bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Body Mass Index (BMI) On Overall Survival in Myeloma
Authors: Kumar Mukesh , Nooka Ajay K. , Langston Amelia , Gleason Charise , Watson Melanie , Collins Hannah , Casbourne Daniela , Boise Lawrence H. , Kaufman Jonathan L. , Waller Edmund K. , Lonial Sagar
Published: October 2019

Abstract 4289

Introduction:

Excess body weight has been identified as a risk factor for developing myeloma. From a meta-analysis of 15 cohort studies evaluating myeloma incidence and 5 others on mortality, the risk of multiple myeloma was significantly elevated among overweight patients and obese patients. Interleukin-6 (IL-6) promotes normal plasma cell development and proliferation of myeloma cells in culture. A polymorphism (allele 174C) in the IL-6 gene promoter is associated with obesity and hence, the hypothesis that overweight may be a surrogate for IL-6 promoter genotypes, and associated with increased risk of plasma cell neoplasms has been proposed. However, data is limited in evaluating the outcomes in these patients, based on BMI. We have evaluated the impact of BMI on overall survival in patients with myeloma.

Methods:

We have evaluated a total of 589 patients that underwent ASCT during the period January 2005 through June 2011 from our myeloma database. The median time of follow up is 50 months in all patients and 23 months post ASCT. We used IBM SPSS version 20 to generate the survival statistics.

Results:

The median patient age at the time of diagnosis and at ASCT is 57 years (range, 26–77) and 59 years (range, 26–78), respectively. Patient characteristics: 55%/45% male/female; 56%/36% white/black; ISS stage I/II/III 38%/19%/43%; ASCT conditioning melphalan 140/200 15%/85%. At a median follow up of 50 months from the time of diagnosis and 23 months from the time of ASCT, 75% of the patients are expected to be alive at 109 months. OS was not statistically different between the groups stratified on gender and race. OS from the time of diagnosis and from the time of ASCT was not statistically significant in patients stratified by BMI 18.5–24.9; 25–29.9; 30–34.9; 35–39.9 and ≥40. However, for the patient group with BMI<18.4 vs. ≥18.5; the OS from the time of diagnosis (75% survival: 55 months vs. 109 months; p=0.018) and from the time of ASCT (median OS: 43 months vs. NR; p=0.017) was significantly different, in favor of the BMI≥18.5 group.

Conclusion:

Our results suggest that patients that received ASCT have significant survival benefit. Higher BMI does not confer a negative impact on the prognosis of obese patients, as they actually have better survival after ASCT. Patients that are underweight (BMI<18.4) have inferior survival as those with higher BMI. This effect is possibly related to cancer cachexia or malnutrition, or increased toxicity from the conditioning regimen.

Disclosures:

Kaufman: Millenium: Consultancy; Celgene: Consultancy; Novartis: Consultancy; Onyx Pharmaceuticals: Consultancy. Waller:Outsuka: Research Funding.
Study On the Recovery of Immune System After Chemotherapy in Pediatric Acute Lymphoblastic Leukemia
Authors: Jiang Qian , Qun Yang , Zheng Huyong
Published: October 2019

Abstract 4299

Background:

Acute lymphoblastic leukemia (ALL) is the most common malignancy in children. Nowadays, the cornerstone treatment of ALL is chemotherapy, which will destroy immune system at the same time of killing leukemic cells. After the cessation of chemotherapy, the immune system will rebuild gradually. Our previous study based on the BCH 93 and BCH 98 protocols showed immune system reconstruction would take 2–3 years after the cessation of chemotherapy. In recent years, the intensive chemotherapy is decreased because of accurate classification of ALL, resulting in much more proper treatment which can both cure the leukemia and reduce side effects, such as immune depression. In this study we investigate the recovery of immune system of ALL patients treated by an update CCLG-ALL-2008 protocol in Beijing Children's Hospital of Capital Medical University.

Methods:

There were 104 ALL patients enrolled in this study. Their immunenoglobins (Ig) and T lymphocyte subtypes in peripheral blood were examined at the time points of 0 month, 12 months and 24 months after cessation of chemotherapy, respectively. The data was analyzed by statistical software of SPSS19.0.

Results:

Immunenoglobins of IgA, IgG, IgM and IgE were lower at the cessation of chemotherapy, and increased significantly at 12 months of stopping treatment. The IgA, IgG, IgM and IgE recovered to normal level in 61.54%, 89.23%, 90.77%, and 89.83% of ALL patients, respectively at 12 months of stopping treatment. The total B cells, T cells, CD4, CD8 and NK cells recovered to normal level in 84.85%, 92.54%, 71.64%, 67.16% and 69.23% of ALL patients, respectively at 12 months of stopping treatment. The level of immunenoglobins and T lymphocyte subtypes were no statistical differences between the time points of 12 and 24 months after cessation of chemotherapy.

Conclusion:

Most of ALL patients rebuild their immune system within 12 months after the cessation of chemotherapy. Current protocol of CCLG- ALL-2008 based on the risk classification is more reasonable that shorten the time of immune reconstruction, compared to the results in published literatures based on our previous protocols of BCH93 and BCH 98.

Disclosures:

Zheng: Beijing Health System High-level Technical Personel Plan: Research Funding.
Low MDR1 and BAALC expression Identifies a New Subgroup of Adult Cytogenetically Normal Acute Myeloid Leukemia with a Favorable Outcome
Authors: Guo Xutao , Chen Feili , Shi Pengcheng , Zha Jie , Liu Bingshan , Zheng Haiqing , Xu Bing
Published: October 2019

Abstract 4312

OBJECTIVE:

Cytogenetically normal acute myeloid leukemia (AML) is a heterogeneous disease, in terms of genetic/molecular abnormalities resulting into marked differences in outcome. We demonstrated that MDR1, BAALC expression was of prognostic significance and high MDR1 expression correlated with a high BAALC expression (r=0.487, P<0.001) in cytogenetically normal AML in the prophase study then we hypothesized that MDR1 and BAALC expression together would better identify the patient's risk profile.

METHODS:

Pretreatment bone marrow samples from 92 cytogenetically normal AML patients were analyzed for MDR1 and BAALC mRNA expression by real-time reverse transcriptase polymerase chain reaction. Patients were divided into low MDR1 and BAALC expression group and combined group (high MDR1 and/or high BAALC expression) according to MDR1 and BAALC levels and were compared for clinical outcome.

RESULTS:

73 cases of 92 cytogenetically normal AML patients got CR after the first block with a CR rate being 79.3%. However, 29 cases of the CR patients relapsed with the relapsed rate being 39.7%. In contrast, Patients with low expression of both MDR1 and BAALC had a higher CR rate (93.3%vs72.6%, P=0.021), lower relapse rate (7.1% vs. 42.5%, P=0.000), longer OS (50.3% vs 17.8%,P=0.001) than high MDR1 and/or high BAALC expression (combined group) in cytogenecally normal AML. Results showed no statistical difference in CR rate (93.3%vs85.7%, P=0.341),relapse rate (7.1% vs. 8.8%, P=0.000) and OS (50.3% vs 63.1%,P=0.431) for cytogenetically normal patients with both MDR1 and BAALC low expression comparing to those with low-risk cytogenetically abnormal.

CONCLUSION:

The combined assessment of BAALC and MDR1 expression can improve treatment stratification in adult cytogenetically normal AML. Low expression of both MDR1 and BAALC identifies cytogenetically normal AML patients with a favorable long-term outcome.

Disclosures:

No relevant conflicts of interest to declare.
Azacitidine in Relapsed and Refractory AML: Efficacy for Patients Relapsing As MDS Post AML
Authors: Ivanoff Sarah , Lemasle Emilie , Gruson Berengere , Merlusca Lavinia , Charbonnier Amandine , Parcelier Anne , Damaj Gandhi , Royer Bruno , Marolleau Jean Pierre
Published: October 2019

Abstract 4332

Background:

Azacitidine (Aza), inhibitor of DNA methyltransferases, plays an important role in epigenetic regulation of gene expression and tumorogenesis, and is active in myeloid neoplasia such as myelodysplasia (MDS) and de novo acute myeloid leukemia (AML). Efficacy of Aza for relapsed and refractory AML has not been so far reported.

Methods:

We report in 2 french centers (Amiens, Rouen) retrospective study, the results of Aza for relapsed or refractory patients. All patients received Aza (75 mg/m2 per day over 7 days for 4 weeks cycles), until progression, and at least one cycle. Leukocyte blood count was < 10109/l. The primary endpoint was overall response rate (ORR), according to IWG 2006: complete response (CR), partial response (PR), stable disease (SD) or progressive disease (PD), hematological improvement (HI). Secondary endpoints were duration of response and overall survival (OS).

Results:

41 patients (26 males and 15 females) with a median age of 59 years (range 28–78) were studied from August, 2007 and November, 2011 (Table): 15 had refractory and 26 relapsed AML. At relapse 11/26 had MDS, defined by blast count <20% and cytological morphology of MDS in bone marrow aspirate, and 15/26 AML.

Patients received in average 6 cycles (1–30): 4 (1–7) for refractory; 7 (1–30) for relapsed (11 (3–30) for MDS post AML; 3 (3–5) for relapsed AML).

Overall, the ORR was 34% (7 CR, 5 RP, and 9 HI). For patients with MDS post AML OR was 82% (7 CR, 2 RP, 7 HI), 13% (1 RP, 1 HI) for relapsed AML, and 13% (2 PR and 1HI) for refractory AML.

The average duration of response was 4 months (0–39) for the 41 patients: 14 months (0–39) for MDS post AML, 0.3 for relapsed AML, and 0.5 for refractory.

Overall survival from diagnosis was 29 months (6.9–87): 38.2 (12–57) for MDS post AML, 30.1 (6.9–87) for relapsed AML and 13.3 (6.9–35.5) for refractory (no significant differences).

Overall survival from initiation of Aza was 9.4 months (1.1–39.2): 22.6 (4.8–39.2) for MDS post relapsed and 3.9 (0.3–11.3) for relapsed AML and 6.2 (1.1–13.3) for refractory patients. The differences are not statically significant probably due to small effective of our study. Contrarily to the others 2 groups, 6 patients (55%) with MDS post AML are alive in CR at the latest report; moreover three of them underwent an allogeneic transplantation.

Conclusion:

Aza seems to efficient for relapsed AML patient, especially for MDS post AML, but inappropriate for refractory patients.

Disclosures:

No relevant conflicts of interest to declare.
Breakthrough Stroke During Rivaroxaban Therapy in a Patient with Non-Valvular Atrial Fibrillation
Authors: Ahn Inhye E , Kim Myungsun
Published: October 2019

Abstract 4370

Introduction.

Rivaroxaban is the first orally active direct factor Xa inhibitor FDA approved for anticoagulation in non-valvular atrial fibrillation. Rivaroxaban has several remarkable aspects-it has less incidence of fatal bleeding and intracranial hemorrhage compared to Warfarin, excellent pharmacodynamic profile as an oral agent (high bioavailability (80–100%) and short half-life (7 to 11 hours), and it is reversible- which highlights Rivaroxaban as a promising future anticoagulant. However, 1.71% of Rivaroxaban arm in ROCKET-AF (New Eng J Med 2011;365:883-91) trial developed breakthrough stroke or systemic embolism while on therapeutic dose. We present the first report of breakthrough stroke in a patient with non-valvular atrial fibrillation anticoagulated with Rivaroxaban.

Case presentation.

A 52-year-old Caucasian male with non-valvular atrial fibrillation presented with aphasia, right-sided weakness and right facial droop. The patient was newly started on Rivaroxaban for two weeks followed by successful elective cardioversion a day prior to admission. One hour prior to admission, the patient was found with global aphasia, right hemianopsia and hemiparesis consistent with left MCA stroke. Ischemic CVA was confirmed with non-contrast CT. The patient underwent emergent cerebral angiogram and successful mechanical thrombolysis (Solitaire) of left MCA embolus. A week later, neurologic condition improved to hemiparetic and independent gait with residual dysarthria and right facial droop. Given his breakthrough embolic event, the choice of anticoaulgation was changed to Dabigatran.

Conclusion.

We have demonstrated the first case of breakthrough stroke while on therapeutic dose of Rivaroxaban in a patient with non-valvular atrial fibrillation. In a subset of patients with ischemic stroke while on therapeutic doses of oral anticoagulants, mechanical thrombolysis is crucial and intravenous thrombolytics should be avoided to minimize risk of intracranial hemorrhage. The effectiveness of alternative anticoagulation after failing Rivaroxaban needs to determined with further studies.

Disclosures:

No relevant conflicts of interest to declare.
The Benefit of RED CELL Antigen Testing in A Transfusion Service of A Large Cancer Center
Authors: Aung Fleur M. , Reddy Virgil M , Lichtiger Benjamin
Published: October 2019

Abstract 4374

Introduction:

DNA-based tests are increasingly being used to predict blood group phenotypes in patients who have been recently transfused to aid in alloantibody identification, to distinguish an alloantibody from an autoantibody, in patients who have received hematopoietic stem cell transplants, when RBCS are coated with immunoglobulin (+DAT) and to detect weakly expressed antigens when the patient is unlikely to make antibodies to antigen- positive RBC's transfused. The molecular bases associated with most antigens have been determined and PCR-assays available for testing RBC antigens are a powerful adjunct to the conventional hemagglutination, the gold standard technique to type RBCs for the presence or absence of blood group antigens.

Purpose:

This retrospective review of Red Blood cell (RBC) antigen testing data was conducted on all patients typed between January 2010 and December 2011 at MD Anderson Cancer Center. The purpose was to review the concordance between the antibody (ies) identified by serologic testing and the antigen negativity from phenotype predicted from DNA analysis for selecting antigen negative RBCs for transfusion. In addition we also wanted to review whether the extended phenotype red cell units transfused to patients with unidentified alloantibody (ies)/RACT and autoantibodies formed additional red cell alloantibody (ies) after transfusion.

Methods:

The technology of BioArray Solutions (Bioarray Solutions, Immuncor, Norcross, GA, USA) and the Human Erythrocyte Antigen (HEA) v1.2 BeadChip was used to assess the genotype and predicted phenotype of patients. We performed RBC antigen testing on all of our recently transfused patients with alloantibody (ies), patients whose red cell antibody panels showed reaction with all red cells tested (RACT) or unidentified antibodies, patients with positive direct antiglobulin test (DAT) and whose elution studies showing RACT or unidentified antibody (ies) and in patients with autoimmune hemolytic anemia.

Results:

Atotal of 912 BioArray tests were performed on 912 patients [444 (49%) males:468 (51%) females; median age 60 years (range 1–92)]. Antibody screen was positive in 424 (46%) and negative in 488 (54%) patients. A DAT was performed in 126 patients with a positive antibody screen to establish immune from non-immune hemolytic anemia. The results are listed in Tables 1,2 and 3.

Conclusion:

We found 100% concordance between the predicted antigen negativity phenotype and the patient's RBC alloantibody (ies) identified by serologic testing. The RBC antigen typing has allowed us to transfuse patients with unidentified antibodies and RACT with extended phenotype matched red cell units preventing the development of additional red cell alloantibody (ies) as well as allowed us to transfuse antigen-negative units preventing patient/blood donor incompatibility issues.

Disclosures:

No relevant conflicts of interest to declare.
Predicting Blood Loss and Transfusion Requirement in Patients Undergoing Surgery for Musculoskeletal Tumors
Authors: Thompson Philip A , May Deborah , Choong Peter , McArdle Scott , Gaskell Mary , Powell Gerard , Liew Danny , Cole-Sinclair Merrole Faye
Published: October 2019

Abstract 4384

The management of musculoskeletal tumors has evolved markedly over the past 30 years, with improvements in surgical techniques, staging and chemotherapeutic advances delivering improved survival and functional outcomes, both in the setting of primary malignant bone tumors and in the palliation of tumors metastatic to bone. Advances in surgical technique as well as pre-operative chemotherapy has meant that most patients are now able to undergo limb-sparing surgery. Whilst clearly desirable to improve functional outcome, limb-sparing surgery is however more technically challenging and is associated with greater blood loss than amputation.

There is a paucity of data concerning blood utilization in the setting of major musculoskeletal tumor surgery. We retrospectively analyzed a database containing 1322 consecutive surgeries in 1222 patients over a 14 year timeframe, performed at a quaternary referral center in Melbourne, Australia. The following factors were significantly associated with high blood utilization: Chordomas, sacral and pelvic tumors, lower starting haemoglobin, tumors >5cm in size and high American Society of Anesthesiologists (ASA) score. The ASA score also clearly correlated with 30 day mortality. Pre-operative planning in these patients is critical to ensure adequate blood supply, minimize wastage and optimize the patient's general health prior to surgery. As such, based on modeling from our data, we propose a maximum surgical blood order schedule based on the presence or absence of the above factors.

Allogeneic blood transfusion, while a potentially life-saving intervention, is associated with major complications, including transmission of infectious agents and immunological complications. Longer-term, transfusion has been reported to result in immunomodulation and potentially a higher risk of tumor recurrence. Clearly, minimization of blood loss and avoidance of un-necessary transfusion is desirable. There is also a lack of data surrounding interventions designed to minimize blood loss in musculoskeletal tumor surgery, especially when compared to similar studies performed in major orthopedic surgery for benign conditions. These would be ideal targets for future randomized studies in the high-risk groups we have identified.

Disclosures:

No relevant conflicts of interest to declare.
CML Shows Characteristic Pattern by Flow Cytometry Analysis of Peripheral Blood
Authors: Scott Kalley , Gorczyca Wojciech
Published: October 2019

Abstract 4430

A total of 75 chronic myeloid leukemia (CML) cases from peripheral blood (40 cases) and bone marrow (BM, 35 cases) with adequate flow cytometry (FC) data, smear, blood cell count, and the presence of t(9;22)/BCR-ABL by fluorescence in-situ hybridization (FISH) studies were analyzed for immunophenotypic pattern. The FC pattern in CML was compared with benign (healthy) controls (blood, 20 cases; BM, 20 cases), blood with reactive neutrophilia (15 cases), myelodysplastic syndrome (BM, 15 cases), and blood with eosinophilia (15 cases). CML showed a characteristic pattern by FC in blood, which can be easily differentiated from reactive neutrophilia or eosinophilia, regardless of WBC count. The identification of distinct population of blasts, basophilia, lack of CD10, CD11b, CD13 and/or CD16 on subset of granulocytes, decreased granularity, and/or aberrant expression of CD56 on granulocytes and monocytes, can be easily identified by routine FC analysis. We suggest using FC analysis of blood as a screening tool for patients with leukocytosis (neutrophilia) with follow-up FISH studies in cases with the phenotypic features suggestive of CML. Patients with confirmed CML diagnosis by FISH will undergo marrow biopsy for differential count including blast and basophil enumeration (to exclude accelerated phase or blast crisis), degree of reticulin fibrosis and cytogenetic studies (for additional chromosomal changes present at diagnosis). This approach, in our opinion, allows to diagnose early (unsuspected) CML and eliminates the need for unnecessary cytogenetic/FISH testing, and especially bone marrow biopsy in patients with reactive leukocytosis or eosinophilia.

Disclosures:

No relevant conflicts of interest to declare.
Is There a Benefit of Adding Rituximab to CHOP in the Overall Survival of Patients with B-Cell Non-Hodgkin′s Lymphoma in a Developing Country ?
Authors: Ruiz-Delgado Guillermo J. , Almaguer David Gomez , Tarin-Arzaga Luz C. , Cantú-Rodríguez Olga , Alarcon-Urdaneta Carlos , Rodriguez-Morales Uxmal , Calderon-García Jackeline , Fernandez-Vargas Omar , Montes-Montiel Maryel , Sanchez-Cardenas Monica , Ruiz-Arguelles Guillermo J.
Published: October 2019

Abstract 4892

Rituximab ( R ) has changed the prognosis of patients with non-Hodgkin′s lymphoma (NHL) in developed countries, but its role has not been analyzed in underprivileged circumstances. One hundred and two patients with NHL treated in a developing country were analyzed: 28 patients with follicular lymphoma (FL) and 74 diffuse large B cell lymphoma (DLCL). Patients were treated upfront with either CHOP or R-CHOP; the decision to employ R depending solely on the ability of patients to defray it. In DLCL, 42 were given CHOP and 32 R-CHOP, whereas in FL 19 were given CHOP and 9 R-CHOP. The impact of the addition of R was found to be more clear in FL than in DLCL. In patients with DLCL, the overall survival (OS) was 87% at 80 months for those treated with R-CHOP and 84% at 145 months for those treated with CHOP (p NS). In patients with FL, the OS was 89% at 88 months for those treated with R-CHOP and 71% at 92 months for those treated with CHOP (p =.05). In a multivariate analysis, other variables were identified to be associated with the OS were IPI and number of cycles in DLCL. It is concluded that rituximab produced a mild positive impact in the OS of patients with FL, but not in those with DLCL. Since the addition of rituximab results in a 36 fold increase in treatment costs, these observations may be important to decide therapeutic approaches in NHL patients living in underprivileged circumstances.

Disclosures:

No relevant conflicts of interest to declare.
Clinical Features of Chinese Refractory Cytopenia of Childhood: Lower Overall Survival and Higher Possibillity of Clonal Evolution Than Nsaa Patients
Authors: An Wenbin , Li Zhanqi , Wang Shuchun , Chen Xiaojuan , Yang Wenyu , Guo Ye , Zhang Li , Zhang Peihong , Zhu Xiaofan
Published: October 2019

Abstract 4947

Objective:

To analyze the clinical features and survival time in patients with refractory cytopenia of childhood (RCC), and to develop a new diagnostic scoring system for RCC.

Methods:

The clinical information, laboratory findings at primary diagnosis and survival time in 97 non-severe aplastic anemia (NSAA) children diagnosed in August, 2000 to June, 2006 were analyzed retrospectively. According to the criteria for RCC proposed in the 2008 edition of the World Health Organization classification of hematopoietic and lymphoid tissues, diagnosis of these NSAA patients was reassessed.

Results:

37/97 cases (38. 1%) were reassessed as RCC, 60/97 cases (61. 9%) were still diagnosed as NSAA. There was no significant differences in distribution of age, gender and chromosomal abnormalities, absolute neutrophil count, PLT, MCV, absolute reticulocyte count, the number of cytopenias and duration from onset to starting treatmeat between RCC and NSAA. All patients received the treatment of CsA and androgen. There was no significant differences in distribution of treatment response. The overall survival (OS) of RCC patients was significantly lower than NSAA patients (P=0. 041) in long-term follow-up. The former had 91. 5% of 6-year prospective survival (PS) and the latter had 96. 5% of 6-year PS. At a median follow-up of 93 months (range 6–138), 2 patients reassessed as RCC progressed into AML-M5 and PNH respectively, while no NSAA patient progressed into clonal diseases. There was significant differences in the distribution of HGB<11g/dl at onset and lymphocyte count<3×109/L, dysplastic megakarycytes≥10% (CD41 monoclonal antibogy immunohistochemical staining) and periodic acid schiff (PAS) staining (+) in bone marrow aspirate between RCC and NSAA (P=0. 034, 0. 025, 0. 032, 0. 029). Compared with NSAA, there was significant differences in the distribution of bone marrow cellularity down to 10% of the normal age matched value, dysplastic erythroblastic island, number of erythroblastic mitoses and micromegakaryocytes in bone marrow biopsy of RCC(P< 0. 0001, 0. 0001, 0. 0001, 0. 0001). Using Logistic regression, a new diagnostic scoring model including increased dysplastic erythroblastic island, having micromegakaryocytes in bone marrow biopsy, and lymphocyte count<3×109/L in peripheral blood was developed. By ROC curve analysis, score≥2 points was identified as a cut-off value with sensitivity of 70. 3% and specificity of 95%.

Conclusion:

RCC patients had similar clinical features with NSAA patients but had lower OS and higher possibillity of clonal evolution. This new diagnostic scoring model had significance to differentiate diagnose RCC and NSAA.

Disclosures:

No relevant conflicts of interest to declare.
Array Comparative Genomic Hybridization: Impact in the Assessment of Plasma Cell Myeloma Genetic Profile and Prognosis
Authors: Leon Annette , Sevilla Deborah , Marino Joseph M , Moreno Kenneth D , Obrera Rev , Feldman Liliya , Fan Yan G , Persad Rajendra , Villamil Cesar , Maya Melissa , Mehta Nirav , O'Dea Denise , Kemel Yelena , Mazumder Amitabha , Weisberger James
Published: October 2019

Abstract 4965

Identification of specific chromosomal alterations in plasma cell myeloma (PCM) is essential for the prognosis and treatment of this disease. Although cytogenetic chromosome and fluorescence in situ hybridization (FISH) analyses are techniques currently used for this purpose, incorporation of array comparative genomic hybridization (aCGH) analysis as described in this study, demonstrates the insufficient power of traditional techniques in characterizing the complex and heterogeneous genetic profile of this group of hematological malignancies. In our cohort, aCGH study of 500 diagnosed PCM patients identified clinically significant genomic alterations in 56% of cases compared to 22% and 32% by chromosome and FISH analyses, respectively. Important findings by aCGH include the presence of hyperdiploid chromosome complement (15 > 9 > 5 > 19 > 11 > 3 > 7 > 21 > 6 > 18 >14) in 35% of cases which is associated with a favorable prognosis, as well as adverse prognostic markers such as hypodiploid chromosome complement (13 > Y >14 > 22> 10 >16 >8), gain of genetic material in chromosome 1q (CKS1B, PDZK1, ANP32E) and losses in chromosomes 1p (CDKN2C, FAM46C), 6q (PRDM1), 12p (ETV6, CDKN1B), and 17p (TP53) observed in 25% of patients. Recurrent alterations identified in 9% of cases only by aCGH include extra copies of genes which could be potentially poor prognostic indicators such as IRF4 (6p25. 3, transactivates MYC oncogene), IL-6 (7p15. 3, growth factor), BRAF (7q34, regulating the MAP kinase/ERKs signaling pathway), CYLC2 (9q31. 1, active cyclin in cell cycle progression from G1 to S phase), and genes located in the telomeric region of chromosome Xq. The results obtained in this study demonstrate the superior resolution and detection rate of aCGH technology in understanding the genetic heterogeneity of PCM as well as the importance of incorporating this methodology into current algorithms for the diagnosis and prognosis of plasma cell disorders.

Disclosures:

No relevant conflicts of interest to declare.
Sensitive Detection of Plasma Cells by Integrated Enrichment in-Column Staining and Analysis
Authors: Kühbach Katja , Herber Manuela , Koehler Dorothee , Schmitz Jürgen , Assenmacher Mario
Published: October 2019

Abstract 4968

In Multiple Myeloma immunophenotyping by multiparameter flow cytometry is a well-established method in clinical research to identify, characterize and enumerate neoplastic plasma cells. In addition, recent clinical studies demonstrated prognostic relevance of MRD detection by multiparameter flow analysis. The high applicability of the flow cytometric analysis compared to patient-specific RT-PCR holds the potential to become an important tool in MRD assessment.

For the identification of plasma cells the detection of more than one antigen is required and for an unequivocal discrimination between normal and neoplastic plasma cells the combination with clonality assessment is mandatory. Here we established a combination of five antibodies for surface staining: CD138-PE, CD38-FITC, CD56-APC, CD19-PE-Vio770, CD45-APC-Vio770 and two antibodies for intracellular staining: Anti Igκ-PerCP, Anti Igλ-VioBlue. The combination of fluorochromes for this 7-color panel was optimized for high resolution of the target population.

To minimize variability among flow cytometric data, it is crucial to standardize the various procedures – in particular in clinical settings. Pre-defined antibody panels and simplified sample handling are major factors in the reduction of variability. Furthermore, in order to obtain maximal sensitivity processing of large cell numbers is required, which is a major obstacle for conventional flow cytometers.

Here we demonstrate that magnetic pre-enrichment of CD138+ in combination with a new protocol for in-column intracellular staining for κ/λ light chains of retained plasma cells allows handling of up 108 cells (fig. 1). Together with the optimized antibody panel this approach offers a unique and valuable tool for characterization of rare plasma cells and offers increased sensitivity during MRD assessment. The MACSQuant® Analyzer offers the possibility to combine magnetic pre-enrichment with intracellular staining and flow cytometric analysis. This minimizes manual handling steps and reduces cell loss due to omission of centrifugation steps usually required for intracellular staining procedures.

The approach of combined cell enrichment, in-column intracellular staining and usage of standardized antibody panels on the MACSQuant Analyzer provides an automated workflow, which is highly suitable for routine detection and characterization of rare plasma cells.

Figure 1 Figure 1.

Disclosures:

Kühbach: MiltenyiBiotec GmbH: Employment. Herber:Miltenyi Biotec GmbH: Employment. Koehler:Miltenyi Biotec GmbH: Employment. Schmitz:Miltenyi Biotec GmbH: Employment. Assenmacher:Miltenyi Biotec GmbH: Employment.
Expression and Clinical Significance of Notch1 On the Membrane of Bone Marrow CD38+CD138+Plasma Cells in the Patients with Multiple Myeloma
Authors: Fu Rong , Zhao Yiran , Shao Zonghong , Wang Honglei , Zhang Tian , Li Lijuan , Liu Hui
Published: October 2019

Abstract 4981

Objective:

To investigate the expression of bone marrow CD38+CD138+, CD38+CD138-plasma cells and the expression of Notch1 on the membrane of them in the patients with multiple myeloma(MM), and explore the importance of Notch signaling pathway in the formation and progression of MM further.

Methods:

Thirty-three MM patients and 15 healthy controls were enrolled in this study. The expression of bone marrow CD38+CD138+, CD38+CD138-plasma cells and the expression of Notch1 on the membrane of them were analyzed by flow cytometry. The expression of Notch1 mRNA of bone marrow mononuclear cells were analyzed by RT-PCR.

Results:

The ratio of CD38+CD138+ plasma cells from 24 newly diagnosed MM patients was (51. 50%±12. 48%) which was significantly higher than CD38+CD138- plasma cells of MM patients (42. 88%±11. 41%)(P=0. 016)and controls 20. 13%±5. 8(P=0. 000). The expression of CD38+CD138+ plasma cells from 24 newly diagnosed MM patients was correlated to the level of malignant plasma cells in there bone marrow(r=0. 546, p=0. 006), serum level of lactate dehydrogenase(LDH)(r=0. 567, p=0. 004), and β2-MG(r=0. 431, p=0. 035). The ratio of Notch1 on the membrane of CD38+CD138+ plasma cells of MM patients was (60. 21%±25. 06%) which was significantly higher than those of CD38+CD138- plasma cells of MM patients 39. 84%±18. 94%(P=0. 000)and controls (38. 34%±19. 39%)(P=0. 004). There was no statistical difference between the two latter groups(P>0. 05). The expression of Notch1 on CD38+CD138+ plasma cells from 24 newly diagnosed MM patients was correlated to the level of malignant plasma cells in there brone marrow(r=0. 914, p=0. 000), serum level of lactate dehydrogenase(LDH) (r=0. 604, p=0. 002), and β2-MG(r=0. 455, p=0. 026). The ratio of Notch1 on the membrane of CD38+CD138+ plasma cells of MM patients who had renal dysfunction was correlated to their abnormal serum creatinine levels. The expression of Notch1 on CD38+CD138+ plasma cells from 17 MM patients who received VD chemotherapy was correlated to the ratio of plasma cell reduction after the first VD chemotherapy(r=0. 842, p=0. 000). The expression of Notch1 mRNA of bone marrow mononuclear from 10 MM patients was (0. 8252±0. 4079) which was significantly higher than those of controls (0. 3759±0. 0813)(p=0. 032).

Conclusion:

Notch1 over expressed on CD38+CD138+ plasma cells with relation to the effects of early VD therapy and long term outcome of MM.

Disclosures:

No relevant conflicts of interest to declare.
Tumor Antigens and T Cell Subpopulations: Is There Correlation Between These Two Possible Targets for Immunotherapy in Multiple Myeloma?
Authors: Braga Walter Moises Tobias , Silva Bruna Raphaeli , de Carvalho Fabricio , Alves Veruska Lia Fook , Colleoni Gisele Wally Braga
Published: October 2019

Abstract 4985

Introdution:

Cancer/Testis Antigens (CTAs) are a promising class of tumor antigens that have a limited expression in somatic tissues (testis, fetal ovary, and placental cells). CTAs are attractive targets for immunotherapy in cancer because the gonads are immune privileged organs and anti-CTA immune response can be tumor-specific. Recently, the importance of microenvironment for development and maintenance of many types of malignancies has been demonstrated. Multiple myeloma (MM) offers a good model to study tumor/microenvironment interactions because it involves a series of genetic alterations and changes in bone marrow microenvironment, favoring the tumor growth and failure of local immune control. A detailed analysis of the balance between Tregs and Th17 lymphocytes seems necessary to design more effective and less toxic types of immunotherapy in myeloma which is still an incurable malignancy.

Aims:

The present study aims to characterize and correlate expression of genes related to tumor cells (CTAs) and genes related to antitumoral control (Treg and Th17 lymphocytes) in patients with MM.

Matherial and Methods:

Forty-six bone marrow samples from newly diagnosed MM patients were analyzed. MAGE-C1/CT-7, MAGEA3/6, NY-ESO-1, LAGE-1 and GAGE expression were analyzed by conventional RT-PCR. CTLA4 and RORyt were used to evaluate Treg and Th17 subpopulations, respectively, by quantitative PCR (qPCR). The analysis of the possible correlation between the expression of CTLA4 and RORyt and MAGE-C1/CT7, MAGEA3/6, NY-ESO, LAGE-1 and GAGE were made using median comparison expression of the first two genes with the positive or negative expression of the CTAs, using the Mann-Whitney test.

Results:

The CTA more frequently expressed in MM was MAGEC1/CT7 (66%). Expression MAGEA3/6, NY-ESO-1, LAGE-1 and GAGE were, respectively: 40%, 23%, 32% and 30%. RORyt presented hipoexpression in 41% of the cases and CTLA4 presented overexpression in 70% of MM cases. The correlation analyses between all the CTAs and the CTLA4 or RORyt did not show any significant result (p> 0. 05).

Conclusion:

We found overexpression of a Treg subpopulation related gene (CTLA4) and hipoexpression of Th17 (RORyt), indicating possible imbalance of T lymphocytes in the tumoral microenvironment. However, it was not possible to establish a correlation between the expression of the most frequently expressed CTAs in MM with Treg and Th17 cells′related genes expression. It suggests that the expression of these genes does not seem to occur in a coordinate manner and its importance as therapeutical target in MM must occur by an independent way (Supported by Fapesp 2010/17668-6).

Disclosures:

No relevant conflicts of interest to declare.
Clinical Significance of Serum Free Light Chain Test in the First Onset of Multiple Myeloma
Authors: Hong Pan , Meng Xiaojian , He Jingsong , Wu Wenjun , Li Yi , Han Xiaoyan , Zheng Gaofeng , Zhao Yi , He Donghua , Shi Jimin , Xie Wanzhuo , Zhang Jie , Ye Xiujin , Huang He , Cai Zhen
Published: October 2019

Abstract 4999

Objective:

Clinical significance of serum free light chain (sFLC) test in the diagnosis, efficacy evaluation, prognosis judgment of first onset of multiple myeloma (MM).

Method:

Medical records of 39 cases of first-onset MM were gathered in our hospital from 1/2010 to 2/2012, including 25 male and 14 female patients with the median age being 63 years old. sFLC-kappa and sFLC-lambda were determined by immune turbidimetric assay, sFLC-kappa/lambda were calculated and the results were compared with that of immunofixation electrophoresis (IFE) as well as serum protein electrophoresis (SPE) for sensitivity assessment. All patients were treated with chemotherapy followed by sFLC detection and efficacy assessment. Patients with a sFLC results higher than median were marked as high-sFLC, the rest were marked as low-sFLC, and survival analysis were made comparing the two groups.

Results:

Of the 39 first-onset MM patients, 38 were detected as sFLC- kappa/lambda positive, that's a sensitivity of 97. 4%, higher than IFE (79. 5%), SPE (53. 8%), and uPE (38. 5%). As condition improves after treatment, sFLC gradually decline and sFLC- kappa/lambda level falls back to normal. Evaluation of efficacy by sFLC results has an accuracy of 77. 27%, significantly higher than that of corresponding M protein test£̈ P=0. 009£©. Follow-up survival analysis showed that the OS time and TTP time of low-sFLC group are significantly higher than those of high-sFLC group (P=0. 037, 0. 009 respectively).

Conclusion:

sFLC detection is a highly sensitive quantitative method, can help the diagnosis of MM, gives a quicker assessment of treatment efficacy, and can predict disease recurrence. When it's the first onset, sFLC level can also roughly forecast the prognosis.

Disclosures:

No relevant conflicts of interest to declare.
MiR-15a, Mir-16-1 and Mir-17-92 Cluster Expression Are Linked to Poor Prognosis in Multiple Myeloma
Authors: Lijuan Chen , Gao Xiao , Run Zhang , Xiaoyan Qu , Sensen Zhang
Published: October 2019

Abstract 5000

Multiple myeloma (MM) is characterized by a profound genomic instability of potential prognostic relevance. Loss of chromosome 13, observed in almost half of patients, negatively affects prognosis. MiR-15a, miR16-1 and miR-17-92 cluster, located on 13q, play important roles in the regulation of cell proliferation, differentiation and apoptosis. Therefore, we investigated a possible correlation of miRNA expression with chromosome 13 deletions (del(13)) and prognosis. We measured the expression of miR-15a, miR16-1 in 70 newly diagnosed MM patients and miR-17-92 cluster in 85 newly diagnosed MM patients by quantitative real-time PCR analyses. MiR-15a, miR-16-1 and miR-17 -92 cluster expression levels are independent of the del(13). High levels of miR-15a, miR-16-1, miR-17, miR-20a and miR-92-1 are associated with shorter progression-free survival (PFS), suggesting poor prognosis. Our data suggest that the expression of specific miRNAs may be contributed to MM prognosis.

Disclosures:

No relevant conflicts of interest to declare.
Bortezomib Could Reduce Cell Viability by Activation of Heme Oxygenase 1 in Multiple Myeloma Cells
Authors: Tibullo Daniele , Giallongo Cesarina , Barbagallo Ignazio , La Cava Piera , Parrinello Nunziatina , Volti Giovanni Li , Palumbo Giuseppe A , Conticello Concetta , Raimondo Francesco Di
Published: October 2019

Abstract 5005

Bortezomib (PS-341, Velcade®) specifically targeting the ubiquitin-proteosome pathway, is the first defined therapeutic proteosoma inhibitor approved by FDA for treatment of Multiple Myeloma (MM). We explored the hypothesis that the activity of Bortezomib in MM cells may involve the heme oxygenase 1 (HMOX1), a enzyme with pleiotropic effects, catalysing the degradation of heme to carbon monoxide, biliverdin and ferrous iron.

After 24h of treatment with Bortezomib at 10nM, the U266 and SKM-M1 MM cell lines showed a decrease of cell viability around 50% and an increase of HMOX1 protein and mRNA expression of 40 fold (p<0. 001). Using a colorimetric assay we observed that Bortezomib increased also the HMOX1enzymatic activity (p<0. 03). To elucidate the significance of the HMOX1 increase after bortezomib treatment, we treated myeloma cells with Bortezomib plus an inducer (Hemin 10uM) or an inhibitor (Tin, Sn-Mesoprotoporphyrin, 10uM) of the enzyme activity. Treatment with Hemin and Tin alone not changed the cell viability. Cell viability was reduced (p<0. 003) by addition of Hemin and was increased (p<0. 002) by Tin addiction with respect to Bortezomib alone.

Conclusion:

Bortezomib increases HMOX1expression and its enzymatic activity and this increase could be an important mechanism of Bortezomib induced cytotoxicity in MM cells. Modulation of HMOX1 activity has effect on Bortezomib induced cell death.

Disclosures:

Off Label Use: Eltrombopag is a thrombopoietin receptor agonist indicated for the treatment of thrombocytopenia in patients with chronic immune (idiopathic) thrombocytopenic purpura (ITP).
Cytogenetic Analysis and Correlation Study in Prognosis of Patients with Multiple Myeloma
Authors: Fu Chengcheng , Xin Xue , Depei Wu
Published: October 2019

Abstract 5006

Multiple myeloma (MM) is a fatal neoplasm characterized by the accumulation of malignant plasma cells within the bone marrow (BM) and the presence of a monoclonal immunoglobulin in the serum and/or urine, it was reported that patients with abnormal metaphases by conventional cytogenetics at diagnosis had active disease and a reduced survival rate compared with those normal metaphase. To further explore the cytogenetic characteristics of multiple myeloma and its correlation with clinical treatment and prognosis, 38 cases of MM were collected and chromosome specimens of bone marrow cells were prepared by 24-hour culture. R-banding technique combined with interphase fluorescence in situ hybridization(i-FISH) was used for karyotype analysis. The impact of different treatments and chromosome aberrations on progression-free survival(PFS) and overall survival(OS) time as well as related prognosis factors of MM were analyzed.

The detected total chromosome aberration rate was 34. 2% (13/38), with chromosome complex aberration 53. 8%(7/13). The detection rate of R-banding technique was 23. 7%(9/38) and 21. 1%(8/38) by FISH. The most frequent chromosome aberration was chromosome 1 abnormality. The median PFS time of abnormal group was 16 months and was not reached of normal group after median follow up for 36 months(P=0. 045). Also, the median PFS time of complex abnormal group was 15 months, and the median PFS time of non-complex abnormal group was not reached(P=0. 012). The median OS time of complex abnormal group was 22 months and the median OS time of non-complex abnormal group was not reached(P=0. 041). Boretizomib or autologous hematopoietic stem cell transplantation prolonged the PFS and OS of the chromosome aberration group compared with traditional chemotherapy, but there were no significant statistical differences between the two groups(P>0. 05).

It was concluded that chromosome complex aberration is the mainly cytogenetic characteristics of multiple myeloma, and multiple numerous and structural aberrations are involved. The patients with chromosome complex aberration have poor prognosis. Chromosome abnormalities detected in metaphases from multiple myeloma (MM) cells have a clear impact on prognosis and response to therapy. Techniques to explore cytogenetic and molecular genetic changes of myeloma cells should be used to improve chromosomal aberration detection to guide clinical treatments and evaluate prognosis.

Disclosures:

No relevant conflicts of interest to declare.
Inhibition of Related JAK/STAT Pathways with Molecular Targeted Drugs Shows Strong Synergy with Ruxolitinib in Chronic Myeloproliferative Neoplasms
Authors: Barrio Santiago , Gallardo Miguel , Arenas Alicia , Ayala Rosa M. , Rapado Inmaculada , Rueda Daniel , Jimenez Ana , Albizua Enriqueta , Burgaleta Carmen , Gilsanz Florinda , Martinez Joaquin
Published: October 2019

Abstract 5054

Objectives:

In the present study we have determined the antitumor effects, molecular mechanisms of action and potential synergies between Ruxolitinib and the molecular targeted drugs Sorafenib, KNK437, Dasatinib and Perifosine, in Philadelphia negative chronic myeloproliferative neoplasms (MPN).

Materials and methods:

The cytotoxic and cytostatic effects of the different compounds were determined in the JAK2V617F positive cell lines HEL and Ba/F3 JAK2V617F EPOR, and in mononuclear and bone marrow CD34 positive cells from 19 MPN patients. The effects of different drugs at the molecular level were analyzed by flow cytometry and western blot. The IC50 and the synergy combination index (CI) were determined with GrafPath Prism software and Calcusyn software respectively.

Results:

Ruxolitinib (IC50PV=15nM), as well as Sorafenib (IC50PV=8uM), KNK437 (IC50PV=100uM) and Perifosine (IC50PV=15uM), was able to inhibit proliferation in cell line models and in cells from MPN patients. Dasatinib (IC50PV=12nM), however, was only effective in patient samples, with similar effects in both MPN patients and control donors. Moreover, Dasatinib, KNK437 and Sorafenib showed a strong synergistic effect in combination with Ruxolitinib (CIPV<0. 3, table 1). The Western blot analysis confirmed that Sorafenib inhibited the activation of ERK and P38, and, as a consequence, of STAT5. Ruxolitinib blocked the ERK and STAT5 activation, Dasatinib blocked SRC and STAT5, and KNK437 decreased the stability of the protein JAK2, reducing its expression.

Conclusions:

The blockage of JAK2 related proliferative pathways has the potential to inhibit cell proliferation in MPNs. Furthermore, the combination of Ruxolitinib with inhibitors targeted against these pathways has a strong synergistic effect. This may be related to a decrease in the activation of the final common effector STAT5.

Disclosures:

No relevant conflicts of interest to declare.