bluuebunny/crossref_metadata_2025 huggingface dataset:
Lineage-Specific Sox7 Expression In Hematopoietic Progenitor Cells Derived From Human Umbilical Cord Blood
Authors: Wan HaiXia , Chow Howard C.H. , Fung Tsz-Kan , Fan Kin-Pong , Wong Anders S.Y. , Liang Raymond , Leung Anskar Y.H.
Published: October 2019

Abstract 4781

Introduction:

The SOX (Sry-related HMG box) genes belong to a family of transcription factors containing a High-Mobility-Group box domain. In an initial screen of SOX genes in human leukemias, SOX7 is uniquely down-regulated in acute myeloid leukemia, myelodysplastic syndrome and chronic myelogenous leukemia but up-regulated in most cases of acute lymphoblastic leukemia. The observation led to the proposition that SOX7 may play a role in lineage differentiation in hematopoiesis. In this study, we examined SOX7 expression in human umbilical cord blood (UCB) with a view to understand its role in hematopoietic lineage specification.

Methods:

Mononuclear cells (MNC) were isolated from UCB and fractionated into CD34+, CD34-, CD34+CD38+ and CD34+CD38- populations by immunomagnetic selection and fluorescence activated cell sorting (FACS). 0.1–0.4 × 106 CD34+ UCB cells were transplanted intravenously into sub-lethally irradiated NOD/SCID mice with or without prior anti-CD122 antibody injection. Donor engraftment was assessed after 7–8 weeks and engrafting human cells were sorted for CD34+, CD33+, CD19+ expression. CD34+ UCB cells were also plated in colony-forming unit (CFU) assay. SOX7 expression was evaluated by quantitative real-time reverse-transcriptase polymerase chain reaction (Q-RT-PCR).

Results:

The relative SOX7 expression in CD34+ and CD34+CD38- UCB cells was 9.15±2.18 and 4.53±1.09 fold higher than in CD34- (P=0.0001) and CD34+CD38+ (P=0.014) respectively. SOX7 expression in CD34+ cells (arbitrarily set as 1.0) was down-regulated upon differentiation in CFU assay (CFU-GM/G: 0.09±0.06 fold, P=0.007; BFU-E: 0.05±0.03 fold, P=0.003). CD34+ cells engrafted into NOD/SCID mice as enumerated by human CD45+ cells in mouse BM (with anti-CD122: 51±6.3%; without anti-CD122: 28±9%; p=0.017) and gave rise to predominantly B-lymphoid (CD19+) (53.1±8.18%) and to a less extent myeloid (CD33+) (2.9±0.54%) differentiated cells. Prior injection with anti-CD122 antibody had no effect on lineage differentiation and the results were pooled. When SOX7 expression in engrafting myeloid and B-lymphoid differentiated cells (arbitrarily set as 1.0) were compared, it was significantly down-regulated in the myeloid series (0.28±0.08 fold, p = 0.036).

Conclusion

SOX7 is selectively expressed in human HSC from UCB. It is preferentially down-regulated during myeloid differentiation both in-vivo and in-vitro. The function of SOX7 during lineage specification and its link to myeloid malignancies is currently investigated in our laboratory.

Acknowledgments

The project was supported by a grant from the strategic Research Theme of cancer stem cells in the HKU.

Disclosures:

No relevant conflicts of interest to declare.
Differential Regulation of Expression of the Anti-Leukemogenic Orphan Nuclear Receptor Nur77 (nr4a1) In HSCs, CLPs, and CMPs
Authors: Shin Yusup , Manjunath Shwetha , Gross Matthew , Land Ruben , Garrett Russell , Emerson Stephen G. , Punt Jenni A.
Published: October 2019

Abstract 4782

The molecular basis for the balance between self-renewal and differentiation that regulates hematopoietic stem cell (HSC) development is poorly understood. Recent studies suggest that levels of expression of the orphan nuclear hormone receptor Nur77 (nr4a1), previously implicated in thymic selection, may play a pivotal role in HSC fate decisions. Previously published quantitative PCR data suggest that murine HSCs express relatively high levels of Nur77 mRNA. Genetic deletion of Nur77 and its close family member Nor-1 (nr4a3) in mice resulted in the development of acute myeloid leukemia (Mullican et al. Nat Med, 13:730;2007), consistent with Nur77's role as an anti-leukemogenic transcription factor, but also suggestive of a role in HSC homeostasis. To more closely investigate the role of Nur77 in normal hematopoiesis, we constructed a transgenic reporter mouse in which GFP is driven by the Nur77 promoter (eGFPNur77). Unexpectedly, we found that fewer than 3% of freshly isolated bone marrow HSCs (Lin-c-Kit+Sca-1+, IL-7R-) and common lymphoid progenitors (CLP = Lin-c-Kit+Sca-1+, IL-7R+), and no common myeloid progenitors (CMP) expressed Nur77 in our novel reporter system. Interestingly, culture of Lin- bone marrow cells with serum-containing medium resulted in a marked increase in Nur77 (GFP) expression among the majority of HSC's and CLP's, but not CMP's. Further studies indicate that a heat-insensitive component of serum, also present in plasma, is responsible for this induction. Taken together with published data, our studies suggest that induction and modulation of Nur77 may play a previously unappreciated role in self-renewal and lymphoid versus myeloid lineage commitment decisions of HSCs. We are currently working to understand the developmental difference in HSC/CLP/CMP responsiveness and to identify the serum component(s) responsible for upregulating Nur77.

Disclosures:

No relevant conflicts of interest to declare.
Lung-Derived Microvesicles Induce Stable Long-Term Epigenetic Changes In Marrow Cells
Authors: Aliotta Jason M , Pereira Mandy , Amaral Ashley , Dooner Mark S , DelTatto Michael , Papa Elaine , Goldberg Laura R , Quesenberry Peter J
Published: October 2019

Abstract 4799

We have previously shown that microvesicles derived from irradiated murine lung cells enter murine bone marrow cells and induce expression of the pulmonary epithelial cell-specific mRNAs Surfactants A-D (Sp-A-D), Aquaporin-5 (Aq-5) and Clara Cell Specific Protein (CCSP). The early mRNA increases are due to direct delivery of mRNA and due to the transfer of a transcription factor to marrow cells (Aliotta JM, et al, Exp Hematol, 2010). The modulated marrow cells produce pulmonary epithelial cell-specific protein and have an enhanced ability to convert into pulmonary epithelial cells after transplantation into lethally-irradiated mice. In the present studies, we evaluated the stability of pulmonary epithelial cell-specific mRNA expression in cytokine-stimulated liquid culture and after in vivo transplantation into lethally-irradiated mice. Whole bone marrow cells isolated from C57BL/6 mice were cultured for seven days opposite C57BL/6 lung fragments, but separated from them by a 0.4 micron cell-impermeable membrane. Marrow cells were then either infused into lethally-irradiated C57BL/6 mice or established in culture with DMEM supplemented with IL-3, IL-6, IL-11 and steel factor. Engrafted mice were sacrificed six weeks after transplantation and real-time RT-PCR was carried out on marrow, liver, spleen and thymus tissue. Expression of Sp-A, B and D, CCSP and Aq-5 were variably seen in all tissues and increases over control ranged from 12-fold to 58-fold. Liquid culture was analyzed every two weeks out to 12 weeks and high levels of expression of different pulmonary epithelial cell-specific mRNAs and proteins (by western blot and Immunohistochemical staining) were observed. In order to determine the mechanism for the increased expression in culture, we established co-cultures of Fischer-344 rat lung opposite C57BL/6 mouse bone marrow cells and, using species-specific primers, determined whether the increased pulmonary epithelial cell-specific mRNA expression at each time point of secondary cytokine-supported culture was rat or mouse-specific. In every instance, the pulmonary epithelial cell-specific mRNA which was detected was mouse-specific, not rat-specific, and thus the long-term expression of lung-specific mRNA in mouse marrow was due to induced transcription by the mouse marrow cells. These data indicate that lung-derived microvesicles induce long-term stable epigenetic changes in marrow cells and suggests that the genetic changes induced by tissue microvesicles in neighboring cells may be biologically important.

Disclosures:

No relevant conflicts of interest to declare.
Long Term Results of Stanford V Regimen and Highly Active Antiretroviral Therapy (HAART) In 59 Patients (pts) with HD and HIV Infection (HD-HIV)
Authors: Spina Michele , Gabarre Jean , Mancuso Salvatrice , Re Alessandro , Schiantarelli Clara , Talamini Renato , Vaccher Emanuela , Tirelli Umberto
Published: October 2019

Abstract 4827

Background:

The introduction of HAART has significantly improved the outcome of pts with HD-HIV. However there are no data on the long term follow-up of HD-HIV pts treated with conventional chemotherapy (CT) regimens. In 2002, we reported the results of a prospective phase II study with the intensive 12-week CT with adjuvant radiotherapy (Stanford V) and concomitant HAART in 59 pts (Spina et al. Blood 2002;100:1984-1988).

Methods:

To analyze the long term outcome of patients included in the Stanford V and HAART protocol.

Results:

The median follow-up is 67months (range 3–156 months The 5-yr overall survival (OS), freedom from progression (FFP), disease free survival and event free survival are 54%, 52%, 60% and 37%, respectively. The 5-year OS is significantly different in pts with an international prognostic score (IPS) >2 in comparison to that of pts with an IPS <3 (84% vs 36%, p= 0.0005). Similarly, the percentages of FFP at 5 years in these groups are 72% and 45% (p= 0.03).

Conclusions:

Our data confirm the long term efficacy of Stanford V regimen in combination with HAART in HD-HIV. However, Stanford V is significantly less effective in pts with IPS>2 and therefore new strategies be tested in this setting.

Supported by AIRC and ISS grants.

Disclosures:

No relevant conflicts of interest to declare.
Deletion of PAX5 Is Frequently Accompanied by CDKN2A Deletion but Not by ETV6-RUNX1 Rearrangement In B-Lineage Acute Lymphoblastic Leukemia
Authors: Kim Miyoung , Jeon Young Jin , She Cha Ja , Hwang Sang Mee , Kang Hyoung Jin , Shin Hee Young , Ahn Hyo Seop , Yoon Sung-Soo , Kim Byoung Kook , Park Myoung Hee , Lee Dong Soon
Published: October 2019

Abstract 4841

Introduction

The PAX5 gene (Paired box gene 5) works as a guardian in B-lymphopoiesis. It activates essential components of B lymphopoiesis and represses transcription of genes for differentiation to other lineage. Thus, abrogation of normal function of PAX5 is believed to contribute the leukemogenesis in B-ALL. In this study, we evaluated the utility of the detection of PAX5 gene alteration in practice in relation to PAX5 immunoexpression, other common cytogenetic changes and survival in childhood and in adult B-ALL.

Materials and Methods

Interphase FISH using PAX5 split signal probe (DakoCytomation, Glostrup, Denmark), and probes for BCR-ABL1, ETV6-RUNX1 and MLL rearrangements, and CDKN2A deletion was performed in 112 CD19+ B-ALL patients (79 children, 33 adults). G-banding and immunohistochemical staining with anti-PAX5 (BC/24, BIOCARE MEDICAL, CA, USA) antibody was also performed.

Result

The incidence of translocation, deletion and amplification was 2.5%, 10.0% and 12.7% in children, and 0.0%, 18.2% and 6.1% in adults, respectively. Alteration of PAX5 gene was more common than BCR-ABL1 and MLL rearrangement in children (8.9% and 5.1%, respectively), and MLL rearrangement in adults (3.1%). None of the children with ETV6-RUNX1 rearrangement (22.8%) showed PAX5 gene alteration. Most cases of the deletion of PAX5 were accompanied by CDKN2A deletion. Positive immunoexpression in PAX5 IHC was observed in 74.4% in children and in 76.9% in adults Either the children with positive or negative PAX5 immunoexpression had PAX5 gene abnormalities. The presence of PAX5 gene alteration or negative immunoexpression of PAX5 did not have prognostic impact.

Discussion

Based on high incidence of structural alteration of PAX5 in dominant leukemic clone, we suggest PAX5 FISH as a useful tool for the diagnosis and the disease monitoring in B-ALL. Frequent CDKN2A deletion, but not ETV6-RUNX1 rearrangement with structural alteration of PAX5 implies the role of PAX5 in determining prognosis of B-ALL. No definite relationship between the structural alteration and immunoexpression of PAX5 might be resulted from allele specific regulation and haploinsufficiency.

Disclosures:

No relevant conflicts of interest to declare.
Analysis of Genetic Alterations In 843 Korean Acute Leukemia Patients by Multiplex RT-PCR Assay (HemaVision) and Karyotyping
Authors: Lim Jihyang , Kim Myungshin , Kim Yonggoo , Han Kyungja , Kim Hee-Je , Lee Seok , Min Woo-Sung , Chung Nak-Gyun , Cho Bin , Kim Hack-Ki
Published: October 2019

Abstract 4845

Background:

The importance of molecular tests has been increased for diagnosis, classification and MRD monitoring in acute leukemia. Recently, multiplex RT-PCR assays (HemaVision) for simultaneous detection of 28 leukemia-associated translocations was introduced and has been used for molecular screening of leukemic patients. We analyzed the results of multiplex RT-PCR assay (HemaVision) and karyotyping in Korean acute leukemia patients.

Methods:

From April 2007 to March 2010, multiplex RT-PCR assay (HemaVision, DNA Technology A/S, Denmark) was performed in 843 Korean acute leukemia cases (AML 566 vs. ALL 277 and adults 596 vs. pediatrics 237) for screening leukemia-associated translocations. All results were compared with chromosome analysis results by conventional karyotyping.

Results:

Three hundreds fifty-one cases (41.6%) were positive in multiplex RT-PCR. In AML cases, 221 cases (39.0%) revealed positive results and RUNX1-RUNX1T1 85 cases (38.5%), PML-RARA 74 cases (33.5%), CBFB-MYH11 25 cases (11.3%) and MLL-related 19 cases (8.6%) were frequently detected. In contrast, 130 cases (46.9%) revealed positive results in 277 ALL cases and BCR-ABL1 68 cases (52.3%), ETV6-RUNX1 28 cases (21.5%), TCF3-PBX1 15 cases (11.5%), MLL-related 10 cases (7.7%) were frequently detected. In comparison to karyotyping, 337 cases (96.0%) revealed abnormal karyotype and 14 cases (4.0%) revealed normal karyotype in 351 cases that revealed positive results by multiplex RT-PCR assay. In 492 cases that revealed negative results by multiplex RT-PCR assay, 278 cases (56.5%) revealed abnormal karyotype and 214 cases (43.5%) revealed normal karyotype. Overall 629 cases (74.6%) were detected genetic alterations by multiplex RT-PCR assay and/or karyotyping.

Conclusion:

Most acute leukemias may have a genetic alteration in their leukemogenesis. Both multiplex RT-PCR assay and karyotyping are essential for screening leukemia-associated genetic alterations.

Disclosures:

No relevant conflicts of interest to declare.
Chromatin Texture and Molecular Features Are Independent Prognostic Factors In AML
Authors: Mello Mariana Rezende Bandeira , Albuquerque Dulcinéia Martins , Albanez Krizzia Borges , Adam Randall L. , Pereira-Cunha Fernanda Gonçalves , Pagnano Katia B. , Costa Fernando Ferreira , Metze Konradin , Lorand-Metze Irene
Published: October 2019

Abstract 4850

Introduction:

The WHO 2008 classification of acute myeloid leukemia (AML) is based on morphology, cytogenetics and molecular features. Among them, mutations and internal tandem duplications of FLT3 in AML with a normal karyotype have been associated to a poor prognosis. Mutated NPM1 in the absence of FLT3-ITD is associated to a favorable. On the other hand, variables of nuclear chromatin texture have been described as independent risk factors in several malignancies (ALL, melanoma and multiple myeloma).

Aim:

To compare the influence on overall survival of the chromatin fractal dimension and molecular features in adult patients with AML.

Patients and Methods:

We analyzed 106 consecutive cases diagnosed at our Institution between 2007 and 2009. Diagnosis was made by bone marrow (BM) cytology and karyotype, and cases were classified by WHO criteria. Genomic DNA was extracted by phenol-chloroform. Genotyping was made with the MegaBACE 1000 equipment and analyzed in the Fragment Profiler v1.2. For detection of the FLT3-TKD mutation, genomic DNA was amplified by PCR followed by restriction analysis. Blasts from the diagnostic BM cytology were digitalized, segmented and nuclear morphometric variables were examined. Their influence on overall survival was analyzed in the Cox model.

Results:

Median age: 52 years; peripheral blood (PB) leukocytes: 24.0×109/l (0.7-281.3). In the univariate analysis were significant: PB leukocyte count (p=0.005), low-risk karyotype (p=0.002), FLT3 ITD+ (p=0.002), FLT3+NPM1- (p=0.029) and “goodness of fit” (R245) of the chromatin fractal dimension (Minkowski) (p=0.03). Age, fractal dimension and methylation status of p15, p16, p57, p73, ER and MDR1 were not significant. In the multivariate analyses including age, PB leukocytes, R245 and mutations, high leukocyte counts (p=0.03) and low R245 (p=0.01) were independent unfavorable and FLT3-NPM1- (p=0.04) and FLT3-NPM1+ (p=0.02) were favorable prognostic variables.

Conclusions:

The blast chromatin texture measured by R245 was an independent prognostic factor together with known risk variables in AML.

Supported by: FAPESP and CNPq

Disclosures:

No relevant conflicts of interest to declare.
Genetic Alterations of Wilms' Tumor 1 (WT1) Gene In Korean Patients with Normal Karyotype Acute Myeloid Leukemia
Authors: Choi Yunsuk , Lee Je-Hwan , Lee Jung-Hee , Kim Dae-Young , Kim Sung-Doo , Lim Sung-Nam , Hur Eun-Hye , Seoul Miee , Lee Young-Shin , Kang Young-Ah , Jeon Mijin , Lee Kyoo-Hyung
Published: October 2019

Abstract 4852

Introduction:

Acquired mutations of the WT1 gene have been reported in 5–10% of normal karyotype AML (NK-AML) patients. Poor prognostic impact of the mutations has been reported, but some studied found no such correlation. Recent report from Germany showed the association of single nucleotide polymorphism (SNP) rs16754 status with clinical outcomes in NK-AML (J Clin Oncol 2009; 28:578). We assessed clinical implication of WT1 gene alterations in Korean patients with NK-AML.

Patients and Methods:

This study included a total of 75 patients with NK-AML. All patients received standard induction chemotherapy (‘7+3’ regimen) at the Asan Medical Center, Seoul, Korea between May 1999 and Dec 2006. WT1 exons 7 and 9 were amplified using polymerase chain reaction (PCR) and purified PCR fragments were directly sequenced. The clinico-laboratory data were retrieved from the AMC Leukemia Registry.

Results:

WT1 mutations were found in four patients (5.3%): two duplication mutations and two insertion mutations. All mutations were in exon 7 and none in exon 9. Two of four patients harboring WT1 mutation failed to achieve complete remission (CR) and died at 3.9 and 7.0 months after diagnosis of AML. One of two patients attaining CR was alive without relapse at 44.0 month and another died in CR at 4.7 months. WT1 SNP rs16754 was AA genotype (WT1AA) in 5 (3.7%), AG (WT1AG) in 29 (38.2%), and GG (WT1GG) in 41 (54.7%). The findings are significantly different from those of German report (WT1AA in 74.3%, WT1AG in 24.1%, and WT1GG in 1.6%) (P<0.0001). No significant difference of baseline clinico-laboratory findings was observed between different genotypes of WT1 SNP rs16754 (WT1AA/AG vs. WT1GG). The CR rates were similar between two SNP groups (WT1AA/AG vs. WT1GG, 85.3% vs. 80.5%, P=0.78). The five-year probabilities of overall (OS) and relapse-free (RFS) survival appeared to be lower in patients with WT1GG genotype, but the differences were not statistically significant (WT1AA/AG vs. WT1GG; OS, 49.3% vs. 36.9%, P=0.43; RFS, 56.3% vs. 42.5%, P=0.64).

Conclusion:

The prognostic impact of WT1 mutation in NK-AML could not be assessed in this study due to low number of patients harboring the mutation, but three of four patients died with relatively short survival duration, suggesting poor prognosis of the patients with WT1 mutation. The frequency of WT1 SNPs rs16754 in our patients was different from that of German report and survivals were not significantly different according to the SNP genotypes.

Disclosures:

No relevant conflicts of interest to declare.
The Activator of Transcription 3 (STAT3) Gene Polymorphisms Are Associated with Treatment Outcomes In Acute Myeloid Leukemia
Authors: Chen Bao-An , Zhong Yue-jiao , Feng Ji-feng , Cheng Lu , Li Yufeng , Qian Jun , Cheng Jian , Bao Wen , Ding Jia-hua
Published: October 2019

Abstract 4858

Objectives:

In the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway signaling pathway, the STAT3 is one of the most prominent predictable factors for cancer and leukemia. STAT3 activation might promote cellular transformation and therefore have an important role in human tumors. This study was aimed to investigate the relationship between the STAT3 polymorphisms and the treatment responses of AML in the Chinese population.

Methods:

We tested three single nucleotide polymorphisms (SNPs) with 130 Acute Myeloid Leukemia (AML) patients. Genomic DNA was isolated from peripheral blood and assayed by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF).

Results:

The results showed there were strong relationship between unfavorable cytogenetic, partial remission (even no remission) and the frequencies of GG genotype in rs9909659 (P=0.01 and 0.03) and the patients less than 45 years old was significant association with GA/AA genotype (P=0.01). But associations were not confirmed about event-free survival and leukemia-free survival in the Chinese population studied.

Conclusions:

It is clear that the AML patients with GG genotype in rs9909659 are not sensitive to standard chemotherapy method

Disclosures:

No relevant conflicts of interest to declare.
Polymorphisms In Signal Transducer and Activator of Transcription 5 (STAT5) and Response to Cytarabine-Based Chemotherapy In Acute Myeloid Leukemia Patients
Authors: Chen Bao-An , Zhong Yue-jiao , Feng Ji-feng , Cheng Lu , Li Yu-feng , Qian Jun , Ding Jia-hua , Gao Feng , Xia Guo-hua , Lu Zuhong , Cheng Jian , Bao Wen
Published: October 2019

Abstract 4859

Objectives:

Signal transducer and activator of transcription 5 (STAT5) protein is one of the concernful part of STATs families. The constitutive STATs activation has been especially showed in transformation by fusion genes in leukemias.

Methods:

In this study, we aimed to investigate whether the genetic polymorphisms in the STAT5 gene are associated with the treatment outcomes of Ara-C-based chemotherapy regimens in Acute Myeloid Leukemia (AML) patients. 152 AML patients in a Chinese sample were enrolled in our study. Peripheral blood samples were analyzed by matrix assisted laser desorption ionisation-time of flight mass spectrometry (MALDI-TOF-MS).

Results:

The results showed that the frequencies of the T/T genotype in rs2293157 and rs1135669 were higher in unfavorable and intermediate group respectively (P=0.023 and P=0.033). The frequency of patients with T/T genotype of rs1135669 was significantly higher in complete remission (CR) group.

Conclusions:

We concluded that the T/T genotype of rs1135669 might be an important marker for therapeutic efficiency of Ara-C-based chemotherapy in AML patients, especially in Chinese population.

Disclosures:

No relevant conflicts of interest to declare.
Rituximab, Endoxan, Ganciclovir, Dexamethasone, Tocilizumab Sequential Treatment In Kaposi's Sarcoma-Associated Herpesvirus (KSHV) -Related Multicentric Castleman Disease. A Case Report with 8 Months Follow-up
Authors: Iuliano Francesco , Mingrone Tecla , Infusino Stefania , Perricelli Alessia , Pomillo Angelo , Luci Maria , Maio Massimo Di
Published: October 2019

Abstract 4872

Clinical history of Kaposi sarcoma-associated herpesvirus (KSHV),-related Multicentric Castleman Disease is characterized by a rapidly progressive and often fatal course. We report a case successfully treated sequentially with a course of 4 infusions of rituximab 375mg/m2 at weekly intervals, Endoxan, 750 mg/m2 as an intravenous infusion once weekly for 2 weeks, intravenous ganciclovir, 5 mg/kg twice daily for 2 weeks and then once daily, dexamethasone 20 mg on days 1–4, 9–12, 17–20 and anti-interleukin-6 receptor antibody (tocilizumab) administered intravenously at a dose of 8 mg/kg every 2 weeks, starting at day 32.

On December 12, 2009, a 54-year-old man came to the medicine department because of a 2-week history of progressive fatigue, wasting, high-grade fever (39°C), profuse sweating, and severe autoimmune hemolytic anemia (4.2 mg/dL). He had been diagnosed with asymptomatic MCD 6 months before admission. Generalized lymphadenopathy and hepato-splenomegaly were evident on physical examination and enlarged lymph nodes in retroperitoneal regions (CT scan). Blood biochemistry showed very high CRP, IL-6, beta 2 microglobulin serum levels, low LDH and albumin, increase polyclonal immunoglobulin, grade 3 thrombocytopenia and neutropenia. Serologic tests (Epstein-Barr virus, hepatitis B and C viruses, cytomegalovirus and human immunodeficiency virus) were negative. On hospital day 10, 250.000 copies of HHV-8 DNA were detected in 1 mL plasma by RT-PCR. The patient achieved CR two months after admission, still maintained at 8 month follow-up. HHV-8 DNA became undetectable over the course of 8 weeks, at which time ganciclovir was discontinued.

Conclusions: In our case the onset of the disease was serious and life-threatening.

Sequential therapy has proven to be able to save the life of the patient acting on different biological targets. To date is the first case treated with this schedule.

Disclosures:

No relevant conflicts of interest to declare.
Comparing the Outcomes of CHOP Chemotherapy Alone with Rituximab Plus CHOP for Hong Kong Chinese Patients with Diffuse Large B-Cell lymphoma
Authors: Law Man Fai , Yip Sze-Fai , Chan Hay Nun , Yeung Yiu Ming , Choi Wai
Published: October 2019

Abstract 4894

Introduction

Diffuse large B-cell lymphoma (DLBL) is the commonest type of lymphoma worldwide. The condition is the same in Hong Kong and it accounts for about 30–40% of lymphoma cases. CHOP (cyclophosphamide, doxorubicin, vincristine and prednisolone) was used for treatment of DLBL for many years. Rituximab, a chimeric anti-CD20 monoclonal antibody was used for the treatment of DLBL for about 10 years. Previous studies have shown the better efficacy of rituximab plus CHOP over CHOP alone, including the remission rate, overall and event-free survival. No previous study compared the efficacy of rituximab plus CHOP with CHOP alone in Hong Kong Chinese.

We conducted a study to compare the outcomes of CHOP chemotherapy alone with rituximab plus CHOP in a local hospital in Hong Kong.

Method

It was a retrospective study from January 1999 to December 2009. Hong Kong Chinese patients with newly diagnosed diffuse large B-cell lymphoma were recruited in the study. They were divided into two groups. One group of patients was given CHOP chemotherapy and the other group was given rituximab plus CHOP. The remission rate, overall survival and event-free survival were compared in the two groups.

Results

62 patients were recruited in the study. Twenty-nine patients with median age of 55 (range 23–77) were given CHOP chemotherapy alone. Thirty-two patients with a median age of 53 (range 20–75) were given rituximab plus CHOP. The baseline characteristics were similar in both groups. There were 41% of male patients in each group. IPI (international prognostic index) score as well as the proportion of patients at stage III and IV disease were similar in both groups.

The complete remission rate was 38% in CHOP alone group and 78% in rituximab plus CHOP group (p=0.002, Fisher's exact test).

The 3-year overall survival rate was 47% in CHOP group and 74% in rituximab plus CHOP group (p=0.043). The 3-year event-free survival was also better in the rituximab plus CHOP group (p=0.026). 19 events (including relapse, progression and death) were observed in the CHOP alone group and 8 events were observed in the rituximab plus chemotherapy group.

The side effects profile was similar in both groups and the rate of infection is comparable in both arms.

Conclusion

This study has shown that the use of rituximab plus CHOP was better than CHOP alone in Hong Kong Chinese patients with newly diagnosed diffuse large B-cell lymphoma. The complete remission rate, overall survival and event-free survival were significantly higher in the rituximab plus chemotherapy group.

Disclosures:

No relevant conflicts of interest to declare.
Survival of Patients with Relapsed Diffuse Large B-Cell Lymphoma In a Community Hospital
Authors: Mirault Jaclyn , Milani Arash Ghanbari , Ghetia Ditina , Yeboah Isaac , Sharma Puja , Colmer Jonas , Cotler Samantha , Topilow Arthur A.
Published: October 2019

Abstract 4902

Introduction: Of the various subtypes of non-Hodgkin's lymphoma, Diffuse Large B-cell Lymphoma (DLBCL) is the most prevalent, making up 30 percent of all non-Hodgkin's lymphoma cases. Information from community hospitals regarding the treatment and survival of relapsed patients is scarce. The study was done to obtain the survival data of patients with a relapse of DLBCL, treated with and without transplant at our center.

Patients and Methods: The tumor registry data at Jersey Shore University Medical Center between 2000–2008 was reviewed. 41 patients had been diagnosed with DLBCL. Of these, the diagnosis in 13 pts represented a relapse of DLBCL. Review of the hospital records showed that the treatments for these relapsed patients had been radiation, rituximab -based chemotherapy, and/or autologous stem cell transplant. Eight were female and five were male, with a median age of 66 years at initial diagnosis (range: 51 to 81 years). The median age was 68 at the time of relapse (range: 52 to 82 years). Survival was measured from the time of the diagnosis of relapse until death from any cause or the date when lost to follow up. Information on the duration of time from initial diagnosis to relapse was also obtained.

Results: Of the 13 patients who relapsed, 3 underwent stem cell transplant, one of whom died at 43 weeks and two are alive after 70 and 336 weeks. Of the 10 who did not have transplant, 6 died at 2, 8, 15, 19, 19 and 38 weeks. The 4 remaining non-transplant patients are alive at 37, 61, 167, and 221 weeks. Of the 13 patients who relapsed, at primary diagnosis, one had been stage I, 3 had been stage II, 3 had been stage III, and 5 had been stage IV. One was of unknown stage. The average time from diagnosis to relapse was 80 weeks.

Conclusion: Although our numbers are small, it does appear that there is a role for second-line treatments in those patients with a relapse of large cell lymphoma that are not undergoing stem cell transplant. Further study of the treatment of non-transplanted patients with relapsed DLBCL appears warranted.

Disclosures:

No relevant conflicts of interest to declare.
The Anti-CD20 Monoclonal Antibody GA101 Displays More Robust Anti-Tumor Activity Versus Rituximab In Waldenstrom's Macroglobulinemia (WM)
Authors: Yang Guang , Hanzis Christina , Verselis Sigitas , Xu Lian , Hunter Zachary , Ioakimidis Leukothea , Sheehy Patricia , Patterson Christopher , Fox Edward , Treon Steven P.
Published: October 2019

Abstract 4904

Background:

Rituximab is an IgG class CD20-directed monoclonal antibody used in the treatment of B-cell malignancies, including WM. We and others have previously demonstrated dependence for IgG class therapeutic antibodies on polymorphisms at FcγRIIIA-158. Approximately half of WM patients express V/V or V/F, and the remainder half express F/F at this polymorphic locus. Patients with WM expressing FcγRIIIA-158 V/V or V/F show improved rituximab single agent activity, as well as attainment of deeper responses (VGPR or CR) with combination Rituximab therapy. GA101 is a novel humanized anti-CD20 antibody with a glyco-engineered Fc domain that exhibits increased Fcg receptor binding and ADCC activity.

Methods:

In this study, we examined the in vitro activity of GA101 and Rituximab against WM cells, and also examined the activity of these antibodies in context of FcγRIIIA-158 polymorphisms. ADCC activity for GA101 and Rituximab was assessed using genotyped healthy donor derived NK cells against BCWM.1 WM cells, as well autologous NK cells against the patient's own lymphoplasmacytic cells. In vitro B-cell depletion and direct cell death induction assays were also performed.

Results:

We observed significantly greater ADCC activity against WM cells for GA101 versus Rituximab in both healthy donor, as well as autologous NK cell assays. GA101 mediated ADCC activity was particularly more robust versus Rituximab in patients expressing FcγRIIIA-158 F/F versus V/V or V/F (Figure 1). In addition, GA101 induced significant direct cell death against WM lymphoplasmacytic cells, as well as in vitro B-cell depletion assays in comparison to Rituximab, which exhibited little direct cell death induction activity. Nuclear translocation of apoptosis inducing factor (AIF) was observed following GA101 by immunofluorescence microscopy.

Conclusions:

GA101 is associated with enhanced ADCC activity relative to Rituximab by NK cells, particularly for those subjects expressing FcγRIIIA-158 F/F. In addition, GA101 demonstrated direct cell death in WM lymphoplasmacytic cells through an AIF mediated caspase-independent pathway. These studies provide the framework for the investigation of GA101 in WM, and suggest particular benefit for those patients who express FcγRIIIA-158 F/F.

Disclosures:

No relevant conflicts of interest to declare.
Rituximab in Lymphoma Associated Auto-Immune Haemolytic Anemia
Authors: Bhargava Amit , Banerjee Nirupma
Published: October 2019

Abstract 4923

The association of Autoimmune hemolytic and Non Hodgkin's lymphoma is a known phenomenon. Cold hem agglutinin disease (CHD), is an uncommon Autoimmune hemolytic anemia, mediated by cold reactive auto antibodies that bind to erythrocyte carbohydrate antigen causing hem agglutination and complement mediated hemolysis. Cold agglutinin–mediated hemolysis occurs at low temperature which may be severe and difficult to treat.

CHD may be primary when it is idiopathic or secondary as in lympho-proliferative disorder of bone marrow associated with clonal proliferation of CD20 B cells that produce monoclonal 1g M cold agglutinin (kappa /lamda). Rituximab has been used as an as a novel treatment option in Autoimmune hemolytic anemia. Rituximab is a genetically engineered chimeric monoclonal antibody that targets the CD20 antigen on B cells.

Rituximab in autoimmune conditions possibly acts by destruction of CD20_clonotypic precursor B cells and/or CD20 plasma cells, thus is effective in controlling immunoglobulin-mediated diseases of B lymphocytes.

We analysed four patients of lymphoma with severe CHD who were successfully treated with the chimeric anti-CD20 monoclonal Rituximab (375mg/ m2) intravenously every three weeks for four to six cycles, till the Agglutinins disappeared from the serum.

Patient and method

In this study patients diagnosed with lymphoma associated CHD were treated with Rituximab with a dose of 375mg/ meter square given every three weeks for four to six cycles, till the primary aim was achieved. The primary aim was correction of anemia, disappearance of cold hem-agglutinins from serum, assessed by complete blood counts and measuring cold agglutinins in serum.

Four such patients were studied, all of them had NHL-DLBCL (IHC proven), with severe hemolysis and jaundice with Hemoglobin <5 gm/ dl. All had presence of cold agglutinins in the serum and were positive for direct and indirect coomb's test.

Response

All the patients showed significant improvement in the symptoms due to anemia and there was a rise in hemoglobin detected within the first week of Rituximab treatment. They achieved normal levels of hemoglobin with four to six weeks of treatment and in three patients cold agglutinins disappeared after completion of five cycles and the fourth patients had it after the sixth cycle. None of the patients was given any transfusion because of incompatibility issues.

Conclusion

Our experience with Rituximab was excellent in this indication and patients had a good overall outcome. Further trials need to be done for more evidence in this regard.

Disclosures:

No relevant conflicts of interest to declare.
Prognostic Factors Associated with Progression of Myelodysplastic Syndromes (MDS) to Acute Myeloid Leukemia (AML) In Patients (pts) Treated with Decitabine
Authors: Fullmer Amber , Garcia-Manero Guillermo , Borthakur Gautam , Kadia Tapan , Kantarjian Hagop , Jabbour Elias
Published: October 2019

Abstract 4956

Background:

About 50% of pts with MDS ultimately progress to AML with no clear pattern in underlying parameters leading to progression. This analysis sought to identify predictive factors in pts with MDS that are associated with progression to AML after treatment with decitabine.

Methods:

In a combined analysis of MDS pts treated with either a 3- or 5-day dosing schedule of decitabine, pts were stratified on the basis of those who progressed to AML (P) versus those who did not (NP). CR rates by IWG2006 criteria for each of the baseline factors were compared by using the chi-square test. Factors that were significant in the univariate analysis (p<0.05) were included in a multivariate analysis. Logistic regression analysis was conducted for progression to AML. The following variables were identified as predictors of progression: study effect; age; secondary or de novo MDS; prior MDS treatment, including growth factor use only and prior chemotherapy; del 5Q or 7; time to diagnosis; baseline ANC (< 500, 500 – 1000, or >1000 U/L); baseline HGB, plt and BM blast count ≤20; FAB classification; ECOG status; and IPSS (low, int-1, int-2). A Cox proportional-hazards analysis was conducted for survival.

Results:

A combined analysis of prognostic factors for MDS was completed in 163 pts with a complete response to decitabine; 37 (22.6%) had progressed to AML and 126 (77.3%) had not. Baseline characteristics in pts that progressed had significantly less time since diagnosis (<3mos), more RAEB, RAEB-t and IPSS classification of Int-2 and high-risk, lower baseline HGB levels and fewer prior treatments. From the multivariate analysis, the following factors were selected as predictors of progression: No history of prior treatment, time from diagnosis of MDS <3 months, hemoglobin levels under 10 g/dL, and a 3-day schedule of decitabine (p=0.005, 95% CI 1.4, 6.8). Survival estimates were determined for Del 5q or 7, baseline HGB <10, plt <50 and the 3-day dosing schedule approached but did not achieve a significant effect (p=0.08).

Conclusions:

MDS pts with a deep anemia (HGB <10), thrombocytopenia (plts <50), and a cytogenetic risk profile that includes a del5 and/or 7 anomaly are at a statistically higher risk of transformation into AML and should be considered for additional treatment options.

Disclosures:

Borthakur: Eisai Inc.: Research Funding. Kantarjian: Novartis: Consultancy, Research Funding; Pfizer: Research Funding; Bristol Myers Squibb: Research Funding. Jabbour: Eisai Inc.: Editorial and statistical support, Honoraria.
Establishment and Characterization of a Novel Human Myeloma Cell Line, AMU-MM1, From a Multiple Myeloma Patient Involving Central Nerve System After Treatment with Bortezomib
Authors: Hanamura Ichiro , Goto Mayuko , Nagoshi Hisao , Taki Tomohiko , Imai Norikazu , Suganuma Kazuto , Goto Mineo , Mizuno Shohei , Takahashi Miyuki , Yamamoto Hidesuke , Hiramatsu Akihito , Wakabayashi Motohiro , Watarai Masaya , Shikami Masato , Imamura Akira , Mihara Hidetsugu , Iida Shinsuke , Ueda Ryuzo , Taniwaki Masafumi , Miwa Hiroshi , Nitta Masakazu
Published: October 2019

Abstract 4991

Bortezomib improved the survival in patients with multiple myeloma (MM), however, cannot cure this disease even when it is combined with autologous stem cell transplant(s) and other new drugs. Myeloma cells cannot be excluded completely from a patient due to drug refractoriness and/or inefficiency of drug delivery in extramedullary sites such as cerebrospinal fluid (CSF). Therefore, it is important to clarify the mechanisms of bortezomib resistance and invasion to central nerve system (CNS) of myeloma for the progress of myeloma therapy.

In this study, we established a novel human myeloma cell line from a myeloma patient involved with CNS and analyzed its characters including sensitivity to bortezomib. The patient had been treated with bortezomib for 1.5 years when her CNS was involved with myeloma. The cells from patient's CSF were cultured in RPMI 1640 supplemented with 20% heat-inactivated fetal bovine serum (FBS) and rhIL-6. After 3 months of culture, cell proliferation became continuous with 10% FBS and without rhIL-6. The cell line, named AMU-MM1 was established and negative for EBV. A doubling time of AMU-MM1 cells was about 48 hours. AMU-MM1 cells were positive for CD38, CD54, CD138 and cytoplasmic kappa chain and negative for CD19, CD20, CD33 and CD56 by flow cytometry analysis while those in patient's bone marrow were positive for CD56. AMU-MM1 showed hypo- and pseudodiploid karyotypes with t(4;14), t(8;13), t(1;19), del13q, amp1q21 and others but without del17p by cytogenetic analyses including FISH. The G322A mutation in the proteosome beta 5 subunit (PSMB5) gene, which is reported as a mutation found in bortezomib-resistant cell lines induced via repeated drug selection, was not detected in AMU-MM1 by direct sequencing. Apoptosis analysis using Annexin V/PI assay indicated that AMU-MM1 was sensitive to bortezomib. Our data suggest that AMU-MM1 was derived from the cells that invaded to CSF in which bortezomib concentration was very low and not resistant to bortezomib, and the downregulation of CD56 might play a role in the pathogenesis of CNS involvement as reported before. In addition, we are now focusing on new chimera or dysregulated genes in t(8;13), t(1;19) and other sites as well.

In conclusion AMU-MM1 is a useful cell line for analysis of mechanisms of CNS involvement and also possibly the acquired resistance to bortezomib.

Disclosures:

No relevant conflicts of interest to declare.
Imatinib Treated Chronic Myelogenous Leukemia Concomitant with Multiple Myeloma. Is There a Relationship?
Authors: Kutner Jose Mauro , Bley Claudia , Perini Guilherme Fleury , Santos Fabio , Rodrigues Morgani , Ribeiro Andreza Alice Feitosa , Hamerschlak Nelson
Published: October 2019

Abstract 5005

Chronic myelogenous leukemia (CML) and multiple myeloma (MM) are hematological neoplasms with involvement of distinct hematopoietic cell lineages. The simultaneous occurrence of CML and MM is an extremely rare event. We report the case of a 46-year-old asymptomatic patient who was diagnosed with CML after an abnormal white blood cell count (90×109 u/L) was found on a routine complete blood cell count. Physical examination showed only splenomegaly. At diagnosis, a monoclonal protein peak was observed on electrophoresis (0.82g/dL). Bone marrow (BM) biopsy was characterized by granulocyte hyperplasia without clonal plasmacytosis. Karyotype showed t(9;22)(q34.1;q11.2) and a BCR/ABL mRNA transcript was detected by RT-PCR. Cytoreduction was initiated with hydroxyurea for 20 days until imatinib 400 mg/day was initiated. Patient achieved a major molecular response (MMR) after 7 months of treatment and remains in MMR. After 12 months from diagnosis, the patient persisted with monoclonal peak, and a new BM biopsy showed 15% monoclonal plasma cells, compatible with IgG lambda Myeloma. No end-organ damage was found, and there was no indication for therapy. Karyotype was 46, XX and a FISH analysis was positive for IgH rearrangement and for del 13q. At the time of this report the patient persists in MMR for CML with a monoclonal peak of 1.2 g/dL. No evidence of MM progression has been observed. These findings indicate coexistence of CML in chronic phase and monoclonal gamopathy of uncertain significance (MGUS), progressing to smoldering MM IgG lambda during IM treatment. At this moment, the association of CML and MM has been considered coincidental, but it is possible that imatinib may induce changes in phenotype of plasma cells which might lead to progression of multiple myeloma.

Disclosures:

No relevant conflicts of interest to declare.
NT-ProBNP and VEGF Serum Evaluation In AL Amyloidosis
Authors: Saltarelli Francesca , Moscetti Alessandro , Bianchi Maria Paola , Bruno Guglielmo , Salerno Gerardo , Corsetti Maria Teresa , Cardelli Patrizia , Ferranti Giovanna , La Verde Giacinto
Published: October 2019

Abstract 5008

AL amyloidosis is a plasmacellular dyscrasia in which fibrillary deposits containing monoclonal immunoglobuline light chains infiltrate tissues causing their dysfunction and failure. Amyloidosis could be classified into rare localized or more frequent systemic forms such as AL amyloidosis. Cardiac dysfunction is a very frequent feature in AL amyloidosis patients. To evaluate heart involvement both in systemic and localized AL amyloidosis, serum levels of pro-BNP (peptide NT-proBNP; 76 amino acids) and VEGF were investigated. NT-proBNP has been described as an useful marker for heart dysfunction and its role as a prognostic factor for patients with systemic AL amyloidosis has been validated. VEGF is a signaling protein that stimulates new blood vessel formation and showed to be a mitogen for vascular endothelial cells. Serum NT-proBNP and VEGF levels were performed in systemic or localized AL amyloidosis patients to evaluate if there is any correlation between them. Blood samples were collected from 8 patients with systemic amyloidosis and from 4 patients with localized amyloidosis. To analyze the results of NT-proBnp and VEGF, Mann-Whitney test and Spearman correlation were performed. Serum NT-proBNP values were significantly increased in systemic disease (p=0.004). VEGF serum levels were also significantly higher in patients with systemic disease compared to the other group (p=0.027). No significant correlation between NT-proBNP and VEGF levels was found. We know that proBNP levels are significantly increased in patients with systemic amyloidosis and cardiac involvement. On the other hand, VEGF serum levels resulted to be increased in systemic disease. Nevertheless, on the basis of our results, we found absence of a significant correlation between NT-proBNP and VEGF increases. In our opinion, such finding could suggest that VEGF based neoangiogenesis is not significantly involved in AL amyloidosis heart dysfunction.

Disclosures:

No relevant conflicts of interest to declare.