bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased Sclerostin Secretion in Multiple Myeloma Results in Stimulation of Osteoclastogenesis and Inhibition of Osteoblastogenesis
Authors: Eda Homare , Santo Loredana , Cirstea Diana D. , Pozzi Samantha , Canavese Miriam , Scullen Tyler A. , Vallet Sonia , Raje Noopur S. 
Published: October 2019

Abstract
Abstract 1819

Objectives:
Osteoblasts (OB) and osteoclasts (OC), are an integral part of the bone microenvironment, and play a crucial role in myeloma growth and survival. Their imbalance results in osteolytic disease and elucidating the mechanisms underlying osteolytic lesions is important not only for the improvement of osteolytic bone disease but also for the treatment of multiple myeloma (MM).
The osteocyte-secreted protein sclerostin, encoded by the SOST gene, is a potent inhibitor of osteoblastogenesis. It is regarded as a good target for osteoporosis treatment, but its role in MM remains to be determined. Our objective was to study the role of sclerostin in MM bone disease and determine if sclerostin directed strategies were a reasonable approach in MM.

Methods and Results:
Sclerostin concentration in patients' blood plasma and MM cell line supernatant stimulated by IL-6, FGF-2, TNFalpha, BMP7 and TGFbeta was detected by ELISA (ALPCO immunoassays). Increased level of sclerostin was detected in MM patient plasma (n=20, median: 4.73 ng/mL, range: 1.5–19.5 ng/mL). Plasma concentrations were significantly higher (p<0.01) when compared to sclerostin concentration in the plasma of leukemia patients (n=3), gastric cancer patients (n=40) and healthy volunteers (n=4). High sclerostin levels were not associated with extent of bone disease but rather correlated with tumor burden (High B2M, creatinine and LDH, and low Hb) suggesting an autocrine loop for sclerostin production. Because sclerostin is derived from mature OB or orteocytes, we measured levels during OB differentiation but we were unable to detect increased levels. We then measured sclerostin levels in RPMI-8226 MM cell line supernatant either alone or stimulated by cytokines D Systems).

Conclusions:
These data demonstrate that increased sclerostin levels in MM patients inhibit osteoblastogenesis and stimulate osteoclastogenesis. Taken together, sclerostin may be good target to inhibit myeloma bone disease and help restore normal bone homeostasis.

Disclosures:
Raje: Celgene: Membership on an entity's Board of Directors or advisory committees; Novartis: Membership on an entity's Board of Directors or advisory committees; Millenium: Membership on an entity's Board of Directors or advisory committees; Astra Zeneca: Research Funding; Amgen: Membership on an entity's Board of Directors or advisory committees; Acetylon: Research Funding.
Prognostication by miRNome-Profiling in Multiple Myeloma
Authors: Seckinger Anja , Meißner Tobias , Benes Vladimir , Schmidt Sabine , Blake Jonathan , Zimmermann Jürgen , Goldschmidt Hartmut , Klein Bernard , Hose Dirk 
Published: October 2019

Abstract
Abstract 1822

INTRODUCTION.
MicroRNAs are an abundant class of small non-protein-coding RNAs that function as negative gene regulators in diverse biological processes including cancer by affecting the stability and translation of mRNAs.

METHODS.
We determined expression of 559 miRNAs by miChip (Exiqon LNA Array probes V9.2) in CD138-purified myeloma cells from previously untreated patients (n=69), normal bone marrow plasma cells of healthy donors (pooled to n=3), and human myeloma cell lines (n=20). For normalization, an invariant-based method was applied. Gene expression profiling was performed using Affymetrix U133 2.0 DNA-microarrays.

RESULTS.
We found 29 miRNAs to be significantly up- and 35 down-regulated in myeloma cells vs. normal bone marrow plasma cells, respectively. Expression of 18 miRNAs was simultaneously significantly (P<.01) associated with event-free survival (EFS) and overall survival (OS), and allowed the delineation of prognostic groups. Of these, miRNAs miR-659 (located at 22q12.1) was significantly lower expressed in myeloma cells compared to normal bone marrow plasma cells. For this miRNA, low expression delineates a group with inferior EFS (median 19.7 months vs. not reached, P<.001) and OS (median 52.9 months vs. not reached, P<.001). This group shows a significantly higher gene expression based proliferation index. In contrast, high miR-590 expression (located at 7q11.23) delineates a group with inferior EFS (median 12.4 vs. 36 months, P<0.001) and OS (median 29.4 months vs. not reached, P<.001). By using Goeman's global test, a significant association of the predicted target gene signatures with survival could be found for both, EFS (miR-590-5p, P<.001; miR-659, P<.001) and OS (P=.009; P=.002).

CONCLUSION.
In conclusion, we demonstrate the prognostic relevance of miRNome profiling in multiple myeloma.

Disclosures:
No relevant conflicts of interest to declare.
The Translation Initiation Factor eIF4E Is Overexpressed in Multiple Myeloma and Is Critical for Myeloma Cell Proliferation and Survival
Authors: Li Shirong , Jin MeiHua , Liu Ailing , Mapara Markus Y. , Lentzsch Suzanne 
Published: October 2019

Abstract
Abstract 1853

Methods:
The translation initiation factor eIF4E is central to protein synthesis in general, and overexpression and/or activation of eIF4E is associated with a malignant phenotype by regulating oncogenic protein translation. Several previous publications indicate that aberrant control of protein synthesis contributes to lymphoma genesis but the exact role of protein translation in multiple myeloma (MM) is less clear. Therefore, understanding the mechanisms that control protein synthesis is an emerging new research area in MM with significant potential for developing innovative therapies. The goal of this study was to determine the role and regulation of eIF4E, as well as the effects of protein translation controlling drugs in MM.

Results:
By western blot analysis as well as RT-PCR we found that eIF4E protein and mRNA levels are significantly elevated (up to 20 fold) in MM cell lines (H929, RPMI-8226, MM.1S and OPM2) and primary myeloma cells compared to normal plasma cells. Silencing of eIF4E gene expression in RPMI-8226 MM cells by a stable and inducible shRNA system significantly decreased viability of myeloma cells (by ∼ 43%) but not of HEK 293 suggesting a higher dependency of MM cells to protein translation. Next we evaluated different drugs including pomalidomide, rapamycin, pp242, 4EGI-1 and ribavirin, that are known to inhibit protein synthesis for their effects on protein translation in MM. By m7GTP pull down assays we evaluated the effects of the different drugs on eIF4E expression and activity. Rapamycin blocked the phosphorylation of 4EBP1 and eIF4E release, and subsequently inhibited eIF4G binding. The compound 4EGI-1 decreased the interaction between eIF4E and eIF4G. Pomalidomide decreased eIF4E protein expression. All drugs inhibited MM cell DNA synthesis measured by 3H-Thymidine incorporation. Treatment with pomalidomide (10uM), rapamycin (40nM), pp242 (10uM), 4EGI1 (50uM) or ribavirin (50uM) for 48h significantly decreased (p<0.05) proliferation by 43–62% indicating that drugs controlling protein translation inhibit MM growth. We also found that all drugs decreased expression of eIF4E dependent targets such as cyclin D1 and c-myc.

Conclusion:
Here we show that eIF4E, a key player in translational control, is highly expressed in MM cells and critical for MM growth and survival. Therefore our study helps to understand the function and regulatory mechanism of eIF4E in MM. Further the evaluation of drugs targeting protein translation provides the basis for the optimization of current MM treatment or to open up new strategies such as targeting protein translation in future MM therapy.

Disclosures:
Lentzsch: Celgene Corp: Consultancy, Research Funding; Onyx: Consultancy; Genzyme: Consultancy; prIME Oncology: Honoraria; Imedex: Honoraria; Clinical Care Options: Honoraria.
Influence of Stem Cell Mobilization After Cyclophosphamide, Thalidomide and Dexamethasone (CTD) Regimen in Patients with Newly Diagnosed Multiple Myeloma
Authors: Ahn Jae-Sook , Yang Deok-Hwan , Jung Sung-Hoon , Bae Soo-Young , Kim Yeo-Kyeoung , Kim Hyeoung-Joon , Lee Je-Jung 
Published: October 2019

Abstract
Abstract 1927

Backgrounds:
CTD regimen has been known as an effective induction therapy in patients with newly diagnosed MM. But, there were inconsistent results for the autologous stem cell yield for transplantation. The aim of present study was to identify the influence of CTD therapy on outcome of peripheral blood stem cell (PBSC) collection.

Methods:
Forty-eight patients received 4 cycles of CTD therapy. Stem cells were mobilized with cyclophosphamide (3.0 g/m2) and G-CSF (10 ƒÝg/kg, daily) or G-CSF alone. Patients failing to collect ≤ 4.0 × 106 CD34+ cells /kg received a second mobilization courses.

Results:
The median age at diagnosis was 56 years (range, 39–69). Median duration from start of CTD therapy to first collection was 4.6 months (range, 3.3–8.7). Forty-four patients were mobilized with cyclophosphamide following with G-CSF and 4 patients with G-CSF alone. The median day of apheresis was 3 days (range, 2–7). The response rate for CTD regimen at mobilization was 10% (5/48) of CR, 25% (12/48) of VGPR and 63% (30/48) of PR. A median number of harvested CD34+ cells was 8.6 × 106 cells/kg. At the first mobilization, 83% (40/48) of patients had been reached the minimal PBSC collection target of ≥ 2.0 × 106 CD34+ cells/kg and 71% (34/48) of patients achieved the collection ≥ 4.0 × 106 CD34+ cells/kg. At the end of second mobilization, 90%(43/48) of patients had yields of at least ≥ 2.0 × 106 CD34+ cells/kg and 77% (37/48) of patients had yields of ≥ 4.0 × 106 CD34+ cells/kg. During mobilization period, three patients were developed grade 3/4 non-hematologic adverse events.

Conclusion:
CTD regimen is an effective induction therapy in patients with newly diagnosed MM showing high response rate and acceptable rate of autologuos stem cell yield without any detrimental effect for the following stem cell collection.

Disclosures:
No relevant conflicts of interest to declare.
Increased Plasma Indoleamine 2,3-Dioxygenase Activity Correlates with the Severity of Acute Graft-Versus-Host Disease
Authors: Xu Jinhuan , Zhang Yicheng 
Published: October 2019

Abstract
Abstract 1985
Indoleamine 2,3-dioxygenase (IDO) is a rate-limiting enzyme for the tryptophan catabolism, which plays an important role in the induction of immune tolerance. To evaluate the expression level of IDO in the patients receiving hematopoietic stem cell transplantation (HSCT) and to identify the possible correlation between IDO activity and acute graft-versus-host disease (aGVHD), we collected blood samples from 96 patients before and after HSCT and 10 healthy volunteers served as controls. The expression of IDO mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) in peripheral blood mononuclear cells of the patients and healthy controls. Reverse-phase high-performance liquid chromatography (HPLC) was performed to analyze the level of kynurenine (Kyn) and tryptophan (Trp) in plasma. The IDO activity was calculated as the ratio of kyn to trp. Plasma IFNgama was measured by standard ELISA. RT-PCR showed that IDO mRNA was detected in 65 of 85(65/85, 76.5%) patients with aGVHD; in patients without aGVHD, only 1 of them express IDO mRNA (1/11, 9.1%); none of 10 healthy volunteers was positive for the IDO expression. The plasma IDO activity was much higher in aGVHD group than those without aGVHD(4.49¡À0.46 vs. 1.69¡À0.9, respectively; p < 0.0001) or the healthy controls(4.49¡À0.46 vs. 1.77¡À0.22, respectively; p < 0.0001). Patients with severe aGVHD (grade III/IV) had significantly higher IDO activity than those with mild aGVHD (grade I/II) (6.74¡À0.58 vs. 2.17¡À0.79, respectively; p < 0.0001). IDO activity was decreased following effective treatment of aGVHD, while fluctuation of plasma IDO was also observed upon the recurrence of aGVHD. The plasma IDO activity was correlated with the IFNgama level(r=0.8816).

Conclusion:
The IDO mRNA was expressed in blood mononuclear cells from patients with aGVHD. The plasma IDO activity was elevated in aGVHD and correlated with severity of aGVHD. In combination with plasma IFNgama, IDO may serve as a potential biomarker for the diagnosis and evaluation of aGVHD following HSCT; intervention of IDO pathway may also provide an alternative way to overcome the steroids resistant aGVHD.

Keywords:
acute graft-versus-host disease, Indoleamine 2,3-dioxygenase, hematopoietic stem cell transplantation

Disclosures:
No relevant conflicts of interest to declare.
Donor-Cell Leukemia After Hematopoietic Stem Cell Transplant: a Retrospective Review of a Single Center's Experience
Authors: Washburn Katie , Selby George B. , Dunn S. Terence , Chakrabarty Jennifer L. Holter 
Published: October 2019

Abstract
Abstract 1997
Donor-cell leukemia (DCL) is a rare complication of allogeneic hematopoietic stem cell transplant (ASCT) with a reported incidence varying from 0.1% to 5.0% of all post-transplantation relapses. However, the incidence is likely to be highly underestimated due to the difficulty in making the diagnosis. In the 1970s and 1980s, diagnosis of DCL relied on cytogenetic and fluorescence in situ hybridization studies in sex-mismatched donor/recipient pairs. Recent advances in molecular-based laboratory methods have considerably improved our ability to identify and confirm DCL in ASCT patients, especially when there is not a sex-mismatch between donor and recipient. Recently, an algorithm has been proposed by Wang et al. 2011 (Am J Clin Pathol 135;525–40) to help make the diagnosis of DCL in ASCT patients who develop overt leukemia or a clonal cytogenetic abnormality using these new advanced techniques. We retrospectively evaluated all patients who underwent ASCT at our center for acute leukemia from 2005 to present. Out of 43 patients, 6 apparent DCLs were documented using short tandem repeat (STR) analysis (12 separate loci examined); each ASCT patient relapsed with the full complement of donor STRs. Our data confirms that the true incidence of DCL is likely underestimated, since within our select population, we had a 14% incidence of DCL. Of note, 1/6 were sex mismatches, 3/6 received cord blood transplants, 4/6 cases were ABO mismatches, and 4/6 cases were AML (Table 1). Our findings concur with sparse available reports in the literature that purport there may be a shorter time to relapse in patients who are undergoing ASCT for malignant reasons; indeed, our average latency period was 14 months as opposed to 24 months found by Wang et al. However, we did not find that a disproportionate number of sex-mismatched donor/recipient pairs developed DCL; in fact, nearly all of our DCLs arose in the setting of a sex-matched pair. The apparent increased incidence in cord blood transplants, sex-mismatched donor/recipient pairs and ABO-mismatched transplants needs to be assessed in a large-scale registry using standard methodologies, which could help identify and define potential causes of DCL.Table 1:Patient Characteristics with Donor-Derived LeukemiaSexAge (years)Leukemia TypeTransplant TypeTime to Relapse (months)Sex MismatchABO MismatchChemotherapy PreparationM22ALLunrelated donor15NoYesalemtuzumabF1AMLrelated sibling14NoNoBu/Cy/Mito1M24AMLunrelated double-cord blood40NoNoTVTG2M20AMLunrelated donor5NoYesCy/TBI3M26AMLunrelated double-cord blood10.5YesYesFlu/Cy/TBI4M26ALLunrelated double-cord blood3.5NoYesFlu/Cy/TBI41. busulfan, cytarabine, mitoxantrone2. topotecan, vinorelbine, thiotepa, gemcitabine3. cyclophosphamide & total body irradiation4. fludarabine, cyclophosphamide & total body irradiation

Disclosures:
No relevant conflicts of interest to declare.
Achievement of Complete Remission After High-Dose Melphalan and Autologous Stem Cell Transplantation Is the Only Important Prognostic Factor in Patients with Multiple Myeloma
Authors: Shin Seunghwan , Kim Jeong-A , Lee Bohee , Min Chang Ki , Kwak Jae-Yong , Yhim Ho-Young 
Published: October 2019

Abstract
Abstract 2018

Background:
High-dose melphalan and autologous transplantation (HDM-AT) is the most effective treatment for survival benefits in multiple myeloma (MM) patients. However, the relationship between the HDM-AT response and long-term outcome is not clear. In this study we examined the relationship.

Method:
We analyzed the clinical data of 137 patients treated with HDM-AT in three institutions between 2000 and 2011. The international Myeloma Working Group Uniform Response Criteria was used to evaluate.

Results:
Out of 137 patients, clinical data of 126 patients were evaluable for treatment response according to the medical records. Their median age was 55 years (range, 36–65 years) and 69 (55%) patients were male. Among the patients who had received induction treatment (IT), 79%(100/126) of the patients were treated with bortezomib-containing regimens. After IT, the response rates were 29% CR, 18% VGPR, 37% PR, 15% SD, and 1% PD. After HDM-AT, response rates were 64% CR, 15% VGPR, 15% PR, 4% SD, and 2% PD. Median follow-up duration was 25 months (range, 5–146 months). 5-year overall survival (OS) and progression-free survival (PFS) were 54% and 43 %, respectively. Multivariate analysis regarding OS showed that CR after HDM-AT compared to ≤CR is an important prognostic factor (5-year OS rate of CR vs ≤CR, 78% vs 29%, respectively, p=0.002). However, CR after HDM-AT is not prognostic factor for PFS.
79 (79/126, 64%) patients showed the CR after HDM-AT. Among the 79 patients, 44 patients did not achieve CR after IT however, turned into CR after HDM-AT; 35 patients already achieved CR after IT. However, there were no significant differences between the two groups of patients (5-year PFS, 44% vs. 45%, P=0.245 and 5-year OS 77% vs. 79%, P=0.690 respectively).

Conclusion:
Achievement CR after HDM-AT is the only important prognostic factor to obtain the survival benefit regardless of response after IT in MM patients.

Disclosures:
No relevant conflicts of interest to declare.
Anticoagulation in Ambulatory Cancer Patients
Authors: Pishko Allyson , Smith Kenneth J. , Ragni Margaret V. 
Published: October 2019

Abstract
Abstract 2071

Background:
Venous thromboembolism (VTE) risk is increased in patients with malignancy, occurring in up to 20%, and is a major cause of cancer mortality. VTE prophylaxis is currently recommended for inpatients with malignancy and in those undergoing surgery (Chest 2008). Despite evidence that low molecular weight heparin (LWMH) has anti-tumor effect and improves short-term survival (Cochrane, 2011), the cost-effectiveness of anticoagulation in ambulatory cancer patients is unknown.

Methods:
We constructed a Markov model to evaluate prophylactic AC in ambulatory cancer patients with no previous VTE, during 4 months of chemotherapy following diagnosis. In the model, AC was enoxaparin. Major and minor bleeding risk, VTE risk, mortality, and other clinical parameters were obtained from the Cochrane 2011 analysis and other published studies. Drug costs were based on average wholesale price. Outcomes included medical costs, effectiveness measured by mortality reduction, quality-adjusted life-years (QALY), and incremental cost-effectiveness ratio (ICER) over a 24-month time horizon.

Results:
Base case results are shown in the Table. Compared with no LMWH, primary prophylaxis with LMWH in ambulatory cancer patients over a 4-month period was associated with a relative mortality risk of 0.92 over 24 months (relative risk reduction of 0.08), at a gain of 0.0484 QALY, for an ICER of $76,922/QALY gained, well within the accepted $100,000/QALY threshold. The results were not sensitive to variation in VTE relative risk (RR) with AC (base case RR 0.55), nor to AC-associated major bleeding (base case RR 1.3) or minor bleeding (base case RR 1.05) risks. Variation of the relative mortality risk with AC to ≥ 0.94, or the cost of AC (Figure) to ≥ $1,429/month (base $1,132), increased the ICER to >$100,000/QALY.

Discussion:
Prophylactic AC appears cost-effective in ambulatory cancer patients during the first four months of chemotherapy. Clinical trials will be needed to confirm the survival benefit of LMWH prophylaxis in ambulatory cancer patients.

Disclosures:
No relevant conflicts of interest to declare.
KLF1 Modulates Delta-Aminolevulinic Acid Dehydratase (ALAD) Gene Transcription in a Context-Specific Manner
Authors: Desgardin Aurelie , Abramova Tatiana , Lin Jenny , Shim Eun-Hee , Cunningham John M 
Published: October 2019

Abstract
Abstract 2094
Krüppel-like factor 1 (KLF1) is a zinc finger-encoding transcription factor that recognizes CACC elements, and is essential for maximal erythroid-specific gene transcription. Several critical mechanisms dependent on KLF1 and required for gene activation have been elucidated, predominantly using the beta-globin locus. KLF1 has been associated with the ordered recruitment of SWI/SNF and RNA polymerase-II complexes, necessary for chromatin remodeling and gene transcription respectively. KLF1 has also been reported to influence erythroid-specific heme biosynthesis. Studies in KLF1-null fetal erythroblasts and a KLF-1 deficient cell line have demonstrated that mRNA levels of the first three enzymes of the biosynthetic pathway are underrepresented. However, although in vitro studies of the rate-limiting enzymes ALAS2 and PBGD suggested a potential regulatory role for KLF1, in vivo studies failed to validate these findings. ALAD is the second enzyme of the pathway. Complete loss of ALAD expression in erythroid cells results in catastrophic events during zebrafish ontogeny. Interestingly, no human erythropoietic defect has been reported as a consequence of aberrant ALAD expression. To extend the analysis of KLF1's regulation of heme biosynthesis, we evaluated KLF1 binding of enzyme regulatory sequences by EMSA and ChIP studies, identifying a KLF1 binding CACC element in the erythroid-specific ALAD promoter. This regulatory element was transactivated specifically by a KLF1 transgene in KLF1-deficient cells. Using a unique 4-OH-Tamoxifen (4-OHT) mediated KLF1-inducible erythroid cell line (K1-ERp), we identified KLF1 as an essential, and early (within 2 hours of induction) activator of transcription of the endogenous ALAD, but not ALAS2 or PBGD genes. Further studies in K1-ERp cells, including DNAseI hypersensitivity and ChIP assays revealed that KLF1 occupancy at the erythroid-specific ALAD promoter triggers a series of molecular events including histone modifications, and enhanced recruitment of the sequence-specific transcription factors, GATA-1, NF-E2 and the TAL-1/SCL multiprotein complex. Importantly, we identified differences in the kinetics of recruitment of the closely related histone acetyltransferases proteins CBP and p300 and the SWI/SNF ATPase Brg1. The latter complex was recruited subsequent to KLF1 binding, although the ALAD promoter was already DNAseI hypersensitive. These results suggest strongly that KLF1 plays a major role in the regulation of heme biosynthesis in erythroid cells. Furthermore, our data challenges a model in which an identical temporal cascade of molecular events are required for transcription at KLF1-dependent promoters.

Disclosures:
No relevant conflicts of interest to declare.