bluuebunny/crossref_metadata_2025_split huggingface dataset:
Relation Between Cereblon Expression and Survival in Patients with Newly Diagnosed Multiple Myeloma Treated with Thalidomide
Authors: Broyl Annemiek , Kuiper Rowan , van Duin Mark , van der Holt Bronno , el Jarari Laila , Bertsch Uta , Zweegman Sonja , Buijs Arjan , Hose Dirk , Lokhorst Henk , Goldschmidt Hartmut , Sonneveld Pieter 
Published: October 2019

Abstract
Abstract 3973

Introduction:
Cereblon (CRBN) expression has been described to be essential for the activity of Thalidomide and Lenalidomide. This suggests that presence and possibly increased level of CRBN expression would be associated with better outcome in Thalidomide/Lenalidomide treated patients. The aim of this study was to evaluate CRBN expression in relation to outcome in patients receiving Thalidomide maintenance.

Patients and methods:
The HOVON-65/GMMG-HD4 trial is a multi-center, phase III trial, comparing Bortezomib in induction and post-intensification vs. conventional chemotherapy and daily Thalidomide 50 mg for 2 years post-intensification in newly diagnosed MM patients. This trial demonstrated that Bortezomib during induction and maintenance improved CR and achieved superior PFS and OS (Sonneveld et al., JCO, July 16, 2012). Gene expression profiling was performed at the start of the trial by Affymetrix U133 Plus 2.0 GeneChip, and was available for 96 patients which started Thalidomide maintenance. CRBN expression levels were based on a combined value of probe sets 218142_s_at and 222533_at. CRBN expression was validated using real-time PCR. All survival analyses were performed in SPSS, with survival time taken from the start of maintenance.

Results:
In patients receiving Thalidomide maintenance, increased CRBN expression was significantly associated with longer progression free survival (p=0.005, hazard ratio = 0.7) and longer overall survival (p=0.04, hazard ratio= 0.7). Using Kaplan-Meier analysis for visualization and using the median expression to define high and low expression, a significant separation was found for PFS (Log rank p=0.009) but not for OS (Log rank p=0.13). No association was observed between CRBN expression and PFS/OS after Bortezomib maintenance (PFS, p=0.4, hazard ratio=1.1; OS, p=0.7, hazard ratio=1.1). Multivariate Cox regression analysis was performed using the covariates ISS, CRBN and high-risk cytogenetics, defined as having del(17p) and/or 1q gain and/or t(4;14). Higher CRBN levels remained significantly related to longer PFS (hazard ratio 0.7, p=0.03), but not OS (hazard ratio of 0.8, p=0.3). High-risk cytogenetics and ISS were both significant in both PFS and OS multivariate models, with hazard ratios of 2.8 and 3.6, for high-risk cytogenetics and 2.5 and 5.5, for ISS stage 3, respectively (p=0.0004, p=0.003, high-risk cytogenetics and p=0.01 and p=0.005, ISS stage 3, respectively).

Conclusion:
These data suggest use of CRBN as a biomarker for thalidomide outcome, but further analysis in other Thalidomide trials is required to validate this finding.

Disclosures:
Lokhorst: Genmab: Consultancy. Sonneveld:Onyx: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; celgene: Honoraria, Research Funding.
DNA Copy Number Changes Have Gene Dosage Effects with Consequent Impact On Disease Biology and Prognosis in Multiple Myeloma
Authors: Samur Mehmet Kemal , Shah Parantu K , Wang Xujun , Huang Norman , Minvielle Stephane , Florence Magrangeas , Avet-Loiseau Herve , Munshi Nikhil C. , Li Cheng 
Published: October 2019

Abstract
Abstract 3984
Copy number alterations, deletions and amplifications, are very frequent in multiple myeloma (MM), however, it is less clear how these alterations affect gene expression. We performed a genome-wide analysis of 170 newly-diagnosed uniformly treated MM patients using high-density SNP arrays and Exon ST 1.0 gene expression arrays, and evaluated how copy number alterations affect gene expression in MM. Using SNP array data just over 40% patients had hyperdiploid MM (HMM) while the rest had non-hyperdiploid MM (N-HMM). We used two-step procedure to identify dosage effect scores of genes. At first, for each gene, percentage of copy number altered samples was calculated. Then for each gene percentage of samples that had dosage effect was calculated. Finally dosage effect score for each gene was calculated as a ratio of dosage effect samples percentage to copy number alteration sample percentage. We show that dosage effect in MM is wide-spread and some chromosomal locations are affected by dosage effect more compared to other locations. The dosage effect tracks can be observed at trisomy chromosomes and chromosome 1q, but most explicitly at chromosome 9, 11, 15 and 19. Also for deleted genes, dosage effect can be mostly observed at chromosome 13 and 16q. Separate analysis of HMM and N-HMM patients also showed that HMM patients have higher dosage effect especially in chromosome 15 compared to the others. In addition, relation between dosage effect and gene expression analysis show that the highly expressed genes have significantly higher dosage effect compared to the lowly expressed genes. Also function enrichment analysis showed that genes involved with crucial biological processes including translation, RNA processing and transcription factor genes are enriched in genes with higher dosage effect. Interstingly, dosage resistant genes are enriched in cell death and GTPase processes. These results help us understand the impact of aneuploidy in MM on global gene expression changes. In conclusion, our analysis identifies concordant and discordant gene expression changes associated with DNA copy number alterations, identifying genes and pathways that may play an important role in myeloma disease behavior as well as prognosis.

Disclosures:
No relevant conflicts of interest to declare.
Diagnostic and Prognostic Implications of Spine Magnetic Resonance Imaging At Diagnosis in Patients with Multiple Myeloma
Authors: Song Ik-Chan , Lee Kyu-Seop , Ryu Hae-Won , Lee Myung-Won , Lee Hyo-Jin , Yun Hwan-Jung , Kwon Soon-Tae , Kim Samyong , Jo Deog-Yeon 
Published: October 2019

Abstract
Abstract 3999
The role of baseline spine magnetic resonance imaging (MRI) in patients with multiple myeloma who have no related symptoms or signs is controversial. In the present study, we prospectively evaluated the diagnostic and prognostic implications of whole-spine MRI taken at the time of diagnosis in patients with multiple myeloma.
Patients who were newly diagnosed with multiple myeloma between 2004 and 2011 at Chungnam National University Hospital were enrolled. Abnormal MRI findings that were not detected in plain radiographs were analyzed and categorized as pathological fractures or extramedullary extension of the plasmacytoma. The bone marrow (BM) infiltration patterns on MRI were classified into five categories: homogenous diffuse, micronodular, macronodular, mixed, and normal appearance of the BM. The prognostic implications of the additional lesions found on MRI and the BM infiltration patterns were analyzed.
A total of 113 patients with a median age of 65 (range, 40μ–μ89) years were enrolled in the study. Pathological fractures that were not detected in the bone survey were found in 26 (23.0%) patients, including three (2.6%) patients with no related symptoms or signs. Extramedullary extension of the plasmacytoma was detected in 22 (19.5%) patients, including 15 (13.3%) with epidural extension of the tumor. Of these 22 patients, 11 (50.0%) had no relevant symptoms or signs. Neither the additional pathological fractures nor the extramedullary extensions found correlated with the international staging system stage (ISS). The presence of pathological fractures did not influence overall survival, whereas extramedullary extension of the plasmacytoma was associated with poor overall survival in multivariate analysis (HR 3.205; 95% CI, 1.430 – 9.845; P=0.042). Of the 113 patients, 35 (31.0%) had a diffuse infiltrative pattern, 39 (34.5%) had a micronodular pattern, 15 (13.3%) had a macronodular pattern, 17 (15.0%) had the mixed type, and 7 (6.2%) appeared normal. The BM infiltration patterns did not correlate with the ISS. During follow-up for a median of 21 (range 1 – 91) months, the overall survival of patients with the mixed type (median 24.0 months; 95% CI, 22.9 – 25.1) was shorter compared to those with other patterns (median 56 months; 95% CI, 48.9 – 63.1) (P=0.030). Survival differences were not found among the other patterns.
In summary, baseline spine MRI reveals not only bone lesions, but also extramedullary extension of plasmacytoma that cannot be detected in plain radiographs, even in patients with no relevant symptoms or signs. In addition, the mixed infiltration pattern on MRI is a predictor of poor overall survival. These results indicate that spine MRI should be included in the initial diagnostic investigations for patients with multiple myeloma.

Disclosures:
No relevant conflicts of interest to declare.
Serum Levels of B-Cell Maturation Antigen Are Elevated in Multiple Myeloma Patients and Correlate with Disease Status and Overall Survival
Authors: Sanchez Eric , Li Mingjie , Kitto Alex , Li Jennifer , Wang Cathy S , Rauch Danielle , Delijani Kevin , DeCorso Kevin , Prajogi Andrew , Chen Haiming , Berenson James R. 
Published: October 2019

Abstract
Abstract 4026

Background:
B cell maturation antigen (BCMA) is a receptor whose expression increases during B-cell development and is found on malignant cells from multiple myeloma (MM) patients; however, it has not been identified in human serum.

Methods:
Following informed consent (Western IRB BIO 001), serum was isolated from MM patients and analyzed with a BCMA enzyme-linked immunosorbent assay. Values represent the mean of triplicate experiments. Our human MM xenograft (LAGκ-2) was grown in SCID mice, and animals were treated with bortezomib (BORT) and cyclophosphamide (CY); tumor volume and BCMA levels were determined. Statistical significance of differences observed was determined using a Student's t test and analysis was determined using GraphPad prism software.

Results:
The serum BCMA levels from newly diagnosed MM patients (n = 58) was much higher (13.26 ng/ml) than among age-matched healthy subjects (n = 45; median 2.57 ng/ml; P < 0.0001) and MGUS subjects (n = 25; median 4.43 ng/ml; P = 0.002). Notably, protein levels were much higher among patients with relapsed or refractory disease (n = 88; median 18.99 ng/ml) compared to those with responsive (> partial response) disease (n = 95; median 3.48 ng/ml; P = 0.0016). Following treatment, patients with responsive disease showed decreases in BCMA levels whereas those with progressive disease showed increases. Additionally, with a median follow-up of 8 months (range, < 0 – 83 months), MM patients (n = 193) with BCMA levels above the median (8.43 ng/ml) showed a shortened survival compared to those with amounts below the median concentration (P < 0.0001). Following treatment with BORT and CY, we also showed a marked decrease in tumor volume and serum human BCMA levels in mice bearing the MM LAGκ-2 xenograft whereas untreated animals showed marked increases in tumor size and serum BCMA.

Conclusions:
This is the first report identifying serum BCMA in any human disease and suggests that these levels may be a novel biomarker for monitoring disease status and therapeutic response in MM patients.

Disclosures:
No relevant conflicts of interest to declare.
Casein Kinase II Modulates Topoisomerase II Alpha Nuclear Export and Drug Sensitivity in Multiple Myeloma
Authors: Turner Joel G , Dawson Jana L , Sullivan Daniel M 
Published: October 2019

Abstract
Abstract 4032
Nuclear export of topoisomerase II alpha (topo IIα) to the cytoplasm results in de novo resistance to topo IIα inhibitors in hematological malignancies. We have previously demonstrated that topo IIα is exported from the nucleus of human multiple myeloma cells by a CRM1-dependent mechanism at densities similar to those in myeloma patient bone marrow (Engel et al, Exp. Cell Res. 295:421-31, 2004). We have also identified the nuclear export signals for topo IIα at amino acids 1017–1028 and 1054–1066 using mutated full-length FLAG-topo IIα protein and immunofluorescence microscopy, (Turner et al, J. Cell Sci. 117:3061-71, 2004). In addition, blocking nuclear export with a CRM1 inhibitor or by using CRM1-siRNA sensitizes myeloma cells to topo II poisons (Turner et al. Cancer Res. 69:6899-905, 2009). Therefore, preventing nuclear export may sensitize myeloma cells to topo II inhibitors.
To determine what signals CRM1-mediated nuclear export of topo IIα we investigated the phosphorylation status of topo IIα, both in the nucleus and in the cytoplasm, using proteomic technologies. Topo IIα was isolated from the nuclei and cytoplasm of human myeloma cells by immunoprecipitation. Purified topo IIα fractions were analyzed by Orbitrap mass spectroscopy. We found that serine 1524 was highly phosphorylated in the cytoplasmic fraction. Using site-directed mutagenesis we converted serine 1524 to an alanine to determine if mutated FLAG-tagged topo IIα export was reduced as compared to wild-type FLAG-tagged topo IIα. Serine 1524 is a CK2 phosphorylation site, therefore, we tested a recently published and highly specific inhibitor of CK2; CX-4945 (Ferguson et al, FEBS Letters 585:104–110, 2011). This drug was used in combination with the topo IIα inhibitor doxorubicin and assayed for apoptosis by caspase 3 cleavage and flow cytometry both in vitro using myeloma cell lines and ex vivo using patient bone marrow mononuclear cells.
Using mass spectroscopy we found that cytoplasmic but not nuclear topo IIα was phosphorylated at serine 1524, a published CK2 motif. Using site-directed mutagenesis we converted serine 1524 to an alanine and found that mutated FLAG-topo IIα export was reduced, as compared to wild-type FLAG- topo IIα. The CK2 inhibitor CX-4945 induced apoptosis in both myeloma cell lines and in myeloma patient bone marrow aspirates. In addition, we found that CX-4945 prevented nuclear export of topo IIα in high-density cells. These data were duplicated using a CK2 specific siRNA to knockdown CK2 expression (90% knockdown). Blocking nuclear export of topo IIα with the CK2 inhibitor CX-4945 or CK2-specific siRNA sensitized drug-resistant myeloma cells to the topo II poison doxorubicin. These data may have potential clinical implications in the treatment of multiple myeloma.

Disclosures:
No relevant conflicts of interest to declare.
Comparison of Outcomes Between Unrelated Donor and Sibling Donor Transplant in Adult Patients with Severe Aplastic Anemia: A Single Center Retrospective Study
Authors: Lee Sung-Eun , Yoon Jae-Ho , Shin Seung-Hwan , Yahng Seung-Ah , Cho Byung-Sik , Eom Ki-Sung , Kim Yoo-Jin , Lee Seok , Min Chang-Ki , Kim Hee-je , Cho Seok-Goo , Kim Dong-Wook , Min Woo-Sung , Park Chong-Won , Lee Jong Wook 
Published: October 2019

Abstract
Abstract 4151
For the patient with severe aplastic anemia (SAA) who do not have a sibling donor and had failed to immunosuppressive therapy, allogeneic stem cell transplantation (SCT) from an unrelated donor is therapeutic option. Although unrelated SCT (u-SCT) has shown less favorable outcomes, there were few studies comparing unrelated donor to sibling donor transplant. The aim of the present study was to evaluate the feasibility of u-SCT in adult SAA patients compared with sibling SCT (s-SCT).
This study examined 157 consecutive patients who underwent allogeneic SCT at our institution between April 2001 and December 2011. In this study, for a homogenous cohort, patients who received second allogeneic SCT were excluded. The diagnosis of acute GVHD or chronic GVHD was made as the NIH published consensus criteria.
There were 71 male and 86 female patients with a median age of 31 years (range, 13–59 years). The median interval from the diagnosis to transplantation was 24 months (range; 1–346 months). The median transfusions prior to SCT were 47 units (range; 4–680 units). Patients with SAA (n=114) or VSAA (n=43) had received SCT from sibling donor (n=82) or unrelated donor (n=75). The conditioning regimen for s-SCT consisted of fludarabine (180 mg/m2) + cyclophosphamide (CY, 100 mg/kg) + ATG (10 mg/kg), and the conditioning regimen for u-SCT consisted of TBI (fractionated, 800 cGy) + CY (100–120 mg/kg) ± ATG (2.5 mg/kg) (Lee JW, et al, Biol BMT 2011, 17:101). GVHD prophylaxis consisted of CsA + MTX in s-SCT and FK506 + mini-MTX in u-SCT, respectively. After a median follow-up of 45 and 50 months for s-SCT and u-SCT, respectively, the 3-year estimated OS rates were 92.7 and 89 % for s-SCT and u-SCT (P=0.650), respectively. 15 patients (7 in s-SCT, 8 in u-SCT) died of transplant toxicities, including in s-SCT, acute GVHD (n=1), infections (n=4), and occurrence of MDS or AML (n=2) and in u-SCT, acute GVHD (n=4), chronic GVHD (n=1), infection (n=1), thrombotic microangiopathy (n=1). Nine patients (8 in s-SCT, 1 in u-SCT) developed secondary engraftment failure. The cumulative incidence of acute GVHD (≥grade II) was 4.9% in s-SCT and 44.0% in u-SCT (P<0.001). The cumulative incidence of chronic GVHD also showed higher in u-SCT than s-SCT (4.2% vs 44.4%, P<0.001).
This study showed that survival of u-SCT is comparable to that of s-SCT. Therefore, early application of u-SCT should be considered in patients with SAA/VSAA in case of availability of well matched donor. Because the incidence of GVHD is significantly higher in u-SCT than s-SCT, further study is mandatory to reduce the incidence of GVHD in u-SCT setting.

Disclosures:
No relevant conflicts of interest to declare.
The Impact of ATG/Alg in Preconditioning On the Allogeneic Stem Cell Transplantation for Patients with Acute Leukemia
Authors: Kato Koji , Atsuta Yoshiko , Ohashi Kazuteru , Fukuda Takahiro , Taniguchi Shuichi , Kanamori Heiwa , Sakamaki Hisashi , Yabe Hiromasa , Murata Makoto 
Published: October 2019

Abstract
Abstract 4185

Background:
Since the clinical implication of anti-lymphocyte globulin (ATG/ALG) in allogeneic stem cell transplantation (allo-SCT) is not fully understood, we tried to identify the clinical impact of ATG/ALG in patients with acute leukemia who received allo-SCT in Japan.

Patients and Methods:
We analyzed patients with ALL (n=5494) and AML (n=8115) who received first allogeneic SCT from 1983 to 2009 with (n=356) or without (n=13253) ATG/ALG. Their stem cell sources were bone marrow (BM, n=9056), peripheral blood (PB, n=1918), and cord blood (CB, n=2575) and they were transplanted at 1st complete remission (CR1, n=5681), 2nd CR (CR2, n=2495), and advanced stages (>CR3, n=5033).

Results:
Five year overall survival (5y OS) of all patients with or without ATG/ALG was 33.6% vs 44.5%, respectively (P<0.001) and multivariate analysis showed that ATG/ALG significantly reduced acute GVHD (P<0.001, HR=1.980) as well as chronic GVHD (P<0.001, HR=1.894). According to stem cell sources, 5y OS with or without ATG/ALG was 35.8% vs 47.5% (P<0.001) in BM, 34.7% vs 37.6% (P=0.067) in PB and 18.3% vs 39.9% (P<0.001) in CB. By multivariate analysis, ATG/ALG significantly reduced A-GVHD (P=0.005, HR=1.565) but decreased OS (P=0.004, HR=0.729) in BM, it reduced A-GVHD (P<0.001, HR=2.376) and C-GVHD (P<0.001, HR=2.691) but lowered engraftment (P=0.046, HR=0.810) in PB, and it increased TRM (P=0.004, HR=0.437) with decreased OS (P=0.011, HR=0.576) in CB. In Haplo transplantation (SCT from 2 or 3 antigens of HLA mismatched family donor, n=337), multivariate analysis showed that ATG/ALG did not affect the relapse, TRM and OS but, it significantly lowered engraftment (P=0.002, HR=0.602), and reduced A-GVHD (P<0.001, HR=2.622) as well as C-GVHD (P<0.001, HR=3.834). In contrast to these results, ATG/ALG did not affect the relapse rate irrespective of stem cell source or diagnosis.

Conclusion:
In allogeneic stem cell transplantation for patients with ALL and AML, ATG/ALG contribute in reducing acute and chronic GVHD without affecting relapse rates but it was a risk factor of OS for patients who received BM or CB.

Disclosures:
No relevant conflicts of interest to declare.
A Phase II Study Using Post-Transplant Cyclophosphamide (PTC) As Graft Versus Host Disease (GVHD) Prophylaxis in Patients Undergoing HLA Matched Sibling Donor Stem Cell Transplant (SCT) for Severe Aplastic Anemia (SAA)
Authors: George Biju , Mathews Vikram , Lakshmi Kavitha M , Ahmed Rayaz , Abraham Aby , Srivastava Alok 
Published: October 2019

Abstract
Abstract 4199
Between September 2010 and June 2012, 15 patients with SAA underwent HLA identical sibling donor SCT using post transplant cyclophosphamide for GVHD prophylaxis. The conditioning regimen consisted of Inj Fludarabine 30 mg/m2/day from day -7 to -2 and Inj Cyclophosphamide 50 mg/kg IV on days -3 and -2. Total Body irradiation (TBI) 200 cGy as single fraction was added on day -1 from December 2011 because of concern regarding graft failure. GVHD prophylaxis consisted of Inj Cyclophosphamide administered at 50 mg/kg/day IV on day +3 and +4 post SCT. G-CSF mobilized peripheral blood stem cells (PBSC) was the graft source.
There were 10 males and 5 females with a median age of 25.9 years (range: 8 – 42). The median time from diagnosis to SCT was 7.5 months (range: 2 – 36). All had 6/6 HLA matched sibling donors except 1 who had a 9/10 matched family donor. The median PBSC cell dose infused was 5.4 × 108 MNC/Kg (range: 2.4 – 8.5) and 9.58 × 106CD34/Kg (range: 5.4 – 17.2). Thirteen patients engrafted (86.6%) with a median time to ANC >0.5 × 109/L of 15.4 days (range: 15–17) and Platelet count >20 × 109 of 16.6 days (range: 12–32). Grade II –IV acute GVHD occurred in 3 patients (23%) at 42, 49 and 68 days post SCT. GVHD was grade II in 2 patients and grade IV in 1 patient. Two responded to a combination of cyclosporine and prednisolone while one patient with grade IV GVHD expired with refractory GVHD at median time of 64 days post SCT. Of the 11 evaluable patients, 4 (36.3%) developed chronic GVHD which was limited in all. Two patients who developed de novo chronic GVHD were managed with prednisolone alone. Overall 7 patients (46.6%) have not required any immunosuppression after SCT while 3 have required immunosuppressive therapy for 114, 127 and 225 days respectively At a median follow up of 11 months (range: 1 – 22), 11 (73.3%) are alive and well including 7 patients who did not require any immunosuppressive therapy following SCT.
In conclusion, the use of post transplant cyclophosphamide as GVHD prophylaxis following sibling donor transplant for SAA is associated with low rates of GVHD. What makes it more attractive is the fact that 46% did not require any immunosuppression post SCT. Larger studies are required to understand the utility of this prophylaxis in sibling donor transplants for aplastic anemia.
This trial is registered in the Clinical Trials Registry India - CTRI/2010/091/001480.

Disclosures:
No relevant conflicts of interest to declare.