bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic Factors for Survival in 60 Patients with Classical Hodgkin Lymphoma From Peru
Authors: Beltran Brady E , Alva Jose C , Morales Domingo , Quinones Pilar , Miranda Roberto N. , Castillo Jorge J
Published: October 2019

Abstract 4863

Introduction:

Hodgkin lymphoma (HL) is an uncommon lymphoma with a good prognosis. However, the characteristics of HL in Peruvian patients have not been previously reported. HL is thought to be related to chronic Epstein Barr virus (EBV) infection, and new biological markers such as EBV, CD68 and FOXP3 expression could be of potential biological interest in this entity. Our study aims to identify prognostic factors for survival in Peruvian patients with classical HL.

Methods:

This study was approved by the IRB at our institution. Between January 2001 and December 2009, patients diagnosed with classical HL were selected for this study. HIV-negative patients were excluded. Clinical data were reviewed retrospectively and patient's biopsies were analyzed for the immunohistochemical expression of CD20, PAX5, CD30, CD15, FOXP3 and CD68. Samples were also analyzed for the presence of EBV-encoded RNA (EBER) using an in situ hybridization (ISH) technique. Clinicopathological characteristics will be presented using descriptive statistics. Overall survival (OS) estimates were calculated using the Kaplan-Meier method and compared using the log-rank test. For the multivariate survival analysis, Cox proportional-hazard regression test was used. P-values <0.05 were considered statistically significant.

RESULTS:

A total of 60 patients with a pathological diagnosis of classical HL were included in our study. The median age was 55 years (range: 18–98 years) with a 7:3 male predominance. Advanced stages (stage 3 and 4) were seen in 52%, ECOG performance status >1 in 15% and B symptoms in 67%. White blood cell (WBC) counts >15,000 cells/mm3 was seen in 7%, lymphocyte count <600 cells/mm3 in 12%, albumin levels <4 g/dl in 70% and hemoglobin <10.5 g/dl in 40% of the patients. FOXP3 expression >25% was seen in 45%, CD68 >25% in 48% and EBER 2–3+ in 53%. ABVD was the preferred therapy, accounting for 90% of the cases and a complete response to ABVD was obtained in 78%. With a median follow-up 34 months, the median overall survival (OS) has not been reached and the 5-yr OS was 80%. In the univariate analysis, ECOG >1 (p=0.0001), elevated LDH levels (p=0.04) and lymphocyte count <600 cells/mm3 (p=0.05) were adverse prognostic factors. Age, sex, B symptoms, WBC, albumin and hemoglobin levels were not associated with survival. In the multivariate analysis, only ECOG >1 (p=0.01) was an independent adverse prognostic factor in Peruvian patients with classical HL.

CONCLUSION:

ECOG is a best prognostic factor in Peruvian patients with a diagnosis of classical Hodgkin lymphoma. CD68 expression, EBV status and FOXP3 expression by the malignant cells were not prognostic factors for OS in our cohort of patients.

Disclosures:

Castillo: GlaxoSmithKline: Research Funding; Millennium Pharmaceuticals: Research Funding.
Cytogenetic Analysis of 1863 Chineses Patients with Chronic Myeloid Leukemia
Authors: Mu Qitian , Ma Qiuling , Wang Yungui , Tong Xiangmin , Chen Zhimei , Lou Jiyu , Cheng Yizhi , Lu Ying , Ni Wangmao , Chen Feifei , Jin Jie
Published: October 2019

Abstract 4888

Background:

Cytogenetic analyses of chronic myeloid leukemia (CML) have been performed previously in a large number of reports, but systematical research based on large sample sizes is seldom available. In order to further elucidate the cytogenetic nature of CML, we analyzed retrospectively the cytogenetic profiles of 1863 Ph/BCR-ABL-positive CML patients from a research center in China.

Results:

Of 1266 newly diagnosed CML patients, the median age was 41 years, which is younger than the median age of diagnosis in western populations. The incidence of additional chromosome abnormalities(ACAs) was 3.1% in newly-diagnosed chronic phase(CP), 9.1% in CP after therapy, 35.4% in accelerated phase(AP) and 52.9% in blast phase(BP), reflecting cytogenetic evolution with CML progression. 5.3% patients harbored a variant Ph translocation. A higher prevalence of ACAs was observed in variant Ph translocations than in classical t(9;22) in the disease progression, especially in BP(88.2% vs. 50%, p=0.002). Moreover, a hyperdiploid karyotype and trisomy 8 were closely correlated with myeloid blast crisis(BC) while a hypodiploid karyotype and monosomy 7 were associated with lymphoid-BC. Among subsets of myeloid-BC, compared with myeloid-BC with granulocytic differentiation or monocytic differentiation, myeloid-BC with minimal differentiation had higher ACAs rate (80% vs.46.8%, p=0.009 and 80% vs. 42.9%, p=0.006).

Conclusion:

CML tends to afflict younger population in China. In the disease progression, the incident of ACAs was higher in variant Ph translocations than in classical t(9;22). Among subsets of myeloid-BC, myeloid with minimal differentiation had distinct cytogenetic features.

Disclosures:

No relevant conflicts of interest to declare.
Biomarker for Benzopyrene Exposure Is Mitochondrial Mass and Mitochondrial DNA Copy in Blood Cells and Hematopoietic Tissues
Authors: Shin Myung-Geun , Kim Hye-Ran , Choi Hyun-Jung , Kim Hwan-Young , Han Dong-Kyun , Baek Hee-Jo , Moon Jai Dong , Kook Hoon , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook
Published: October 2019

Abstract 4889

Benzopyrenes are well known pollutants and carcinogens. They can intercalate into DNA and interfere transcriptions, resulting in causing various human diseases. However, biomarkers of benzopyrene toxicity have not been comprehensively studied in blood and leukemia cells. The current study was investigated to discover biomarkers for benzopyrene exposure in blood cells and leukemia cell lines. Peripheral blood, peripheral blood hematopoietic stem cell and leukemia cells (THP-1, K562, Molt-4 and HL-60) were cultured in RPMI 1640 media with adding 0, 50, 100 and 200μM of benzopyrene. Viability and apoptosis were assessed by tryptophan blue dye exclusion test and flowcytometry using annexin V. Hydrogen peroxide was measured using enzyme immunoassay. Mitochondrial mass, membrane potential and mitochondrial DNA (mtDNA) copy number were measured using MitoTracker Green, Red probes and real time PCR, respectively. The number of cell remained constant for three weeks culture. Viability of four cell lines disclosed significant decrease after two weeks of benzopyrene treatment. Apoptosis was increased in time- and dose-dependent manner after two weeks of benzopyrene treatment. Mitochondrial contents and membrane potentials were dramatically increased in three-week culture at dose dependent manner. Hydrogen peroxide level was significantly elevated after two weeks treatment of benzopyrene compared to non-benzopyrene treatment group. The number of mtDNA copy increased gradually after exposure to benzopyrene. These results indicated that increased mitochondrial mass and mtDNA copy number were biomarkers for direct exposure of benzopyrene in blood cells and hematopoietic tissues.

Disclosures:

No relevant conflicts of interest to declare.
CCL19 Induced Responses Are Differentially Regulated by Atypical Chemokine Receptor CRAM and Its Classical Chemokine Receptor CCR7 in B-CLL Cells
Authors: Burger Nathalie , Haerzschel Andrea , Leick Marion , Hartmann Tanja Nicole , Catusse Julie , Burger Meike
Published: October 2019

Abstract 4896

Introduction:

Chemokines are known to play an important role in the migration and survival of B-CLL cells. The non-signalling chemokine receptors, including DARC, D6 and CCX-CKR, have recently been shown to be involved in chemokine clearance and activity regulation. The human chemokine receptor CRAM is the most recently identified member of this atypical group. CRAM is expressed on B cells in a maturation-stage dependent manner, and to variable degrees on B-CLL cells. We have recently shown that it competitively binds CCL19 and that this binding is not followed by classical chemokine responses. CCL19 and its signalling receptor CCR7 are centrally involved in B cell localisation and maturation within the secondary lymphoid tissues. CCR7 is also highly expressed on B cells from CLL patients and mediates migration towards its ligands CCL19 and CCL21 which have been shown to be present at higher concentrations in serum of patients with lymphadenopathia compared to patients without. In this study we investigate the influence of CRAM on the CCL19 dependent responses of B-CLL cells and potential correlations to clinical data with a specific focus on lymphadenopathia.

Results:

We demonstrate that B cells from patients with B-CLL present high, but variable degrees of CCR7 and CRAM expression. Patients with compared to patients without lymphadenopathia show a higher CRAM expression level whereas the CCR7 expression is not significantly different. In single samples showing extremly high CRAM expression the migration towards CCL19 is reduced compared to patients with lower CRAM expression. These observations confirm results in the B-CLL cell line MEC-1 showing increased migration toward CCL19 when CRAM expression is reduced using CRAM-siRNA. On the other hand, CRAM seems to be a chemokine presenter as we can show that it does not degrade its chemokine ligand but presents it on the surface of polarised cell layers. Thus, we assume that CRAM plays a role for cell migration, possibly transmigration and cell localisation within lymph nodes of B-CLL cells.

Conclusions:

We show that CRAM can act as an integrator of different recruitment and activation factors. It is associated to CCR7 driven recruitment of B cells by regulating CCL19 availability. Expression of CRAM differs in B cell malignancies for which CCL19 and CCL21 have already been shown to be implicated in lymphadenopathia. We therefore suggest that CRAM is an additional player in the localisation and differentiation/maturation processes of malignant B cells of B-CLL patients.

Disclosures:

No relevant conflicts of interest to declare.
Erythrokinetics in Cooley’s Anemia
Authors: STURGEON PHILLIP , FINCH CLEMENT A.
Published: October 2019

Blood production and destructions have been measured in four patients with Cooley’s anemia. Methods employed included the determination of erythroid/ myeloid ratio of the marrow, reticulocyte count, plasma iron turnover and red cell utilization, Cr51 survival and fecal urobilinogen. Rates of production obtained by these measurements have been compared to normal.

Patients with Cooley’s anemia have been shown to have an increased turnover of hemoglobin constituents comparable to the maximal response seen in other hemolytic anemias. There is, howvever, a marked decrease in maximal delivery of erythrocytes to the peripheral blood amounting to about 50 per cent in the mildly anemic patients and 85 per cent in severely anemic patients. The rate of destruction of circulating erythrocytes was similar in the three patients studied. The severity of anemia was therefore largely related to the production defect.

It was concluded that the defect in Cooley’s anemia is not in total hemoglobin synthesis, but in the fabrications of circulating erythrocytes, which in turn have the associated manifestations of hypochromia, increased percentage of fetal hemoglobin and shortened survival time.
Mixed Agglutination of Leukocytes and Erythrocytes in Relation to Studies of Leukocyte Antigens
Authors: BAKEMEIER RICHARD F. , SWISHER SCOTT N.
Published: October 2019

Serologic experiments are described in which was studied the agglutination of leukocytes from donors representing various ABO, Rh, and Lewis erythrocyte groups an certain canine blood groups, in the presence of corresponding antisera.

Appropriate mixing of antisera with leukocytes and erythrocytes from donors of different groups was seen to produce clumping of leukocytes which did not conform to the reactions of the erythrocytes from the leukocyte donor. When viewed under phase microscopy, certain of these leukocyte clumps, which appeared homogeneous with ordinary light microscopy were found to be clumps of leukocytes mixed with ghosts of erythrocytes reacting with the antibody present.

Factors contributing to this apparently immunologically non-specific clumping were the presence of complement-fixing, potentially hemolytic antibody and thermolabile components of serum. A possible relationship with erythrophagocytosis is suggested.

These observations indicate that certain results of this and other "leukoagglutination" technics, which have been interpreted as demonstrating the presence of A and B antigens on human leukocytes, deserve re-evaluation and emphasize the importance of developing methods of preparing homogeneous leukocyte suspensions.
The Postnatal Synthesis of Fetal Hemoglobin. I. Some Studies in Newborn Goats Using Radioactive Fe59 and Glycine-2-C14
Authors: VISSER H. K. A. , HUISMAN T. H. J. , WOLDRING M. G.
Published: October 2019

Postnatal synthesis of fetal and adult hemoglobin was studied with radioactive iron (Fe59) and glycine-2-C14 in newborn goats.

At birth about 60 per cent of Hb-F is present in newborn goats; after 60 days only about 5 per cent is found. Two goats were treated with "exchange transfusions" with blood of the mother. Using an alkali denaturation method the fetal hemoglobin was not demonstrable during the first few days after this treatment, but 20 days later about 20 per cent was present in circulating blood.

Maxima of radioiron-specific activity in adult hemoglobin occurred in untreated newborn goats at the 12th day, of C14 at the sixth day; in "exchange goats" for radioiron at the 8th day. The fetal hemoglobin maxima of specific activity occurred in the untreated goats resp. after 32 and 24 days, in "exchange goats" after 8 days.

The reappearance of Hb-F in "exchange goats" and the specific activity of Hb-F are direct indications for a synthesis of this component after birth in this species.
Physicochemical Properties of Circulating Red Blood Cells of Lethally X-irradiated Mice Treated with Rat Bone Marrow
Authors: MAKINODAN TAKASHI , ANDERSON NORMAN G.
Published: October 2019

1. Two months after injection of rat bone marrow into lethally X-irradiated mice (950 r-RBM mice), 100% of the circulating RBC were serologically of the rat type, indicating that the surface molecular configuration of RBC from these experimental mice are of the rat type.

2. The hemoglobin was found to be also very much like the rat type in its ease in crystallization, its alkali denaturation property, its electrophoretic property, and its tendency to form a paracrystalline state at low temperature.

3. These cells possessed dual osmotic properties; the relative hemoglobin concentration released when cells were lysed in water was more comparable to the rat type, but its temperature dependency was more comparable to the mouse type.
Desoxyribonucleic Acid Synthesis by Bone Marrow Cells in Vitro
Authors: THOMAS E. DONNALL , LOCHTE HARRY L.
Published: October 2019

The incorporation of C14-formate into thymine is described as a method for measuring desoxyribonucleic acid synthesis by bone marrow cells in vitro. The method is satisfactorily reproducible and permits measurement of DNA synthesis over short periods of time with small amounts of bone marrow.

Normal serum was found to be essential for rapid rates of DNA synthesis and could not be replaced by supplemented media. The rate of DNA synthesis was almost constant during the first 4-6 hours of incubation, but fell off rapidly thereafter. The ability to promote DNA synthesis was found to differ from one normal serum to another. The optimal pH for DNA synthesis was approximately 7.8.

Because DNA synthesis is vitally concerned with cell duplication, measurement of the rate of its synthesis should be a valuable tool for measuring viability of those marrow cells young enough to proliferate.
The Mode of Action and Nature of a Heat Stable Plasma Erythropoietic Factor
Authors: RAMBACH WALTER A. , ALT HOWARD L. , COOPER JOHN A. D.
Published: October 2019

Boiled filtrates of "anemic" rabbit plasma have been studied by physicochemical means, and the response to their injection into the rat has been followed utilizing tracer and morphologic technics. It has been demonstrated that the erythropoietic activity of these filtrates is associated with an alpha-2 globulin, most probably a mucoprotein; and that this material produces, in the rat, an increase in the rate of red cell division and maturation, and an acceleration of release of these cells from the bone marrow and accessory organs of hemopoiesis.
Preparing Standard Solutions of Cyanmethemoglobin
Authors: CROSBY WILLIAM H. , HOUCHIN DONALD N.
Published: October 2019

The method is described whereby solutions of cyanmethemoglobin were prepared for distribution as a National Standard for the calibration of methods for clinical hemoglobinometry. The characteristics of the solutions are given.
The Pseudo-L.E. Cell Phenomenon, with Report of a Case
Authors: KURNICK N. B.
Published: October 2019

An example of leuko-phagocytosis secondary to metastic bronchogenic carcinoma is presented. The transitory development of leukoagglutination and leukophagocytosis (pseudo-L.E. cell phenomenon) led to the erroneous diagnosis of systemic lupus erythematosus. The differentiation between the L.E. cell phenomenon and the pseudo-phenomenon is discussed.
Electron Microscopy of Formed Elements of Normal Human Blood
Authors: GOODMAN JOSEPH R. , REILLY EMMETT B. , MOORE ROGER E.
Published: October 2019

The history of the development of practicable electron microscope technics for examination of the formed elements of the blood was reviewed. A method found to yield good reproductions of ultrathin sections of blood cells was presented in considerable detail. The various phases of the technic were critically evaluated.

The morphologic characteristics of the various formed elements of normal peripheral blood were defined in association with sample electron micrographs. Ultramicroscopic detail of neutrophils, eosinophils, basophils, lymphocytes, monocytes, platelets and erythrocytes was discussed.
The Separate Existence of the Pituitary Erythropoietic Hormone
Authors: VAN DYKE DONALD C. , SIMPSON M. E. , CONTOPOULOS A. N. , EVANS H. M.
Published: October 2019

Although it is recognized that a number of factors are involved in the mechanism controlling red blood cell production, the anterior pituitary furnishes one factor of importance. This pituitary factor is apparently a hormone distinct from other trophic hormones. It is allied chemically only to ACTH and yet both biological and chemical evidence indicates that the pituitary erythropoietic hormone and ACTH are not the same. The major problem in further elucidating the role of the pituitary in the control of erythropoiesis resolves itself into developing a satisfactory method of preparation of erythropoietic hormone which will separate it from ACTH. It is true that no ACTH preparation has been made which does not possess some erythropoietic activity, and no pituitary erythropoietic hormone has been prepared which is entirely devoid of ACTH. Nevertheless, the proportion of these two activities in different preparations has varied so greatly as to assure their separate existence. Autolytic digestion consistently reduces the ACTH activity, while erythropoietic activity is retained. It has been found that oxycellulose is a better adsorbent of ACTH than of erythropoietic hormone. These observations add chemical evidence to the accumulated biological evidence establishing the separate existence of these two pituitary principles.
TYK2-STAT1 Pathway Positively Regulates BCL2 Gene Expression in T-Cell Acute Lymphoblastic Leukemia
Authors: Sanda Takaomi , Tyner Jeffrey W , Gutierrez Alejandro , Ngo Vu N , Glover Jason M , Chang Bill H , Yost Arla J , Weng Andrew P , Ma Wenxue , Tatarek Jessica , Fleischman Angela G. , Ahn Yebin , Yang Yandan , Zhou Wenjun , Neuberg Donna S. , Moriggl Richard , Müller Mathias , Kelliher Michelle , Jamieson Catriona , Gray Nathanael S , Staudt Louis M. , Druker Brian J. , Look Thomas
Published: October 2019

Abstract 1470

To discover oncogenic pathways that are characteristically deregulated in T-cell acute lymphoblastic leukemia (T-ALL), we performed RNA interference screens both in T-ALL cell lines and primary specimens. We found that the JAK tyrosine kinase family member, TYK2, and its downstream effector, STAT1, are each required for the survival of T-ALL cells. To identify the effector molecules downstream of the TYK2-STAT1 pathway in T-ALL, we analyzed global gene expression profiles in TYK2-dependent T-ALL cell lines after silencing of TYK2 or STAT1. As expected, gene set enrichment analysis revealed that genes downregulated by TYK2 knockdown were generally also downregulated by knockdown of STAT1. Importantly, we found that expression of the anti-apoptotic gene BCL2 was significantly downregulated after silencing of both TYK2 and STAT1. Analysis by quantitative PCR of additional T-ALL cell lines revealed that silencing of TYK2 resulted in significant reductions of BCL2 mRNA expression in multiple TYK2-dependent cell lines. Expression of the wild-type but not the kinase-dead TYK2 protein was sufficient to rescue BCL2 protein expression and to prevent apoptosis after knockdown of endogenous TYK2, indicating that the tyrosine kinase activity of TYK2 is required for BCL2 upregulation. Similarly, expression of the shRNA-resistant wild-type STAT1A protein partially rescued BCL2 protein expression and prevented apoptosis, while a variant of STAT1A (Y701F) that is incapable of becoming phosphorylated on a requisite tyrosine residue did not rescue BCL2 levels. Taken together, our findings indicate that aberrant activation of a TYK2-STAT1 pathway upregulates BCL2 expression in T-ALL cells, and that the T-ALL cells develop pathway dependence, in that they require these sustained high levels BCL2 expression for survival.

Disclosures:

No relevant conflicts of interest to declare.
NCI Common Toxicity Criteria and Mortality After Chemotherapy for Acute Myeloid Leukemia (AML)
Authors: Veatch Joshua R. , Sandhu Ravinder K , Shannon-Dorcy Kathleen , Pagel John M. , Becker Pamela S. , Petersdorf Stephen , Appelbaum Frederick R. , Estey Elihu H.
Published: October 2019

Abstract 1479

Abnormal values for creatinine, bilirubin, AST, and ALT following chemotherapy, in particular in phase 1 trials, are interpreted using NCI common toxicity criteria. By convention, grade 3 or 4 toxicity leads to dose reduction while grade 1 or 2 toxicity does not. However, grade 3 – 4 toxicity is typically asymptomatic, calling into question the relevance of these criteria. In an attempt to provide a more empirical basis for decisions we analyzed the relation between (a) grades of NCI toxicity for bilirubin, creatinine, ALT, and AST following chemotherapy for AML and (b) death. Specifically we examined 752 courses of chemotherapy in 533 patients delivered at the University of Washington or Fred Hutchinson Cancer Research Center for newly- diagnosed or relapsed/refractory AML between 2002 and 2012. Patients had at least 2 sets of renal and liver function test values but were excluded if either pretreatment bilirubin or creatinine was greater than 2.0 to better approximate a population in an early phase trial. We compared initial and maximal lab values in the first 28 days in the 40 courses that resulted in death before 28 days, and in the 712 courses that did not (6% death rate; the death rate per patient was 7%). Maximum lab values for creatinine were strongly related to mortality, with grade O having a 1.0% mortality rate and with each increasing grade having a statistically significant increase in mortality over the immediately preceding level: grade I: 5%, grade II: 22%, grade III or IV: 50%. Bilirubin had a significant increase in mortality at grade II (10.0%, p=.009 relative to grade O) and grade III-IV (15.4%, p=.0007 relative to grade O). AST was also associated with increased mortality at grade II (9.5%, p=.04 relative to grade O) and grade III-IV (15.6%, p =.0006 relative to grade O). ALT was not correlated to mortality, even at grade III-IV (9.5% versus 5.3% at grade 0, p=0.23). Although these correlations obviously do not imply causation, it might be argued that grade II toxicity for creatinine, associated with a 4-fold increase in mortality, should result in dose reduction. In contrast, it might be argued that dose reduction should not occur for grade III/IV ALT toxicity, which correlated poorly with mortality. In any event our data suggest that if the principal purpose of toxicity grading is to guide subsequent dose reduction to prevent death, the NCI toxicity criteria should be revisited.

Disclosures:

No relevant conflicts of interest to declare.
CNS Relapse in Acute Promyelocytic Leukemia: Incidence, Management and Outcome
Authors: Kohrt Holbrook E , Coutre Steven
Published: October 2019

Abstract 1489

Background:

A standardized approach to the diagnosis, prognosis and therapy of patients with APL and CNS involvement is limited by the rare nature of the complication. We report the incidence, risk factors, management and outcome of APL with CNS involvement at Stanford University and review the published cases in the literature during the prior three decades.

Methods:

Between 1983 and Jan 2012, 7 cases of APL with CNS involvement were identified at Stanford University and 65 in the literature during the same interval. All pts in the Stanford series received ATRA and anthracycline containing induction regimens followed by consolidation with anthracycline and cytarabine (7 pts), plus ATRA (5 pts), ATO (1 pt), or IT prophylaxis (1 pt). Five pts received maintenance with ATRA containing regimens in all cases (2 pts relapsed prior to starting maintenance). Median follow-up was 6 years.

Results:

Seventy-two relapses involving the CNS were seen after 1 to 50 months (median 8 months) and 61 (84.7%) occurred in CR1 with 8 (11.1%) in CR2, 2 (2.8%) in CR3 and 1 (1.4%) in CR4. Forty-seven (66.2%) were accompanied by bone marrow involvement, 14 (29.8%) molecular and 33 (70.2%) hematological. Median age was 35y with 9 pediatric pts (range 1.3 to 20y) and 63 adult pts (range 22 to 69y). Median WBC count was 28,100 cells/mm3 (range 1,100 to 162,000 cells/mm3) with 61.4% >10 and 6.8% >100 cells/mm3. PML/RARα isoform was reported in 29 pts with 37.9% bcr1, 3.4% bcr2, and 58.6% bcr3. Five pts presented with concurrent CNS-localized hemorrhage. Therapy included methotrexate (88.9%), cytarabine (80.6%), arsenic trioxide (ATO, 47.2%), and anthracycline (11.1%) containing regimens with 88.9% receiving intrathecal therapy. Eighty-six percent received cranial radiation with dose ranging 18 to 30 Gy. Autologous or allogeneic transplantation followed salvage therapy in 30.8% and 5.6% of pts, respectively. Fifty-four (75.0%) pts were salvaged successfully, including 88.2% of pts receiving ATO, however 18 (33.3%) pts later relapsed, 4 (22.2%) of which were salvaged with additional treatment.

Conclusion:

CNS relapse is a rare complications of APL with the majority of cases occurring in pts with WBC >10 cells/mm3, and in first CR. CNS relapse was frequently associated with concurrent bone marrow relapse and approximately one-third occurred with only molecular involvement. A preventative role of IT prophylaxis could not be determined due to only 1 pt receiving prophylaxis. A high percentage of pts salvaged with ATO achieved CR, supporting its therapeutic efficacy and ability to cross the blood brain barrier. Incorporation of ATO in first line therapy for APL may reduce the risk of CNS relapse.

Disclosures:

No relevant conflicts of interest to declare.
Reactivating PP2A by FTY720-Loaded Lipid Nanoparticles, a Novel Therapeutic Strategy in Acute Myeloid Leukemia
Authors: de Mendoza Ander Estella-Hermoso , Castello-Cros Remedios , Cirauqui Cristina , Pippa Raffaella , Moreno Isabel , Blanco-Prieto Maria J , Odero Maria D
Published: October 2019

Abstract 1511

Protein phosphatase 2A (PP2A) is a human tumor suppressor that inhibits cellular transformation by regulating the activity of several signaling proteins critical for malignant cell behavior. Our group has shown that PP2A activity is reduced in acute myeloid leukemia (AML), and that restoration of PP2A activity in AML cells blocks proliferation, induces caspase-dependent apoptosis, and affects AKT and ERK1/2 activity. It has been proposed that FTY720 suppress PP2A activity by directly interacting with PP2AA-C dimers, and possibly interfering with the SET-PP2A interplay. Reactivation of PP2A by FTY720 in vivo restricts leukemogenesis in different types of hematological malignancies; however, its antitumor effects in AML remain underexplored. Here, we developed lipid nanoparticles (LN) encapsulating FTY720, and evaluated its pharmacokinetic profile and biodistribution in mice, and its antileukemic effects in AML in vitro and in vivo. FTY720-LN show suitable size (around 210 nm) and encapsulation efficiency (nearly 75%) for the oral and intravenous administration of the compound. Blood concentration of FTY720 decreased rapidly 2 hours after the intravenous administration of FTY720-LN; however, orally administered LN provided both a sustained release of the drug over four days and a high distribution in tissues, significantly increasing the accumulation of FTY720 in spleen, compared to the administration of the same dose of FTY720 solution and intravenous administration of FTY720-LN. In vitro studies revealed that FTY720 and FTY720-LN restore PP2A phosphatase activity in AML cells, leading to cell growth arrest, induction of caspase-dependent apoptosis and decrease in phosphorylation of PP2A targets AKT and ERK. Moreover, in vivo daily intraperitoneal administration of FTY720 in RAG2−/− γc−/− mice bearing an AML xenograft tumor reduced significant tumor growth. Of note, oral administration of FTY720-LN every three days was as efficient in reducing AML xenograft tumor growth as a daily oral administration of FTY720. These results provide the first evidence for the potential use of lipid nanoparticles encapsulating FTY720 as a therapeutic agent in AML.

Disclosures:

No relevant conflicts of interest to declare.
Functional Characterization of HLA SNPs Associated with Risk of Follicular Lymphoma
Authors: Skibola Christine F. , Conde Lucia , Smith Martyn T. , Sille Fenna C.M.
Published: October 2019

Abstract 1551

Background:

Genome-wide association studies (GWAS) suggest an important role of genetic variation in the etiology of follicular lymphoma (FL). Multiple susceptibility alleles in the human leukocyte antigen (HLA) Class I and II regions on chromosome 6p21.33–32 have been identified for FL, including rs10484561 (OR = 1.95, P = 1.12×10−29) and rs2647012 SNP (OR = 0.64, P = 2×10−21) (Conde et al., Nat Gen, 2010; Smedby et al. Plos Gen, 2011). Moreover, high-resolution HLA typing using next-generation sequencing has identified a number of HLA alleles associated with FL including the extended haplotype DRB1*01:01—DQA1*01:01—DQB1*05:01 that is in complete linkage disequilibrium with rs10484561 (D'=1), and DRB1*15-DQA1*01-DQB1*06, which is highly correlated with rs2647012 (D'=1) (Skibola et al., Tissue Antigens, 2012). We hypothesized that these SNPs or protein variants may alter immune responses to specific antigens and influence tumor development through effects on gene expression or protein structure. To gain a further understanding of the mechanisms involved, here we explored the effects of HLA alleles on expression at the transcriptional and protein level.

Methods:

HLA-DRB1, HLA-DQB1 and HLA-DQA1 mRNA expression and protein levels were measured by RT-qPCR and flow cytometry, respectively, in 40 lymphoblastoid cell lines (Coriell) where wild type and variant rs10484561 and rs2647012 genotypes were compared.

Results:

HLA-DQB1 gene expression changes were significantly associated with rs2647012 genotypes, where the presence of the variant (protective) allele correlated with higher HLA-DQB1 transcript levels. Flow cytometry with HLA-specific antibodies and pan-HLA antibodies (against DR/DQ/DP) also confirmed that the variant rs2647012 allele was associated with higher HLA-DQB1 protein levels. No significant changes were observed in HLA-DRB1, HLA-DQB1 or HLA-DQA1 transcript or protein levels when comparing rs10484561 wild type versus variant alleles.

Conclusions:

We did not detect significant differences in transcript or surface protein levels based on rs10484561 genotypes. These findings suggest that the increased risk of FL associated with rs10484561 that is in high LD with the HLA-DRB1*0101—DQA1*0101—DQB1*0501 extended haplotype is not due to transcriptional modulation, but may be due to effects on protein structure. On the other hand, the protective rs2647012 variant was associated with higher HLA-DQB1 mRNA and protein expression, suggesting that rs2647012 or a SNP in LD influences FL risk through effects on gene expression. We are currently testing the influence of additional FL risk alleles in the 6p21.3 region on HLA gene and protein expression, as well as investigating whether FL risk alleles alter HLA expression in other cell types.

Disclosures:

No relevant conflicts of interest to declare.
Prognositc Inpact of Tumor-Infiltrating FOXP3 + Regulatory T Cells in DLBCL Treated with R-CHOP
Authors: Goto Naoe , Tsurumi Hisashi , Shibata Yuhei , Mabuchi Ryoko , Nakamura Nobuhiko , Nakamura Hiroshi , Kitagawa Junichi , Kanemura Nobuhiro , Hara Takeshi , Moriwaki Hisataka
Published: October 2019

Abstract 1555

Background:

Tumor-infiltrating immune cells perform important functions in host immune reaction against tumor cells including diffuse large B cell lymphoma (DLBCL). Recently, variable tumor-infiltrating cells were reported to give a influence for prognosis, for example, regulatory T cell (T reg), cytotoxic T cell, macrophage and mast cell etc. Among these microenvironmental cells, we focused on Treg cell, and we assessed the distribution and prognostic significance of these cells in DLBCL. The forkhead/winged helix transcription factor 3 (FOXP3) is a transcriptional factorshown to be the key control gene in the development and function of Tregs both in mice and humans.

Patients and Methods:

We examined samples from 94 patients (54 men and 40 women; median age, 70 years) at diagnosis who were prospectively enrolled between 2002 and 2008. All patients treated with R-CHOP(rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisolone). The pattern of FOXP3 protein expression was evaluated using immunehistochemistry in paraffin-embeded tissue samples. In addition, these samples were stained with antibodies for CD10, bcl-6 and MUM-1 via tissue microarray to classify into subgroups.

Results:

The median percentage of FOXP3+ cells was 91/mm2 (range 4–2100 /mm2). Patients with poor performance status (PS), and high serum lactate dehydrogenase (LDH) showed lower numbers of FOXP3+ cells. (PS; p= 0.014, LDH; p=0.0048) Patients with high counts of FOXP3+ cells (>90/mm2) have better prognosis than those of low counts (5 year (5-y) overall survival (OS); 72.1%, 49.7% p=0.024, respectively). Although no prognostic difference was observed between GCB type and non-GCB type (5-y OS: GCB 71.2%, non GCB 53.1%, p=0.12), low counts of FOXP3+ cell and non-GCB type patient was poorer prognosis than high counts and non GCB type. (low 5-y OS 31.2%, high 5-y OS 69.8% p=0.02).

Conclusion:

Increased count of FOXP3+ tumor-infiltrating cell might predict better prognosis of DLBCL.

Disclosures:

No relevant conflicts of interest to declare.
Pro-Angiogenic Mir-296 Is Frequently Overexpressed and Is Associated with Advanced Stage Disease in Diffuse Large B-Cell Lymphoma
Authors: Borges Natália Morais , de Conti Marina Lourenço , de Andrade Tathiana Azevedo , Macedo Marina Petaccia , de Souza Begnami Maria Dirlei Ferreira , Alves Antônio Corrêa , Colleoni Gisele Wally Braga
Published: October 2019

Abstract 1570

Introduction:

MicroRNAs (miRNAs) are a class of endogenous short non-coding RNAs that control gene expression by acting on target mRNAs for promoting either their degradation or translational repression. Some of them have involvement in regulating various aspects of angiogenesis, including proliferation, migration and morphogenesis of endothelial cells, which are important in regulating cardiovascular development and cancer. The term “angiomiR” is used to define the miRNAs that control angiogenesis. They are classified into pro-angiomiRs, those that promote angiogenesis, and anti-angiomiRs, those that inhibit angiogenesis. The identification of angiomiRs as the key to regulating angiogenesis has opened new paths in the treatment of vascular and oncology diseases.

Aims:

This study aims to analyze the expression angiomiRs in diffuse large B-cell lymphoma (DLBCL) and to correlate them with clinical and histological features to identify possible biomarkers and prognostic factors.

Patients and Methods:

We studied 93 samples of de novo DLBCL diagnosed between 2000 and 2010. All the cases were HIV-negative. MicroRNAs were obtained from paraffin embedded tumor samples using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE Tissues (Applied Biosystems). Four angiomiRs (miR-378, miR-296, miR-210 and miR-126) were analyzed. RNU44 and U18 were used as endogenous controls for quantitative PCR (TaqMan®Small RNA Assays). We set a threshold of a 1.5-fold difference in angiomiRs expression compared to controls (palatine tonsil).

Results:

miR-378, miR-296, miR-210 and miR-126 overexpression were observed in 43%, 47%, 22% and 5%, respectively. Considering that miR-378 and miR-296 were frequently overexpressed in DLBCL, we further analyzed the following variables: age (<=60 versus >60 years), Ann Arbor Staging System (I-II versus III-IV), International Prognostic Index (0–2 versus 3–5), DLBCL classification (NOS versus subtypes), tumor origin according to Hans (2004) algorithm (GCB versus non-GCB). We observed higher median miR-296 expression in DLBCL classified as stage III-IV (p = 0.0415, Mann-Whitney). For the other variables, were did not find any statistically significant difference between groups.

Conclusions:

miR-296, that directly decreased the levels of hepatocyte-growth factor regulated tyrosine kinase substrate (HGS) and indirectly upregulate VEGFR2 and PDGFRβ, was overexpressed in almost 50% of de novo DLBCL and was associated with advanced stage disease. Our study brings new information about the role of microRNA importance in DLBCL development and can be explored as prognostic and therapeutic target for patients suffering from this prevalent malignancy (Supported by FAPESP 2010/17668-6).

Disclosures:

No relevant conflicts of interest to declare.
Lymphoma Occurring Over the Age of 90: Clinical Presentation and Outcome
Authors: Trebouet Adrien , Marchand Tony , Lemal Richard , Gyan Emmanuel , Broussais Florence , Guillermin Yann , Monjanel Hélène , Salles Gilles , Le Gouill Steven , Boulanger Stephanie , Leguyader Sandra , Collignon Albert , Rossille Delphine , Thieblemont Catherine , Lamy Thierry , Houot Roch
Published: October 2019

Abstract 1582

Background

Lymphoma occurring in very elderly patients (i.e. >90 years of age) is not uncommon and its incidence is expected to increase due to prolongation of life expectancy. Its management raises therapeutic as well as ethical considerations. However, no reports are currently available in this population of patients rendering their management particularly challenging. The purpose of our study is to describe the clinical presentation, management and outcome of patients diagnosed with lymphoma at the age of 90 or over.

Methods

In a retrospective, multicenter study, we analyzed 234 patients with Non-Hodgkin (NHL, n=227) and Hodgkin (HL, n=7) lymphoma diagnosed at the age of 90 or over, between 1990 and 2012.

Results

Histologic incidence of lymphoma subtypes was evaluated using 3 epidemiologic cancer registries, accounting for 128 patients. In this cohort, 55% of the patients presented with aggressive NHL, 42% with indolent NHL and 3% with HL. The most frequent histologies were diffuse large B cell lymphoma (46%), follicular lymphoma (9.5%), lymphoplasmacytic lymphoma (8%) and marginal zone lymphoma (5.5%).

Clinical characteristics, management and outcome were collected for 169 patients among several institutions. At diagnosis, median age was 92 [range 90–100]. Most patients were female (66%) and still living at home (73%). Charlson index was low (i.e. ≤2) in 68% of the patients. Forty-five percent of the patients had a low (i.e. ≤1) performance status (PS).

The majority of the patients (63.5%) received a treatment (Fig. 1).

Median overall survival (OS) for the entire cohort was 6.9 months. OS was significantly shorter in patients with aggressive lymphoma (OS = 5.2 vs 19.2 months, p<0.001, Fig. 2), poor (i.e. ≥2) PS (OS = 4.1 vs 16.6 months, p=0.001), advanced (i.e. III-IV) Ann Arbor stage (OS = 2.3 vs 7.1 months, p=0.04), low (i.e. ≤36g/L) albumin level (OS = 4.6 vs 13.8 months, p=0.002) and in the absence of treatment (OS = 4.5 vs 9 months, p=0.05).

Causes of death were: lymphoma in 58% of cases, anti-lymphoma treatment in 8.5% of cases and other causes in 33.5% of cases.

Conclusions

Lymphoma occurring over the age of 90 is more frequently aggressive than indolent. Most of the patients die of lymphoma rather than from toxicities of the treatment. This suggests that a subset of patients may benefit from adapted therapies in spite of their very advanced age. Management of these patients may be guided by prognostic factors such as histologic subtype, PS, Ann Arbor stage and albumin level.

Disclosures:

No relevant conflicts of interest to declare.
Survival and Clinicopathological Characteristics of EBV-Positive Diffuse Large B-Cell Lymphoma
Authors: Beltran Brady E , Cotacallapa Erick , Castillo Jorge J
Published: October 2019

Abstract 1588

Background:

EBV-positive diffuse large B-cell lymphoma (EBV+ DLBCL) of the elderly is a provisional entity included in the 2008 WHO Classification of Lymphomas. Diagnostic criteria include age >50 years, DLBCL morphology and EBV expression in lymphomatous cells. However, these criteria are evolving as several patients are <50 years and a specific cut-off for the percentage of EBV expression has not been defined. The goal of this retrospective study is to evaluate clinical and pathological characteristics of EBV+ DLBCL from Peruvian patients.

Methods:

Between January 2002 and January 2012, all patients meeting criteria for EBV+ DLBCL were included in the analysis. Patients with evidence of immunosuppression were excluded. All cases re positive for the presence of EBV-encoded RNA (EBER) by in situ hybridization, and CD20 and/or PAX-5 expression by immuno-histochemistry. Clinical data were reviewed retrospectively and patient's biopsies were analyzed for the expression of BCL6, CD10, CD30 and MUM-1/IRF4 using a tissue microarray (TMA) technique. The overall survival (OS) curves were calculated using the Kaplan-Meier method, and compared using the log-rank test.

Results:

A total of 43 EBV+ DLBCL patients are included in this study. The median age was 73 years (range 25–95 years). Four patients (9% ) were <50 years. The male:female ratio was 2.2:1. B symptoms occurred in 59%, ECOG >21 in 60%, advanced stage (III/IV) in 58%, elevated LDH levels in 44%, and lymphocyte count <1000/uL in 35%. The International Prognostic Index (IPI) score was 0–2 in 39% and 3–5 in 61% of the patients. Extranodal disease occurred in 20 patients (46%): stomach (n=3), tonsil (n=3), pleura (n=2), palate (n=2), cecum (n=2), bone marrow (n=2), ileum (n=1), bone (n=1), skin (n=1), lung (n=1), meninges (n=1), breast (n=1) and peritoneum (n=1). Three patients had central nervous system involvement (7%), one at presentation and two at relapse. Based on the Hans classification, 76% had non-germinal center profile. Ki67 expression was >80% in 53% of the patients. Eleven evaluated patients had a c-myc-negative status. Chemotherapy was received in 75% of the cases due to poor performance status. The overall response rate with conventional chemotherapy was 46%, with complete response in 39%, partial response in 7%, and no response in 54%. The median survival was 7.5 months. The Oyama score was: 0 factors (13%), 1 factor (47%), and 2 factors (40%) with median OS of 41, 11 and 1.5 months respectively (p=0.07). A lymphocyte count <1000/uL was a prognostic factor for OS (p=0.001).

Conclusions:

Based on our study, which is the largest cohort in Latin-America, EBV+ DLBCL is an aggressive entity with frequent extranodal disease and poor response to conventional chemotherapy. The overall survival remains poor. Lymphopenia, as defined as lymphocyte count <1000/uL, appears as a prognostic factor for OS.

Disclosures:

No relevant conflicts of interest to declare.
Preferential Acquision of N-Glycosylation Sites in the VDJ Region in Germinal Center B-Cell-Like Difusse Large B-Cell Lymphoma
Authors: Alcoceba Miguel , Sebastián Elena , Balanzategui Ana , Marín Luis , Montes-Moreno Santiago , Flores Teresa , Puig Noemí , Sarasquete M. Eugenia , Chillón M. Carmen , Jiménez Cristina , Corral Rocío , González-Barca Eva , Caballero M. Dolores , Miguel Jesús F. San , García-Sanz Ramón , González Marcos
Published: October 2019

Abstract 1589

Introduction:

Acquired potentially N-glycosylation sites are produced by somatic hypermutation (SHM) in the immunoglobulin (Ig) variable region. This phenomenon is produced in ∼9% of normal B-cells and seems to be related to certain B-cell lymphoproliferative disorders (B-LPDs) such as follicular lymphoma (FL, 79%), endemic Burkitt lymphoma (BL, 82%) and diffuse large B-cell lymphoma (DLBCL, 41%). These data suggest that new potential N-glycosylation sites could be related to germinal center B (GCB)-LPDs. By contrast, in other B-LPDs, such as chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), MALT lymphoma, Waldenström macroglobulinemia (WM) or multiple myeloma (MM), these modifications have not been analyzed in deep.

Aims:

To evaluate the acquisition of potential N-glycosylation sites in B-LPDs, including immunohystochemical DLBCL subtypes (GCB and non-GCB) and specific non-GCB-LPDs, such as hairy cell leukemia (HCL), splenic marginal-zone lymphoma (SMZL), CLL, MCL, ocular extranodal marginal zone lymphoma (OAEMZL), MM and WM.

Patients:

A total of 953 sequences (203 from our group and 750 previously published sequences) of B-LPDs were included. Diagnosis distribution was as follows: DLBCL (n=235), MCL (n=235), CLL (n=166), MM (n=96), OAEMZL (n=82), SMZL (n=68), WM (n=38) and HCL (n=33).

Methods:

Acquired N-glycosylation sites were counted according to the sequence Asn-X-Ser/Thr, where X could be any amino acid except Pro. Natural motifs in germline sequences of IGHV1–08, IGHV4–34 e IGHV-5a were not considered. Fisher test was used to perform comparisons between groups. To distinguish DLBCL biological subtypes (GCB and non-GCB DLBCL), Hans' algorithm was used.

Results:

A total of 83 out of the 235 DLBCL cases acquired at least a new N-glycosylation site, a higher value than in normal B-cells (35% vs. 9%, p<0.0001). Higher incidence of these motifs in the group of GCB as compared to non-GCB DLBCL were observed (52% vs. 20%, p<0.0001). Those cases diagnosed of HCL, CLL, MCL, MM, WM, OAEMZL and SMZL presented a reduced number of new N-glycosylation sites, showing similar values than normal B-cells (range 3–18%, p=ns).

Conclusions:

We described for the first time the pattern of N-glycosylation in HCL, SMZL, OAEMZL and in the immunohystochemical DLBCL subtypes, where the GCB-DLBCL showed a higher number of new N-glycosylation sites with respect to non-GCB DLBCL and other non-GCB-LPDs. The presence of novel N-glycosylation sites in FL, BL and in GCB-DLBCL strongly suggests that these motifs are characteristic of the germinal center B-LPDs.

Disclosures:

No relevant conflicts of interest to declare.