bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Relevance of IDH1 and IDH2 Mutations and Single Nucleotide Polymorphism (SNP) rs11554137 in Patients with Acute Myeloid Leukemia Treated with Stem Cell Transplant in First Complete Remission
Authors: Abalkhail Hala , Chaudhri Naeem A. , Walter Claudia Ulrike , Al Zahrani Hazzaa , Nounou Randa , Altombakty Ola , Almhareb Fahed , Mohamed Said , Rasheed Walid , Alsharif Fahad , Aljurf Mahmoud , Albitar Maher
Published: October 2019

Abstract 4643

Background:

Mutations in the genes encoding for the cytosolic isocitrate dehydrogenase 1 (IDH1) and the mitochondrial version of this enzyme (IDH2) have been reported in acute myeloid leukemia (AML) and other types of cancer. Presence of these mutations in AML correlated with more aggressive disease in some studies, but other studies did not find significant correlation with outcome when patients were treated with intensive chemotherapy. The polymorphism at rs11554137 in IDH1 was reported to correlate with inferior outcome in cytogenetically normal AML patients. Most of the studies evaluating the prognostic relevance of IDH1, IDH2 and the rs11554137 polymorphism were reported in patients treated with standard chemotherapy. The prognostic significance of these markers is not fully studied when AML patients are treated with intensive chemotherapy followed by allogeneic hematopoietic stem cell transplantation (HSCT). We studied the prognostic value of the IDH1, IDH2, and rs11554137 polymorphism in patients with AML treated with HSCT in first complete remission (CR1).

Methods:

Samples from 69 patients with AML were analyzed for mutations in IDH1, IDH2, and the SNP rs11554137 by direct sequencing. All patients were diagnosed with AML, treated with chemotherapy followed by HSCT in CR1. They included intermediate (N=42) and adverse (N=27) cytogenetic risk groups.

Results:

The R132H and the SNP C to T change at rs11554137 (silent G105) in IDH1 were detected in 5 (7%) and 6 (9%) of 69 AML patients. The IDH2 mutations (N=5; 7%) included R140Q, 172K, and T169A. Patients with IDH1 or IDH2 mutations did not differ significantly in their overall survival, event free survival, or time to relapse from those without mutation. In addition, the rs11554137 SNP polymorphism did not correlate with outcome in this group of AML patients. We also looked at the combination of mutations as compared with cytogenetic risk and found no difference in survival or event free survival based on IDH1 or IDH2 mutations or rs11554137 polymorphism in the intermediate cytogenetic risk group.

Conclusion:

The data suggests that HSCT after intensive chemotherapy in CR1 may neutralize the negative prognostic impact of IDH1 and IDH2 or the rs11554137 SNP polymorphism. However, the number of cases is relatively small and further studies with larger number of cases are needed.

Disclosures:

No relevant conflicts of interest to declare.
Bortezomib Inhibit Acute Graft-Versus-Host Disease Via Shifting the Combination of T Cells Subpopulations
Authors: Weng Jianyu , Lin Shaoze , Lai Peilong , Lv Meikun , Du Xin , Ling Wei , Guo Rong , Luo Chengwei
Published: October 2019

Abstract 4709

Objective:

Acute graft-versus-host disease (aGVHD) confines the wider application of allogeneic bone marrow transplantation (allo-BMT), but recently studies indicate that it is possible to reduce the incidence and severity of aGVHD while preserving the GVT by using bortezomib. In current study, we explored the changes of T cell subsets after allo-BMT administrated with bortezomib immediately, in order to establish the mechanism about bortezomib attenuation aGVHD.

Materials and Methods:

BALB/c mouse were injected of 0.5 mL PBS containing C57BL/6 2×107 nucleated BM cells plus 1×107 splenocytes followed a single dose of lethal total body irradiation (TBI, 0.7 Gy/min, 8.0 Gy) with or without bortezomib at 1.0 mg/kg. The level of CD4+ CD25+ Foxp3+ regulator T cells is quantified by flow cytometry, and the cytokine level of IL-2 and IL-4 is quantified by ELISA.

Results:

Bortezomib remarkably reduce aGVHD severity and prolonged the surviving time. Along with bortezomib injection, the level of CD4+ CD25+ Foxp3+ regulator T cell is significantly increased, the cytokine level of IL-2 is decreased but IL-4 is increased.

Conclusion:

Bortezomib inhibit aGVHD through shifting the combination of T cell subpopulations with up regulation CD4+CD25+ Foxp3+ regulator T cells lead to reset Th1/Th2 cytokine balance.

Disclosures:

No relevant conflicts of interest to declare.
Erythropoietin Treatment Is Associated with Decreased Blood Glucose Levels: A Preliminary Study in Hematologic Patients
Authors: Oster Howard S , Gvili Moran , Katz Odelia , Hoffman Michael , Neumann Drorit , Mittelman Moshe
Published: October 2019

Abstract 4793

Introduction:

Erythropoietin (EPO) is the major hormone which enhances proliferation and maturation of the red cell lineage, and its recombinant form (rHuEPO) is used extensively to treat various types of anemia. rHuEPO has also been found to exert effects in other organ systems, and our previous work has demonstrated an immunomodulatory role for EPO. Recently, we have also found that mice exposed to high levels of EPO (either rHuEPO injections or transgenic mice overexpressing human EPO), have significantly lower levels of blood glucose than those of their respective controls (Katz et al., J Endocrinol 2010;205:87-95). The current retrospective study was designed to determine whether rHuEPO treatment in hematologic patients, is associated with decreased blood glucose levels.

Methods:

Patients receiving rHuEPO were examined, comparing glucose levels (morning blood tests, assumed to be fasting) while on rHuEPO treatment to those off treatment. All patients served as their own controls. To test the association between rHuEPO treatment and blood glucose levels, we employed a mixed-model repeated-measures analysis of variance (ANOVA).

Results:

The charts of 19 patients were reviewed to determine the starting date of rHuEPO and the levels of blood glucose in relation to rHuEPO treatment. Mean age: 77 (range: 54–93). Thirteen patients had myelodysplastic syndrome, and six had multiple myeloma. Two patients had diabetes mellitus. Average glucose levels (mean±95%CI) without rHuEPO treatment were 116.07±4.98. Glucose measurements were available for a median of 9.23 (interquartile range: 7.90–16.80) months after the initiation of rHuEPO treatment. The average glucose level over that period of time was 101.77±4.86 (p<0.0001). The two diabetic patients also demonstrated a trend towards reduced serum blood glucose and lower HbA1C while being treated with rHuEPO.

Conclusions:

Treatment of hematologic patients with rHuEPO is associated with significantly lower blood glucose levels, and might serve in the future to improve glucose control in anemic patients with hyperglycemia. Further studies with both diabetic and non-diabetic patients are currently underway to clarify this association.

Disclosures:

No relevant conflicts of interest to declare.
Adequacy of Vaccine Documentation Against Encapsulated Bacteria in a Population of Children with Sickle Cell Disease
Authors: Freiberg Andrew S. , Singleton Antea C
Published: October 2019

Abstract 4853

Immunization status is often used as an indicator of improved quality of health care and access in pediatric populations. The increased life expectancy in the general pediatric population has largely been attributed to the prevention of infectious disease through immunizations, and this fact is even truer for individuals with sickle cell disease. Adherence to recommended vaccination schedules against encapsulated bacteria in patients with sickle-cell anemia has not been well studied in any population. In order to determine documented compliance rates based on the current recommendations of the Department of Health and Human Services Center for Disease Control Immunization Schedules, a retrospective chart review was performed on 117 patients diagnosed with sickle cell anemia (HbSS) followed at the Penn State Hershey Children's Hospital Regional Pediatric Sickle Cell Center, through the year 2009. Since the vaccinations were given by each patient's primary provider(s), we attempted to obtain documentation of immunizations given by these providers, particularly against encapsulated bacteria. For 35 of the 117 patients, we were unable to obtain any forwarded documentation of any vaccinations. The vaccination schedules of the remaining 82 patients were audited for coverage against encapsulated bacteria. Only 16 of the 82 patients (19.5%) with forwarded documents demonstrated complete documented compliance, defined as having complete coverage against H. influenzae, N. meningitides, and S. pneumococcus. Of the remaining forwarded charts, 63 patients (76.8%) demonstrated some form of partial documented compliance, but four patients (4.9%) did not have documentation of vaccinations against any encapsulated bacteria despite documentation of receiving other vaccinations. Full documented compliance as indicated by forwarded records was most noted in children between the ages of 0–11.9 years (14/16 of patients with complete compliance). The proportion of adolescents without any forwarded documentation was larger than the younger age group; however, this difference did not reach statistical significance (p=0.36). Of the 82 patients whose primary care physicians forwarded documents, significantly more (14 of 34) adolescents (≥12 years of age) than children aged 0–11.9 years (8 of 48) were missing documentation of more than half of their vaccines (p=.014). These results indicate deficient documentation of vaccination coverage against encapsulated organisms, particularly in the adolescent population. To determine the clinical implications of these results, further studies detailing the demographics, socioeconomic status and correlations of such characteristics to morbidity and mortality should be completed.

Disclosures:

No relevant conflicts of interest to declare.
Glucocorticoid-Induced Proliferation and Differentiation of Untreated Pediatric AML Cells
Authors: Haarman Eric , Cloos Jacqueline , Zwaan Christian M. , de Haas Valerie , Reinhardt Dirk , Creutzig Ursula , Kaspers Gertjan J.
Published: October 2019

Abstract 4873

The aim of the present study was to contribute to a more rational use of glucocorticoids (GC) in pediatric AML, by studying the relationship between in vitro GC responsiveness of AML cells, cell biological features and prognosis in 116 pediatric AML patients treated with GC-containing protocols (AML-BFM-87/93, DCOG-ANLL 87/93). In vitro GC resistance and GC-induced proliferation were determined by the 4-days MTT assay, GC-induced differentiation by morphology. Resistance to prednisolone (PRD) was expressed as the LC50 value, the concentration lethal to 50% of leukemic cells. Fifteen percent of the AML samples were somewhat sensitive to GCs (LC50<150 ug/ml), with an overrepresentation in that subgroup of FAB type M1 samples. Six out of 14 AML FAB type M1 samples were somewhat sensitive as compared to only 11 out of 101 non-FAB type M1 samples (p=0.002). However, none of the AML samples were highly sensitive to GC (LC50 < 0.1 ug/ml), in contrast to what is commonly observed in ALL cells (Kaspers et al., Blood 1998). Moreover, GC-induced AML cell proliferation was observed in 26% of samples, more frequently in FAB type M5 (p=0.01). Patients whose AML cells proliferated in vitro upon GC exposure had a lower 10-year event-free survival (32 % SE 5.5% versus 49% SE 8.5%, p=0.04). Such a difference was not found for patients with or without spontaneous proliferation. GC-induced differentiation was not observed in vitro. Multivariate analysis including BFM risk-group classification, FLT3 internal tandem duplications (FLT3/ITD) and GC induced proliferation, showed that only FLT3/ITD was an independent prognostic factor (P = 0.01). In conclusion, GCs are rarely highly cytotoxic towards AML cells, but do induce proliferation in a significant part of AML samples. That phenomenon is undesirable, and it is associated, though not independently, with a worse treatment outcome. We thus advice to use GC as antileukemic or anti-emetic drug with great reluctance in pediatric AML.

Disclosures:

No relevant conflicts of interest to declare.
Clinical and Experimental Features of Natural Killer Cell Leukemias in a Chinese Population: A Single Center Experience of 10 Years
Authors: Zhang QiGuo , Ou-Yang Jian , Zhou Rong-Fu
Published: October 2019

Abstract 4892

Leukemia of natural killer (NK) cell lineage is rare which affects Asian population preferentially. Here 10 Chinese patients with NK cell leukemia were retrospectively evaluated. Among them 7 man and 3 women, the middle age was 47 years (range 22–57 years). Aggressive NK cell leukemias were finally diagnosed in 7 patients, myeloid/NK cell precursor acute leukemias in 2 patients and NK-CLPD in 1 patient. Pronounced extramedullary involvements (breast mass, multiple subcutaneous nodes and eye rectus muscle mass) were the main manifestation in 2 myeloid/NK cell precursor acute leukemias and 1 aggressive NK-cell leukemia. Clonal cytogenetic abnormality (46, XX, 16P-) was only found in 1 myeloid/NK cell precursor acute leukemia patient. Epstein-Barr virus test was performed in 8 patients and 3 were Positive (all were aggressive NK-cell leukemias). The overall survival of 4 cases of aggressive NK-cell leukemias combined with hematophagocytic syndrome were less than 1.3 months. During following up, 8 patients died of disease, 1 NK-CLPD patient and 1 aggressive NK-cell leukemia patient treated by allo-BMT were still alive. The former NK-CLPD, who was in indolent course, survived 60+ months. The later patient was first diagnosed as NK-CLPD but with lymphoadenopathy and splenomegaly, the metabolic activity of spleen by F-18 FDG uptake was in normal range, lymph node pathology showed reactive change and PB EBV-DNA copy was negative, later hematophagocytic syndrome occurred and EBV-DNA copy became positive. The lymph node pathology were reexamined by FISH method and scattered EBV positive cells were found, so the exact diagnosis from beginning should be occult aggressive NK cell leukemia, soon matched sibling donor BMT was performed and the patient got complete remission and the EBV-DNA copy became negative and lived well for 10+ months. Natural killer cell neoplasms are rare and most in aggressive clinical course, novel therapeutic regimens including allogeneic bone marrow transplantation should be investigated to improve outcome of this disease.

Disclosures:

No relevant conflicts of interest to declare.
Paroxysmal Cold Hemoglobinuria with Positive Treponema Immobilization Test. Report of Case
Authors: MILIé N.
Published: October 2019

A case of paroxysmal cold hemoglobinuria is described without a familial or personal history of syphilis. The patient, a 34 year old woman, shows no clinical evidence of syphilis. Numerous serologic tests for syphilis including treponemal immobilization are consistently positive. Despite therapy with penicillin and cortisone the hemolytic disease remains unchanged.
Siderocytes and the Spleen
Authors: CROSBY WILLIAM H. , BENJAMIN NAOMI R.
Published: October 2019

1. Blood containing high concentrations of siderocytes was transfused into four recipients with spleens and into two who lacked a spleen. In the circulation of recipients with spleens the siderin granules rapidly disappeared while the transfused red cells remained. In the recipients who had no spleen both the granules and the red cells remained.

2. It is concluded that the spleen is somehow able to bring about the removal of siderin inclusion bodies without destroying the red cells that contain them.
Further Observations on the Effect of Cortisone and Thyroxin on the Blood Picture of Hypophysectomized Rats
Authors: MEINEKE HOWARD A. , CRAFTS ROGER C.
Published: October 2019

In the article "Further Observations on the Effect of Cortisone and Thyroxin on the Blood Picture of Hypophysectomized Rats" by Howared A. Meineke and Roger C. Crafts, appearing in the January 1957 issue of BLOOD, the amount 5 ml. (page 12, line 17) should read 0.5 ml.
The Life Span of the Red Blood Cell and the Red Blood Cell Volume in the Chicken, Pigeon and Duck as Estimated by the Use of Na2Cr51O4
Authors: RODNAN GERALD P. , EBAUGH FRANKLIN G. , FOX M. R. SPIVEY , CHAMBERS DORIS M.
Published: October 2019

Eight to 25 per cent of intravenously injected Na2Cr51O4 binds firmly with erythrocytes of the chicken, pigeon and duck. Calculation of the maximum life span of these avian red cells was made from the disappearance time of circulating radioactivity. The maximum life span of the chicken erythrocyte was found to be 35 days, of the pigeon erythrocyte 35-45 days, and the duck erythrocyte 42 days. Comparing the life span of avian erythrocytes with those of other animal species, the rate of red cell turnover in the mammals, birds, and reptile (turtle) was found to correlate directly with basal heat production per kilogram body weight.

Using erythrocytes tagged with Na2Cr51O4 in vitro, the total red blood cell volume was found to be 17-24 ml. per Kg. body weight in the rooster, 9-12 ml. per Kg. in the hen, 25-31 ml. per Kg. in the duck, and 31-34 ml. per Kg. in the pigeon. These values proved somewhat lower than those obtained from the indirect estimates of red cell volume, using plasma volume figures and periphera blood hematocrit.
Pernicious Anemia without Gastric Atrophy and in the Presence of Free Hydrochloric Acid
Authors: HARRIS-JONES J. N. , SWAN H. T. , TUDHOPE G. R.
Published: October 2019

A case is described of Addisonian pernicious anemia in a girl aged 16 years, associated with a normal gastric mucosa and free hydrochloric acid in the stomach. Partial, but significant deficiency of intrinsic factor activity was demonstrated in her gastric juice by the use of Co56-labeled vitamin B12.
The Effect of Psychological Stress Procedures on the Prothrombin Time of Rats
Authors: MOGENSON G. J. , JAQUES L. B.
Published: October 2019

Forced jumping in the Lashley jumping apparatus, maintenance in cages with an electrified water source, transportation, sound-induced seizures, and electroconvulsive treatment were used in studying the effects of psychological stress on the prothrombin time of the albino rat. An increased prothrombin time occurred at certain times following forced jumping, sound-induced seizures, and electroshock. Frequently the altered coagulability was reflected in a greater heterogeneity of variance due to some animals having a longer prothrombin time and others shorter. The changes in the prothrombin time following stress were more pronounced when the prothrombopenic drug, Dicumarol, was administered. This was evident from two indices, prothrombin time and mortality.
An Abnormal Hemolytic System Associated with Leukemia and Other Disseminated Malignant Diseases
Authors: CROSBY WILLIAM H. , BENJAMIN NAOMI R.
Published: October 2019

1. In the blood of some people with leukemia and other disseminated neoplastic diseases it is possible to demonstrate an abnormal hemolytic mechanism by incubating sterile blood in vitro at 37 C. for 24 hours. There is a latent period of 6 to 8 hours before hemolysis begins. Hemolysis can be inhibited by an excess of glucose or by low pH and also by removal of calcium. The heat stability of the system indicates that the plasmal abnormality is not enzymatic.

2. It is suggested that hemolysis is due to a reaction between ionic calcium and some component of the erythrocyte. The nature of the plasma abnormality that permits the reaction is not known.

3. The significance of this phenomenon in the pathogenesis of anemia in neoplastic disease has not been established.
Association of A313g Glutathione S-Transferase P1 (GSTP1) Inborn Polymorphism with Susceptibility to De Novo MDS
Authors: Zachaki Sophia , Stavropoulou Chryssa , Daraki Aggeliki , Kalomoiraki Marina , Kollia Panagoula , Aleporou Vassiliki , Pantelias Gabriel , Kanavakis Emmanuel , Sambani Constantina
Published: October 2019

Abstract 1445

Models for the pathogenesis of myelodysplastic syndromes (MDS) imply the role of individual genetic variations in genes involved in detoxification mechanisms. GSTP1 enzyme plays a key role in detoxification of a variety of electrophilic compounds, such as benzo [a]-pyrene and other polycyclic aromatic hydrocarbons (PAHs), chemotherapy drugs and products of oxidative stress. GSTP1 acts through a common mechanism of conjugating reactive oxygen species (ROS) with glutathione, enabling their detoxification and elimination and thus defending tissues against DNA damage. The corresponding gene is subject to a single-nucleotide polymorphism (A313G) leading to abolished enzyme activity. Thus, individuals homozygous for the variant G allele (G/G) have a lower conjugating activity than individuals homozygous for the wild type A allele (A/A), while heterozygotes (A/G) display intermediate activity. The aim of the present study was to evaluate whether the GSTP1 polymorphism influences susceptibility to MDS and/or promote specific chromosomal aberrations.

We conducted a case-control study in 310 de novo MDS patients and 370 unrelated healthy controls using both a conventional PCR-RFLP assay and a novel Real-Time PCR genotyping method using hybridization probe technology. The GSTP1 gene status was also evaluated in relation to patients' characteristics and chromosomal abnormalities.

Comparison of the genotype distribution between controls and MDS cases revealed a significantly higher frequency of the variant genotypes (heterozygotes A/A and homozygotes G/G) among MDS patients, as compared to controls (p<0.0001, χ2=31.167, df=2). The most marked statistical difference between MDS patients and controls was observed between the wild-type (A/A) and the homozygous variant genotype (G/G), since subjects carrying the G/G variant genotype showed a 4.1-fold increased risk of MDS prevalence than subjects carrying the wild-type A/A genotype (p=0.000, χ2=30.5, d.f.=1, OR=4.098, 95%CI=[2.433–6.897]). Allele frequencies distribution analysis between patients and controls, showed that MDS patients exhibited a 1.9-fold increased risk of carrying at least one variant G allele, as compared to the controls (p<0.0001, d.f.=1, OR =1.9, 95%CI=[1.48–2.34]). There was no association between the GSTP1 polymorphism and gender or any specific cytogenetic subgroup, while stratification of patients according to age showed a differential GSTP1 genotype distribution (p=0.007). Our results, derived from the larger series of primary MDS cases tested for the GSTP1 genetic background, reveal an increased incidence of the GSTP1 variant genotypes among MDS patients, providing evidence for a potential pathogenetic role of the GSTP1 polymorphism on de novo MDS risk.

Disclosures:

No relevant conflicts of interest to declare.
Strong Cell Surface Expression of the Toll-Like Receptor Homolog CD180 Identifies Circulating Cells of Marginal Zone Lymphoma From Other B-Cell Malignancies
Authors: Miguet Laurent , Baseggio Lucile , Lennon Sarah , Felman Pascale , Fornecker Luc , Galoisy Anne-Cecile , Mayeur-Rousse Caroline , Herbrecht Raoul , Eischen Alice , Cianferani Sarah , Mauvieux Laurent
Published: October 2019

Abstract 1542

Despite typical features described for marginal zone lymphoma (MZL), the distinction of MZL from other small mature B-cell leukemias/lymphomas (in particular atypical chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL) or lymphoplasmacytic lymphoma (LPL) may be still difficult due to the lack of immunological positive markers of MZL and before complementary analysis (histology, karyotype). In order to find new markers, we conducted proteomic analyses on plasma membrane microparticles derived from malignant cells and identified the Toll-like receptor homolog CD180 protein as a candidate marker of MZL. This protein, also called RP105, is a leucin-rich repeat type 1 membrane protein with high extra-cellular homology to the LPS receptor. The relevance of this marker was evaluated in peripheral blood B-cells from 25 normal controls and 91 patients (MZL, n=30; CLL, n=30; MCL, n=16 and LPL, n=15) by flow cytometry.

As already described by Porakishvili et al, normal B-cells displayed higher CD180 expression than CLL B-cells (MFI=5940+/− 1708 and MFI=150 +/− 794.4, respectively p<0.001, Mann-Whitney test). We observed also a low CD180 expression in MCL (1292 +/− 1084, p<0.001) and LPL (1443+/− 905.5, p<0.001), not different from CLL and significantly lower than control B-cells (see figure). On the contrary, staining of MZL was not different from controls (7856 +/− 1030, p>0.05), but significantly higher than CLL, MCL and LPL. In addition, among the lymphomas derived from MZ B-cell, CD180 expression was statistically higher in splenic diffuse red pulp lymphoma (SDRPL) than MZL. Intra-cellular staining of CD180 showed a significant positivity in tumoral B-cell tested (CLL, MCL, MZL), suggesting that the protein is produced but its expression at the cell surface is impaired. The reasons of this alteration are not known. Furthermore, cross-linking of this receptor induced a stronger increase of CD86 expression at the cell surface in MZL than in control B-cells and in CLL B-cells, suggesting an activated NF-kB pathway.

These results strongly suggest that CD180 may be the first positive immunological marker for MZL, able to distinguish MZL from CLL, MCL, LPL in blood samples. Large prospective studies are needed to precise its diagnosis impact and its expression in the different subtypes of MZL.

Figure: CD180 expression of normal and malignant B-cells. The box plot represents the median of mean fluorescence intensity (MFI), 25/75 percentiles and extreme values. Figure:. CD180 expression of normal and malignant B-cells. The box plot represents the median of mean fluorescence intensity (MFI), 25/75 percentiles and extreme values.

Disclosures:

No relevant conflicts of interest to declare.
Prognostic Impact of Fc Gamma-Receptor Polymorphisms and Efficacy of Rituximab in Children and Adolescents with Mature Aggressive B-NHL
Authors: Burkhardt Birgit , Yavuz Deniz , Zimmerman Martin , Schieferstein Jutta , Kabickova Edita , Attarbaschi Andishe , Lisfeld Jasmin , Bonn Bettina R. , Reiter Alfred , Damm-Welk Christine
Published: October 2019

Abstract 1547

Recent studies in adult lymphoma patients indicated a correlation of polymorphisms of Fc gamma-receptors (FcγRs, encoded by the respective FCGR genes) and response to rituximab or outcome. FcγRIIIa is expressed on natural killer (NK) cells and macrophages representing the most important antibody-dependent cellular cytotoxicity (ADCC) effector cells. It is reported that the FcγRIIIa-158 valine (V) isotype has a higher affinity to human immunoglobulin G1 than the phenylalanine (F) isotype and that cells with FcγRIIIa-158V mediate ADCC more efficiently. Besides FCGR3A, a polymorphism of FCGR2A resulting in either histidine (H) or arginine (R) at amino acid position 131 was analyzed in clinical trials for its prognostic impact on rituximab response but the results remained inconclusive.

The trial B-NHL BFM Rituximab tested the efficacy of rituximab in pediatric patients (pts) with mature aggressive B-NHL. In this context the impact of FCGR-polymorphisms was analyzed in the current project.

Patients were registered and treated according to protocol B-NHL BFM Rituximab with a single dose rituximab (375 mg/m2 n= 88 and 700 mg/m2 n=40) at day one, rituximab response evaluation at day five, followed by standard chemotherapy. The FCGR3A-158V/F polymorphism and the FCGR2A-131H/R polymorphism were assessed.

Of a total of 128 pts, 105 were evaluable for rituximab response at day 5 with 47 responders and 58 non-responders (response rate 45%).

Rituximab response rate was 26% and therefore significantly lower in pts with FcγRIIa-131R/R isotype (n=23, 6 responders) compared to the response for FcγRIIa-131H isotype carriers (H/H n=34, 19 responders and H/R n= 45, 22 responders; response rate 52%). Concerning the FCGR3A polymorphism, the F/F genotype was highly significantly associated with favorable response rate of 70% (F/F n=33, 23 responders) compared to the response rate in FcγRIIIa-158V isotype carriers (V/V n=9, 2 responders and V/F n=63, 22 responders; response rate 33%). Response rate for FCGR3A-158F/F genotype was favorable for both, low-risk-R2 (14/21 responders) and higher-risk-R3/R4 (9/12 responders) pts.

In the current context of rituximab response evaluation after an upfront window, the FcγRIIIa-158 phenylalanine (F) isotype is associated with favorable response in pediatric pts with aggressive B-NHL, but not the high-affinity FcγRIIIa-158 valine isotype. These are conflicting results compared to published data for lymphoma trials in adults and the in vitro data on FcγRIIIa-158. This raises the question whether ADCC is a relevant mechanism of action in the current patient population or whether other factors might impact the interaction of NK cells, macrophages, neutrophils and lymphoma cells.

Disclosures:

No relevant conflicts of interest to declare.
Decreased Brain FDG Uptake in Patients with Diffuse Large B Cell Lymphoma (cold brain) Is Associated with High Risk Disease and Predicts Poor Response to R-CHOP Chemotherapy
Authors: Raghavendra Meghana
Published: October 2019

Abstract 1593

Background:

Decreased 2-deoxy-2-(18F)fluoro-D-glucose (FDG) uptake by brain tissue, (henceforth “cold brain”) has been occasionally described in the literature in patients with bulky solid tumors. There is currently no data describing brain FDG uptake on positron emission tomography (PET) scan in patients with diffuse large B cell lymphoma (DLBCL) and its association with clinical characteristics as well as prognosis.

Method:

We retrospectively analyzed clinical data from 110 patients with histologically confirmed DLBCL diagnosed between November 1, 2004 and December 31, 2011. Initial staging PET scans prior to treatment were reviewed. Brain FDG uptake on PET scan was analyzed by an expert nuclear radiologist. Qualitative determination of cold versus normal brain activity was made relative to usual distribution and metabolism. Secondarily, a region of interest cursor was placed on the right basal ganglia for a quantitative standard uptake value (SUV) measurement. In cases where measurement of the right basal ganglia was not possible, the cerebellum was used. Patient characteristics were compared using Fisher's exact test. Survival analyses were performed using Kaplan-Meier curves and compared using log-rank test. Cox proportional regression model was used for multivariate analyses.

Results:

We included 110 patients in the study. Twenty patients (18%) had cold brain by initial PET scan. The cold brain subgroup had a median age of 72.5 years (range, 46 to 85) and 55.0% were males. Cold brain was associated with higher rates of stage III/IV disease (80.0% vs 52.2%; P = 0.026), International Prognostic Index (IPI) score of ≥3 (85.2% vs. 36.7%; P = 0.0001), and > 1 extranodal sites of involvement (50.0% vs. 26.7%; P = 0.059) compared to the non-cold brain subgroup.

Among patients who received rituximab/cyclophosphamide/doxorubicin/vincristine/prednisone chemotherapy (R-CHOP), those with cold brain had a lower complete response rate (44.0% vs. 86.0%; P = 0.030), shorter progression-free-survival (PFS) (4.5 months vs. 9.1 months; P = 0.06), but similar overall survival (OS) (47.9 months vs. 58.4 months; P = 0.26). After adjusting for IPI, patients with cold brain seemed to have a shorter PFS compared to those without cold brain, although this was not statistically significant (hazard ratio: 3.13; 95% CI: 8.20-1.22; P = 0.096).

Conclusion:

In our study, cold brain was a phenomenon frequently observed among patients with newly diagnosed DLBCL. It was associated with higher risk disease, lower complete response rate to R-CHOP chemotherapy, and potentially shorter PFS. Further studies in larger patient population are needed to determine its true prognostic significance.

Disclosures:

No relevant conflicts of interest to declare.
Primary Mediastinal B Cell Lymphoma Treated with CHOP-Like Chemotherapy with or without Rituximab: 5-Year Results of the Mabthera International Trial Group (MInT) Study
Authors: Witzens-Harig Mathias , Ho Anthony D , Kuhnt Evelyn , Trneny Marek , Rieger Michael , Österborg Anders , Pettengell Ruth , Stevens Robert , Gill Devinder S , Walewski Jan , Loeffler Markus , Pfreundschuh Michael
Published: October 2019

Abstract 1612

Purpose

The aim of this subgroup analysis of the MInT study was to evaluate the impact of chemotherapy and rituximab in primary mediastinal B cell lymphoma (PMBCL) in comparison to other diffuse large B-cell lymphoma (DLBCL). Extended follow-up was needed to establish long-term effects.

Patients and Methods

Eligible for the randomized open-label MInT study were patients aged 18–60 years with DLBCL who had 0–1 risk factors according to age-adjusted International Prognostic Index (aaIPI), stage II-IV disease, or stage I disease with bulk. Patients were randomly assigned to six cycles of CHOP-like regimens with or without rituximab. Consolidating radiotherapy was given to sites of primary bulky disease.

Results

Of 824 patients enrolled, 87 had PMBCL and 627 other types of DLBCL. Rituximab increased the rates of complete remission (unconfirmed) in both PMBCL (from 54% to 80%; p =.015) and DLBCL (from 72% to 87%; p<.001). In PMBCL rituximab virtually eliminated progressive disease (PD) (2.5% vs 24%; p =.006), whereas without rituximab PD was more frequent in PMBCL than in DLBCL (24% vs 10%; p =.023). With a median observation time of 62 months for PMBCL and 73 months for DLBCL, the 5-year event-free survival was improved by rituximab for PMBCL (79.1% vs 47.3%; p =.011) and for DLBCL (76.9% vs 59.7%; p <.001). Furthermore, 5-year progression-free survival was improved by rituximab for PMBCL (89.8% vs 60.1%; p=.006) and DLBCL (81.1% vs. 67.8%; p <.001). Overall survival benefit was similar for DLBCL (92.0% vs 80.9%; p <.001) and PMBCL (90.2% vs 78.3%; p =.234).

Conclusion

Addition of rituximab to 6 cycles of CHOP-like chemotherapy improved long-term outcome for young patients with PMBCL and aaIPI 0–1 and eliminated differences in outcome between PMBCL and DLBCL.

Disclosures:

No relevant conflicts of interest to declare.
Transfusion Requirement, Spleen Size and Donor Type Are Strong Predictors of Outcome in Patients with Primary Myelofibrosis Undergoing an Allogeneic Stem Cell Transplant
Authors: Bacigalupo Andrea P , Dominietto Alida , Pozzi Sarah , Geroldi Simona , van Lint Maria Teresa , Raiola Anna Maria , Barosi Giovanni
Published: October 2019

Abstract 1725

Patients with myelofibrosis undergoing an allogeneic stem cell transplant (SCT), can be classified as low or high risk based on spleen size, transfusion history, and donor type (BMT 2010;45:458). In the present study we wished to validate this scoring system, based on a larger number of patients and longer follow up.

We have now analyzed 70 patients with myelofibrosis and a median age of 51, who underwent an allogeneic SCT in our Unit. The median follow up for surviving patients is 4.4 years (range 0.4–17 years). 33 patients had 0–1 unfavourable predictors (low risk) and 37 patients had 2 or more negative predictors (high risk).

Results.

The overall actuarial 10 year survival is 41%. In multivariate COX analysis independent unfavourable factors for survival were red blood cell (RBC) transfusions > 20 (RR 3,9; p=0,007), a spleen size >22 cm (RR 2,8; p=0.01) and an alternative donor (RR 3,4;p=0.001). Donor and patient age and gender, splenectomy and interval diagnosis transplant were not predictive.

The actuarial 10 year survival of patients with no risk factors (n=14), is 100%, with 1 risk factor (n=19) 47%, with 2 factors (n=27), 30%, and with 3 negative predictors (n=10) 0%.The actuarial 10 year survival of low risk (0–1 risk factors) and high risk patients (2–3 risk factors) is respectively 66% and 20%(p=0.0001): the difference is due both to a higher transplant related mortality for high risk patients (38% vs 9%, p=0.005) and a higher relapse related death (35% vs 21%; p=0.1). Peripheral blood CD34+ cell counts, pre-transplant, was a strong predictor of survival in univariate, but not in multivariate analysis.

Conclusions.

Patients with myelofibrosis undergoing an allogeneic SCT, can be classified as low or high risk based on spleen size, transfusion history, and donor type. This scoring system may be useful to identify patients who would most benefit of the transplant procedure.

Disclosures:

No relevant conflicts of interest to declare.
Germline MPLW515R Mutation in a Family with Isolated Thrombocytosis
Authors: Vilaine Mathias , Gourain Victor , Cleyrat Cédric , Feron Delphine , Girodon Francois , Morineau Nadine , Harutyunyan Ashot , Kralovics Robert , Hermouet Sylvie
Published: October 2019

Abstract 1764

Background:

Myeloproliferative neoplasms (MPNs) and related hereditary diseases form a group of chronic hematological diseases that represent a useful model of cancer development. Identification in 2005 of an activating somatic mutation (V617F) in the JAK2 gene in >50% MPN patients was of major importance in the understanding of these diseases. Later, somatic mutations of W515 in exon 10 of MPL were reported for 5–10% MPN patients with essential thrombocythemia (ET) and primary myelofibrosis (PMF), whereas certain germline mutations in the MPL and JAK2 genes (MPLS505N and JAK2V617I respectively) were associated with hereditary thrombocytosis. Here, we report the first cases of germline MPLW515 mutation (R515) in the context of familial, hereditary thrombocytosis.

Methods:

A father and daughter presented with isolated thrombocytosis. Their JAK2, THPO and MPL genes were analyzed by direct sequencing on genomic DNA extracted from whole blood, CD3+ lymphocytes, buccal swabs and hair follicles. Whole blood DNA was also analyzed on Affymetrix 6.0 SNP arrays. Impact of MPLW515R mutation on thrombopoietin (Tpo) signalling was determined by comparison with wild-type and mutated (W515K/L/A) MPL in the Ba/F3-EpoR and K562 cell lines using transient transfection.

Results:

A 36 year old woman presented with chronic isolated thrombocytosis (628–780.109 Plts/L), known for >10 years. There was a familial history of isolated thrombocytosis for the father (74 years old; 646.109 Plts/L) and the father's mother (deceased). The JAK2V617F mutation was not detected in these patients; all other blood tests and bone marrow examination were normal. The two patients presented no evidence of MPN, no splenomegaly, no hepatomegaly, no cause of secondary thrombocytosis and no history of thrombosis. The daughter has two pregnancies without complication. A germline T1588C (W515R) mutation in exon 10 of MPL was found in all tested tissues; no other mutation was found in the JAK2, MPL and THPO genes. Affymetrix 6.0 SNP array analysis revealed no genomic abnormality except for a chromosome Y deletion for the father. Transient transfection in Ba/F3-EpoR and K562 cell lines showed that R515 was activating compared to wild type Mpl, as assessed by the constitutive and Tpo-induced tyrosine phosphorylation of Stat5, Stat3 and Akt. However, R515 was less activating than the K/L/A515 mutants.

Conclusion:

We report the first germline MPLW515R mutation, found associated with isolated hereditary thrombocytosis. W515R increases Mpl activity but to a lesser extent than the somatic MPLW515K/L/A mutations observed in the context of MPNs. Thus the presence of a clearly activating JAK2 or MPL mutation is not sufficient for MPN development. Further investigation of this family might reveal why MPLW515R does not lead to MPN.

Disclosures:

No relevant conflicts of interest to declare.
The Metabolic Signature of CLL: Enhanced Glucose Metabolism in A Subset of High-Risk CLL Patients
Authors: Liu Tingyu , Weinberg J. Brice , Davis Evan D. , Volkheimer Alicia D. , Rathmell Jeffrey
Published: October 2019

Abstract 1785

Background:

Chronic lymphocytic leukemia (CLL), the most common hematopoietic malignancies in adults, remains incurable, and new approaches to treatment are needed. Previous research has defined certain molecular characteristics of CLL (e.g., cytogenetic abnormalities, dysregulated signaling pathways, and alterations of apoptotic proteins) that improve our understanding of this disease and provide prognostic information. In addition to these molecular signatures, it is now clear from multiple settings that changes in cellular metabolism can impact the survival and promote the growth and proliferation of malignant cells. Unlike many cancers, however, CLL cells can have a very low rate of proliferation. Because targeting cancer metabolism may attack a fundamental feature of cancer biology, we sought to characterize the metabolic program of CLL in hopes of establishing new prognostic markers and identifying novel treatment targets. We evaluated the glycolytic and lipid metabolism profiles of B-CLL cells from low- and high-risk CLL patients.

Methods:

We collected blood from CLL patients seen at the Duke Center for CLL and enrolled in IRB-approved protocols at the Duke University and Durham VA Medical Centers. CLL lymphocytes were isolated using negative selection yielding greater than 95% purity of CLL lymphocytes. Standard prognostic testing was done as we have described before. Glut1 expression was detected by immunoblot. Glycolytic rate and lipid oxidation rate was measured by using 3H-labled glucose or palmitic acid respectively as described in the literature. Cell viability was measured by propidium iodide exclusion assay.

Results:

CLL cells exhibited higher expression of the major glucose transporter Glut1 compared to naïve B cells purified from healthy donors (n=20 for CLL and n=3 for normal). CLL cells from a subset of high-risk patients (unmutated IGVH; total n=6) had higher glycolytic rate compared to those from low-risk (mutated IGVH; total n=11) patients. In contrast, no significant change in lipid oxidation was observed between high-risk (IGVH unmutated; n=2) to low-risk (IGVH mutated; n=4) patients. CLL cells were also sensitive to glycolysis inhibitor 2-deoxyglucose (2-DOG), with 2-DOG (5uM) mediating cytotoxicity in vitro.

Conclusion:

We establish for the first time that purified, malignant CLL cells overexpress the glucose transporter Glut1, and that the glycolysis inhibitor 2-DOG is cytotoxic for CLL cells in vitro. In addition, our results demonstrate that cells from high-risk CLL patients have enhanced glucose metabolism relative to leukemia cells from low-risk patients. Therefore, glucose metabolism in these malignant B cells may represent a novel prognostic marker and a novel therapeutic target in this incurable disease.

Disclosures:

No relevant conflicts of interest to declare.
Heparanase Expression Correlates with Bone Destruction in the Patients with Multiple Myeloma
Authors: Wei Shi , Innis-Shelton Racquel , Nan Li , Ruan Jian , Sollie Rebecca S , Reddy Vishnu V B , Yang Yang
Published: October 2019

Abstract 1813

Multiple myeloma is an incurable malignancy, and excessive bone destruction is a major cause of morbidity in myeloma patients. However, the biologic mechanisms involved in the pathogenesis of myeloma-induced bone disease are poorly understood. Heparanase, an enzyme that cleaves the heparan sulfate chains of proteoglycans, is upregulated in a variety of human tumors, including myeloma. In the present study, bone marrow biopsies from 40 myeloma patients were stained with antibodies raised against heparanase, RANKL (an osteoclastogenic cytokine), OPG (a decoy receptor for RANKL), TRAP (a marker of osteoclastogenesis) and osteocalcin (a marker of osteoblastogenesis). The radiologic studies for bone lesions of these patients were also recorded. We analyzed the correlations between heparanase expression in bone marrow myeloma cells with (1) the numbers of TRAP positive osteoclasts, (2) RANKL and OPG expression in myeloma cells and osteoblastic cells, (3) the numbers of osteocalcin positive osteoblasts in bone marrow, and (4) the presence/absence of lytic bone lesions. We found a positive correlation between heparanase expression and RANKL expression as well as the numbers of TRAP positive osteoclasts in myeloma and bone marrow cells, but no correlation was found between the expressions of heparanase and OPG in bone marrow cells (myeloma cells do not express OPG). In contrast, heparanase expression was negatively correlated with the numbers of osteocalcin positive osteoblasts. Taken together, these data suggest that heparanase expression by myeloma cells promotes osteoclastogenesis and at same time inhibits osteoblastogenesis. Clinical data show that 92% of patients with high level of heparanase had one or more lytic bone lesions, while only 63% of patients with median∼ low levels of heparanase had bone lesions (p<0.0001). In summary, enhanced heparanase expression in myeloma cells promotes bone resorption and inhibits bone formation; these events contribute to the uncontrolled bone destruction that is characteristic of myeloma. These data provide novel insight into the mechanisms driving myeloma bone disease and suggest that heparanase inhibitors are valid therapeutic targets for the treatment of multiple myeloma.

Disclosures:

No relevant conflicts of interest to declare.
The RNA Editase ADAR1 Promotes Malignant Progenitor Reprogramming in Chronic Myeloid Leukemia.
Authors: Jiang Qingfei , Crews-Robertson Leslie , Barrett Christian L , Chun Hyn-June , Court-Recart Angela C. , Goff Daniel J , Minden Mark D. , Sadarangani Anil , Rusert Jessica , Dao Kim-Hien , Morris Sheldon R , Goldstein Lawrence S. B. , Marra Marco A. , Frazer Kelly A , Jamieson Catriona
Published: October 2019

Abstract 2772

While advanced malignancies in Chronic Myeoloid Leukemia (CML) are diverse in phenotype, they often exhibit stem cell properties including enhanced survival, quiescence and self-renewal potential. The molecular etiology of human progenitor reprogramming into self-renewing leukemia stem cells (LSC) has remained elusive. While DNA sequencing has uncovered spliceosome gene mutations that promote alternative splicing and portend leukemic transformation, isoform diversity may also be generated by aberrant RNA editing mediated by adenosine deaminase acting on dsRNA (ADAR) family, which have been shown to promote an embryonic transcriptional program and regulate fetal and adult hematopoietic stem cell (HSC) self-renewal as well as stem cell responses to inflammation. In this study, whole transcriptome sequencing of normal, chronic phase (CP) and functionally validated blast crisis (BC) chronic myeloid leukemia (CML) progenitors revealed increased inflammatory pathway gene expression in concert with BCR-ABL amplification, enhanced expression of interferon-responsive ADAR1 and a propensity for increased A-to-I RNA editing during CML progression. Mechanistic studies demonstrated that lentivirally enforced ADAR1 p150 expression induced expression of the myeloid-skewing transcription factor PU.1 and skewed cell fate towards granulocyte-macrophage progenitors - the initiating LSC population in BC CML. Moreover, lentiviral ADAR1 knockdown reduced BC LSC self-renewal capacity in RAG2−/−gc−/− mice. These data shed new light on the role of ADAR1-directed RNA editing in myeloid progenitor reprogramming and self-renewal potential of malignant progenitors that drive disease progression and therapeutic resistance in CML, and provide a compelling rationale for developing ADAR1-based LSC detection and eradication strategies.

Disclosures:

No relevant conflicts of interest to declare.