bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mutational Profiling of Multiple Myeloma Bone Marrow Aspirates As a Clinical Tool for Personalized Treatment of Myeloma
Authors: Mahindra Anuj , Dias-Santagata Dora , Borger Darrell , Santo Loredana , Scullen Tyler A. , Yee Andrew , Eda Homare , Cirstea Diana , Iafrate Anthony John , Raje Noopur 
Published: October 2019

Abstract
Abstract 3990

Introduction:
Multiple Myeloma (MM) is increasingly being recognized as a heterogeneous disease. Previous studies have reported various oncogenic mutations in myeloma. Recent genome sequencing studies have revealed activating mutations of BRAF kinase in 4% of patients, a potentially druggable target, underscoring the importance of identifying these and other potential oncogenic mutations. However, a validated methodology for clinical assessment of mutation profiles is lacking. Applying such technologies will be increasingly important to accelerate development of targeted therapies toward tumor-specific oncogenic pathways. Utilizing a high-throughput genotyping platform we evaluated the commonly described somatic mutations in bone marrow aspirates in patients diagnosed with MM.

Methods:
We developed a CLIA (Clinical Laboratory Improvement Amendments) validated, highly sensitive multiplexed PCR-based assay (SNaPshot) to simultaneously identify 70 genetic loci frequently mutated in 15 cancer genes. This assay has been used at our institution for over 3 years for tumor genotyping and to help guide therapeutic decisions for patients with solid malignancies. To test the value of this approach in patients with MM, we first validated the methodology on MM cell lines followed by testing patient derived bone marrow samples.

Results:
The detectable mutations in the 8 MM cell lines tested include: MMIR: KRAS 35G>C, G12A mutation; MMIS: KRAS 35G>C, G12A mutation; H929: NRAS 38G>A, G13D mutation; LR5: KRAS 35G>C, G12A mutation; RPMI8226: TP53 743G>A, R248Q mutation;U266: no mutations identified; OPM2: NRAS 182A>T, Q61L mutation; OPM1: TP53 524G>A, R175H mutation.
Thirty six bone marrow aspirates from patients with MM have been analyzed to date. Tweny five percent of patients had a detectable mutation, with KRAS mutation in 17%, NRAS in 5% and TP53 in 3%. Additional data in a larger cohort is currently being analyzed and will be presented.

Conclusion:
Oncogenic driver mutations can be detected by SNaPshot from routinely-collected bone marrow aspirate samples. Experience from lung and breast cancer studies indicates that while the type and frequency of mutation varies somewhat by tumor phenotype, the common oncogenic mutations are often found in isolation. Broad tumor mutational analysis will be helpful to efficiently tailor personalized therapies based on individual tumor genetic profiles.

Disclosures:
Dias-Santagata: Bio-Reference Laboratories, Inc.: Consultancy. Borger:Bio-Reference Laboratories, Inc: Consultancy. Iafrate:Bio-Reference Laboratories, Inc: Consultancy. Raje:Eli-Lilly: Research Funding; Amgen: Research Funding; Acetylon: Research Funding; Millennium: Consultancy; Celgene: Consultancy; Onyx: Consultancy.
Disulfiram, an Clinically Used Anti-Alcoholism Drug Has the Potential to Target Leukemia Stem and Progenitor Cells in a Copper-Dependent Manner
Authors: Xu Bing , Wang Shiyun , Chen Feili , Shi Pengcheng , Zha Jie , Zhang Yanyan , Guo Xutao , Wang Weiguang 
Published: October 2019

Abstract
Abstract 4088

Backgrounds
Acute myeloid leukemia(AML) is a hierarchical disease initiating from a rare population of cells known as leukemia stem cells (LSCs), which are typically enriched in CD34+CD38- cells and presumed responsible for the relapse and refractory of AML. Moreover, current regimens may not effectively discriminate between normal and malignant cells. For this reason, it is important to identify therapies that can specifically target the LSC population without affecting normal cells. Disulfiram (DS) is an anti-alcoholism drug that has recently been indicated to show cytotoxic to multiple cancers including acute myeloid leukemia (AML) and the antineoplastic activity was enhanced in the present of copper (Cu). In the present study, we investigated the effect of DS/Cu on LSCs and further explored its mechanism.

Methods and Results
CD34+CD38- leukemia stem cell (LSC) enriched subpopulations were sorted from both KG1a cell lines and primary AML bone marrow or peripheral blood mononuclear cells (n=6) by fluoresce-activated cell sorting (FACS) analysis. Using MTT cell proliferation assay and Annexin-V/PI staining assay, We demonstrated that DS/Cu inhibited proliferation and induced apoptosis in CD34+CD38−KG1a cells (IC50= 0.788± 0.451 μM at 24h). With the increasing concentrations of DS (DS=0.05, 0.5, 5, 50μM), the apoptotic proportion increased from 7.2% to 89.5% at 24h. Apoptosis was also observed in CD34+CD38- primary AML cells and the exposure to DS/Cu (DS=0.01, 0.1, 1μM;Cu=0.5μM clearly inhibited the growth of AML-colony-forming units (CFUs) for both CD34+CD38-LSC enriched subpopulations (AML-CFUs decreased from 34.2% to 0% in KG1a cells), but was relatively sparing to normal hematopoietic progenitors. Further more, using flow cytometric analysis, western blot and RT-PCR, we identified that the change in redox status and redox-dependent signaling events play a crucial role in DS/Cu-induced apoptosis. We showed that DS/Cu(DS= 0.625,1.25,2.5,5μM, Cu=1μM) increased reactive oxygen species (ROS) and activated its downstream apoptosis-related SAPK/JNK pathway in association with blockade translocation of Nrf2 and expression of Nrf2-regulated genes in CD34+CD38−KG1a cells. Notably, blockade of ROS by glutathione precursor N-acetylcysteine (NAC)(10mM) strongly diminished DS/Cu mediated lethality and restored Nrf2 nuclear translocation and blocked JNK activation. Additionally, consistent with the ROS accumulation, we also seen that translocation of RelA/p65 and the expression of NF-κb-related gene, associated with abnormal apoptotic response of LSCs, were significantly inhibited by DS/Cu.

Conclusion
Taken together, we concluded that DS/Cu might selectively eradicate LSCs by induction of oxidatibe stress and blockade the NF-κb pathway and offers a potential therapeutic option in AML.

Disclosures:
No relevant conflicts of interest to declare.
Use of the Immunoproteasome Subunit LMP7 Inhibitor to Ameliorate Gvhd in an MHC-Matched miHA-Disparate Murine Model
Authors: Zilberberg Jenny , Dziopa Eugenia , Kirk Christopher J , Korngold Robert 
Published: October 2019

Abstract
Abstract 4106

Background:
Therapeutic targeting of the ubiquitin-proteasome pathway is currently used for the treatment of multiple myeloma. The immunoproteasome however, is a distinct class of proteasome found predominantly in monocytes and lymphocytes, and it is known to regulate antigen presentation on class I major histocompatibility complexes (MHC-I). ONX 0914 is a LMP7-selective peptide-epoxyketone proteasome inhibitor that is currently in pre-clinical development for the treatment of inflammatory diseases. In the current study, we evaluated the feasibility of using ONX 0914 in the B10.BR→CBA MHC-matched (H2k), minor histocompatibility antigen (miHA)-disparate bone marrow transplantation (BMT) model to decrease the incidence of GVHD. This well established allogeneic BMT model is relevant to the MHC-matched sibling donor or completely matched unrelated donor clinical situation.

Methods:
To evaluate the effects of ONX 0914 on GVHD, we tested different compound regimens in the model. CBA recipient mice were lethally irradiated (11 Gy, split dose, 4 h apart) and transplanted with 2×106 anti-T cell depleted B10.BR bone marrow cells (ATBM) alone, or in combination with 1×107 enriched B10.BR T cells (GVHD control and ONX 0914 treated groups). ONX 0914 was administrated in full (s.c., once/day at 8 mg/kg) or as a split dose (half dose, twice/day approximately 6 hr apart).

Results:
Treatment with the ONX 0914 compound significantly improved the survival of mice compared to control GVHD recipients when dosing was performed once/day for three consecutive days (0,1,2 post-BMT) [p = 0.02]. Alternating dosage (days 0,2,4) or daily drug treatment (days 0–4) provided comparable amelioration of GVHD in this model.

Conclusion:
Our preclinical findings suggest that the immunoproteasome subunit LMP7 is potentially a novel therapeutic target in GVHD as demonstrated by the improved survival rate of recipient mice treated with ONX 0914 in the B10.BR→CBA BMT murine model.

Disclosures:
Zilberberg: Onyx Pharmaceuticals: Research Funding. Dziopa:Onyx Pharmaceuticals: Research Funding. Korngold:Onyx Pharmaceuticals: Research Funding.
Effectiveness of Ursodeoxycholic Acid As Prophylactic Agent Against Hepatic Sinusoidal Obstruction Syndrome in Patients Undergoing Allogeneic Hematopoietic Cell Transplantation for Treatment of Hematological Malignancies
Authors: Ng Hong Yen HY , Chong Mui Fong MF , Song Jielin JL , Hwang William YK , Linn Yeh Ching , Loh Yvonne SM , Ho Aloysius YL , Goh Yeow-Tee YT 
Published: October 2019

Abstract
Abstract 4165

Introduction:
Hepatic sinusoidal obstruction syndrome (SOS) is a major and potentially fatal complication of hematopoietic cell transplantation (HCT). In May 2008, a risk-adapted SOS prophylaxis guideline was implemented for patients undergoing allogeneic HCT in Singapore General Hospital. Patients were stratified into 3 risk categories according to conditioning regimen and risk factors including preexisting hepatitis/cirrhosis or hepatic dysfunction. Low risk patients did not receive routine prophylaxis, and patients at moderate or high risk received oral ursodeoxycholic acid 250 mg TDS starting before conditioning till D+21 of HCT.

Objective:
This study aimed to evaluate the efficacy of ursodeoxycholic acid in preventing hepatic SOS in patients undergoing allogeneic HCT. In addition, additional risk factors associated with SOS were also identified.

Method:
A retrospective study of patients who underwent allogeneic HSCT between Jan 2005 and May 2012 was carried out where medical records were reviewed. Outcomes were compared between patients who received ursodeoxycholic acid as prophylaxis (from May 2008 to May 2012) against historical controls (from Jan 2005 to Apr 2008). The primary outcome was the incidence of SOS. Secondary outcomes included SOS-related mortality, and liver function post HCT.

Results:
A total of 202 patients were included with 129 patients in the study group and 73 historical controls. The incidence of SOS was 14% in the study group vs 23.3% in historical controls (p=0.121). In the moderate risk category, a statistically significant reduction of 12.7% in SOS occurrence was associated with routine prophylaxis (12.7% vs 25.4%, p=0.039). Among high risk patients, the incidence was 27.3% in the study group and 10% in controls, p=0.586. SOS-related mortality was comparable between the 2 groups. Elevation in liver function tests (including bilirubilin, ALT, AST, ALP, and GGT) post HCT was significantly lower in patients receiving ursodeoxycholic acid as prophylaxis. Prior HCT was identified as risk factor associated with SOS (OR, 9.082; 95% CI, 2.57 – 32.089; p=0.01).

Conclusion:
The use of ursodeoxycholic acid as prophylaxis resulted in a significant reduction in SOS occurrence in patients at moderate risk for this complication. However, the high incidence of VOD among high risk patients underscores the need for more effective prophylactic strategies in this population.

Disclosures:
No relevant conflicts of interest to declare.
Single-Infusion of Donor Early Apoptotic Cells (ApoCell) As Prophylaxis of Graft-Versus-Host Disease in Myeloablative HLA-Matched Allogeneic Bone Marrow Transplantati
Authors: Mevorach Dror , Reiner Inna , Or Reuven 
Published: October 2019

Abstract
Abstract 4176
Because of its potent immunomodulatory effect together with presumably high safety profile, apoptotic cells infusion (ApoCell) was tested in addition to cyclosporine and methotrexate, as prophylaxis of graft-versus-host disease (GVHD) in HLA-matched myeloablative allogeneic bone marrow transplantation (alloBMT) from a related donor. We conducted a phase I//IIa clinical trial where we treated 13 patients (median age, 37 years; range, 20–59 years) with advanced hematologic malignancies (7 patients with ALL, 5 patients with AML (4 patients with MDS that transformed to AML and one patient with AML de novo), and 1 patient with CML that received conventional myeloablation with 35, or 70, or 140, or 210×106 cell/kg of donor ApoCell, on day -1 of transplantation. Ten serious adverse effects were reported with all being not related (seven) or unlikely to be related (3) to ApoCell infusion. Out of hundreds adverse effects, only three were reported as possibly related to ApoCell infusion (with no definite or probable adverse effects). The non-relapse mortality at day 100 and 180 after transplantation was the same, 7.7%. The overall survival at 100 and 180 days after transplantation was 92% and 85%, respectively. The incidences of acute grades II through IV GVHD for all 13 patients and for the two higher doses were 23% and 0% (six patients), respectively. Biomarker ratio levels including tumor necrosis factor receptor I (TNFR1), hepatocyte growth factor (HGF), interleukin 2 receptor alpha (IL-2Ra), and interleukin 7 (IL-7), supported the clinical data. These results suggest that single-infusion high-dose donor early apoptotic cells in HLA-matched myeloablative alloBMT is a safe and potentially effective single-agent prophylaxis of acute GVHD occurring after myeloablative conditioning (clinicaltrials.gov no. NCT00524784).

Disclosures:
Mevorach: Enlivex: Consultancy, Employment, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties. Reiner:Enlivex: Employment.
SOCS1 Mutation Subtypes Predict Divergent Outcomes in DLBCL Patients
Authors: Lennerz Jochen K , Schif Birgit , Kohler Christian W , Bentink Stefan , Kreuz Markus , Melzner Ingo , Trümper Lorenz , Löffler Markus , Spang Rainer , Moeller Peter 
Published: October 2019

Abstract
Abstract 419
Suppressor of cytokine signaling 1 (SOCS1) is frequently mutated in Hodgkin, primary mediastinal and diffuse large B-cell lymphomas (DLBCL). In the primary mediastinal B-cell lymphoma line MedB-1, mutated SOCS1 abnormally stabilizes phospho-JAK2, thereby enhances STAT signaling leading to continuous proliferation. Here, we evaluated the prognostic value of SOCS1 mutations by full-length gene sequencing of SOCS1 in 154 comprehensively characterized DLBCL cases. By sequence analysis, we identified 90 SOCS1 mutations in 16% of lymphomas. We defined two distinct subtypes with respect to putative mutational consequences: those predicting the full-length (minor) and a truncated protein (major), respectively. Neither the SOCS1 mutation group, nor minor/major subgroups can be distinguished by clinical phenotype; however, assignment of four established expression-based classifiers revealed significant associations of SOCS1 major cases with germinal center- and specific pathway activation pattern signatures. Above all, SOCS1 major cases had an excellent overall survival, even better than the GCB-like subgroup (see Figure). SOCS1 minor cases had a dismal survival, even worse than the ABC gene signature group (see Figure). SOCS1 mutation subsets retain prognostic significance in uni- and multivariate analyses. Thus, if a SOCS1 mutation is present, the mutation type is an important single gene prognostic biomarker in DLBCL.

Disclosures:
No relevant conflicts of interest to declare.
T Cell Repertoire Diversity After Umbilical Cord Transplant Predicts Mortality From Infection
Authors: Delaney Colleen , Emerson Ryan O , Milano Filippo , Sherwood Anna , Papermaster Adrienne , Guthrie Katherine A. , Carlson Christopher S , Warren Edus H. , Robins Harlan 
Published: October 2019

Abstract
Abstract 4202

Background
In a transplant study run at Fred Hutchinson Cancer Research Center, 34 patients with high risk hematological malignancies underwent a myeloablative procedure and were subsequently transplanted with double umbilical cord blood units. Peripheral blood samples were collected from each patient before myeloablation, and at 28, 56, 100, 180, and 360 days post-transplant.

Methods
At each time point, we used the immunoSEQ platform to perform high-throughput sequencing of rearranged T Cell Receptor (TCR) loci. Using immunoSEQ data, we were able to track the presence and frequency of individual TCR clones in each patient across time-points, as well as measuring the diversity of the TCR repertoire as a whole. We correlated our measure of TCR repertoire diversity with clinical outcomes in this cohort.

Results
The study produced two primary results. First, using the ability to track clones, the reconstituting TCR repertoire is shown to oscillate wildly with nearly complete turnover of the T cell repertoire occurring at least monthly after CB transplant. The largest T cell clones present in each blood draw drop below detection within weeks, contrasting with control data in which the top clones in healthy patients are not only observed in multiple subsequent time-points, but remain at high frequency. The second result is a test of the hypothesis that diversity of the T cell repertoire is a measure of immunocompetence, as a clinical application of high-throughput sequencing. Of the 34 patients, six died between Day 100 and Day 360 of non-relapse causes. At both Day 56 and Day 100, the diversity of the T cell repertoires of those six patients were far lower than the T cell repertoire diversity values of the remaining patients (P-value = 0.015).

Conclusions
We have demonstrated that the reconstitution of clinical immunity in cord blood transplantation patients is characterized by a highly unstable T cell compartment with very rapid turnover of T cell clones. Despite the transience of individual T cell clones, however, by two months after transplant T cell repertoire diversity as measured by high-throughput TCR sequencing accurately predicts risk of non-relapse mortality.

Disclosures:
Emerson: Adaptive Biotechnologies: Employment, Equity Ownership. Sherwood:Adaptive Biotechnologies: Employment, Equity Ownership. Carlson:Adaptive Biotechnologies: Consultancy, Equity Ownership, Patents & Royalties. Robins:Adaptive Biotechnologies: Consultancy, Equity Ownership, Patents & Royalties.
Clinical Characteristics and Outcome of Isolated Extramedullary Relapse in Acute Leukemia Following Allogeneic Stem Cell Transplantation: A Single Institutional Analysis
Authors: Shi Jimin , Meng Xiaojian , Luo Yi , Tan Yamin , Zhu Xiaoli , Zheng Gaofeng , He Jingsong , Zheng Weiyan , Xie Wanzhuo , Li Li , Ye Xiujin , Zhang Jie , Cai Zhen , Lin Maofang , Huang He 
Published: October 2019

Abstract
Abstract 4211
Isolated extramedullary relapse (EMR) of acute leukemia (AL) is a rare occurrence although it appears to be more common following allogeneic stem cell transplantation (allo-SCT). To characterize what has been observed in isolated EMR, we investigated a total of 287 consecutive AL patients (144 acute myeloid leukemia, 138 acute lymphocytic leukemia and 5 acute mixed lineage leukemia) who underwent allo-SCT.
Forty-seven (16.4%) patients experienced relapse after allo-SCT. 12 cases (4.2%) experienced relapse of extramedullary sites without concomitant bone marrow involvement. Isolated EMR accounted for 25.5% of the overall initial relapse. The time to relapse after allo-SCT was longer in the extramedullary sites than in the marrow (median, 10 months vs. 5.5 months, P<0.05,). Sites of EMR varied widely including central nervous system, skin, bone, pelvis and breasts. The variables considered for univariate analysis included donor gender, age, primary disease, disease status, cytogenetics/molecular abnormalities, preconditioning regimen, donor type, HLA match, aGVHD and cGVHD. The variables that showed significant correlation with isolated EMR were the cytogenetics abnormalities at diagnosis. The prognosis for patients who develop EMR remained poor but was relatively better than that after bone marrow (BM) relapse (overall survival, 10 vs. 18 months, P<0.05). Compared with local or single therapy, patients treated with systemic in combination with local treatment could yield favorable prognosis. Three patients survived 63, 55 and 49 months after transplant respectively, of whom two received DLI with subsequent chemotherapy and (or) irradiation and one had surgery with subsequent chemotherapy.
In conclusion, we observed a significant number of isolated EMR of AL after allo-SCT and intensive approaches combined of local and systemic therapy can produce favorable response which may cure a percentage of these patients.

Disclosures:
No relevant conflicts of interest to declare.
Hematopoietic Stem Cell Transplantation and Secondary Primary Malignancies in Taiwan
Authors: Liu Chia-Jen , Chen Kuo-Wei , Hu Yu-Wen , Hong Ying-Chung , Huang Yu-Chung , Chiou Tzeon-Jye , Tzeng Cheng-Hwai 
Published: October 2019

Abstract
Abstract 4219

Objectives
Hematopoietic stem cell transplantation (HSCT) is a curative strategy for many hematological disorders. The improvement of HSCT may lead to longer overall survival of patients with catastrophic illness and the risk of secondary cancer development become an emerging issue in long-term survivors.

Patients and Methods
We conducted a nationwide population-based study of 1,881 patients with hematologic diseases undergoing HSCT between January, 1997 and January, 2007 using Taiwan's National Health Insurance Research database. Performing HSCT to treat non-hematological diseases were excluded. All patients were followed until solid cancer development, death, or the end of 2010. We did not put hematological malignancies as endpoint because solid tumors and hematological malignancies might have different carcinogensis mechanism. We used standardized incidence ratios (SIRs) to compare patterns of cancer incidence in patients with those of the general population. Multivariate analysis was undertaken using Cox proportional-hazards regression using a forward selection, likelihood ratio model to identify independent predictors of cancer development among patients after HSCT.

Results
We observed a total of 8,753.87 person-years in this study. Patients received HSCT had a significant increased risk of developing any kind of cancer (SIR 1.77, 95% confidence interval [CI] 1.15 – 2.62; p = 0.011). Specifically, patients after HSCT had increased cancer incidence of head and neck cancer (SIR 3.96, 95% CI 1.90 – 7.29; p < 0.001) and bone and soft tissue sarcomas (SIR 10.08, 95% CI 1.22 – 36.42; p = 0.035). In subgroup analyses, cancers were more likely to develop in patients aged 0 – 19 years (SIR 22.31, 95% CI 4.60 – 65.20; p < 0.001), and those who survived more than 5 years after HSCT (SIR 3.30, 95% CI 1.96 – 5.22; p < 0.001).

Conclusion
Our study demonstrates an increased incidence of cancer development in patients after HSCT, especially malignancies of head and neck and also bone and soft tissue sarcomas. Patients aged 0 – 19 years and those survived more than 5 years after HSCT have a higher incidence of developing cancer comparing to normal population. HSCT long-term survivors should therefore be monitored more carefully for cancer development and targeted with preventive intervention strategies.

Disclosures:
No relevant conflicts of interest to declare.