bluuebunny/crossref_metadata_2025_split huggingface dataset:
Igh Translocation with Undefined Partners Is Associated with Superior Outcome in Multiple Myeloma Patients
Authors: Mao Xue-Han , Xu Yan , Yan Yuting , Zhuang Junling , Fan Huishou , Sui Weiwei , Liu Jiahui , Deng Shuhui , Yi Shuhua , Qiu Lugui , An Gang 
Published: November 2020

Background: In multiple myeloma (MM), impact of specific chromosomal translocations involving IgH (14q21 locus, including t(4;14), t(11;14), and t(14;16) etc.) has been explored extensively. However, over 15% MM patients harboring IgH translocation with undefined partners have long been ignored.
Methods: A prospective non-randomized cohort study with a total of 715 newly-diagnosed MM cases was conducted, 13.6% of whom were t(14;undefined) positive. The whole cohort was divided into four groups: no IgH split (47.7%); t(14;undefined) (13.6%); t(11;14) (17.6%); and t(4;14) or t(14;16) group (21.1%).
Results: Median OS for the four groups were 84.2, not reached (NR), 58.7, and 44.2 months respectively, with p values for t(14;undefined) vs. no IgH split, t(11;14), and t(4;14)/t(14;16) groups of 0.197, 0.022 and 0.001, respectively(Figure B).In bortezomib-based group, the survival advantage gained by t(14;undefined) group was much more significant compared to t(11;14) and t(4;14)/t(14;16) groups. Importantly, t(14;undefined) turned out to be an independent predictive factor for longer OS of MM patients in multivariate analysis, especially in the context of bortezomib-treatment. Similar results were also observed in the PUMCH external validation cohort (Figure C).
Conclusion: Collectively, our data confirmed and externally validated the favorable prognosis of the t(14;undefined) groups, especially in the era of novel agents.

Disclosures
No relevant conflicts of interest to declare.
Characterization of the Immunoglobulin Heavy- and Light-Chain Repertoires in a Single Reaction
Authors: Lowman Geoffrey , Toro Michelle , Chang Jayde , Pickle Loni , Yang Chenchen , Looney Timothy 
Published: November 2020

Background
B cell repertoire analysis by next-generation sequencing (NGS) has shown particular utility in the field of hematological oncology research. Some advantages provided by NGS-based techniques include a lower limit-of-detection and simpler paths to standardization compared to flow-based methods, and the elimination of specifically designed primers often required for qPCR-based methods. Owing to primer-primer interactions and incompatibility of reaction conditions, current multiplex PCR assays require separate PCR reactions to survey each immunoglobulin chain (IGH, IGK, IGL), often leading to a longer time-to-answer for samples in which no marker is initially detected. We have developed an assay for receptor analysis based on Ion AmpliSeq technology to circumvent these issues, allowing the effective use of up to thousands of primers in a single reaction. The highly multiplexed, pan-clonality NGS assay provides for efficient detection of IGH, IGK, and IGL chain rearrangements in a single reaction.
Methods
We developed a single primer panel targeting the framework 3 (FR3) portion of the variable gene and the joining gene region of heavy- and light-chain loci (IGH, IGK, IGL) for all alleles found within the IMGT database, enabling readout of the complementary-determining region 3 (CDR3) sequence of each immunoglobulin chain. To maximize sensitivity, we included primers to amplify IGK loci rearrangements involving Kappa deletion and C intron elements. To evaluate performance, we conducted clonality assessment and limit-of-detection testing used gDNA from a total of 45 research samples representing common B cell malignancies. We included samples derived from peripheral blood, bone marrow, and FFPE-preserved tissues at input levels ranging from 100ng to 2µg. Finally, we further characterized the samples via a separate AmpliSeq-based multiplex PCR assay targeting rearranged TCRB and TCRG chains. Sequencing and clonality analysis was performed using the Ion GeneStudio S5 System and Ion Reporter 5.16.
Results
Clonality assessments carried out using gDNA collected from both cell line and clinical research samples (CLL, B-ALL, Multiple Myeloma, Burkitt's Lymphoma, NHL, and DLBCL) show a &gt;90% overall positive detection rate. Assessment of linearity-of-response and limit-of-detection was carried out using cell lines diluted in PBL to between 10-3 and 10-6 by mass. The multi-receptor assay performs as expected, with linear response to the cell line frequency across the range tested, including the ability to detect clones of interest at 10-6.
Conclusions
These results demonstrate the robustness of our newly developed Ion AmpliSeq-assay for B cell receptor heavy and light chains. We expect this assay to simplify the workflow for clonality assessment and rare clone detection in B cell malignancy research.
For research use only.

Disclosures
No relevant conflicts of interest to declare.
CD19 CAR-T Cells Treatment Conferred a Sustained Remission in Patients with Chemotherapy-Refractory MRD in B-ALL
Authors: Lu Wenyi , Wei Yunxiong , Cao Yaqing , Xiao Xia , Li Qing , Lyu Hairong , Jiang Yili , Zhang Huan , Wang Jia , Li Xin , Jiang Yanyu , Juanxia Meng , Ting Yuan , Zhu Haibo , He Xiaoyuan , Jin Xin , Sun Rui , Sui Tao , Liu Kaiqi , Zhao Mingfeng 
Published: November 2020

Background: The persistence or recurrence of minimal residual disease (MRD) after chemotherapy predicts relapse of B-cell acute lymphoblastic leukemia (B-ALL). CD19-directed chimeric antigen receptor-modified T (CD19 CAR-T) cells have shown excitable response in B-ALL. Recently, some studies have shown that ALL patients with lower burden had higher CR rate and lower risk of CRS after CAR-T therapy (Park, et al, NEJM. 2018; 378(5):439-448. Lee, et al, Blood.128 (22):Abstract 218.). However, its role in chemotherapy-refractory MRD-positive B-ALL remains unclear. Here we aimed to assess the effectiveness and safety of CD19 CAR-T in MRD-positive B-ALL patients.
Methods: Since January 2018, a total of 14 B-ALL patients with persistent (n=8) or recurrent (n=6) MRD were enrolled in the CAR-T clinical trials (ChiCTR-ONN- 16009862 and ChiCTR1800015164). The patients were from two different clinical trials about CAR-T. If patients were treated in an MRD positive state, they would be included in this analysis. All the patients received one or more infusions of autogenous CD19 CAR-T.
Results: Median age was 37.5 (13-62) years and 7 patients were female. The median dose of infused CAR-T cells was 6.78´106cells/kg, and 5 patients received more than one infusion. After one cycle of CAR-T infusion, 12 patients achieved MRD-negative remission, leading a response rate of 85.7%. Of the subgroup of 5 Ph-positive patients who subsequently underwent transplantation, 2 patients died due to transplant-related toxic effects, whereas the other 3 patients all currently alive without leukemia. Of the subgroup of 9 Ph-negative patients, 8 patients did not undergo subsequent transplantation (Figure 1). Three patients finally suffered CD19-positive relapse and 1 patient suffered CD19-negative relapse. Importantly, 4 patients (50%) are in ongoing molecular remission without transplantation, with a duration of response averaging 22.9 months (range: 12.1-28.6 months).
The most frequent adverse events were fever and hematopoietic toxicities. Ten patients (71.4%) had grade 1 or 2 cytokine release syndrome (CRS) and no patients died of CRS. At a median follow-up time of 599.5 days (range: 172-915 days), the probability of 2-year relapse-free survival and 2-year overall survival was 61.2%±14.0% and 78.6%±11.0%, respectively.
Conclusion: In conclusion, pre-emptive CD19 CAR-T treatment is an effective and safe approach and may confer a sustained remission in B-ALL patients with chemotherapy-refractory MRD.

Disclosures
No relevant conflicts of interest to declare.
Three-Dimensional Telomere Analysis Using Teloview® Technology Identifies Smouldering Myeloma Patients with High Risk of Progression to Full Stage Multiple Myeloma in a Proof of Concept Cohort
Authors: Louis Sherif , Rangel-Pozzo Aline , Knecht Hans , Mai Sabine 
Published: November 2020

Multiple Myeloma (MM) remains incurable in spite of the recent development of several advanced therapeutic modalities. Similar to other incurable diseases, the early diagnosis and treatment of MM patients may lead to better prognosis and increased survival rates. Multiple myeloma follows two asymptomatic precursor lesions, monoclonal gammopathy of undetermined significance (MGUS) and smouldering myeloma (SMM) [1]. In current clinical practice patients diagnosed with MGUS or SMM are monitored but not treated. MGUS and SMM may progress to the full stage of multiple myeloma at any time without clinically detectable signals. While 1% of MGUS patients progress to full stage MM every year, 10% of SMM patients progress to full stage MM every year [1]. In this study, we report for the first time that the 3-dimensional telomeres analysis using the TeloView® software platform is able to identify SMM patients with high risk of progression to full stage MM. The prospective study we conducted included a proof of concept cohort of total 21 SMM patients, 16 patients that remained stable for over 5 years, and 5 patients progressed to full stage multiple myeloma within 1 to 3 years from point of diagnosis. The disease progression of high risk SMM patients was confirmed clinically by MM caused morbidity. TeloView® analysis of the 2 SMM patient-groups revealed with high significance (p &lt;0.001) distinct telomeres profiles of the stable patients versus the patients who progressed to full stage MM across 5 independent telomeric parameters measured by TeloView®. The study was conducted blindly on the diagnostic specimens suggesting the ability of TeloView® analysis to stratify SMM patients at point of diagnosis. The results we report have the potential, for the first time, to guide evidence-based decisions to treat SMM patients with high risk of progression, addressing a critical unmet clinical need in the management of MM. Follow up studies including expanded cohorts are needed to validate the results of this study, and to confirm the utility of TeloView® technology to predict the progression of SMM patients on the level of the individual patient.
References: Boutros M. et al Genomic Profiling of Smouldering Multiple Myeloma Identifies Patients at a High Risk of Disease Progression. J Clin Oncol. 2020 Jul 20;38(21):2380-2389

Disclosures
Louis: Telo Genomics Corp.: Consultancy, Current equity holder in publicly-traded company. Knecht:Telo Genomics Corp.: Consultancy, Current equity holder in publicly-traded company. Mai:Telo Genomics Corp.: Consultancy, Current equity holder in publicly-traded company.
Bortezomib and Rituximab in Newly Diagnosed Adolescent and Adult CD20-Positive Philadelphia (Ph) Negative Precursor B-Cell Acute Lymphoblastic Leukemia: A Phase II Study
Authors: Goli Vasu Babu , Jain Hasmukh , Tembhare Prashant , Shetty Dhanlaxmi Lalit , Bonda Avinash , Nayak Lingaraj , Patkar Nikhil , Subramanian Papagudi Ganesan , Sengar Manju 
Published: November 2020

Background:
The expression of CD20 in precursor B-cell ALL has been associated with poor outcomes. The addition of rituximab with intensive chemotherapy in this subset of ALL has led to improvement in the event-free survival. Further increment in outcomes require novel approaches. Bortezomib is an active drug in relapsed ALL as well as has synergistic activity with rituximab in B-cell lymphomas; thus the addition of bortezomib to rituximab and chemotherapy may improve the outcomes in CD20-positive precursor B-cell ALL.
Methods:
We conducted a phase II study to test the activity of bortezomib and rituximab in combination with a paediatric inspired regimen during induction therapy in newly diagnosed adolescent and adults (&gt;14 years of age) with CD20-positive, Philadelphia (Ph)-negative precursor B-ALL, with bone marrow measurable residual disease (MRD) negativity at the end of induction (EOI) as the primary endpoint.
Results:
From December 2017 through August 2019, a total of 35 patients were enrolled. A total of 70.99% patients achieved EOI MRD negative status. MRD negative rates improved to 88% post consolidation. There was no significant increase in toxicity with addition of bortezomib and rituximab to standard chemotherapy. The incidence of neuropathy was 26% (&lt;grade 3). After a median follow up of 18 months, event-free survival and overall survival were 81.8% and 84.7%, respectively.
Conclusion:
The combination of bortezomib, rituximab and paediatric-inspired ALL regimen is active and well tolerated in in de-novo CD20 positive Ph-negative precursor B-ALL.

Disclosures
No relevant conflicts of interest to declare.
Supplementation of Barium to Cold-Stored Platelets Improves Platelet Quality over 7 Days of Storage
Authors: Reddoch-Cardenas Kristin M , McIntosh Colby , Calhoun Cheresa , Barrera Gema , Lizarraga Stephanny , Bynum James , Cap Andrew P. 
Published: November 2020

Background:
Hemorrhage is responsible for the majority of potentially survivable deaths amongst military and civilian sectors. Early interventions to manage bleeding during hemorrhagic injury, including administration of platelets (PLTs), are crucial for decreasing mortality. Cold-stored PLTs are an attractive treatment for active bleeding as they provide greater functional preservation and a reduced risk of bacterial sepsis than conventional room temperature stored PLTs. However, cold storage causes a rise in PLT intracellular calcium, subsequent activation of signaling pathways, and often results in aggregate formation. In this study, we hypothesized that storing PLTs in the cold with barium, a divalent cation chelated by citrate anticoagulant, would improve PLT quality.
Methods:
Apheresis PLTs in 100% plasma were collected from healthy donors (n=4). Briefly, 10 mL of PLTs were sterilely aliquotted and added to 15 mL mini-bags containing 0- (Control), 3-, or 6-mM BaCl2. Individual mini-bags were used for each timepoint and condition. Bags were incubated for 15 min at room temperature prior to long-term storage in a walk-in refrigerator (1°-6°C). Samples were assayed on days 1 (the day after collection) and 7 of storage for PLT metabolism, count, clot formation, clot retraction, and activation (phosphatidylserine, mitochondrial depolarization, GPIb, and P-Selectin). Data were reported as means±SD.
Results:
Compared to Control at day 7, BaCl2-treated bags exhibited higher PLT counts (Control: 422±215 x103 PLTs/µL; 3 mM BaCl2: 468±222 x103 PLTs/µL; 6 mM BaCl2: 697±194 x103 PLTs/µL), less PS exposure (Control: 6.3±3.5%; 3 mM BaCl2: 5.8±3 %; 6 mM BaCl2: 5.6±2.8 %), and less mitochondrial membrane depolarization. Additionally, glucose consumption and lactate production were lower in barium-supplemented PLTs. GPIb expression and P-Selectin was comparable amongst all groups. Hemostatic function, as assessed via thromboelastography and clot retraction, was not negatively affected by addition of BaCl2. In most assays, cold storage of PLTs with the higher concentration of BaCl2 (6 mM) seemed to confer the greatest advantages.
Conclusion:
Supplementation of barium to PLTs provides a promising avenue for transfusion medicine since PLT count is better preserved and activation is decreased compared to normal refrigerated storage. Future studies are warranted to determine the effects of barium supplementation on other aspects of cold-stored PLT function (e.g. procoagulant activity and PLT adhesion under physiologic flow) and with respect to alternate storage media (e.g. PLT additive solutions).

Disclosures
No relevant conflicts of interest to declare.
Therapeutic Effect of a Traditional Chinese Medicine Compound Mixture Compound Kushen Injection in Treating Multiple Myeloma
Authors: Jiang Hongqiang , Tu Honglei , Jin Yanxia , Wu Xianjin , Luo Ziyi , Chen Yanling , Zhang Dongdong , Wu Balu , Wei Yongchang , Yang Yi , Zhou Fuling 
Published: November 2020

Abstract
Background: Multiple myeloma (MM) is a hematology malignant disease originated from B-cell line and still incurable. Compound Kushen Injection (CKI) as a Traditional Chinese Medicines are promising agents in our previous research for treating cancer. The effect of CKI on multiple myeloma was still unknown.
Methods: In vitro experiment, flow cytometry was used to evaluate effect of CKI on multiple myeloma cells. Optofluidic chip was applied to detect effect at single-cell level. And in vivo RPMI-8226 GFP+ B-NSG mouse model was built to assess the role of CKI in multiple myeloma treatment.
Results: CKI inhibited MM cells proliferation of and increased its apoptosis rate. And the cell cycle of MM cells was also arrested by CKI treatment. In contrast, CKI has few toxic effects on mesenchymal stem cells (MSCs) and MC3T3 cells. At the single-cell level, MM cells was died in time and dose dependent manner. Transcriptome find that the expression of MYC and TERT in CKI-treated RPMI-8226 cells was significantly down-regulated and confirmed by qRT-PCR and Western blot. Overexpression of TERT can partly reverse the inhibition effect of CKI on RPMI-8226 cells. B-NSG mouse was injected with GFP+ RPMI-8226 cells through caudal vein, and the disease was partially alleviated by decreased tumor burden in the CKI-treated group. Furthermore, it is surprising that in animal models with myeloma bone disease, the bone mass was higher in CKI treatment group than control.
Conclusions: CKI inhibits MM cells through the MYC/TERT signaling pathway and improve the quality of life of MM mouse. Our findings provide preclinical evidence to show that CKI could be a promising candidate in human MM therapy.

Disclosures
No relevant conflicts of interest to declare.
Vitamin D Levels Are Associated with a Dichotomous Distribution of Hematological Parameters in a Large Cohort of Otherwise Healthy Subjects
Authors: Sevindik Omur Gokmen , Olmuşçelik Oktay 
Published: November 2020

Introduction
Vitamin D deficiency is generally associated with a slowed hematopoiesis and the final outcome is varying degrees of peripheral blood cytopenia(s). The net effect of increased levels of Vitamin D is not well defined. In this study we tried to document the effect of varying levels of Vitamin D on hematological parameters.
Material and Methods
Whole patient registry of Medipol Complexes screened and a total of 12709 patients who met the pre-defined inclusion criteria enrolled in the study. Two datasets formed, first including all patients (Dataset A), second including patients with no CBC abnormality and not having any nutritional deficiency (Iron, Vitamin B12 and Folic acid).
Results
There was a statistically significant correlation between major CBC indices showing a maximum for the second pre-defined (according to quartiles 1st group included patients with a Vitamin D below 10 ng/ml, 2nd group included Vitamin D between 10-20 ng/ml, 3rd group included vitamin D between 20-30 ng/ml and the 4th group included patients who had a Vitamin d level above 30 ng/ml) Vitamin D group. This trend was statistically significant for major hematological parameters and post-hoc analysis also revealed a positive trend through Group 2 (10-20 ng/ml). Regression analysis also documented that patients who were classified under group 2 possessed a more potent hematopoiesis when compared to others even when adjusted for age, gender and baseline CRP values (Figure 1).
Conclusion
Our study reports a statistically significant correlation between major hematological parameters and Vitamin D levels in a particularly large patient population who lacks a significant confounder like chronic illnesses or conditions which can contribute and potentially serves as a bias. Even more, the group who has a maximal capacity of hematopoiesis are the ones harboring relatively normal but not high levels of Vitamin D.
Figure

Disclosures
No relevant conflicts of interest to declare.
DNA Methylation-Mediated Silencing of Mir-204 Enhances the Occurrence of T-Cell Acute Lymphoblastic Leukemia By Upregulating MMP-2 and MMP-9 through the NF-Κb Signaling Pathway
Authors: Chen Dabing , Xiao Tingting , Lin Dandan , Zhu Haojie , Xu Jingjing , Luo Xiaofeng , Ren Jinhua , Zheng Zhihong , Chen Zhizhe , Hu Jianda , Yang Ting 
Published: November 2020

Background : MicroRNAs (miR) are non-coding RNAs that play a role in regulation multiple functions in different cell types. Previous studies have shown that miR-204 is downregulated in T-ALL. We previously reported that matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) gene polymorphisms may be associated with the risk of T-cell acute lymphoblastic leukemia (T-ALL). The present study aims to decipher the role of miR-204 and MMP-2/MMP-9 in T-ALL occurrence to guide the diagnosis and treatment of T-ALL in the clinics.
Methods: Expression of miR-204 was determined in the bone marrow and peripheral blood samples from 70 T-ALL patients and 70 healthy volunteers by real-time quantitative PCR (RT-qPCR). Bisulfite sequencing PCR (BSP) was used to detect the DNA methylation levels of the miR-204 promoter region in T-ALL patients and T-ALL cell lines.The effect of miR-204 on cell proliferation was evaluated with the cell counting kit-8 solution (CCK-8) assay and by Hoechst and PI double staining. The binding site of miR-204 on IRAK1 was predicted by the Primer Premier 5.0 and the defined binding sequences were used to construct luciferase-tag plasmids. The regulation of IRAK1 expression by miR-204 was evaluated by RT-qPCR and Western blot analysis. With the purpose to confirm the role of MMP-2 and MMP-9 in the occurrence of T-ALL, we investigated the effect of related proteins on T-ALL cells using Western blot. To determine that miR-204 affects the occurrence of T-ALL disease by regulating the NF-KB signaling pathway, RT-qPCR and Western Blot were used for verification.
Results: DNA methylation directly affects the miR-204 expression in the promoter region when T-ALL developed. Moreover, overexpression of miR-204 inhibited the proliferation and enhanced the apoptosis of T-ALL cells. Notably, overexpression of miR-204 inhibited IRAK1, which in turn inhibited the proliferation and enhanced the apoptosis of T-ALL cells. Furthermore, IRAK1 enhanced the expression of MMP-2 and MMP-9 through phosphorylation of of p65 NF-κB, and miR-204 modulated MMP-2 and MMP-9 expression through the IRAK1/NF-κB signaling pathway.
Conclusion s : Our results demonstrate that in T-ALL cells, DNA methylation-mediated silencing of miR-204 regulates the expression of MMP-2 and MMP-9 through increased transcription of IRAK1, and activation of the NF-κB signaling pathway. These data provide a potential mechanism for the role of MMP-2 and MMP-9 in the occurrence of T-ALL. Further studies will be needed to demonstrate whether demethylation of miR-204 may be a promising treatment for T-ALL.

Disclosures
No relevant conflicts of interest to declare.