bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of High Dose Chemotherapy/Autologous Stem Cell Transplantation for Salvage Treatment for Relapsed Peripheral T Cell Lymphoma.
Authors: Cho Yoojin , Kang Myoung Joo , Choi Dae Ro , Kim Eun Kyoung , Kim Shin , Lee Dae Ho , Kim Sang We , Huh Jooryung , Lee Jeong Shin , Suh Cheolwon
Published: October 2019

Abstract 4363

Introduction

Although the role of high dose chemotherapy (HDCT) and autologous stem cell transplantation (ASCT) in the treatment of aggressive lymphoma has been established, optimal second line treatment, including high-dose therapy with autologous stem cell support, is not well established in patients with peripheral T cell lymphoma (PTCL). The purpose of this retrospective study was to compare the outcome of salvage chemotherapy only with that of HDCT and ASCT in patients with relapsed disease after having reaching a complete response (CR).

Methods

We retrospectively investigated the outcome of salvage treatments for patients with relapsed PTCL who had reached a CR.

Results

Between January 1999 and February 2009, 39 patients (15 peripheral T cell lymphoma; 14 extranodal natural killer/T cell lymphoma; 5 anaplastic large cell lymphoma; and 6 angioimmunoblastic T-cell lymphoma) were identified to be eligible for this analysis in our institution. Among them, 25 were treated with only salvage chemotherapy and 14 were treated with HDCT/ASCT. The salvage chemotherapy regimens were ESHAP given in 52% and DHAP given in 12%. Also the conditioning chemotherapy regimens for autologous stem cell transplantation (ASCT) were ESHAP given in 50% and DHAP given in 14.2%. The median number of chemotherapy regimen was 3. The overall response rate of salvage chemotherapy was 92% with a CR rate of 50%. With median follow-up time of 20.7 months, there were no statistical differences between the two groups in terms of progression-free survival and overall survival (p=0.794, and p=0.390, respectively).

Conclusion

There were no significant differences between the two groups in terms of survival outcome. Therefore, considering the dismal outcome in relapsed PTCL patients even after treated with HDCT/ASCT, incorporation of novel therapeutics into a treatment regimen should be considered.

Disclosures:

No relevant conflicts of interest to declare.
Primary Granulocytic Sarcoma:A Allo-Transplantation Case Report and Literature Review.
Authors: Li Xiaolin , Fu Jie , Xue Yan , Yang Yang , Wang Yang , Zhang Cixian
Published: October 2019

Abstract 4368

Objective

To investigate the effect of allogeneic transplantation in treating primary granulocytic sarcoma.

Methods

We report the diagnosis and treatment of one patient with primary granulocytic sarcoma and review the related articles.

Results

Firstly appeared as one isolated tumor in breast, the disease was diagnosed by pathological methods with immunochemistry positive stain in MPO, CD68(KP-1) and bacteriolysis enzyme. After radical operation on sick breast, the patient was treated with four courses chemotherapy according to that of acute granulocytic leukemia,then followed by matched sibling transplantation. After 6 months follow-up, the patients is still in complete remission.

Conclusion

If having suitable donor, patient with primary granulocytic sarcoma shoud be treated with allogeneic transplantation.

Disclosures:

No relevant conflicts of interest to declare.
Spontaneous in Vitro Death of CLL Cells Can Be Prevented by Mitochondrial Stabilization Via CD160 Signaling.
Authors: Liu Feng-Ting , Jia Li , Farren Timothy , Giustiniani Jerome , Bensussan Armand , Gribben John G. , Agrawal Samir G
Published: October 2019

Abstract 4372

B-cell chronic lymphocytic leukemia (CLL) is an incurable disease, which is at least partly attributable to the majority of cells being in the G0/G1 phase of the cell cycle and expressing high levels of anti-apoptotic Bcl-2 family proteins. Despite their prolonged survival in vivo, CLL cells rapidly undergo spontaneous apoptosis in vitro, suggesting that survival signals in vivo have been lost in in vitro culture conditions. CD160, a glycosylphosphatidylinositol-linked surface antigen, was found to be expressed by CLL cells. In normal NK and T-cells, CD160 mediates cellular growth and activation, but its role in CLL is unclear. Using monoclonal antibodies to CD160 (CL1-R2 or BY55 - non cross blocking) led to increased expression of Bcl-2, Bcl-xL and Mcl-1 anti-apoptotic proteins and protected CLL from spontaneous apoptosis in vitro - mean cell viability increased from 66.8 to 79.4% (n = 17, p = 0.02). These CD160-mediated events were also accompanied by decreased cytochrome C release and prevention of mitochondrial membrane potential collapse, indicating stabilization of both inner and outer mitochondrial membrane integrity. PI3K/AKT signalling is a well known survival pathway in cancer cells and in normal lymphocytes CD160 has been shown to act via PI3K/AKT. Activation of CD160 in CLL led to phosphorylated AKT, while inhibition of PI3K by wortmannin completely blocked AKT phosphorylation and CD160-mediated protection from apoptosis.

In summary, the activation of CD160 protected CLL cells from spontaneous cell death in vitro via a PI3-kinase/AKT pathway. This improved survival was also associated with increased Bcl-2, Bcl-xL and Mcl-1 expression and preservation of mitochondrial function.

Disclosures:

No relevant conflicts of interest to declare.
Vascular Endothelial Growth Factor (VEGF) Acts Via Auto- and Paracrine Mechanisms as a Critical Microenvironmental Factor for the Survival of Chronic Lymphocytic Leukemia (CLL) Cells.
Authors: Gehrke Iris , Paesler Julian , Gandhirajan Rajesh Kumar , Razavi Regina , Poll-Wolbeck Simon Jonas , Filipovich Alexandra , Uhrmacher Sabrina , Erdfelder Felix , Hertweck Magdalena , Hallek Michael , Kreuzer Karl Anton
Published: October 2019

Abstract 4376

The major pathophysiological characteristic of chronic lymphocytic leukemia (CLL) is the accumulation of malignant, immuno-incomptetent B-cells, which is commonly known as the apoptotic block. Recently, it was speculated that vascular endothelial growth factor (VEGF) might be involved in this process. Whereas most of the available studies are solely descriptive, functional data are largely missing. The aim of this study was therefore to describe the effect and functional background of VEGF on CLL cells for the identification of potential strategies for a targeted CLL therapy. We identified an autocrine VEGF feedback loop in CLL cells, but not healthy B-cells. Recombinant human (rh) VEGF stimulation lead to increased expression of the anti-apoptotic proteins Mcl1 and XIAP, but was not sufficient to prolong survival. When CLL cells were cocultured with the bone marrow derived stromal cell line HS5, survival was significantly enhanced 28.5 ± 6.5% after 72h relative to monoculture, whereas healthy B-cells did not profit from coculture (-0.5 ± 4.7%). HS5 cells produce and secrete high amounts of VEGF themselves. When cocultured with HS5 CLL cells also showed an increased VEGF expression, indicating the existence of a paracrine VEGF feedback loop. The actual relevance of VEGF in the coculture mediated survival support was proofed by siRNA experiments. When VEGF expression and secretion was downregulated in HS5 cell by siRNA, the survival advantage CLL cells obtained from the coculture was reduced down to that in monoculture. Also the addition of a VEGF-neutralizing monoclonal antibody largely reversed the survival supporting effect of the coculture, clearly indicating the critical role of VEGF in bone marrow stromal cell-mediated CLL cell survival. We further found rhVEGF to induce upregulation and activation of STAT3 via Tyr705-phosphorylation and a subsequent expression of STAT3 targets such as Cyclin D1 and BclXL. VEGF-stimulation further induced downregulation of the tumor suppressor RB1 and E2F1, which act as proapoptotic factors. Vice versa inhibition of VEGF by using selective VEGF-R inhibitors reduced both, Tyr705 phosphorylation and expression of STAT3 target genes. We therefore, propose a dual mechanism of VEGF-mediated CLL cell survival support via upregulation of the potent oncogene STAT3 and downregulation of the tumor suppressor RB1/E2F1. Hence, VEGF might function as a potential new therapeutic target to overcome the apoptotic block in CLL cells.

Disclosures:

Hallek: BayerScheringAG: Honoraria, Research Funding.
Chromosome Study On Chronic Lymphocytic Leukemia Using CpG- Oligodeoxynucleotide as Immunostimulant Agent.
Authors: Wu Yafang , Xue Yongquan , Yao Li , Jiang Hui , Zhang Jun , Shen Juan , Pan Jinlan , Wang Yong , Bai Shuxiao , Chen Suning
Published: October 2019

Abstract 4383

Objective

To verify whether CpG-oligodeoxynucleotide (CpG-ODN) can raise the detection rate of the karyotypic abnormalities in chronic lymphocytic leukemia (CLL).

Methods

The bone marrow (BM) or peripheral blood (PB) cells from 57 cases of CLL were collected and cultured with CpG-ODN DSP30+interleukin-2 (IL-2), phytohemagglutinin (PHA), pokeweed(PWM) or IL-2, respectively. 5 days later cells were harvested for chromosome preparation. Karyotypic analysis was performed using R banding technique. Panel fluorescence in situ hybridization (FISH) was carried out on 19 cases of CLL with normal karyotype using following probes: Cen12, D13S25, Rb1, ATM, P53, MYB and IgH. Genomic DNA from 21 cases of them was extracted from BM or PB. The immunoglobulin variable heavy chain (IgVH) were amplified by polymerase reaction (PCR) and sequenced. CD38 and ZAP70 expressions in leukemic cells were determined by flow cytometry (FCM).

Results

The detection rate of karyotypic abnormalities in CpG-ODN+IL-2 group (43.85%) was obviously higher than PHA (15.09%), PWM (17.31%) and IL-2 (3.13%) groups (P<0.01). 52 karyotypic abnormalities were found. Among them, trisomy12 (+12) or +12 with other abnormalities were the most common, while translocations were the most frequent structural abnormalities including 3 unbalanced and 11 balanced translocations among which 7 had 14q32 rearrangements. 13 cases showed one or more abnormalities on FISH examination including trisomy12 and P53 deletion each in one case, IgH rearrangement and partial deletion each in one case, 13q14.3 deletion in 11 cases of which 5 cases also had Rb1 deletion, one case had Rb1 partial deletion. No cases with ATM or MYB deletions were found. IgVH mutation was detected in 10/21 cases by PCR and sequencing. FCM showed 10/45 cases expressed CD38, while 11/27 cases expressed ZAP70. Among 26 cases examined for CD38 and ZAP70 expressions simultaneously, 5 cases were CD38+ZAP70+, 13 cases CD38-ZAP70-, 6 cases CD38-ZAP70+, 2 cases CD38+ZAP70-, respectively. Statistics disclosed a correlation between complex karyotype and IgVH without mutation, but no association between karyotype and CD38 or ZAP70 expression.

Conclusion

CpG-ODN immunostimulation can obviously raise the detection rate of abnormal karyotypes, especially balanced and unbalance translocations in CLL. FISH is important complement to conventional karyotypic analysis, the combination of both methods can provide more comprehensive genetic information for CLL.

Disclosures:

No relevant conflicts of interest to declare.
TNF-α and MDM2 Polymorphism in Pathogenesis of Chronic Lymphocytic Leukemia.
Authors: Bakirov Bulat , Warshavsky Anton , Sgibneva Anastasiya , Karimov Denis
Published: October 2019

Abstract 4390

Introduction

Apoptosis dysfunction one of the main process in pathogenesis of CLL. One of the apoptosis regulators is cytokine TNF- α. In normal condition in cells expressed protein p53 and main tumor suppressor MDM2. The high expression of protein MDM2 is oncogenic factor. The gene of protein MDM2 placed on the long arm 12 chromosome (12q14.3-12q15). Our aim was studying of frequencies of polymorphic variants of genes TNA-α and MDM2.

Patients and Methods

We observed 133 patients with CLL (male-75, female-58). The median age was 49. The control group was 196 healthy donors from the same region, same ethnic group, age 47 (male-107, female-89).

Results

In patient group in polymorphic locus -308G>A gene TNFα genotype GG met often than in control group (82% and 65% accordingly, χ2=10,674; p=0,002). On the contrary genotype GA authentically often met in group of healthy individuals (33 %) whereas its frequency in group of CLL patients was 17 % (χ2=8,837; p=0,004). Frequency of allele G a polymorphic locus-308G> A gene TNF-α in group of CLL patients it is authentic more than in control group (91 % and 81 % accordingly, χ2=10,411; p=0,002). Distinctions also were observed on frequencies of allele A, in control group 19 % and 9 % and in the group of CLL patients (χ2=10,411; p=0,002). On frequencies of genotype AA of authentic distinctions it is not revealed. Thus, as a result of research it has been shown that markers of risk of CLL development are genotype GG (OR=2,468; 95 % CI 1,452-4,194) and allele G (OR=2,244; 95 % CI 1,384-3,638) of polymorphic locus-308G> A gene TNF-α. Opposite stability markers are genotype GA (OR=0,432; 95 % CI 0,252-0,741) and allele A (OR=0,446; 95 % CI 0,275-0,724). The analysis of distribution of frequencies of a polymorphic locus 309T> G gene MDM2 in patients group and group of healthy individuals has shown statistically authentic distinctions, on allele T, frequency in group of CLL patients was 54 % and in group of healthy individuals was 63 % (χ2=4,141; p=0,042; OR=0,679; 95 % CI 0,475-0,972). Also in allele G 46 % in patient group against control group - 37 % (χ2=4,141; p=0,042; OR=1,474; 95 % CI 1,03-2,108).

Conclusions

Thus, the cited data of researches testifies to the important role of genes TNF α, MDM2 in CLL development. In particular genotypes GG and allele G in a polymorphic locus-308G> A gene TNF-α and allele G in a polymorphic locus 309T> G gene MDM2 have appeared as markers of the raised risk in CLL development. On the contrary genotype GA and allele A in a polymorphic locus-308G> A gene TNF α and allele T in a polymorphic locus 309T> G gene MDM2 are the markers of the raised stability to CLL development.

Disclosures:

No relevant conflicts of interest to declare.
MK886 Induced Apoptosis in HL-60 Cells Via Inhibition mPGES-1 Expression and Down-Regulation Prostaglandin E2 Synthesize.
Authors: Li Yiqing , Yin Songmei , Xie Shuangfeng , Nie Danian , Ma Liping , Wang Xiuju , Wu Yudan
Published: October 2019

Abstract 4824

Recent studies have shown that prostaglandin E2 (PGE2) may play a key role in the tumorigenesis and tumor development. Membrane-bound prostaglandin E2 synthase-1 (mPGES-1), an inducible enzyme that acts downstream of cyclooxygenase (COX) and specifically catalyzes the conversion of prostaglandin H2 (PGH2) to PGE2, was over-expression in a variety of solid tumor cells and tissues such as nonsmall-cell lung cancer, colon carcinoma, gastric carcinoma and breast cancer. MK886, a small molecular inhibitor, is a reasonable potency as an inhibitor of mPGES-1 in vitro experiment. In this study, we examined effects of MK886 on expression of mPGES-1 and PGE2 synthesis in human acute myeloid leukemia cell line (HL-60), observed cell proliferation and apoptosis after 24-h treatment with MK886, and tried to explore the possible mechanisms by checking some protein belong AKT cell singling pathway such as P-AKT, Bax and Bcl-2. We found that the expression levels of mPGES-1 mRNA and protein were higher in HL-60 cells than in normal mononuclearcells (MNC). MK886 inhibited mPGES-1 mRNA and protein expression and reduced PGE2 secretion in HL-60 cells in a dose-dependent manner. The cell proliferation was inhibited and the IC50 was 132.16μmol/L. With the increase of MK886 concentration, the cell apoptosis rate assayed by flow cytometry increased and the apparent apoptotic bodies increased when staining by Hoechst 33258. After treated with MK886 for 24h, protein was extracted and assayed by western blot. The results showed that the expression levels of P-AKT, Bcl-2 and c-myc decreased while the Bax protein expression increased in a dose-dependent manner. The caspase-3 activity, determined by colorimetric detection, also increased dose-dependently. These results indicated that mPGES-1 over-expressed in leukemia cell line HL-60, MK886 could induce apoptosis in HL-60 cells via reducing mPGES-1 expression and PGE2 synthesis dose-dependently, thereby regulate the AKT pathway including Bcl-2 family and the activity of caspase-3. It suggested that mPGES-1 inhibitor might emerge as an important therapeutic tool for leukemia treatment.

Disclosures:

No relevant conflicts of interest to declare.
Aspirin Induces Apoptosis in Human Leukemia Cells Independently of NF-κB and MAPKs through Alteration of the Mcl-1/Noxa Balance.
Authors: Cosialls Ana M , Iglesias-Serret Daniel , Piqué Maria , Barragán Montserrat , Santidrián Antonio F , González-Gironès Diana M , Coll-Mulet Llorenç , de Frias Mercè , Fernandez de Sevilla Alberto , Pons Gabriel , Gil Joan
Published: October 2019

Abstract 4825

Aspirin and other non-steroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in most cell types. We examined the mechanism of aspirin-induced apoptosis in human leukemia cells. Our results show that aspirin induced apoptosis in leukemia Jurkat T cells independently of NF-κB. Although aspirin induced p38 MAPK and c-Jun N-terminal kinase (JNK) activation, selective inhibitors of these kinases did not inhibit aspirin-induced apoptosis. We studied the regulation of Bcl-2 family members in aspirin-induced apoptosis. The mRNA levels of some pro-apoptotic members, such as BIM, NOXA, BMF or PUMA, were induced by aspirin. However, none of these pro-apoptotic proteins increased and the levels of Mcl-1 protein were reduced. Interestingly, in the presence of aspirin the protein levels of Noxa remained high. This alteration of the Mcl-1/Noxa balance was also found in other leukemia cell lines and primary chronic lymphocytic leukemia cells (CLL). Furthermore, in CLL cells aspirin induced an increase in the protein levels of Noxa. Knockdown of Noxa or Puma significantly attenuated aspirin-induced apoptosis. These results indicate that aspirin induces apoptosis through alteration of the Mcl-1/Noxa balance.

Disclosures

No relevant conflicts of interest to declare.
Reversal of Multidrug Resistance by Magnetic Fe3O4 Nanoparticle Copolymerizating Daunorubicin and mdr1-shRNA Expression Vetor in Leukemia Cells.
Authors: Chen Bao-An , Mao Peipei , Cheng Jian , Gao Feng , Ding Jia-Hua , Gao Chong , Zhang Wei , Xu Cui-Rong , Xu Wen-Lin , Sun Xin-Chen , Chen Ning-Na , Li Xiao-Mao , Wang Xue-Mei
Published: October 2019

Abstract 4829

Object

In many instances, Multidrug resistance (MDR) is mediated by increased expression at the cell surface of the MDR1 gene product, P-glycoprotein (P-gp), a 170-kD energy-dependent efflux pump. The aim of this study was to investigate the potential benefit of combination therapy with magnetic nanoparticle of Fe3O4 (MNP(Fe3O4)) and mdr1-shRNA Expression vetor.in K562/A02 leukemic cells.

Methods

To synthesis short hairpin RNA (shRNA)aiming divectly at the target sequence,we choice the 3491-3509,1539-1557and 3103-3121 nucleotide of mdr-1 mRNA as targets. Cloning in the plasmid vetor PGCSilencer-U6-neo-GFP, The recombinant plasmid vetors were called for PGY1-1,PGY1-2 and PGY1-3.The recombinant plasmid vetors were transfected into the cell 1ines K562/A02 by lipofection. After transfected 48 hours,the inhibition of mdr-1mRNA expression and the expression of P-gp was detected by realtime–PCR and Weston-blot, screening the recombinant plasmid vetor which has the most greatest mdr-1 gene inhibition ratio is PGY1-2.Analysis of the reveral ratio of multidrug resistance, the concentration of DNR and the content of mdr-1 gene and P-gp in K562/A02 cell line.

Results

The combination of daunorubicin (DNR) with either MNP(Fe3O4) or PGY1-2 exerted a potent cytotoxic effect on K562/A02 cells, while MNP(Fe3O4) and PGY1-2 cotreatment can synergistically down regulation the expression of mdr-1 gene and the expression of P-gp(P<0.05). The transfection efficiency was 20%; the concentration of DNR in K562/A02 cell line was obviously elevated (P<0.05);the multidrug resistance index of K562/A02 cell line was obviously decreased (P<0.05).

Conclusion

MNP(Fe3O4) and PGY1-2 cotreatment can synergistically reveral multidrug resistance. Thus our in vitro data strongly suggests a potential clinical application of MNP(Fe3O4) and PGY1-2 combination on CML.

Disclosures

No relevant conflicts of interest to declare.
Over-Expression of IGF-IR in Malignant Clonal Cells in Bone Marrow of Myelodysplastic Syndromes.
Authors: He Qi , Li Xiao , Zhang Zheng , Zhang Qingqiao , Xu Feng , Yang Lianping , Tao Ying , Liu Yizhi
Published: October 2019

Abstract 4832

To explore the expression level of insulin-like growth factor-1 receptor (IGF-IR) in malignant clone cells of myelodysplastic syndromes (MDS).

Method

Fluorescence in situ hybridization (FISH) and immunochemistry (APAAP, alkaline phosphatase anti-alkaline phosphatase) were used together to detect the expression of IGF-IR in the bone marrow cells of 26 MDS patients with known abnormal karyotypes.

Result

The average IGF-IR expression level on the surface of clone cells from the 26 MDS cases was markedly elevated compared to the corresponding level in normal cells (78.2±13.7% vs 14.1±14.0%,P <0.0001). The percentages of malignant clone cells in all 26 MDS cases were significantly correlated with the respective percentages of IGF-IR positive nucleated cells (r = 0.909; P <0.0001). No significant difference in the IGF-IR expression level on the clone cells were observed either between high- and low-risk MDS patients or among MDS patients with different abnormal karyotypes.

Conclusion

IGF-IR might be taken as a marker of clone cells in MDS because of its propensity to cause malignant proliferation.

Disclosures

No relevant conflicts of interest to declare.
Analysis of TCR Vgamma and Vdelta Repertoire and Clonality of T Cells in MDS Patients.
Authors: Geng SuXia , Du Xin , Li Yangqiu , Weng Jianyu , Zhong Liye , Wu Suijing , Guo Rong , Lu Zesheng
Published: October 2019

Abstract 4833

Myelodysplastic syndromes (MDS) are clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis and frequent progression to acute myeloid leukemia. The pathophysiology of these syndromes remains poorly explained. The immune system also seems to contribute to the progressive cytopenias observed in MDS in some cases. We investigated the frequency and the discriminative value of TCR gamma and delta CDR3 clonality using RT-PCR and genescan technique in 40 MDS patients (24 male, 16 female),which included refractory anemia(n=1), refractory anemia with with ringed sideroblasts(n=6), refractory anemia with excess of blasts I (n=16)and II (n=17) respectively. The median age was 60 years (range 15-84).The results showed that 35(87.5%) MDS patients expressed all three TCR Vgamma subfamilies. Four(10%) and one(2.5%) MDS patients expressed two and one TCR Vgamma subfamilies respectively. Clonal expansion of T cells in some Vgamma subfamilies could be identified in 31 patients. The clonal expansion of Vgamma2 subfamily were identified most frequently. All the Vdelta subfamilies were absent in 2 MDS patients. The remaining 38 (95%) MDS cases expressed 0-6 Vdelta subfamilies. Vdelta1 and Vdelta2 were expressed most frequently followed by Vdelta8. While only 3(7.5%) patients expressed Vdelta5 subfamilies. Clonal expansion of Vdelta T cells were found in all MDS patients. The most frequent clonal expansion T cell was Vdelta3. In addition, 12 patients expressed monoclonal T cell Vdelta subfamily. Monoclonal expanded T cells were found in Vdelta3, Vdelta4, Vdelta6, Vdelta7 and Vdelta8 subfamilies. In summary,we found that MDS patients showed TCR Vgamma and delta skew distribution and clonal expansion. The absent of TCR Vdelta subfamily was more evident than TCR Vgamma subfamily. These findings provided further evidence that T cell mediated immune processes were a feature of MDS.

Disclosures

No relevant conflicts of interest to declare.
Polish Myelodysplastic Syndromes Registry- Report of 15 Months of Activity.
Authors: Dwilewicz-Trojaczek Jadwiga , Madry Krzysztof , Stella-Holowiecka Beata , Mital Andrzej , Szmigielska Anna , Obara Agata , Sikorska Anita , Nita Ewa , Mendrek Wlodzimierz , Wiater Elzbieta , Zalewska Magdalena , Biedron Monika , Jachalska Anna , Wasilewska Ewa , Wojciechowska Malgorzata , Kopacz Agnieszka , Nowicki Adam , Kolkowska Agnieszka , Guzicka-Kazimierczak Renata , Pedziwiatr Maria , Sedzimirska Mariola , Nowak Ewa , Blasiak Anna , Baranska Ewa , Soroka-Wojtaszko Maria , Gornik Slawomir , Kruger Witold , Katinas Katarzyna , Salamanczuk Zoriana , Sokolowska Bozena , Jedrzejczak Wieslaw W. , Deptala Andrzej
Published: October 2019

Abstract 4848

Background

Myelodysplastic syndrome(MDS) belong to the most common hematological diseases however epidemiological data on MDS are sparse. Until 2008 there were no data about epidemiology of MDS in Poland.

Methods

From 03.2008-05.2009 we have registered 966 patients in Polish MDS Registry. We have included only alive patients of various time of diagnosis. Patients from 22 centers were diagnosed according to WHO 2001 criteria.

Results

There were 508(53%)males and 458(47%) females. Median age at diagnosis was 70(range 19-99). Under 50 were 83(9%) cases with preponderance of females- 51 cases( males 32cases), between 50-70 there were 353(41%) cases, half of the patients-432(50%) were above 70( 247 males and 185 females).Prior chemotherapy and/or radiotherapy had 37((3,8%) patients. Distribution of MDS subtypes was as follows: RA-170(20%) cases, RARS-58(7%), RCMD-244(28%), RCMD-RS-18(2%), RAEB-1-120(14%), RAEB-2-169(19,5%), 5q- -40(4,6%), MDS-U-44(5%).In 103(10%) subtype was not done. Karyotype was available in 276(28%) cases. Cytogenetic risk groups were: low risk-182(68%), intermediate-52(20%) and high risk-33(12%). The most frequent cytogenetic results were: normal karyotype 44%, isolated 5q deletion 19%, complex karyotype 6%, 5q deletion + another one change 3% and 5q deletion with at least 2 changes 3%. According to IPSS risk groups low risk was found in 61( 22%) of cases, intermediate-1 -130(48%), intermediate-2-47(17%) and high risk in 31(11,5%). Median values of Hb was 9,1 g/dL, plts 129 G/L, ANC 1,7 G/L. RBC transfusion dependent were 429(44%) patients and platelet transfusion dependent were 100( 11%) pts. At least 2 U/month RBC transfusion requirement was 140(14%) patients. Serum ferritin level was assessed in 530 cases-171 of them( 32%) had higher than 1000μg/L level.

Conclusions

We have observed predominance of females among MDS patients under 50. Half of the patients had RA or RCMD subtype. Isolated 5 q deletion was the most frequent cytogenetic abnormality. Forty four percentage of patients was RBC transfusion dependant. Serum ferritin level was significantly elevated in 32% of assessed patients at the moment of MDS diagnosis.

Disclosures

No relevant conflicts of interest to declare.
Study On the Clonal Origin of Dysplastic Cells by Fluorescence in Situ Hybridization in Myelodysplastic Syndromes.
Authors: Zhang Jun , Xue Yongquan , Pan Jinlan , Wu Yafang , Shen Juan , Wang Yong , Liu Dandan , Chen Suning
Published: October 2019

Abstract 4858

Objective

To determining the clonal origin of dysplatic cells in Myelodysplastic syndromes (MDS) .

Methods

Karyotypic analyses of bone marrow cells using R-banding technique were carried out to determine the chromosomal abnormalities. Interphase fluorescence in situ hybridization (FISH) and morphologic analysis of bone marrow aspirates were performed in the same cells to investigate the clonal origin of dysplatic cells in 8 MDS patients.

Result

All patients had clonal karyotypic abnormalities: simple abnormality in 1 patient, complex abnormalities in 6 patients, coexistent of two unrelated clones in 1 patient. Most of dysplastic cells in 7 of 8 MDS patients derived from neoplasia clone while 1 patient had a reverse result,no matter what cell lineage was involved. Some of non-dysplastic cells of all patients derived from malignant clone; in 7 patients, the proportion of dysplastic cells in malignant clone were significantly higher than that of non- malignant clone.

Conclusion

Most of dysplastic cells in MDS derived from malignant clones, while the minority of them derived from non-malignant clones. Thus, it is reasonable to expect that in most cases myelodysplasia is present in malignant clone and can be taken as an important diagnostic evidence for MDS.

Disclosures

No relevant conflicts of interest to declare.
MD.
Authors: Gul Zartash , Galili Naomi , Tageja Nishant , Raza Azra
Published: October 2019

Abstract 4863

THE SECONDARY TRANSFORMATION OF MDS INTO MYELOFIBROSIS IS INDEPENDENT OF THE JAK2 MUTATION

Introduction

The exact significance of the JAK2 V617F mutation in MDS is unclear. It has previously been suggested that MF could be associated with MDS as one phenotype of myeloproliferative disorders. Ohyashiki et al in 2005 showed that two of six patients with MDS terminating in MF had the mutation at the time of MF, while no MDS patient without MF had the JAK2 V617F mutation, suggesting that MDS patients with the JAK2 V617F mutation may be responsible for secondary MF in MDS patients. We conducted the following experiments to further probe the role of JAK2 mutation in the pathology of MDS with myelofibrosis at our institution.

Methods

DNA samples of 8 patients in our institution who had MDS with subsequent myelofibrosis were isolated. Genomic DNA isolated using DN easy tissue kit(Qiagen) was amplified by PCR. After confirmation of DNA presence on 1 % agarose gel and purification using QIA quick PCR purification kit (Qiagen), gene sequencing was done. This sequencing was then analyzed for G to T mutation V617F in JAK2.

Results

We found no evidence of the JAK2 mutation in any of the eight samples.

Discussion

Given the fact that nearly half the patients with myelofibrosis carry the JAK2 V617F mutation, the possibility of an association between those patients who have myelodysplasia and subsequent myelofibrosis needs to researched. Our cohort which consisted of 8 patients showed no evidence for any such correlation. This evidence confirms the presence of different pathophysiological mechanisms for phenotypic expression of MDS and primary MF and also leads us to suggest that the expression of JAK2 is not the driving mutation in the pathogenesis of MDS progression to myelofibrosis.

Disclosures

No relevant conflicts of interest to declare.
Pulmonary Alveolar Proteinosis as a Terminal Complication in a Case of Myelodysplastic Syndrome with Idic(20q-).
Authors: Xue Yongquan , Han Yongsheng , Li Tianyu , Chen Suning , Zhang Jun , Pan Jinlan , Wu Yafang , Wang Yong , Shen Juan
Published: October 2019

Abstract 4864

Secondary pulmonary alveolar proteinosis (PAP) is a rare lung disease which was reported in 13 cases of myelodysplastic syndromes (MDS). A dicentric isochromosome of deleted chromosome 20q i.e., idic(20q-) is a newly recognized rare but recurrent cytogenetic anomaly which has been described in thirty cases of MDS. Recently we encounted an interesting MDS patient with idic(20q-) and secondary PAP. At presentation she was a forty-year-old women manifesting pancytopenia and dysplasia involving two cell lineages which were compatible with refractory cytopenia with multilineage desplasia. Chromosome analysis using R-banding technique showed a karyotype of 46,XX,-20,+a small metacentric chromosomal marker. FISH analysis proved this marker to be a dicentric isochromosome of a deleted 20q. Three yaeas after presentation her disease was complicated by secondary PAP which was confirmed by chest computed tomographic scan and the thoracoscopic lung biopsy which revealed the characteristic periodic acid Schiff stain-positive materials filling the alveoli. After prednisolone and intibiotics freatment, her symptoms had temporary improvement. At last she died of respiratory failure with a survival of 45 months. As we know, no MDS patients with idic(20q-) and secondary PAP have been reported in the literature. Thus, this patient is not only the thirty-first MDS case with idic(20q-), but also the first MDS case with idic(20q-) and secondary PAP.

Disclosures

No relevant conflicts of interest to declare.
Analysis of Familial Dysglobulinemia: a Multicenter IFM Study.
Authors: Dumontet Charles , Galia Perrine , Leleu Xavier , Avet-Loiseau Herve , Sobol Hagay
Published: October 2019

Abstract 4891

Multiple myeloma is a haematological malignancy which rarely occurs in a familial context. MGUS corresponds to the existence of a clone secreting a monoclonal immunoglobulin and can in some patients evolve to multiple myeloma. Although recurrent cytogenetic abnormalities have been described in tumor cells of sporadic cases of multiple myeloma, there are presently no identified genetic mechanisms which contribute to myelomagenesis, in particular in familial cases. The identification and study of families with several cases of dysglobulinemia should contribute to the determination of genetic factors predisposing to multiple myeloma. The aim of this study is to identify genetic factors predisposing to familial dysglobulinemia, thanks to the establishment of a DNA bank. Peripheral blood samples were collected both from patients with dysglobulinemia and from healthy family members and lymphoblastoid lines established for further studies. For this purpose, we have initiated a multicenter study aiming to identify families with several cases of dysglobulinemia, including both multiple myeloma and/or Monoclonal Gammapathy of Undetermined Significance (MGUS). This effort is supported by a French National Programme Hospitalier de Recherche Clinique (PHRC) and the French National Institute for Cancer Research (INCa)., and involves the French multicentric Myeloma groups, Intergroupe Francophone du Myélome (IFM) and Myeloma Autograft Group (MAG), in coordination with the Réseau des Hémopathies Familiales. This study was activated in Octobrer 2007 and as of April 2009, 72 families with at least two cases of dysglobulinemia (median 2, range 2-5) have been identified. Among 166 cases of dysglobulinemia, 100 were diagnosed as multiple myeloma, 49 as MGUS and the remainder as light chain amyloidosis or Waldenstrom's disease. The sex ratio was 1:1 for all groups. The median range at inclusion in this study was 64 years (range 33-85 years). The heavy chain subtype distribution was classical, with a majority of IgG cases. 42% of cases corresponded to cases involving parents and offspring and 54% corresponded to cases involving siblings. This study is ongoing in order to collect samples from 100 families and to initiate genetic analyses to identify predisposing factors.

Disclosures

No relevant conflicts of interest to declare.
Effect of Artesunate On Inhibiting Proliferation and Inducing Apoptosis of SP2/0 Myeloma Cells through Affecting NFκB p65.
Authors: LI Shihui , PAN Ling , Xue Fang , Cheng Zhiyong , Yang Xiaoyang , Guo Xiaoling , Wang Suyun
Published: October 2019

Abstract 4893

The initial treatment of multiple myeloma (MM) experienced a paradigm shift, in the past decade, with the introduction of novel agents such as thalidomide, lenalidomide and bortezomib, leading to improved outcomes. High dose therapy and autologous stem cell transplantation remains an important therapeutic option for patients with multiple myeloma eligible for the procedure. However, most of these treatment regimens are too expensive for Chinese patients. Therefore, we investigated the effects of artesunate, which is commonly used in the treatment of severe malaria, on inhibition of proliferation and induction of apoptosis of a mouse myeloma cell line SP2/0. The growth inhibition of SP2/0 cell proliferation induced by artesunate (ART) treatment was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and the rate of apoptosis and cell cycle changes induced by ART were analyzed by flow cytometry. ART induced morphology changes of apoptosis in SP2/0 cells, as observed by light and transmission electron microscopy. Additionally, DNA laddering, which is a hallmark of apoptosis, was observed by agarose gel electrophoresis of DNA harvested from SP2/0 cells treated with ART. The levels of nuclear factor kappa B p65 (NFκB p65) protein in nucleus and the inhibitor of NFκB (IκBα) in the cytoplasm were measured by western blot analysis and ELISA to evaluate NFκB p65 transcription activity indirectly. The results show that artesunate inhibited the proliferation and induced apoptosis of SP2/0 cells in a dose- and time-dependent manner. Artesunate also increased the proportion of SP2/0 cells in G0/G1 phase, while decreased the proportion of cells in G2/M or S phase. Additionally, artesunate treatment decreased the level of NFκB p65 protein in the nucleus, while increased the level of IκBα protein in the cytoplasm. The present result is the first report to show that artesunate may be useful in the treatment of multiple myeloma.

Disclosures

No relevant conflicts of interest to declare.
Three-Month Cumulative Incidence of Thromboembolism and Bleeding After Periprocedural Anticoagulation Management of Patients with Active Cancer.
Authors: Tafur Alfonso J , Wolny Edyta , McBane Robert , Sutkowska Edyta , Slusser Joshua P , Hodge David O , Heit John A. , Wysokinski Waldemar E
Published: October 2019

Abstract 491

Introduction:

Chronically-anticoagulated patients with active cancer (Ca) often require temporary interruption of warfarin for an invasive diagnostic or therapeutic procedure. While such patients may be at increased risk for periprocedural thromboembolism and bleeding, there are virtually no estimates of these outcomes among Ca patients or the most appropriate periprocedural anticoagulation management. The aims of this study were to estimate rates of these outcomes among Ca patients and to test active cancer and “bridging” low molecular weight heparin (LMWH) as potential predictors of these outcomes.

Patients and Methods:

In a retrospective cohort study, all chronically-anticoagulated patients referred to the Mayo Clinic Thrombophilia Center for periprocedural anticoagulation management over the 11-year period, 1997-2007, were followed forward in time for the outcomes of venous and arterial thromboembolism (VTE; ATE), major bleeding and vital status within 3 months of consultation. Warfarin was stopped 4-5 days prior to the procedure and patients received bridging LMWH according to the estimated risk of thromboembolism and bleeding. All outcome events were centrally adjudicated using pre specified criteria.

Results:

The total cohort (n=2517) included 500 patients with Ca. Chronic anticoagulation indications included prior VTE (n=1033), mechanical heart valve (n=669), atrial fibrillation (n=530) and other (mainly vascular bypass; n=285). 65% and 64% of patients with and without Ca received bridging LMWH, respectively. Prior VTE was more common among Ca patients compared to those without CA, both in the bridged (56% vs 37%; p<0.001) and the non-bridged (48% vs 39%; p<0.05) groups. The 3 month rate for the composite of VTE, ATE or bleeding was higher among Ca patients compared to patients without Ca (5% vs 2%; p=0.004) due to higher post procedure rates of VTE (1% vs 0.2%; p<0.001) and major bleeding (4% vs 1.7%; p=0.009); the post procedure ATE rates were low in both groups (0.40% vs 0.45%). For the entire cohort, patients receiving bridging LMWH had a significantly higher rate of post procedure bleeding (5% vs 1%; p<0.05). Finally, there were significantly more deaths in Ca patients compared to non-Ca patients (5% vs. 1%; p<0.001).

Conclusions:

The three-month rates of VTE, major bleeding and death among patients with Ca in whom anticoagulation is temporarily interrupted for an invasive procedure is significantly higher than in patients without Ca, especially the major bleeding rate. Use of heparin increases this rate even farther, suggesting that bridging should be used with caution in patients with active cancer.

Disclosures:

No relevant conflicts of interest to declare.
GCS-100 Targets Apoptosis and Cell Cycle Regulation in Myeloma Cells.
Authors: Streetly Matthew , Maharaj Lenushka , Joel Simon P , Cotter Finbarr E
Published: October 2019

Abstract 4923

Introduction

Myeloma is characterised by abnormalities of multiple cellular pathways that result in apoptosis resistance, aberrant signalling and deregulated cell cycle. GCS-100 is a modified citrus pectin that has demonstrated galectin-3 inhibitory activity. It has recently been reported to have clinical activity against CLL (Cotter et al. ASCO 2009) and has previously been observed to have pre-clinical anti-myeloma effect even in the context of bortezomib resistance (Chauhan et al. Canc Res 2005). We have elucidated the mechanisms of action of GCS-100 in the context of myeloma cell biology.

Methods

The effects of GCS-100 on myeloma cell proliferation, cell cycle, apoptosis induction, cell death and cell signalling were examined in representative cell lines and primary myeloma cells. Cell viability assay, flow cytometric assessment of apoptosis, DNA content, mitchochondrial transmembrane potential and immunoblotting assessment of protein expression were assessed following GCS-100 exposure.

Results

Exposure of RPMI8226, U266 and OPM2 myeloma cell lines to GCS-100 confirmed that it inhibits proliferation and induces apoptosis with activation of both caspase-8 and -9 pathways. GCS-100 exposure was also associated with accumulation of cells in sub-G1 and G1 phases of cell cycle and a dose and time dependent loss of mitochondrial potential. Primary myeloma cells treated with GCS-100 had reduced viability both in isolation and in a stromal cell co-culture model. Examination of key anti-apoptotic and pro-apoptotic proteins revealed that Mcl-1 and Bcl-xL levels were reduced by GCS-100 and this was accompanied by a rapid induction of pro-apoptotic Noxa. Bcl-2, Bax, Bak, Bim, Bad, Bid and Puma remained unchanged. Pan-caspase inhibition abrogated Mcl-1 but not Bcl-XL reduction and inhibited loss of mitochondrial transmembrane potential. GCS-100 treatment also upregulated the cell cycle inhibitor p21Cip1 with concurrent reduction of the procycling proteins cyclin E2, cyclin D2 and CDK6. Furthermore, there was a reduction in signal transduction in the form of reduced activated IκBα, IKK and Akt as well as prevention of upregulated IκBα and Akt following stimulation with appropriate cytokines suggesting a potential microenvironment effect.

Conclusion

GCS-100 is a potent modifier of myeloma cell biology in a manner suitable for novel myeloma therapy.

Disclosures

No relevant conflicts of interest to declare.
Effects of Natural Compound EBSC-26 On Human Multiple Myeloma Cells.
Authors: Zhou Guangbiao , Liu Ying , Cheng Yongxian
Published: October 2019

Abstract 4924

Background

Human multiple myeloma (MM) is an incurable hematological malignancy at present, and screen for novel therapy remains an urgent need. The objective of this study was to assess the efficacy of natural compound EBSC-26 on multiple myeloma cells.

Methods

Inhibition of cell growth and proliferation of MM cell lines by compounds were assessed by WST-8 [2-(2-methoxy-4-nitrophenyl)-3-4-nitrophenyl)-5-(2,4- disulfophenyl)-2H-tetrazolium, monosodium salt] which allows sensitive colorimetric assays for the determination of the number of viable cells. Effects of compounds on cell cycle progression were analyzed by using flow cytometry. Apoptosis was evaluated by analysis of Annexin V. Microtubules were detected by immunofluorescence staining and confocal microscopy. Western blot and semi-quantitative/quantitative RT-PCR were performed to test protein/gene expression.

Results

EBSC-26 with a purity of up to 99.5%, was extracted from Centipeda minima (L.), a compositae plant used for the treatment of cold, nasal allergy, diarrhea, malaria, and asthma in China. We found that EBSC-26 suppressed proliferation/growth of U266, RPMI8226, dexamethasone-sensitive and resistant MM.1 cells, and induced apoptosis of these cells in a dose- and time-dependent manner. It synergized with Bortezomib and Doxorubicin in inhibition of MM cell proliferation. EBSC-26 overcame the protective effects of interleukin-6 and insulin-like growth factor-1 on multiple myeloma cells. It down-regulated interleukin-6-induced phosphorylation of STAT3 and insulin-like growth factor-1-induced phosphorylation of AKT. Moreover, EBSC-26 caused polymerization of microtubules, and induced G2/M arrest MM cells. Interestingly, an important G2/M-phase regulator, cyclin B1 was dramatically increased by EBSC-26 at protein level in a dose-dependent manner. EBSC-26 also decreased the phosphorylation of CDC2 at tyrosine 15.

Conclusions

These results suggest that EBSC-26 alone may have a potential in the treatment of multiple myeloma, and a combination of this agent with other compounds might provide further benefits.

Disclosures

No relevant conflicts of interest to declare.
Bortezomib, Doxorubicin and Dexamethasone (PAD) Combination Therapy for Chinese Relapsed or Refractory Multiple Myeloma.
Authors: Zhang Yongqing , Gao Guangxun , Wang Jishi , Chen Xiequn
Published: October 2019

Abstract 4948

Objective

To investigate the efficacy and safety of PAD (bortezomib, doxorubicin and dexamethasone) combination therapy for Chinese relapsed or refractory multiple myeloma (MM).

Methods

31 patients with relapsed or refractory MM received two to eight 21-days cycles of PAD: comprising an intravenous bolus of bortezomib 1.3 mg/m2 (P1,N=13) or 1.0 mg/m2 (P2,N=18) on days 1, 4, 8, and 11, doxorubicin 10mg per day on days 1 to 4, along with dexamethasone 40mg on days 1-4. Response to PAD was evaluated according to International Myeloma Working Group Criteria (IMWG 2006), toxicity was graded according to NCI CTCAE v3.0.

Results

25 patients (80.6%) achieved at least a partial response (PR), including complete response (CR) in 9 patients (29%), very good partial response (VGPR) in 7 patients (22.6%), PR in 9 patients (29%) and stable disease (SD) in 4 patients(12.9%), progression of disease (PD) in 2 patients (6.5%); median time to progression was 9.2 months, the median courses to achieve at least PR was 1.6(1-3) cycles, all of 7 patients with extramedullary plasmacytoma achieved at least PR after the first cycle of PAD, extramedullary plasmacytoma disappeared with 1-2 cycles of PAD. The efficacy was independent of traditional prognostic factors such asβ2-MG, Albumin,LDH and Hemoglobin which have previously influenced response to traditional chemotherapy. 1.5 year OS (Overall survival)of CR+VGPR group and PR group were 87.5% vs 46.7% (P=0.09). ≥PR response rate (CR +VGPR +PR) of P1AD and P2AD were 84.6% VS 77.8% (P= 0.501), CR+VGPR rate of P1AD and P2AD were 53.8% vs 50.0% (P=0.561 ). 1 year PFS(Progession-free survival) of P1AD and P2AD were 61.2% vs 55.6%(P=0.638), there were not difference between P1AD and P2AD in response rate(P= 0.501) and 1 year OS (P=0.872). Adverse events included thrombocytopenia in 15 patients ( 48.4% ), leukopenia in 8 patients(25.8%), peripheral neuropathy in 6 patients (19.4% ), varicella herpes zoster in 7 patients (22.6%), fatigue in 11 patients (35.5%) and diarrhea in 5 patients (16.1%), Thrombocytopenia and peripheral neuropathy of P1AD and P2AD were 46.2% vs 11.1%( P=0.037)and 53.8% vs 11.1%(P= 0.014).Common adverse reactions could be controlled with routine supportive treatmemt, one patient (3.2% ) died from respiratory failure during his fifth P1>AD.

Conclusions

PAD should be considered an appropriate treatment for Chinese relapsed or refractory MM, especially for MM with extramedullary plasmacytoma, its efficacy were independent of traditional prognosis factors, bortezomib dose reduction may reduce toxicities of PAD while retaining the efficacy.

Disclosures

No relevant conflicts of interest to declare.