Li Xiao
Published: October 2019

Abstract 4747

Aim

Use human leukocyte antigen –A2 (HLA-A2) positive human dendritic cell (DC) derived DEX to support NY-ESO-1 antigen and polyI:C to increase the proliferation of specific cytotoxic T lymphocyte (CTL) in transgenic mice, and increase its anti-tumor effect.

Methods

Mature dendritic cells derived from peripheral blood mononuclear cells (PBMC) are isolated from the blood from healthy adults with positive HLA-2A. By centrifugation and membrane ultrafiltration, EXO is extracted from the supernatant of DC secretions. Transgenic C57 mice were immunized by human derived tumor testis antigen NY-ESO-1/EXO with or without polyI:C. Mice were sacrificed four weeks after immunization, and spleen cells were isolated and underwent function test. The experiments include: antigen specific CTL proliferation efficiency was tested by dimerization experiment; the antitumor effect for K562 cells and melanoma were tested under different ratio between effecter cells and target cells (0:1, 10:1, 50:1 and 100:1).

Results

Dimerization experiment indicated that the effect of DEX/TSA+PolyI:C was (1.98±0.79)%. The effect of DEX/TSA was (1.61±0.58)%. The antitumor effect for the ratio of 0:1, 10:1, 50:1 and 100:1 by DEX/TSA:PolyI:C were (11.14±1.36%) A (14.17±0.62%) A (15.71±2.48%) A (24.31±2.91%) for K562 cells; The antitumor effect for DEX/TSA group (12.23±2.25%)A(13.10±1.57%)A(15.27±2.93%)A(19.87±2.72%)for K562 cells; With ratio 10:1 and 100:1, the antitumor effect of DEX/TSA +PolyI:C is better than DEX/TSA group (P<0.05). Whereas, with increased ratio between effecter cells and target cells, there is no significant improvement on antitumor effect for control cells.

Conclusion

It is promising to combine DEX/TSA derived from healthy human blood with positive HLA-A2 and PolyI:C as a new subcellular antitumor vaccination.

Disclosures:

No relevant conflicts of interest to declare.
Whole Brain Radiation Alone Brings the Favorable Outcomes for AIDS Related Primary Central Nervous System Lymphoma (PCNSL) Especially with Good Performance Status in the HAART Era.
Authors: Nagai Hirokazu , Odawara Takashi , Ajisawa Atsushi , Hagiwara Shotaro , Uehira Tomoko , Uchiumi Hideki , Yotsumoto Mihoko , Miyakawa Toshikazu , Watanabe Akira , Konishi Mitsuru , Saito Seiji , Takahama Souichirou , Tateyama Masao , Okada Seiji
Published: October 2019

Abstract 4751

Background

Primary central nervous system lymphoma (PCNSL) is one of an acquired immunodeficiency syndrome (AIDS)-defining illness and is the second common cerebral mass lesion after toxoplasmosis in HIV infected population. The introduction of highly active antiretroviral therapy (HAART) significantly improved the survival of AIDS related PCNSL. But, the standard procedure to manage these patients still remains to be established. We performed the nationwide retrospective survey to elucidate the actual status and the significant prognostic variables of AIDS-related PCNSL in the HAART era.

Methods

All AIDS related PCNSL newly diagnosed at 12 regional center hospitals for HIV/AIDS in Japan during the period 2002-2008 were registered. The data about diagnostic modalities, CD4 counts, performance status (PS), HAART regimens, treatment modalities, and clinical outcomes were collected. A multivariate and univariate Cox regression analysis was performed to assess the effects of treatment and the clinical variables on OS.

Results

Twenty four AIDS related PCNSLs were registered. Median age was 41 (21-60) year old, and male gender was 96%. Ten patients were diagnosed as PCNSL based on imaging features and positive EBV DNA PCR of cerebrospinal fluid without brain biopsy. HAART was administrated in 88%. Twenty two patients were treated by radiotherapy alone, and only one cases were received the combined modality treatment. Multivariate analysis revealed that the significant predictor of improved survival was completion of whole brain radiotherapy (WBRT) with a total dosage more than 30 Gy (P=0.017). Furthermore, 9 of 10 patients with good PS (0-2) group were alive with CR (all were received WBRT), nevertheless 64% of poor PS (3 or 4) group (9/14) died sometimes with short clinical course. The estimated 3yrs overall survival rate of each group was 100% and 38% respectively (p=0.0198). The incidence and grade of radiation related leukoencephalopathy was limited in good PS group.

Conclusion

Radiotherapy alone with HAART is shown to be the most common treatment for the AIDS-related PCNSLs, but the role of systemic chemotherapy should be elucidated. Some AIDS-related PCNSLs, especially with good PS, achieved the durable remission by WBRT with the curative intent dose, in the HAART era.

Disclosures:

No relevant conflicts of interest to declare.