bluuebunny/crossref_metadata_2025_split huggingface dataset:
Chromosomal Abnormalities in Bone Marrow Lymphocytes of MM Patients
Authors: Marun Carina S. Debes , Belch Andrew , Pilarski Linda M. 
Published: October 2019

Abstract
Abstract 4045
Multiple Myeloma (MM) is characterized by accumulations of chromosomal abnormalities throughout the course of disease. Recurrent IgH translocations such as t(4;14) or deletion (del)13 occur during early stages of disease or perhaps as originating events. Other abnormalities appear to be acquired late in the disease process, after development of frank MM, including p53 deletion (Ch17) and amplification of CKS-1B (Ch1). We have previously identified drug resistant clonotypic B lymphocytes in MM that have malignant characteristics and can xenograft MM to immunodeficient mice.
Using an automatic scanning system we scanned bone marrow (BM) cytospin slides stained with May-Grünwald Giemsa to identify lymphocytes and to determine whether or not they share the same chromosomal abnormalities that are found in autologous plasma cells (PC). For 200 MM patients, we performed interphase FISH using commercial probes, to detect deletion of chromosome 13, any translocations in 14q32, t(4;14)(p16;q32),t(11;14)(q13;q32), deletion of p53 and a custom probe to detect amplification of CKS-1B. For patients harboring a given chromosomal abnormality in their PC, we found the identical abnormalities in BM lymphocytes for 35% of patients having del13, 30% having t(4;14) and 23% having t(11;14). Likewise we detected amplification of locus 4p16, 11q13, 14q32, trisomy 1 and trisomy 17 in 45%, 15%, 30%, 37.5% and 44% respectively in the lymphocytes of patients demonstrating those same abnormalities in the PC. In a more limited series of experiments using immuno-FISH, we observed that BM lymphocytes with chromosomal abnormalities expressed CD20. In contrast, amplification of CKS-1B or p53deletion were undetectable in lymphocytes from patients having these abnormalities in their PC. We are currently analyzing the impact of abnormalities present in BM lymphocytes with regards to the clinical parameters and survival. An update of these results will be given at the meeting. Given the compelling evidence that B cells comprise a clinically important compartment of the MM clone, this work suggests that fundamental genetic events shared by all compartments of the MM clone include recurrent IgH translocations and Ch13 deletions. However, further clonal expansion occurring during disease progression and after relapse may be restricted to the PC population, characterized by acquisition of p53 deletions and CSK1B amplification. Furthermore in overt MM, clonal expansion may simultaneously occur on at least two levels, firstly that of the B cells which persistently give rise to PC, and secondly that of PC themselves as they continue clonal expansion in the absence of contributions from the B cells. Abnormalities such as del13 or IgH translocations characterize early events in the disease while p53del or amplification of CKS-1B may occur as progression events restricted to PC.

Disclosures:
No relevant conflicts of interest to declare.
Feasibility and Efficacy of ABVD In Elderly Hodgkin Lymphoma Patients: Analysis of Two Randomized Prospective Multicenter Trials of the German Hodgkin Study Group (HD10 and HD11)
Authors: Böll Boris , Görgen Helen , Fuchs Michael , von Tresckow Bastian , Engert Andreas , Borchmann Peter 
Published: October 2019

Abstract
Abstract 418
About 20% of all patients with first diagnosis of Hodgkin Lymphoma (HL) are older than 60 years. They have a poor prognosis mainly due to an increased toxicity of chemo- and radiotherapy. Although resulting in better disease control, aggressive regimens as BEACOPP are not feasible in this cohort of patients due to a treatment related mortality of up to 20%. Therefore, ABVD is considered treatment of choice for elderly HL patients, although prospective studies are lacking and current concepts mostly rely on cohort analyses. We therefore analyzed feasibility, safety and outcome of patients older than 60 years with early favorable- or early unfavorable-stage HL treated with 4 cycles of ABVD within the HD10 and HD11 trials of the GHSG. Sixty-eight and 49 elderly patients with a median age of 65 and 64 years were treated in HD10 and HD11, respectively. Early termination of protocol therapy was documented in 18% of HD10 patients, but only in 8% of HD11 patients, resulting in a lower relative total chemotherapy dose (RCD) in HD10. The relative dose intensity (RCD divided by total relative chemotherapy duration) was much lower in both studies compared to younger patients, due to more toxicity-related therapy delays and dose reductions, as 4 cycles of ABVD caused WHO grade III/IV toxicities in 67% (HD10) and 69% (HD11). Overall efficacy was significantly lower than in younger patients with an overall response rate of 90% in HD10 and 92% in HD11. The rate of relapsing patients was the same as in younger patients in HD11 (14%), whereas in HD10 it was much higher in the elderly (12%) which was mainly due to late relapses. Overall 22% and 37% of the patients died in HD10 and HD11, respectively (median observation time: 92 months). Besides other causes as cardiovascular disease (7%) or secondary neoplasia (5%), there was a high rate of deaths due to insufficient HL-control (5%) and treatment-related toxicities (5%). The 5-year PFS estimates for elderly patients were 79% (95% CI, 67% to 87%) in HD10 and 69% (95% CI, 54% to 80%) in HD11 compared to 96% (95% CI, 93% to 97%) and 86% (95% CI, 83% to 88%) for younger patients in HD10 and HD11, respectively. In conclusion, 4 cycles of ABVD are effective in elderly HL patients; however, treatment-related toxicity is high.

Disclosures:
No relevant conflicts of interest to declare.
Effect of CMX001 on Levels of DNA Virus In Patients After Allogeneic Stem Cell Transplantation.
Authors: Gabriel Don A. , Whitley Julia , Rao Kamakshi , Horst Charles van der , Serody Jonathan , Coghill James , Sarantopoulos Stefanie , Armistead Paul , Wood William , Shea Thomas C. 
Published: October 2019

Abstract
Abstract 4550
Latent viral infections occurring in the post bone marrow transplant period remain a significant clinical problem. New and effective antiviral drugs (AVD) are needed to provide adequate therapy for infected patients. We report the use of a new oral AVD, CMX001 (Chimerix Inc. Durham, NC) to treat DNA virus infections in six severely ill patients.Pt #123456Adeno Blood (PCR Cycles)Pre: 30.6Pre: 33.1Post: 37.3Post: NegBK polyoma virus Urine (log copies/mL)Pre: 4.89Pre: 4.86Pre: 6.54Pre: 9.06Post: 7.88Post: 5.99Post: 7.55Post: 7.16BKV BloodCleared after 1 dosePre: No valueCleared at 6 doses;Post: detectableHHV6 Blood (PCR Cycles)Pre: 42Post: 39.9Dose (mg/kg) biw22444 reduced to 32StatusDied GVHDDied GVHDDied GVHD + SepsisDied GVHDAliveAlive
All patients had grade 2 or greater gut graft versus host disease (GVHD) during or after CMX001 treatment. The Table shows viral levels before and after treatment with CMX001 twice/wk. Pt 1's AV cycle number fluctuated and never cleared. A higher cycle number indicates less virus. Pt 2 cleared AV from blood after first dose, but urine, stool, gut biopsies remained AV positive. Pt 3's HHV6 plasma PCR levels fluctuated, but the pericardial fluid levels had cleared at autopsy. Pt 4 did not have pre dosing blood BKV PCR levels taken and his subsequent blood BKV low level was measurable (below the positive limit level). Interestingly, however, the patient's BKV related hemorrhagic cystitis (HC) symptoms resolved after 4 doses of CMX001. Pt 5's BKV HC symptoms also resolved after 4 doses of drug, and his positive blood viral level cleared after 6 doses. Pt 6 discontinued drug because of abdominal pain and the need for NPO, but his hemorrhagic cystitis symptoms had abated. No specific toxicities were documented in these severely ill patients with gut GVHD, but gut pain and transient thrombocytopenia were observed. In summary, one of 2 pts with AV cleared their blood. BKV was not cleared from urine, but 2/4 cleared BKV from blood. One patient with HHV6 did not clear blood DNA viral levels, but did clear pericardial fluid. In both cases of HC, symptoms resolved. Thus, CMX001 shows promise as new antiviral drug given its ability to decrease DNA viral levels in this small group of critically ill patients.

Disclosures:
No relevant conflicts of interest to declare.
Cytogenetic Abnormalities In the Spleen In CLL Patients.
Authors: Norin Stefan , Wallblom Ann , Aggarwal Mohit , Sander Birgitta , Kimby Eva 
Published: October 2019

Abstract
Abstract 4618

Background:
In CLL the prognostic importance of cytogenetic abnormalities in bone marrow or peripheral blood is well-known. However data is lacking on the frequency and importance of the aberrations in other tissues such as lymph nodes and spleens.

Aims:
To develop a method to assess the chromosomal aberrations commonly seen in CLL (13q-, 11q-, 17p- and +12) on paraffin-embedded tissue sections from spleens and to relate the findings to data on abnormalities found in bone marrow or peripheral blood.

Material and methods:
24 out of 61 CLL patients splenectomized between 1990–2009 have so far been analyzed. Paraffin-embedded sections were obtained from stored tissue material from spleen. Fluorescence in situ hybridization (FISH) was used to detect cytogenetic abnormalities and the results were compared with analyses made on blood or bone marrow. Monosomy 12, an abnormality rarely occurring in CLL was used as a surrogate marker to avoid false-positive results for deletions due to incomplete nuclei. In all but one case this was below 20%. Therefore the cut-off for the deletions (13q-, 11q- and 17p-) were set to 30%. For +12 we used 5%. Data on flow cytometry for quantification of the CLL clone in the spleen was available in 13 patients with a median of 71% clonal cells (range 11–86%). Indications for splenectomy were splenomegaly (n=13), AIHA (n=8) or ITP (n=3). All but five patients had received chemotherapy before splenectomy including purine analogs (n=7), alemtuzumab (n=1) or both (n=2) The remaining patients were treated with alkylators and/or anthracycline-containg regimens.

Result:
In all but one patient, FISH results from the splenic sections could be obtained with cytogenetic aberrations detected in 18 of 23 patients (78%). In 11 patients a single abnormality was detected, whereas multiple aberrations were present in 7 cases. The most common aberration was 13q- (70%), followed by 17p- (22%), 11q- and +12 (17% each) (Table 1). In 8 cases FISH samples from blood/bone marrow were available from time points both pre- and post- splenectomy and 6 additional cases had FISH data before splenectomy. All FISH abnormalities affecting >20% CLL cells in blood or bone marrow were also found in the spleen. In contrast an additional 17p clone was found in one patient and in two patients the 13q- and 17p- clones were significantly larger in the spleen. Remarkably one patient had both homo- and heterozygous 13q- clones in blood and in the spleen. In repeated blood samples nine years after splenectomy only a heterozygous clone remained.

Conclusion:
FISH on paraffin-embedded sections is a useful tool for the evaluation of genetic abnormalities. Clonal evolution does appear to occur in the spleen in CLL and with splenectomy selected clones might be obliterated.

Disclosures:
Kimby: Roche, Bayer-Schering, Mundipharma: Membership on an entity's Board of Directors or advisory committees, lecturer.
Can Human Umbilical Cord Mesenchymal Stem Cells Inhibit Acute Graft-Versus-Host-Disease In Murine Allogeneic Bone Marrow Transplantation?
Authors: Guo Juan , Li Xiao , Chang Chunkang 
Published: October 2019

Abstract
Abstract 4703

Objective:
The purpose of this study was to investigate the therapeutic potential of Human umbilical cord mesenchymal stem cells (HUCMSCs) from normal donors for preventing acute GVHD following allogeneic bone marrow transplantation (BMT).

Methods:
In this study, a new method of separating and culturing HUCMSCs was used. The human umbilical cord tissue was digested by collagenase II for 2h. And after being passaged to P3, more HUCMSCs could be obtained. In the control group, after being lethally irradiated, DBA/2(H-2Kd) mice were transferred bone marrow (BM) and splenocytes (SC) from C57BL/6 (H-2Kb) mice. Recipients in the HUCMSCs-treated group were adoptive transferred HUCMSCs, BM and SC at day 0 and were transferred only HUCMSCs at day 3 again. The two groups were compared in weight, survival, histopathological specimens, lymphocyte subgroups and serum cytokine analysis.

Results:
In vitro, IFN-γ treated HUCMSCs have a strong inhibitory effect on the proliferation of CD4 + T cells (P<0.05), but untreated HUCMSCs cannot obviously inhibit the proliferation of CD4 + T cells (P>0.05). IDO is involved in the immunosuppression of HUCMSCs. In the HUCMSCs-treated group, 50% of the mice survived over 30 days after BMT, but in the control group all mice died within 18 days. The mice treated with HUMSCs exhibited light symptoms of aGVHD after day 14. There were higher IFN-γ and IL-12 levels by day 7 in serum of mice that received HUCMSCs compared to those without HUCMSCs-treatment, while the IL-4 levels showed no significant difference between the two groups. The proportion of CD3+NK1.1+NKT cells in splenocytes of HUCMSCs-treated group is higher in day 21 compared with in day 7.

Conclusion:
Using the new method of digesting human umbilical cord tissue by collagenase II for 2h, we obtained stable and pure HUCMSCs. IDO is involved in the immunosuppression of HUCMSCs. After the xenogeneic transplantation of HUCMSCs to the aGVHD model of mice, HUCMSCs can regulate cytokines such as IFN-γ, IL-12 to increase CD3+NK1.1+NKT cells.

Disclosures:
No relevant conflicts of interest to declare.
An Open-Label, Single Ascending Dose Phase I Study of F-627, a G-CSF Dimer, In Healthy Male Subjects
Authors: Yan Xiao Qiang , Hodsman Peter G , Huang Zhi Hua , Sun Rou Yun , Sun Nai Chau , Huang Yu Liang 
Published: October 2019

Abstract
Abstract 4722
The management of severe neutropenia in cancer patients after high-dose chemotherapy remains a challenge, although recombinant human G-CSFs including filgrastim, lengogratim and pegfilgrastim have been widely used. We developed F-627, a mammalian expressed recombinant human G-CSF dimer to treat severe neutropenia. In a cyclophosphamide-induced neutropenia monkey model, F-627 significantly reduced the duration and lessened the severity of neutropenia compared to monkeys treated with G-CSF or pegylated G-CSF. We report the phase I results of F-627 in healthy male subjects receiving ascending single dose at 30, 60, 120 and 240 ug/kg by subcutaneous injection. The primary objective of the phase I study was to assess the safety and tolerability of F-627. The secondary objective was to evaluate the pharmacokinetics and pharmacodynamics (PK/PD) responses. F-627 was well tolerated in healthy male volunteers. Dose-dependent PD responses including increased WBC, ANC and CD34 levels in peripheral blood were demonstrated. These results indicate that the mechanism of action of F-627 is similar to the monomer recombinant hG-CSFs, while the mode of action of F-627 is different with unique PK/PD properties in human.

Disclosures:
No relevant conflicts of interest to declare.
Heart Function Disorders In Sickle Cell Disease
Authors: Heinen Charlotte , Fijnvandraat Karin , Peters Marjolein , Heijboer Henriette , Blom Nico , Kuipers Irene 
Published: October 2019

Abstract
Abstract 4815
Abstract

Background:
Sickle cell disease (SCD) is a hereditary anemia characterized by chronic hemolytic anemia and vaso-occlusive episodes, associated with a high susceptibility for infections and damage to all vital organs. Cardiologic effects of SCD are dilatation and hypertrophy of the left ventricle and pulmonary hypertension. The prevalence of these cardiac complications in children with SCD living in the Netherlands is unknown. The role of hemolysis in this process is still unclear. Therefore, the objectives of this study are: 1) to evaluate the prevalence of cardiac abnormalities in children with SCD living in the Netherlands, 2) to analyse the association between cardiac abnormalities and the severity of anemia and hemolysis.

Methods:
Cross-sectional study, including 92 children with SCD (genotype HBSS, HBSC or HBS-beta 0) treated at the Emma Children's Hospital AMC. None of the children had cardiac abnormalities unrelated to SCD. Echocardiograms and electrocardiograms were performed in a stable state. All cardiac values were corrected for BSA. Echocardiographic values 20% above or under the average of an age matched control group were considered abnormal. Laboratory hematological parameters were obtained within one year of the cardiac imaging. All statistical analyses were performed with SPSS 18 software, using Pearson correlations and T-tests. Forced-entry multivariate regression analysis was conducted with hematological parameters, age and SCD genotype as independent variables and cardiologic parameters as dependent variables.

Results:
The study group had a mean age of 13 years (SD 5,5) and contained 49 boys (53,3%). All children had a normal shortening fraction. Almost half of the children (39%) had a dilated left ventricular internal diameter in diastole and systole. Dilatation was inversely correlated with the Hb level. When analysed with multivariate analysis, both Hb and bilirubin were independently associated with left ventricular dilatation, suggesting that hemolysis deteriorates cardiac structure by a mechanism separate from the effect of anemia. There was no association between the left ventricular dilatation and age.

Conclusion:
Pediatric sickle cell patients have significant dilatation of the left ventricle, an abnormality that is significantly and independently associated with the severity of anemia and bilirubin levels, but in spite retain a normal shortening fraction.

Disclosures:
No relevant conflicts of interest to declare.
Study of the Quantity and Function of Regulatory T Cells In the Hemocytopenic Patients with Positive BMMNC-Coombs Test
Authors: Fu Rong , Chen Jin , Shao Zonghong , Wang Jun , Li Lijuan , Liu Hui 
Published: October 2019

Abstract
Abstract 4875

Objective
To investigate the quantity and function of regulatory T cells in the IRP, then explore the significance of Treg cells in the pathogenesis of IRP. Methods Sixty-two patients with IRP and twenty-four healthy donors were enrolled in this study. The ratios of CD4+CD25+/CD4+ and CD4+CD25+CD127low/CD4+ in bone marrow were examined with flow cytometry. The levels of IL-2, ATGF-β were tested with ELISA. The expressions of FoxP3 and Galectin-10 mRNA in BMMNC were measured by semiquantitive RT-PCR. Results The levels of IL-2, ATGF-β in bone marrow of untreated or recovered IRP patients (5.64±1.70, 6.19±2.53; 1.79±0.67, 1.86±0.76) were significantly lower than them of normal controls (7.91±3.71,2.48±0.94) (p<0.05); The ratio of CD4+CD25+/CD4+ cells in bone marrow of untreated IRP patients (22.46±9.47) was significantly lower than that of recovered IRP patients or normal controls (27.10±7.08, 30.59±8.58) (p<0.05); The ratio of CD4+CD25+CD127low/CD4+ cells in bone marrow of untreated IRP patients (7.18±2.72) was significantly lower than that of recovered IRP patients or normal controls (9.07±4.67, 10.44±3.24) (p<0.05). The relative mRNA expressions of FoxP3 and galentin-10 were 0.34±0.25, 0.69±0.51, 0.82±0.65 and0.66±0.11, 0.74±0.11, 0.76±0.09 in three groups, respectively. The expressions of the two factors in untreated IRP patients were significantly lower than them in recovered IRP patients or normal controls (p<0.05). Conclusions There exist abnormalities in quantity and function of Treg cells in IRP patients which might play important role in the pathogenesis of IRP.

Disclosures:
No relevant conflicts of interest to declare.
Cytoprotective Autophagy Induction by Imatinib Mesylate In Non-BCR-ABL Expressing Cells
Authors: Miyazawa Keisuke , Ohtomo Tadashi , Moriya Shota , Naito Munekazu , Kuroda Masahiko , Itoh Masahiro , Tomoda Akio 
Published: October 2019

Abstract
Abstract 4937
Autophagy is an intracellular degradation system that delivers cytoplasmic constitutions to the lysosome. Intracellular proteins and organelle are sequestered by the autophagosomes, then delivered to the lysosome and degraded. It has been reported that combining imatinib mesylate (IM) with an autophagy inhibitor such as chloroquine resulted in enhanced elimination of CML clones in vitro and in vivo (Bellodi C et al. J. Clin. Invest. 2009). We here report that IM induces the cytoprotective effect in non-BCR-ABL expressing cells along with autophagosome formation.
Treatment with IM resulted in an increased viable cell number of non-BCR-ABL expressing leukemia cell lines by inhibiting spontaneous apoptosis. IM did not show any effects on cell-cycle progression. When HL-60 cells were cultured in the presence of 3–30 μM of IM for 24–96 hr, up to a 1.6-fold increase in the viable cell number was observed in a dose-dependent manner. Furthermore, when HL-60 cells were cultured either under serum depletion or at lower concentrations of FBS, the cytoprotection by IM became more pronounced as compared with the cells cultured with 10% FBS. As well as in BCR-ABL expressing CML cell lines, electron microscopy revealed an increased autophagosomes after IM treatment in non-BCR-ABL expressing cells. Cytoprotection with autophagosome formation by IM was observed in various leukemia and cancer cell lines as well as normal murine embryonic fibroblasts (MEFs). IM attenuated the cytotoxic effect of cytosine arabinoside and bortezomib in leukemia cells.
Inhibition of autophagy by knockdown of LC3 gene using shRNA in HL-60 as well as complete knockout of atg5 in the Tet-off atg5-/- MEF system attenuated the cytoprotective effect of IM. This indicates that the effect is at least in part dependent on autophagy. However, the cytoprotection by IM was not medicated through supression of ROS production via “mitophagy” or reduction of ER stress via “ribophagy”. The cytoprotection with autophagy was still detectable in the HL-60 cells after knockdown of c-abl gene and in an ABL deficient-HL-60 cell line. This indicates that the proapoptotic function of ABL kinase is not involved. In addition, this effect was still observed in the presence of cycloheximide, suggesting that the effect is not mediated through translation process.
Although the target tyrosine kinase(s) of IM remains unclear, our data provide novel therapeutic possibilities of targeting tyrosine kinase(s) for induction of cytoprotective autophagy.

Disclosures:
No relevant conflicts of interest to declare.