bluuebunny/crossref_metadata_2025 huggingface dataset:
Intravenous Gammaglobulin (IVIg) Therapy Prevents Antibody-Mediated Transfusion Related Acute Lung Injury (TRALI) by Directly Inhibiting Recipient Neutrophil Activation in a Murine Model
Authors: Semple John W. , Kim Michael , Hou Jing , Lee Young Jin , Tabuchi Arata , Kuebler Wolfgang M. , Lazarus Alan H
Published: October 2019

Abstract 42

Transfusion-related acute lung injury (TRALI) is one of the leading causes of transfusion-related fatalities and most TRALI reactions are thought to be caused by donor antibodies. Because intravenous gammaglobulin (IVIg) therapy has been shown to be beneficial in many antibody-mediated disorders, we hypothesized that IVIg would be effective in treating antibody-mediated TRALI. TRALI was induced in CB.17 SCID mice by injection of an anti-murine major histocompatibility complex (MHC) class I antibody (34-1-2s). The mice were treated with doses of IVIg either prophylactically 18 hours before TRALI induction or after the first symptoms of TRALI were observed (within 3 minutes after 34-1-2s injection). Body temperatures, lung edema, mortality and neutrophil activation were assessed. In IVIg non-treated mice, injection of 34-1-2s caused significantly reduced body temperatures indicating systemic shock within 2 minutes post injection. Pulmonary neutrophil accumulation and mortality occurred within 1 hour post 34-1-2s injection and mortality reach 25% by 2 hours post administration. When bone marrow neutrophils were purified after 34-1-2s administration, they produced significantly more reactive oxygen species compared with 34-1-2s non-treated mice. In contrast, if the mice were pre-treated with either 1 or 2 g/kg IVIg 18 hours before 34-1-2s-TRALI induction, they were completely protected from systemic shock, lung edema and mortality. The prophylactic in vivo IVIg treatment also significantly reduced 34-1-2s-induced neutrophil ROS production. IVIg's effect on ROS production appeared to be due to a direct contact mechanism as titrations of IVIg significantly inhibited the ability of neutrophils to produce ROS in vitro. In mice treated with IVIg 2–3 minutes after 34-1-2s-TRALI induction (when symptoms were first observed), there was also significant protection against the lung damage and mortality. In contrast, IVIg treatment was not as effective against acute lung injury induced by acid inhalation. The results suggest that the treatment of antibody-mediated TRALI reactions with IVIg can effectively prevent pulmonary reactions and mortality and it does so by directly interfering with 34-1-2s-induced neutrophil activation.

Disclosures:

No relevant conflicts of interest to declare.
Pseudotumor Cerebri Associated with All-Trans Retinoic Acid Treatment During Induction Therapy for Acute Promyelocytic Leukemia: A Case Report and Literature Review
Authors: Holmes Dylan , Aboulafia David M.
Published: October 2019

Abstract 4264

Pseudotumor cerebri (PC) is a rare disorder characterized by elevated intracranial pressure with normal cerebrospinal fluid composition, normal cranial imaging findings, and symptoms and signs of increased intracranial pressure. A number of medications have been linked to PC including vitamin A and its derivative all-trans-retinoic acid (ATRA). Herein, we describe the case of a 38-year-old woman who was diagnosed with PC during standard induction treatment for acute promyelocytic leukemia (APL). Her neurologic complaints of severe headache and diplopia improved slightly after ATRA was discontinued and she was treated with acetazolamide, but symptoms did not abate until fluconazole was held five days later. Through a literature review, we identified 41 case reports of APL patients with ATRA-associated PC, 23 of which provided details of clinical presentation and follow-up. The median age at diagnosis of PC was 27 for females and 16 for males; the female-to-male ratio was 1.3:1. All 23 patients complained of headache; papilledema was present in all 21 instances (100%) where a retinal exam was described. ATRA was held due to PC in 20 cases (87%), and neurologic symptoms resolved without further intervention in 7 of 20 (35%). The remaining 13 patients were treated with diuretics (11 of 13 cases, 85%), high-volume lumbar punctures (5, 38%), corticosteroids (4, 31%), and/or analgesics (3, 23%). Including our index patient, eight patients were rechallenged with ATRA after neurologic improvement, and 5 (63%) required further interventions to resolve recrudescent symptoms. We identified one other patient whose neurologic complaints persisted despite dose reductions in ATRA. This patient improved only after fluconazole was discontinued. Because both ATRA and fluconazole are metabolized by the cytochrome P-450 system, we suggest that if taken concurrently, they may exacerbate the risk of PC. Physicians should be aware of this interaction between ATRA and fluconazole and consider discontinuing both fluconazole and ATRA in APL patients who develop PC.

Disclosures:

No relevant conflicts of interest to declare.
Anti-Vascular and Anti-Tumor Effects of the Vascular Disrupting Agent ASA404 (DMXAA) in Human Acute Leukemia Xenograft Models
Authors: Schaefer Christopher W , Lal Deepika , Vethanayagam R Robert , Spernyak Joseph A , Seshadri Mukund , Wang Eunice S.
Published: October 2019

Abstract 4293

Acute leukemia growth and expansion within the marrow microenvironment is linked to increased vascularization. Vascular delivery of chemotherapy drugs within the marrow space may also be an important mediator of anti-leukemic activity. We examined the effects of a tumor -vascular disrupting agent (tumor-VDA), ASA404 (DMXAA; Vadimezan), against human acute leukemia alone and in combination with anthracycline therapy. Exposure of human acute leukemia cells (HEL, Raji) to ASA404 (200–1000 mM) in vitro for up to 72 hours resulted in no evidence of direct cytotoxicity. However, contrast-enhanced magnetic resonance imaging (CE-MRI) performed in mice bearing subcutaneous HEL xenografts revealed an early increase in vascular permeability 4 hours after treatment with a single dose of ASA404 (25 mg/kg). This was followed by significant vascular disruption at 24 hours after therapy as determined by tumor immunohistochemistry. In systemic leukemia xenograft models established with luciferase-transfected HEL and Raji cells, continuous ASA404 treatment (20 mg/kg twice weekly) slowed overall systemic leukemia disease progression (as determined by whole animal bioluminescent imaging) in association with marked marrow vessel dilation, loss of vascular patency, and marked erythrocyte extravasation within the marrow microenvironment. These effects were accompanied by 20–30-fold elevations of circulating systemic TNF-alpha levels in ASA404 versus vehicle treated animals. Combining in vivo ASA404 treatment with anti-human VEGF antibody (bevacizumab) therapy further inhibited leukemia growth as compared with vehicle- or single-agent treated controls. However, concomitant administration of ASA404 with doxorubicin chemotherapy did not improve anti-leukemia effects and was in fact less effective than single agent therapy. In summary, our results demonstrate that ASA404 therapy is capable of mediating in vivo anti-tumor, anti-vascular and immunomodulatory effects in human acute leukemia models. Further strategies to modulate the marrow environment by combining a vascular disrupting agent with other biological agents for acute leukemia therapy may be warranted. However, given the time-dependent effects of VDAs such as ASA404 on tumor vascular function, combining VDAs with chemotherapy is likely to require further optimization of sequence and schedule of administration to ensure adequate marrow drug delivery and maximal therapeutic efficacy in hematological malignancy.

Disclosures:

No relevant conflicts of interest to declare.
Detection of CD55- and CD59-Negative Immature Reticulocytes May Improves Sensitivity/Specificity to Identify a Minor Population of PNH-Type Cells in Patients with Myelodysplastic Syndrome/Aplastic Anemia
Authors: Obara Naoshi , Satoh Shoko , Okoshi Yasushi , Chiba Shigeru , Ninomiya Haruhiko
Published: October 2019

Abstract 4373

[Background]

Paroxysmal nocturnal hemogloblinuria (PNH) is a hemolytic disease characterized by complement-sensitive red blood cells (RBC). Sometimes, a tiny amount of PNH-type, i.e., CD55- or CD59-negative, red blood cells as well as neutrophils can be detected in peripheral blood from patients with myelodysplastic syndrome (MDS)/ aplastic anemia (AA). It has been reported that the presence of PNH-type cells is correlated with a good response to immunosuppressive therapy foe these disease. However, the ratio of PNH-type RBCs to normal RBCs is always underestimated because PNH-type red blood cells have a shortened life span in the circulation, and there are some patients who have very low number of circulating granulocytes.

[Metods]

Peripheral blood was obtained from 37 patients with MDS or AA and 30 normal subjects. Mononuclear cells were then stained with antibodies against CD55, CD59, and CD71, and analyzed by a flowcytometer. Subsequently, CD55 and CD59-negative and CD71-positive reticulocytes, representing an immature fraction of reticulocytes, were identified. Sensitivity and specificity were compared for the detection of PNH-type immature reticulocytes with those evaluated by a PNH-type RBC- and granulocyte-detecting method.

[Results and discussion]

All 30 normal subjects were negative when the cut-off value for CD55 and CD59-negative and CD71-positive reticulocytes was set at 0.008%. With this cut-off value, PNH-type immature reticulocytes were detected in 18 out of 37 MSA/AA patients. When the cut-off values for PNH-type RBCs and granulocytes were set at 0.005% and 0.01%, respectively, in accordance with the original report by Sugimori, et al, 16 and 18 cases among 37 MDS/AA patients were positive for PNH-type RBCs and granulocytes, respectively. All the 16 cases positive for PNH-type RBCs were also positive for PNH-type granulocytes and immature reticulocytes, while 2 cases negative for PNH-type RBCs were evaluated to be positive for both PNH-type granulocytes and immature reticulocytes. All the cases positive for granulocytes were also positive for immature reticulocytes, and vice versa. These observations indicate that the method to detect minor population of PNH-type immature reticulocytes is feasible and comparable with that to detect PNH-type granulocytes, and could be superior to the PNH-type RBC-detection method in terms of sensitivity, without reducing specificity. If the evaluation of PNH-type granulocytes is difficult, PNH-type immature reticulocytes may be a good alternative to be evaluated.

Disclosures:

No relevant conflicts of interest to declare.
Efficacy and SAFETY of Dasatinib 50 MG Once Daily DOSE In PATIENTS with CHRONIC PHASE CML WHO Failed IMATINIB
Authors: Yassin Mohamed A. , El-Ayoubi Hanadi R. , Kamzoul Rami T
Published: October 2019

Abstract 4440

Background:

Dasatinib is multi-targeted kinase inhibitor and it is approved therapy for patients with imatinib-resistant or imatinib-intolerant CML or newly diagnosed CML (chronic phase). The recommended dosing regimen is Dasatinib 100 mg once daily (QD), based on results from the CA180-034 dose-optimization study in patients with CML-CP and resistance or intolerance to prior therapy and it is active against the vast majority of BCR/ABL mutations apart from T315I and can overcome emerging resistance.

Aims:

to assess the efficacy and safety of 50 mg once daily dose of Dasatinib in achieving complete hematological remission (CHR),complete cytogenetic remission(CCYR) and major molecular response (MMR) in patients with CML-CP and resistance or intolerance to prior imatinib therapy.

Patients and Methods:

nine adult patients (aged ≥18 years) with prior imatinib failure were enrolled in this study after informed consent, between September and November 2009 and followed for 18 months. Received Dasatinib 50 mg once daily as second line therapy and were evaluated at baseline,3,6, 12, and 18 months as per European leukemia net(ELN) guidelines for response CHR, CCYR, MMR mutation analysis was not done because it is not available in our centre. Patients with baseline pleural effusion or ejection fraction less than 50% were excluded. Chest x-rays were performed at baseline and at 6 months, or more frequently if indicated clinically. Pleural effusions were graded according to CTCAE v3.0 criteria: grade 1, asymptomatic; grade 2, symptomatic, ≤ 2 therapeutic thoracenteses; grade 3, symptomatic requiring supplemental oxygen, > 2 therapeutic thoracenteses; grade 4, life-threatening, hemodynamic instability.

Results:

All patients achieved complete hematological as well as complete cytogenic response within three months and remains so at 6 and 12 months assessment, 18 months assessment for molecular response showed that 8 out of 9 achieved major molecular response and one patient achieved complete molecular response, grade two or more toxicity were not seen in any patient. Anemia and neutropenia Grade ≥ 2 were not observed in any patient, thrombocytopenia grade one was observed in one patient. Non-hematologic adverse drug reactions like fluid retention; including pleural effusion, nausea vomiting, myalgia, muscle inflammation and rash were not observed in any patient.

Conclusion:

In patients with chronic phase chronic myeloid leukemia who failed first generation TKI (Imatinib). Dasatinib 50 mg once daily is safe, effective and capable of achieving complete hematological and cytogenetic remissions well as major molecular response‘ with better toxicity profile keeping in consideration the small number of patient’s further studies are needed to confirm this result.

Disclosures:

No relevant conflicts of interest to declare.
Induction Therapy with Novel Agents Improves Relapse-Free Survival After Autologous Stem-Cell Transplantation in Multiple Myeloma
Authors: Eder Sandra , Lamm Wolfgang , Gruber Michaela , Schauer Beatrice , Ackermann Jutta , Gisslinger Heinz , Drach Johannes
Published: October 2019

Abstract 4494

Introduction:

Multiple myeloma (MM) is still an incurable disease. Initial treatment depends on age, risk stratification and co-morbidities. High-dose therapy followed by autologous stem-cell transplantation (ASCT) is the standard of care in transplant eligible patients.

In recent years, novel agents became a promising role in newly diagnosed, relapsed myeloma as well as in maintenance treatment. Bortezomib, a proteasome inhibitor, and thalidomide or lenalidomide, immunomodulatory agents have been found to be superior to chemotherapy regimens.

Methods:

121 adult patients (50 female, 71 male) with a median age of 53.5 years at diagnosis underwent autologous stem-cell transplantation at the Medical University of Vienna between June 1985 and September 2010. Median age at ASCT was 57.4 years.

46 patients received novel therapeutic agents, such as VDT (bortezomib, dexamethasone, thalidomide), Rd (lenalidomide, low dose dexamethasone) and VD (bortezomib, dexamethasone), whereas 75 patients were treated on the basis of chemotherapy schedules, e.g. VAD-based (vincristine, doxorubicin, dexamethasone).

Results:

Progression-free survival (PFS) after ASCT was 7.9 months (6,4 – 9,3, 95% CI) after treatment with chemotherapy, whereas it was increased to 21,6 months (5 – 38,1, 95% CI) after initial treatment with either bortezomib and/or thalidomide/lenalidomide (p= 0.001).

Post-induction VGPR or better was superior with new agents compared to chemotherapy, 34,1% versus 11,9%, respectively. Post-transplant VGPR or better was as high as 68% after initial therapy with novel agents prior to transplant, whereas VGPR or better was reached in 48.6% in patients receiving chemotherapy.

Conclusion:

Novel agents significantly prolong relapse-free survival in patients eligible for autologous stem-cell transplantation. Previous studies showed improved remission duration with thalidomide or lenalidomide maintenance after ASCT. The data of maintenance therapy after ASCT at our department will be presented at the meeting.

Disclosures:

No relevant conflicts of interest to declare.
Design, Synthesis and Biological Evaluation of A Boron Containing Retinoid As a Novel Therapeutic Agent for Acute Promyelocytic Leukemia
Authors: Schinke Carolina D. , Das Bhaskar C , Goel Swati , Bhagat Tushar , Nischal Sangeeta , Minucci Saverio , Kabalka George , Gallagher Robert E. , Evans Todd , Platanias Leonidas C , Verma Amit
Published: October 2019

Abstract 5008

Cure rates of acute leukemia remain poor emphasizing the need to for novel therapies. All trans retinoic acid (ATRA) is an effective therapeutic agent in a subtype of acute myeloid leukemia and relieves transcriptional repression induced by the PML-RAR oncoprotein by binding to the retinoic acid receptor. Even though ATRA is effective, the treatment course is characterized by a high rate of toxicity and ATRA resistance is seen in some cases of acute promyelocytic leukemia. In an attempt to improve outcomes, we devised a methodology for creation of boronic acid and other newer retinoic acid analogues. Our lead compound, MA-21 was generated by replacing the terminal carboxyl group of ATRA with a boronic ester using Wittig reactions. Computation modeling revealed that MA-21 can fit in the RARa pocket and can form increased covalent bonding with cysteine residues within the receptor. As opposed to other synthetic retinoids, the addition of a boron atom resulted in significantly enhanced cytotoxicity in leukemic cell lines, even those that were resistant to ATRA. MA-21 at 1uM dose led to significant reduction in proliferation of ATRA sensitive NB4 APL cell line (1.8 fold decrease after 96hrs, p= 0.028) as well as in ATRA resistant cell lines NB4.007/6 (3.3 fold decrease after 72hrs and 2.1 decrease after 96hrs, p values of 0.018 and 0.046) and NB4.306 (2.6 fold decrease after 96hrs, p= 0.032). MA-21 was able to induce these effects by inducing significant G2/M cell cycle arrest and not by increased apoptosis or cellular differentiation. Cell cycle was assessed by Flow Cytometry after 96hrs of incubation and showed a significant increase in G2/M percentage in the ATRA sensitive and resistant cell lines compared to DMSO (NB4 cell line- 1.35 fold increase, p= 0.035; NB4.007/6- 1.35 fold increase, p= 0.015 and NB4.306- 2 fold increase, p= 0.023). Thus, we demonstrate novel synthetic methodology to synthesize boron containing novel retinoids and demonstrate the potential of these compounds as therapeutic agents in resistant leukemias.Figure1:Structures of ATRA and MA-21Figure1:. Structures of ATRA and MA-21

Disclosures:

No relevant conflicts of interest to declare.
POTENTIAL ROLE of 8-COLOR FLOW CYTOMETRY for CHARACTERIZATION of MDS with Del(5q) ABNORMALITIES
Authors: Oelschlaegel Uta , Sockel Katja , Mohr Brigitte , Thiede Christian , Ehninger Gerhard , Platzbecker Uwe
Published: October 2019

Abstract 5020

Four-colour based flow cytometry (FCM) of bone marrow (BM) samples has become a valuable tool in the diagnostics of patients (pts) with myelodysplastic syndrome (MDS). It is still unknown, whether aberrations detected by FCM differ between certain cytogenetic subgroups. By using an 8-color (5-tube) FCM diagnostic panel we aimed to investigate the antigen patterns of MDS pts with del(5q) abnormalities and compared these to MDS pts with non-del(5q) MDS.

BM samples of healthy donors (n=19) and 57 MDS pts including 28 pts (15 RCMD; 3 RAEB-1; 8 RAEB-2; 1 RAEB-t; 1 CMMoL-1) with del(5q) MDS (single n=15) and 29 pts (1 RCUD; 2 RA±RS; 17 RCMD; 2 RAEB-2; 7 sAML including 1 RAEB-t) with non-del(5q) MDS were investigated. FCM procedure was performed at a FACS Canto II (200,000 events) and standardized including the analysis of all samples after overnight storage at room temperature and erythrocytes lysis prior to antibody staining. Thresholds were set according to median ± 2SD and/or ½ log differences compared to healthy BM. Aberrations were scored according to the FCSS by Wells et al. 2003 and modified by Van de Loosdrecht et al. 2008.

In del(5q) MDS several phenotypic characteristics differed to non-del(5q) pts. Of note, lineage infidelity (CD2, CD5, CD7, CD19, CD56) was detectable in significantly fewer del(5q) pts compared to non-del(5q) MDS (in granulocytes: 8% vs. 33%, p=0.040 and in monocytes: 24% vs. 56%, p=0.027). Thus, especially aberrant CD56 expression contributed to these results with only small subpopulations being positive in del(5q) vs. often more than 50% of aberrant granulocytes/monocytes in non-del(5q) MDS pts. Additionally, the lymphoid-to-granulocytes/monocytes-ratios were higher in del(5q) (p=0.029 and 0.025) and hypogranularity was more pronounced in those pts (p=0.05). Intensity of expression of CD14 on monocytes as well as CD71 on granulocytes showed significant differences (2478 vs. 4402, p=0.035 and 2098 vs. 1552, p=0.007). Most of the above mentioned differences could also be confirmed if only RCMD pts of both groups were considered (e.g. lineage infidelity in granulocytes: 0% vs. 36%, p=0.041). Furthermore, trends were seen for a lower CD45-lymphocytes-to-blasts-ratio (4.8 vs. 5.4; p=0.09), higher CD33 intensity on blasts (2416 vs. 1673, p=0.08), and higher qualitative CD36 expression on granulocytes (3.2% vs. 2.5%, p=0.0619) as well as quantitative expression on monocytes (1940 vs. 663, p=0.06). Finally, monitoring of aberrations by FCM allowed for minimal residual disease monitoring in del(5q) MDS pts. Interestingly, the decreased CD19 expression on blasts seemed to be stable independent from the extent of treatment response.

Eight-colour based FCM is a valuable tool in the diagnostics of MDS. It might also be able to distinguish del(5q) MDS from other MDS subtypes.

Disclosures:

No relevant conflicts of interest to declare.
The Prognostic Implications of Nucleated Red Blood Cells in Myelodysplastic Syndrome
Authors: Munoz Javier , Younes Mohamad A. , Hammoud Marwa , Khanshour Ammar , Ortega Rosa Michel , Alhyari Mohammad , Yaghmour George , Kuriakose Philip
Published: October 2019

Abstract 5058

Background:

The presence of elevated nucleated red blood cells (NRBC) in the peripheral blood smear points towards the presence of myelodysplastic syndrome (MDS) or myelophthisis. The intent of our study was to assess the prognostic implication of the presence of increased NRBC in patients with MDS.

Methods:

Retrospective single institution chart review over a period of 10 years from 2000 to 2010. Our study population comprised 288 diagnosed cases of MDS whose diagnoses were made by bone marrow (BM) examination.

Results:

174 patients were male (60.4%) and 114 were female (39.6%). Average age at diagnosis of MDS was 71.92 years. 204 patients were Caucasian (70.8%), 69 patients were African-American (23.9%), 1 patient was Hispanic, 2 patients were Asian, and race was not documented in 12 patients. 117 patients had intermediate-1 IPSS (40.6%), 79 had low IPSS (27.4%), 54 patients had intermediate-2 IPSS (18.7%), 26 patients had high IPSS (9.0%), and 12 patients had unknown IPSS. 131 patients (45.5%) had undetectable NRBC in their peripheral smear prior to their initial diagnostic BM examination. 13 out of these 131 patients (9.92%) progressed to acute myeloid leukemia (AML). 157 patients (54.5%) had one or more NRBC in their peripheral smear prior to their diagnostic BM examination. 27 out of these 157 patients (17.19%) progressed to AML. Survival in days was available in 59 patients. 19 out of 59 patients had undetectable NRBC in their peripheral smear prior to their initial diagnostic BM examination with an average survival of 1,256.2 days. Only one patient in this group underwent allogeneic matched unrelated peripheral stem cell transplantation (PSCT) with a survival of 2,532 days. 40 out of 59 patients had one or more NRBC in their peripheral smear prior to their diagnostic BM examination with an average survival of 850.4 days. Only 3 patients in this group underwent allogeneic matched PSCT (2 related and 1 unrelated) with an average survival of 924.3 days.

Conclusions:

The presence of nucleated red blood cells in the peripheral blood smear of patients with MDS seems to be a marker of more aggressive disease, higher rates of progression to AML and portends worse survival. To our knowledge this is the first study documenting the prognostic implication of NRBC in the peripheral circulation of patients with MDS.

Disclosures:

No relevant conflicts of interest to declare.
4-1BBL Reverse Signaling Enhances Proliferation of Human Multiple Myeloma Cell Line U266
Authors: Wang Juan , Ju Songguang , Fu Chengcheng , Depei Wu
Published: October 2019

Abstract 5074

Multiple myeloma is an incurable plasma cell malignancy. Unraveling the intricate pathobiology and exploitation of novel therapeutic approaches are needed. 4-1BBL, a member of the tumor necrosis factor family, cross-links its receptor, 4-1BB, and co-stimulates T cell activities. Recent studies confirm that 4-1BB-4-1BBL interaction generates bi-directional signals. 4-1BBL has been detected on many tumor cells, and many tumor microenvironmental cells express 4-1BB molecule. However, whether the reverse signaling through 4-1BBL participates the pathology of multiple myeloma is unknown. In this study, we identify the constitutive expression of 4-1BBL on U266, LP1, MY5 and OPM2 MM cell lines as 89.2%, 57.9%, 77.9% and 78.7%. Then we select U266 cell to explore the biological effect of 4-1BBL reverse signaling and its underlying mechanism. Triggered by anti-4-1BBL mAb (1F1), cell counting and MTT assay both demonstrated that U266 cell proliferation increased dramatically. Intracellular cytokine staining showed that treatment of cells with 1F1 also increased the production of IL-6. When neutralizing anti-IL-6 mAb was added to the culture medium, 1F1-induced U266 cell proliferation was effectively inhibited. These effects of 4-1BBL require immobilization of 1F1 but not soluble form. Taken together, 4-1BBL reverse signaling promotes U266 cell proliferation remarkably while increasing the production of IL-6. Moreover, neutralizing anti-IL-6 mAb inhibits U266 cell proliferation stimulated by mAb 1F1, supporting that 4-1BBL reverse signaling promotes the proliferation of U266 cell through increasing the production of IL-6.

Disclosures:

No relevant conflicts of interest to declare.
Epigenetic Dysregulation of the Tissue Inhibitor of Metalloproteinases-2 Gene in Multiple Myeloma
Authors: Galm Oliver , Jost Edgar , Schubert Claudia , Herman James G. , Brümmendorf Tim H , Wilop Stefan
Published: October 2019

Abstract 5089

Background:

Multiple myeloma (MM) is a B-cell neoplasm characterized by the accumulation of malignant plasma cells in the bone marrow (BM) as well as abundant BM angiogenesis. Aberrant methylation of CpG islands near gene promoter regions is the most widely studied epigenetic abnormality in human malignancies and is associated with loss of gene expression. There is increasing evidence for a role of the tissue inhibitor of metalloproteinases 2 (TIMP2) gene as a tumor suppressor. Overexpression of TIMP2 may result in decreased invasive potential, suppression of tumor growth and vascularization as well as inhibition of angiogenesis. We could previously show that TIMP2 may become hypermethylated in association with transcriptional silencing in indolent and aggressive non-Hodgkin's lymphomas. Recently, downregulation of TIMP2 was also reported in MM, and this is thought to contribute to increased BM angiogenesis. In this study, we determined the methylation status of the promoter-associated CpG island of TIMP2 in primary MM samples and investigated correlations between TIMP2 methylation and clinical parameters.

Methods:

Methylation of the promoter-associated CpG island of TIMP2 was analyzed by methylation-specific polymerase chain reaction (MSP) in samples from 76 MM patients (median age 65 years [range 40–94]; 44 males, 32 females; 71 BM samples, 5 peripheral blood samples). Overall survival curves were plotted according to the method of Kaplan and Meier and compared using the log-rank test. Correlations between variables were analyzed using the Fisher's exact two-sided test and independent t-test, respectively.

Results:

MSP analysis revealed that there was aberrant methylation of the TIMP2 promoter region in 4/76 MM patient samples. We found an association of TIMP2 hypermethylation with plasma cell leukemia (p<0.0001) and decreased platelet counts (257.1 G/l vs. 111.5 G/l; p=0.04). There was a trend towards an inferior overall survival in MM cases with a hypermethylated TIMP2 gene, but this difference was not statistically significant (p=0.11).

Conclusions:

Our study shows that promoter hypermethylation of TIMP2 is a novel epigenetic event in MM that may contribute to disease progression and could serve as a prognostic biomarker. Further studies are warranted to elucidate the functional consequences of epigenetic dysregulation of TIMP2 in the pathogenesis of MM. Additionally, the increasing evidence for the role of DNA methylation changes in MM may serve as a basis for the use of epigenetically targeted therapeutic approaches in malignant plasma cell disorders.

Disclosures:

Herman: MDxHealth: Consultancy.
Lenalidomide Plus Bortezomib and Dexamethasone (RVD) Followed by Lenalidomide Plus Dexamethasone As Salvage Treatment for Myeloma Patients in First Relapse After Autologous Stem Cell Transplantation. Single Institution Experience
Authors: Belhadj Karim , Amraoui Kamel , Safar Violaine , Gaillard Isabelle , El Gnaoui Taoufik , Dupuis Jehan , Luciani Alain , Molinier-Frenkel Valérie , Rahmouni Alain , Haioun Corinne
Published: October 2019

Abstract 5143

The RVD regimen has shown a promising activity in patients with relapsed/refractory multiple myeloma (MM). In the present study, we report preliminary results on 14 patients in first relapse after autologous stem cell transplantation (ASCT) treated by VRD followed by a maintenance treatment with revlimid-dexamethasone (Rev-dex) in responder-patients.

Treatment was as follows: RVD included lenalidomide 15 mg per day (d) d1 through d14, bortezomib 1 mg/m2 on d1,4,8 and 11 and dexamethasone 20 mg on d1,2,4,5,8,9,11 and 12 of 21-d cycles with the objective to deliver at least 6 cycles. A maintenance treatment by Rev-dex with lenalidomide 25 mg d1 to d21 plus dexamethasone 20 mg weekly of d-28 cycles, delivered until progression was planned in patients achieving at least partial response to RVD.

Patients' characteristics at relapse: median age was 65 years (range 59 to 70), median time to relapse following ASCT before RVD was 26 months (range 4 to 86). All pts were naive for lenalinomide and 8 for bortezomib. Translocation t(4;14) was observed in 4 patients and was associated with del(17p13) in one case.

Patients received a median number of 6 cycles of RVD (range 3 to 8). Overall response rate was 71% and 43% of the patients achieved very good partial or better response. Discontinuation of RVD was due to disease progression in 4 cases. During RVD treatment, 4 patients (29%) experienced grade 3–4 adverse events: neutropenia n =2, thrombocytopenia n=1, thrombosis n=1. Following RVD, 10 patients received Rev-dex maintenance.

With a median follow-up of 15 months (range 6 to 29), 11 patients are alive free of evolutive disease. Four patients progressed during VRD: one with solitary plasmocytoma was efficiently treated by radiotherapy, the 3 other patients received anthracycline or platin based regimens but were refractory and 2 of them died. One patient progressed during maintenance and was efficiently treated by melphalan-cyclophosphamide-dexamethasone. Among the 4 patients with high-risk cytogenetic abnormalities, one CR, one PR and 2 progressions were observed during RVD. The first patient remains in CR 16 months after the beginning of salvage treatment.

Conclusion:

RVD followed by Rev-dex maintenance is a promising therapeutic option in MM patients in first relapse after ASCT with low toxicity profile.

Disclosures:

No relevant conflicts of interest to declare.
Incidence of the JAK2V617F Mutation In Mexican Patients with Myeloproliferative Neoplasms (MPNs)
Authors: Ignacio Gregorio , Arana-Trejo Rosa María , Gónzalez Verónica , Hérnandez Maria Paula , Lugo Yolanda , Solis Luis , Castellanos Jesús , Celestino Samuel , Váldez Carlos , Rivas Ramón
Published: October 2019

Abstract 5180

Introduction:

The V617F mutation in JAK2 gene has been described in approximately 50–90% of patients with ET, MF AND PV [essential trombocythaemia, idiopathic myelofibrosis and policithemia vera]; but has also been reported, albeit at a lower frequency, in patients with other myeloid malignancies such as atypical CML, CMML, AML, MDS, JMML and CNL. A single G>T base substitution in exon 12 results in the conversion of valine to a phenylalanine aminoacid at position 617 of the JAK2 gene. The identification in this study were using techniques such as allele-specific PCR, RFLP-PCR and direct sequencing; for to determine the incidence in Mexican patients with MPNs.

Patients and Methods:

The JAK2V617F mutation was determined in 88 patients and 5 normal blood samples for healthy individuals as controls. About the patients, 60 were cataloged like MPNs and 28 patients with features suggestive of MPNs vs CML. Samples for bone marrow or peripherical blood were taken either at time of diagnosis of MPNs or during treatment with cytoreductive or anti-thrombotic agents. DNA and RNA were extracted using the QIAamp DNA and RNeasy mini kit (Qiagen) and amplified by the three techniques mentioned for JAK2V617F and by nested RT-PCR for BCR/ABL.

Results:

The five normal blood samples for controls were negative for JAK2V617F mutation and to BCR/ABL. Patients had median age 65 years (47–85 years old), 46% male and 54% female. In de overall patients: 60 patients with MPNs all were BCR/ABL negative and 20 (33%) had JAK2V617F. In the 28 patients with likely MPNs vs CML, 23 were BCR/ABL positive/JAK2 negative, two had the coexistence of both genetics defects [BCR/ABL+ and JAK2V617F+] and 3 BCR/ABL and JAK2 negative. Finally the patiens with JAK2V617F+, were 12 ET, PV 1, MF 2, CML 2, and 5 continued like MPNs.

Discussion:

The incidence of the JAK2V617F in this study for MPNs patiens were 33% and the incidence varied between MPNs subtype. Less than ten cases of BCR/ABL+ CML with JAK2V617F have been published; we report two patients with the coexistence and we agree with previous reports that screening for JAK2V617F mutation should be considered in any BCR/ABL+ CML patients and the clinical outcome will be define in long period.

Disclosures:

No relevant conflicts of interest to declare.
Orbital Non Hodgkin Lymphomas. Features and Evolution of 63 Patients From Two Oncological Centers in Buenos Aires, Argentina: Marie Curie Hospital and Henry Moore Institute
Authors: Dragosky Marta O , Marquez Marisa , Luchetta Graciela P , Alcaraz Susana , Annetta Isabel , Corzo Ariel , Jozami Cecilia , Gil Deza Eduardo , Zerga Marta
Published: October 2019

Abstract 5201

Orbital Non Hodgkin Lymphoma (NHL) represents 2 to 5% of extranodal NHL and 8 to 11% of orbital tumors. Published reports usually comprise a reduced number of patients.

Out of a total of 668 extranodal NHL treated in the period 1991–2010, the orbital locations accounted for 63 patients (10.6%).This high value is due to selected referral from ophthalmologic centers. The diagnosis was made by cytology or surgical biopsy and immunohistochemistry. Staging included CT or MRI of the orbit and brain and bone marrow biopsy.

Results:

Age range:17 to 86 years (mean 60), only 5 < 40 years. The ratio M/F: 1/0.9.Location:right orbit: 50%,left: 36%, bilateral: 14%. Conjunctiva: 44.5%, orbit soft tissue: 23.8%, eyelid: 22.2%, lachrymal gland: 9.5%.

Initial symptoms: orbital tumor: 47.6%, proptosis: 26.9%, ptosis: 11.1%, eyelid edema: 6.3%, diplopia: 4.7%, epiphora: 3.2%.

Histology subtypes: MALT: 50.8%, lymphocytic: 30.1% LDGCB: 6.3%, follicular 6.3%, lymphoplasmacytic: 4.7%, angiocentric T: 1.6%. Time to diagnosis was above 12 months in 63.5%.

Treatment in 49 patients (77.7%). Chemotherapy (CT) (CVP, CHOP, CLB) plus radiotherapy (RT) in 57,14 %, CT in 28.57%, radiotherapy: 12.24%, Rituximab:.28% (12.24% + QT, 2.04% alone).

Complete remission (CR) was obtained in 87.77% and PR in 12.23%. Relapse in 16 (32.65%), local homo-or contralateral 9 (56.25%), systemic 6 (37.50%), 1 (6.25%) in breast (MALT). Died in progression: 4 (25%). The 46,93% of patients had survivals of 5 to 10 years, and 12.5% 11 to 15 years.

Conclusions

The outstanding features are: presentation in 5* to 6* decade, low grade subtypes, initial symptoms: painless mass and proptosis, prevalent location in conjunctiva and eyelid. Is remarkable the delay until diagnosis, due to its low frequency and low specificity of the initial symptoms. The high rate of complete remission obtained with chemotherapy and radiotherapy in frontline and relapse and prolonged survival, are characteristics that determine the good prognosis of this location of extranodal NHL.

Disclosures:

No relevant conflicts of interest to declare.
Role of FDG-PET/CT in Detecting Lymphomatous Bone Marrow Involvement in Patients with Newly Diagnosed Diffuse Large B-Cell Lymphoma
Authors: Hong Junshik , Lee Yukyung , Kim Seog Gyun , Hwang Kyung Hoon , Park Soon Ho , Jeong Jihoon , Kim Kyung-Hee , Ahn Jeong Yeal , Park Sanghui , Park Jinny , Lee Jae Hoon
Published: October 2019

Abstract 5204

Backgrounds:

As a bone marrow bopsy (BMB) is a painful and invasive procedure with a restrictive reliability as only a limited area of the bone marrow (BM) can be evaluated, role of FDG-PET/CT to demonstrate lymphomatous BM involvement as an alternative or at least a complementary to BMB is an area of interest. Several previous studies exist but most of them included heterogeneous types of lymphomas with various treatments.

Patients and methods:

To evaluate the role of FDG-PET/CT in detecting BM involvement, pre-treatment bilateral BMBs and FDG-PET/CT scans of 89 patients with diffuse large B-cell lymphoma (DLBCL) treated with standard immunochemotherapy, rituximab-CHOP were reviewed and analyzed. Uptake more than liver parenchyma intensity on FDG-PET/CT was interpreted as 'with a possibility' of involvement. The final interpretation on the possibility of BM involvement in each patient was reported after discussion among three nuclear medicine physicians and results of BMB were blinded at the time of FDG-PET/CT review. Fourteen patients (15.7%) had lymphomatous involvement based on BMB (BMB+) and 17 patients (19.1%) had the possibility of BM involvement on FDG-PET/CT (FDG-PET/CT+). Seventy-two patients (80.8%) had concordant results between BMB and FDG-PET/CT (7 patients were positive for both and 65 patients were negative for both), but 17 patients (19.2%) had a discordant interpretation (7 patients were BMB+ and FDG-PET/CT-, and 10 for BMB- and FDG-PET/CT+; table 1). Although BMB+ patients had an inferior 2-year EFS (37.0% vs. 79.8%, p < 0.001) and OS (36.3% vs. 81.0%, p < 0.001) compared to BMB- patients, no differences in EFS (62.6% vs. 72.7%, p = 0.185) and OS (59.4% vs. 78.0%, p = 0.146) were shown between FDG-PET/CT+ and FDG-PET/CT- patients. Six of 7 patients with BMB+ and FDG-PET/CT+ had a diffuse involvement on FDG-PET/CT whereas 9 of 10 patients with BMB- and FDG-PET/CT+ had a focal BM involvement on FDG-PET/CT (table 2). Six of 7 patients with diffuse involvement on FDG-PET/CT were BMB+ whereas only 1 of 10 patients with focal BM involvement on FDG-PET/CT were BMB+ (table 2). It is likely therefore that patients with diffuse BM involvement on FDG-PET/CT had higher probability for BMB+ and they might have poorer survival than those with focal BM involvement.

Conclusion:

The results suggest that FDG-PET/CT had a limited value to detect BM involvement in patients with DLBCL. It may not be justified to upgrade patient's Ann Arbor stage to IV according to focal hypermetabolic BM lesion on FDG-PET/CT. Until additional results on the role of FDG-PET/CT in detecting BM involvement available, FDG-PET/CT should be used as an adjuvant rather than an alternative in detecting BM involvement in patients with newly diagnosed DLBCL.

Disclosures:

No relevant conflicts of interest to declare.
Gemcitabine, L-Asparaginase, Ifosfamide, Dexamethasone and Etoposide (GLIDE) Regiment In Advanced or Refractory Aggressive NK/T Cell Lymphomas: A Pilot Study
Authors: Ji Jie , Liu Ting , Xiang Bing , Jia Yongqian
Published: October 2019

Abstract 5208

Background:

Extra nodal NK/T cell lymphoma, nasal type (ENKL) with advanced stage and aggressive NK-cell leukemia (ANKL) are highly aggressive neoplasm with a dismal clinical outcome. Both of these diseases presented with rapid progressed clinical course and resistant to chemotherapy regimens such as CHOP(cyclophosphamide, doxorubicin, vincristine and prednisone). Expression of multi-drug resistance (MDR) associated P-glycoprotein of NK/T cell neoplasms which actively exports many anti-tumor agents outside the tumor cells is one of the well-studied mechanisms. Our new chemotherapeutic regimen GLIDE (Gemcitabine, L-asparaginase, Isofosfomide, Dexamethasone and Epotoside) showed promising results.

Patients and methods:

From 2010–2011, 3 cases of newly diagnosed ENKL,1 case of newly diagnosed ANKL, and 3 cases of refractory ENKL were enrolled. Former treatments included at least 2–6 cycles of chemotherapy (CHOP or L-asparaginase containing regimens) and local radiotherapy. All of these patients received chemotherapy of GLIDE regimen(Gemcibabine 1000mg/m̂2 D1,8; L-asparaginase 6000u/m̂2 D4,6,8,10,12; Isofosfomaide 1000mg/m̂2 D1-3;Dexamethasone 20mg D1-4; Epotoside 100mg/m̂2 D1-3).The stage of all patients was IV. IPI scores were all in high risk. All patients provided informed written consents and this trial was registered at http://www.chictr.org (ChiCTR-TNC-10000782)

Results:

The median age was 41 years (range, 23–54 years);and male (n = 2) to female (n = 5) was 1:2.5. 4 patients finished all 6 cycles, with 3 in CR and 1 in PR; 1 patient died of progression of disease after 2 cycles; 1 patient died of infection in nCR and 1 patient just finished cycle 2 with PR. The OR (CR+nCR+PR) was 85%. The major toxicity of GLIDE were grade 3/4 leucopenia (70.1%, 22 of 31 cycles), and infection (36.3%, 12 of 31 cycles) with 1 death. The non-hematologic toxicity was tolerable, and the hepatic toxicity-associated with the use of L-asparaginase was frequent (42.9%, 3 of 7 patients). However, the majority of the hepatic toxicities were grade 1 or 2.

Conclusion:

GLIDE chemotherapy can be an effective treatment strategy with acceptable toxicity in advanced stage or refractory ENKL and ANKL.

Disclosures:

No relevant conflicts of interest to declare.
Relative Survival of Older Patients with Indolent Lymphoma–Analysis From the SEER Database
Authors: Prabhakar Mamatha , Kanapuru Bindu , Jemal Ahmedin , Hesdorffer Charles , Ershler William , Longo Dan L. , Yates Jerome W
Published: October 2019

Abstract 5214

Background:

Indolent lymphomas account for 35–40% of Non-Hodgkin's lymphomas (NHL). The treatment choices, and as a result, the overall outcome of these indolent lymphomas seem to be changing with the introduction of chemoimmunotherapy. However, the impact of these new treatment approaches on the survival in older patients has not been specifically studied.

Methods:

We used data from the Surveillance, Epidemiology and End Results database to determine survival for older patients diagnosed with indolent lymphomas. We compared trends in survival between 1977–86, 1987–96 and 1997–2006 in men and women, for three age groups 65–74, 75–84 and 85+ years. Survival rates were calculated up to 9 years post-diagnosis.

Results:

Between 1977 and 2006, survival rates increased for both men and women in all three age groups. The greatest improvement in survival was seen at 5 years. Between 1977–86 and 1997–2006, 5-year survival rates increased in men/women by 21%/22%, 23%/29% and 16%/24% in the 65–74, 75–84 and 85+ age groups respectively. Survival gains increased with each decade in all age groups for both sexes with the most marked improvements between 1987–1996 & 1997–2006. Women in the first two groups consistently demonstrated a better survival than men. The oldest old men (85+) had the lowest survival rate with the majority of the deaths occurring in the first year of diagnosis.

Conclusions:

Survival for older patients has increased considerably in the interval between 1977–2006 with the largest improvement seen in the 75–84 year old age group. Larger increases in the survival rates between 1987–1996 and 1997–2006 might be accounted for by the benefits of immunotherapy with the introduction of rituximab in 1998. The increased early deaths noted in men 85+, could be related to treatment toxicity. While earlier treatment is being advocated for patients diagnosed with indolent lymphoma, the data we present imply that more careful consideration should be given to the selection of patients above the age of 85 for such treatment. Male-female differences in survival are interesting and further study would seem important to elucidate the causes.

Disclosures:

No relevant conflicts of interest to declare.
Conditional C-Myc Inactivation Impacts on Chromosomal Translocation Mechanisms and B Lymphomagenesis
Authors: Yan Catherine T. , Muniandy Parames , Yang Xiaoli
Published: October 2019

Abstract 5238

The requirement for Myc oncogenes in cellular transformation and tumor progression in human cancers has been studied extensively. Others and we have shown that the spontaneous development and progression of B lymphomas involving translocations that activate c-Myc can be faithfully recapitulated in model systems. However, for other oncogenes that are the predominant translocation targets in hematopoietic malignancies in humans, such as Bcl6, animal models that spontaneously promote translocations involving them are still lacking. Importantly, the molecular basis for the recurrence of particular oncogene translocations remains undefined. We have shown that a breakdown in DNA double strand break (DSB) repair resulting from conditionally inactivating the essential non-homologous end-joining (NHEJ) repair gene XRCC4 in p53 (XcP) deficient mature B cells reproducibly leads to the development of mature B lymphomas. These lymphomas, termed CD21/XcP, routinely harbor a clonal translocation that fuses IgH switch (S) regions to exon 1 of c-Myc, which we showed are formed by the aberrant repair of AID-induced DSBs during IgH Class Switch Recombination (CSR). These lymphomas harbor also a second clonal translocation that fuses the Igl locus into various chromosomal locations, including c-Myc, which our studies showed were formed by aberrant V(D)J recombination. Juxtaposition of Ig heavy chain (IgH) sequences to the c-Myc oncogene is one of the most extensively modeled and well-studied translocations. It is a hallmark of Burkitts Lymphoma, a subset of other B lymphomas. By limiting or eliminating c-Myc via conditional c-Myc inactivation in peripheral naïve and GC B cells in wildtype, p53 deficient and combined XRCC4 and p53 deficient settings, our studies reveal new insights into normal, functional requirements for C-myc in peripheral B lymphocytes, selection mechanisms underlying its activation in B lymphomagenesis, and the identification of new oncogenic translocation partners resulting from its inactivation in the CD21/XcP model.

Disclosures:

No relevant conflicts of interest to declare.