bluuebunny/crossref_metadata_2025_split huggingface dataset:
Early Enhancement of IgM Expression and Signaling Capacity during Ibrutinib Therapy in CLL Patients
Authors: Drennan Samantha , Chiodin Giorgia , D'Avola Annalisa , Johnson Peter W , Trentin Livio , Packham Graham , Steele Andrew J , Stevenson Freda K , Forconi Francesco 
Published: October 2019

Abstract
In chronic lymphocytic leukemia (CLL), B-cell receptor (BCR) signaling is key to survival and proliferation, and can be effectively targeted by the Bruton's tyrosine kinase (BTK) inhibitor ibrutinib. BCR surface IgM (sIgM) levels and signaling capacity are low but variable in CLL. The variability has clinical consequences and relatively high sIgM in circulating cells is associated with aggressive disease, a higher proliferative component and higher levels of adhesion and migration receptors (D'Avola, Drennan et al, Blood 2016).
The aim of this study was to investigate the expression and function of sIgM and other surface markers on the circulating tumor cells of CLL patients following one week of ibrutinib therapy. Expression of markers associated with the BCR complex (IgM, IgD, CD19), and with adhesion or migration (CD38, CD49d, CXCR4) was assessed by flow cytometry in 13 CLL patients prior to and following one week of therapy. Signaling capacity was measured by immunoblotting following sIgM or CXCR4 stimulation. N-glycosylation status of the µ-heavy chains was measured by immunoblotting after biotinylation of cell surface proteins to determine IgM maturation (Krysov et al, Blood 2010).
Following one week of treatment basal activity of BTK was significantly reduced compared with pre-treatment (P=0.002), and anti-IgM induced BTK auto-phosphorylation was completely suppressed, confirming full inhibition of BTK in vivo. Expression of sIgM, but not of sIgD, increased dramatically (P=0.005), with little change in the other receptors. The increase was similar in the tissue-egressed CXCR4dimCD5hi cells and in the CXCR4brightCD5low long-resting cells of the circulation. Increased sIgM expression associated with the fully N-glycosylated form of sIgM, characteristic of the absence of antigen engagement. Also, it correlated with increased capacity to phosphorylate SYK following anti-IgM stimulation (r=0.91, P<0.0001). Conversely, CXCL12-induced CXCR4 signaling, measured by ERK phosphorylation, was fully suppressed.
Whilst ibrutinib induces a rapid and persistent redistribution of CLL cells into the blood, early suspension is typically associated with reduction of the lymphocytosis, nodal progression and short survival. These data reveal a dangerous early phenomenon during ibrutinib in which IgM expression has increased in the circulating cells, resulting in enhanced IgM-mediated signaling capacity through SYK. CXCR4 inhibition may prevent migration into tissue during ibrutinib and avoid (super)antigen encounter, but cells could be at risk of increased IgM mediated proliferation and disease progression if ibrutinib is withdrawn at this stage.

Disclosures
Packham: Aquinox Pharmaceuticals: Research Funding; Karus Therapeutics: Other: Share Holder & Founder. Steele:Portola Pharmaceuticals: Honoraria.
3' Untranslated Region (UTR) Alterations Are Frequently Targeted By MM-Related Mirnas and Affects the Clinical Outcome
Authors: Samur Mehmet K. , Singh Irtisha , Shih-Han Lee , Sperling Adam Samuel , Fulciniti Mariateresa , Tai Yu-Tzu , Parmigiani Giovanni , Leslie Christina S. , Mayr Christine , Munshi Nikhil C 
Published: October 2019

Abstract
More than half of human genes use alternative cleavage and polyadenylation to generate alternative 3' untranslated region (3'UTR) isoforms that play a role in gene expression regulation. The 3' untranslated region (3'UTR) of mRNA contains elements that play regulatory roles in polyadenylation, localization, translation efficiency, and mRNA stability. Although relative contributions of different regulatory mechanisms remain poorly understood, several recent studies showed that alterations in 3'UTRs might affect protein localization as well as their interactions with other proteins.
Here, we obtained global measurements of 3' UTR sequencing and RNA-seq in MM and normal plasma cells to study the effects of 3'UTR alterations in MM. Whole transcriptome sequencing (RNAseq) data from 420 uniformly treated newly diagnosed MM samples were compared with 18 normal plasma cell by using dynamic analyses of alternative PolyAdenylation and results were compared with alternative 3'UTR isoforms measured using 3'-seq, an established quantitative 3' end sequencing method from 15 samples pilot study.
We observed ~10000 different isoforms with a median frequency of 297 (range 1-2495) altered 3'UTRs per sample. We observed 563 isoforms that have distal alternative poly A (APA) site and 449 isoforms that have proximal APA site compared to normal plasma cells in at least 10% of our patients. Enrichment analysis showed that short UTR genes are significantly targeted by miR-506, miR-133, miR-130, miR-27, miR-181 and miR-200 (FDR q-value < 0.05). We also observed that expression of 90 genes significantly increased between those samples with shorter UTR vs. regular UTR. Longer UTR genes are enriched in mir-124, mir-186, mir-200, mir-302, mir-495 and expression of 123 genes were significantly downregulated between long UTR and others. There were no tumor suppressor genes and 8 oncogenes in short UTR genes and 4 tumor suppressor and 14 oncogenes in longer UTR genes.
Our results were confirmed using the 3'seq study which showed that 170 genes have longer UTR in MM compared to normal plasma cells and 187 genes have shorter UTR. ~60% of longer and ~65% of shorter UTR genes reported by 3'seq were also observed with RNAseq. Furthermore, we focused on common genes reported by both high-throughput sequencing methods and analyzed any connection between outcome and UTR shortening or lengthening. We observed 26 genes with altered UTR regions have impact on PFS or OS.
In conclusion, we report significant alternate UTR usage, including intronic UTRs in MM affecting the disease biology and clinical outcome. Our data suggests the need to further investigate the molecular impact of alternate UTR usage and its relationship with miR in myeloma.

Disclosures
No relevant conflicts of interest to declare.
Mesenchymal Stem Cells (MSC) from Patients with Multiple Myeloma Promote Myeloid Cells to Become Granulocytic-Myeloid-Derived Suppressor Cells (G-MDSC) with Immunosuppressive, Bone Resorption and Pro-Angiogenic Activity
Authors: Giallongo Cesarina , Tibullo Daniele , Parrinello Nunziatina Laura , La Cava Piera , Camiolo Giuseppina , Caporarello Nunzia , Anfuso Carmelina Daniela , Puglisi Fabrizio , Conticello Concetta , Brundo Maria Violetta , Romano Alessandra , Di Raimondo Francesco 
Published: October 2019

Abstract
Purpose
A well-recognized feature of MM is the intimate relationship between plasma cells and bone marrow microenvironment, which is mainly composed of MSC, endothelial cells, immune cells and extracellular matrix. G-MDSC accumulate in the tumor microenvironment during tumor development promoting tumor growth and immunosuppression.
Aim
Analyzing MSC from MGUS, Smoldering myeloma (SMM) and MM patients in promoting G-MDSC generation.
Methods
Human peripheral blood mononucleated cells (PBMC) isolated from healthy subjects (HS) were cultured alone and with HS- (n=10), MGUS- (n=10), SMM- (n=4) or MM-MSC (n=14)(1:100). After 6 days, G-MDSC were isolated using anti-CD66b magnetic microbeads and the phenotype(CD11b+CD33+CD14-HLADR-) was confirmed by cytofluorimetric analysis.
Results
Only G-MDSC educated by SMM- and MM-MSC co-cultures (MSCed-G-MDSC) were able to suppress T cell proliferation when cultured with normal lymphocytes (p<0.001) compared to G-MDSC control (isolated from PBMC cultured in medium alone). SMM- and MM-MSCed-G-MDSC significantly up-regulated Arg1, NOS2, TNFα and CEBPA, a transcription factor promoting suppressive phenotype. Since also the angiogenic factor BV8 was significantly up-regulated, we next investigated the pro-angiogenic effect in vitro co-culturing MSCed-G-MDSC with Human Brain Microvascular Endothelial Cells (HBMEC) (1:2). After 5 h, we observed that MM-MSCed-G-MDSC were able to increase both tube length and number of branch points compared to G-MDSC control (p<0.05). Moreover, MM-MSCed-G-MDSC were able to digest bone matrix in vitro (p<0.01).
Adding Bortezomib (5 nM), Lenalidomide (10 μM) or Pomalidomide (1 nM) during co-culture with MM-MSC, isolated G-MDSC showed a significant reduction of pro-angiogenic and bone resorption activity (p<0.05) but did not lose immunosuppressive ability.
Conclusion
MSC play a key role promoting tumor microenvironment transformation in SMM and MM patients.Indeed, only SMM- and MM-MSC and not MGUS-MSC are able to activate myeloid cells in G-MDSC with immunosuppressive, pro-angiogenic, and bone resorptionactivity.

Disclosures
No relevant conflicts of interest to declare.
Global Genomic Analysis of Newly Diagnosed t(4 ;14) Multiple Myeloma Reveals a Specific Mutational Spectrum and Identifies PKD2 As a Potential Therapeutic Target
Authors: Song Xiu Ly , Szalat Raphaël , Talbot Alexis , Nguyen HaiVu , Samur Mehmet K. , Mosen-Ansorena David , Munshi Nikhil C. , Minvielle Stephane , Avet Loiseau Herve , Cuccuini Wendy , Fermand Jean-Paul , Arnulf Bertrand , Bories Jean-Christophe 
Published: October 2019

Abstract

In Multiple Myeloma (MM), the t(4;14) translocation is associated with a poor outcome. However, beside this translocation, the genetic events which determine the adverse evolution of the disease and the resistance to treatments remain elusive.
In this study we performed whole exome or RNA sequencing analysis of samples from 65 newly diagnosed t(4;14) MM. We found that NRAS, KRAS, MAPK and FGFR3 are frequently mutated (12%, 9%, 13.8%, and 20% respectively). Overall, the FGFR3/RAS/BRAF/MAPK genes were mutated in 36 cases (54%). There was a negative correlation between mutations in FGFR3 and those occurring in NRAS, KRAS and BRAF as expected from the mutually exclusive occurrence of mutations in these genes. In addition to alterations in TP53 and DIS3, we found marked elevated frequency of mutations in PRKD2 (10.7%), ATM/ATR (10.7%) and MYCBP2 (7.6%), reduced frequency in FAM46C (1.5%) and no mutation in TRAF3 and CCND1. Mutations in ATM/ATR were strongly associated with the MB4-2 breakpoint (Bp) (p = 1.62 10-4) and significantly correlated with mutations affecting genes coding for members of the MAPK family. We observed a positive correlation between non-silent mutations in PRKD2 and the MB4-1 or MB4-3 Bp (p = 1.3 10-2). Of note, PRKD2 mutations are exclusively found in 3 t(4;14) MM cell lines and among the 84 MM sequenced by Bolli et al. (1), none of the non t(4;14) patient were mutated in PRKD2, indicating that this genetic lesion is associated with t(4;14) MM.
In the NCI-H929 t(4;14) MM cell line, which is mutated for PRKD2, encoding the PKD2 serine/threonine kinase, we observed elevated levels of phosphorylated PKD2. Furthermore, inhibition of PKD, decreased PKD2 phosphorylation and triggered reduced proliferation and apoptosis of MM cell lines and fresh plasma cells from patients in vitro. These results define a specific mutational landscape for t(4;14) MM and identify PKD2 as a potential therapeutic target in MM patients.
Altogether, these results define a specific mutational landscape for t(4;14) MM and identify PKD2 as a potential therapeutic target in MM patients.
Reference
1. Bolli, N., Avet-Loiseau, H., Wedge, D.C., Van Loo, P., Alexandrov, L.B., Martincorena, I., Dawson, K.J., Iorio, F., Nik-Zainal, S., Bignell, G.R., et al. (2014). Heterogeneity of genomic evolution and mutational profiles in multiple myeloma. Nat Commun 5, 2997.

Disclosures
Munshi: Janssen: Consultancy; Takeda: Consultancy; Celgene: Consultancy; Amgen: Consultancy; Merck: Consultancy; Pfizer: Consultancy; Oncopep: Patents & Royalties.
Prospective, Multicenter Clinical Trial of Everolimus As Primary Therapy in Waldenstrom Macroglobulinemia (WMCTG 09-214)
Authors: Treon Steven P , Meid Kirsten , Tripsas Christina K. , Heffner Leonard T. , Eradat Herbert , Badros Ashraf Z , Xu Lian , Hunter Zachary , Yang Guang , Patterson Christopher J , Gustine Joshua , Castillo Jorge J. , Matous Jeffrey V. , Ghobrial Irene M. 
Published: October 2019

Abstract
Purpose: Everolimus inhibits mTOR, a component of PI3K/AKT pro-survival signaling triggered by MYD88 and CXCR4 activating mutations in Waldenstrom's Macroglobulinemia (WM).
Experimental design: We evaluated everolimus in a prospective, multicenter study of 33 symptomatic, previously untreated WM patients. Intended therapy consisted of everolimus (10 mg/day) until progression or unacceptable toxicity. Dose de-escalation was permitted. The study was registered at www.clinicaltrials.gov(NCT01470196).
Results: At best response, median serum IgM levels declined from 4,440 to 1,360 mg/dL (p<0.0001); median hemoglobin rose from 10.8 to 12 g/dL (p=0.001); and median bone marrow disease burden declined from 75% to 52.5% in serially biopsied patients. The ORR and major response rates were 72.7% and 60.6%, respectively. Among genotyped patients, non-responders associated with wild-type MYD88 and mutated CXCR4 status. Median time to response was 4 weeks. Discordance between serum IgM levels and BM disease burden was common. With a median follow-up of 13.1 (range 1.6-64.6 months), median time to progression was 21 months, and 33 months for major responders. Discontinuation of everolimus led to rapid serum IgM rebound in 7 patients, and symptomatic hyperviscosity in two patients. Toxicity led to treatment discontinuation in 27% of patients, including 18% for pneumonitis.
Conclusions: Everolimus is active as primary therapy in WM. IgM discordance is common, and treatment cessation can often lead to rapid serum IgM rebound. Pneumonitis also appears more pronounced in untreated versus previously treated WM patients. The risks and benefits of everolimus should carefully be weighed against other primary WM therapy options.

Disclosures
Treon: Janssen: Consultancy; Pharmacyclics: Consultancy, Research Funding. Heffner:Celgene: Research Funding; Millennium: Research Funding; AbbVie: Research Funding; Pharmacyclics: Research Funding. Eradat:AbbVie: Consultancy, Honoraria, Research Funding, Speakers Bureau; Genentech: Consultancy, Honoraria, Speakers Bureau; Gilead: Consultancy, Honoraria, Research Funding, Speakers Bureau; Pharmacyclics: Consultancy, Honoraria. Castillo:Millennium: Research Funding; Janssen: Honoraria; Otsuka: Consultancy; Pharmacyclics: Honoraria; Biogen: Consultancy; Abbvie: Research Funding. Ghobrial:Takeda: Consultancy, Honoraria; Amgen: Consultancy, Honoraria; Noxxon: Consultancy, Honoraria; Celgene: Consultancy, Honoraria; BMS: Honoraria.
Two Doses of Antithymocyteglobulin As Graft-Versus-Host Disease Prophylaxis in Haploidentical Allogeneic Stem Cell Transplantation: A Multicenter Prospective Study
Authors: Lin Ren , Wang Yu , Huang Fen , Fan Zhiping , Zhang Shen , Yang Ting , Xu Yajing , Liang Xinquan , Ji Yu , Xu Na , Xuan Li , Ye Jieyu , Sun Jing , Huang Xiaojun , Liu Qifa 
Published: October 2019

Abstract
Background
Graft-versus-host disease (GVHD) is an important complication after haploidentical hematopoietic stem cell transplantation (haplo-HSCT). Antithymocyteglobulin (ATG) has been widely used for GVHD prophylaxis. However, the delay of T-cell reconstitution causes by ATG may increase the risk of opportunistic infections. In this multicenter prospective study, we compared the outcomes of the patients undergoing haplo-HSCT who received 7.5 mg/kg ATG or 10 mg/kg ATG as GVHD prophylaxis.
Methods
A total of 368 consecutive patients undergoing haplo-HSCT were randomized in 5 hospitals between May 2013 and March 2016. ATG with a total dosage of 7.5 mg/kg were administered in 184 patients and 10 mg/kg ATG were administered in 181 for GVHD prophylaxis. Three patients did not received allocated intervention and transplantation due to leukemia relapse before transplant or toxicity of conditioning regimens. The primary endpoint was the incidence of acute GVHD grade II to IV on day 100. Noninferiority of 7.5mg/kg ATG against 10mg/kg ATG was established if the difference of the 95% confidence interval (CI) in acute GVHD grade II to IV between the two groups was within 15%.
Results
The incidences of acute GVHD grade II to IV on day 100 were 31.5% and 28.2% in 7.5 mg/kg and 10 mg/kg ATG (P=0.485), respectively. The difference between these two groups was 3.3% with 95% CI of -6.1% to 12.7%, which indicated that 7.5 mg/kg ATG was not inferior to 10 mg/kg ATG. The incidences of acute GVHD grade III to IV were similar in the two groups (8.2% in 7.5 mg/kg group vs. 5.0% in 10 mg/kg group, P=0.220). No difference in chronic GVHD was found in the two groups (21.2% in 7.5 mg/kg arm vs. 19.8% in 10 mg/kg arm, P=0.748). More patients developed EBV viremia in 10 mg/kg group (37.5±3.8%) than those in 7.5 mg/kg group (25.2±3.3%, P=0.018). The 2-years overall survival were 69.5±4.3% and 66.7±4.0% for 7.5 mg/kg and 10 mg/kg group (P=0.356). And the 2-year disease-free survival were also comparative in the two groups (66.3±5.7% in 7.5 mg/kg arm vs. 66.6±3.8% in 10 mg/kg arm, P=0.288).
Conclusion
Compared with 10 mg/kg ATG, the 7.5mg/kg ATG applied in haplo-HSCT appears not to increase GVHD and might reduce the risk of EBV infection after transplantation.

Disclosures
Lin: National Natural Science Foundation of China 81400141: Research Funding; National Natural Science Foundation of China 81270647: Research Funding; Science and technology planning project of Guangdong Province 2014B020226004: Research Funding; The project of health collaborative innovation of Guangzhou City 201400000003-4: Research Funding.
Bortezomib Based Induction Is Superior to Thalidomide Based Induction in Reducing Early Relapses Following Upfront HDM: an Analysis By the Asian Myeloma Working Group
Authors: Gopalakrishnan Sathish Kumar , Ong Shin-Yeu , Kim Seok Jin , Kim Hae Su , Lee Ji Yun , Lim Sung Hee , Chen Yunxin , De Mel Sanjay , Ooi Melissa G. , Surendran Shilpa , Lin Adeline , Tan Daryl , Durie Brian G M , Chng Wee Joo , Kim Kihyun 
Published: October 2019

Abstract
Background: Multiple myeloma is a heterogeneous disease with certain genetic features associated with worse outcomes. The benefit of Bortezomib for high risk genetic patient subgroups remains unclear in an Asian population. We performed a retrospective analysis to compare bortezomib-based induction versus thalidomide-based induction in Asian patients who were treated with upfront autologous transplant.
Methods: We retrospectively analyzed outcomes of 240 patients who received autologous transplant in two centers in Singapore (n=167) and one center in Korea (n=73) after bortezomib-based (n=120) versus thalidomide-based induction (n=120) between 2006 and 2014. Patients were staged according to International Staging System (ISS), and responses were defined according to International Myeloma Working Group criteria. Early relapse was defined as relapse within 12 months post-transplant. High risk cytogenetics included the presence of 17p13 deletion, t(14;16), or t(4;14) by FISH.
Results: Baseline characteristics (age, gender, ISS, cytogenetics) were not significantly different between groups receiving bortezomib versus thalidomide induction. Bortezomib induction was associated with improved rates of complete response (CR) post-induction (40% vs 25%, p=0.013) and post-transplant (52% vs 48%, p=0.038). Median overall survival (OS) was prolonged with bortezomib (53 months vs 27 months, p=0.027). Multivariate analysis adjusted for baseline characteristics showed bortezomib reduced the risk of early relapse (HR 0.23, 95% CI 0.09-0.57, p=0.001), and improved overall survival (OS) (HR 0.2, 95% CI 041-0.93, p=0.021). Factors independently associated with poorer OS were high risk cytogenetics (HR 1.77, p=0.011), failure to achieve very good partial response (VGPR) or better post-induction (HR 1.70, p=0.005), early relapse (HR 6.20, p<0.001), but not advanced ISS stage or hypodiploidy karyotype. In subgroup analysis, patients with high-risk cytogenetics (n=46) who received bortezomib had improved OS (HR 0.39, CI 0.17 to 0.93, p=0.033), but patients with standard risk cytogenetics did not have significant OS benefit with bortezomib (HR 0.71, p=0.139).
Conclusion: Our results suggest that despite upfront autologous stem cell transplantation, patients benefit from bortezomib versus thalidomide induction. Furthermore, bortezomib may be beneficial in overcoming the adverse prognostic effect of high-risk cytogenetics.

Disclosures
Durie: Janssen: Consultancy; Takeda: Consultancy; Amgen: Consultancy.
A District General Hospital Experience of Patients Receiving Apixaban, Dabigatran, Rivaroxaban or Warfarin for Anticoagulation
Authors: Raiman Lewis , Antbring Richard H , Abu-Sitta Abed , Al-Refaie Faris N 
Published: October 2019

Abstract
Introduction
Novel oral anticoagulants (NOACs) are becoming increasingly more prevalent as a preventative measure for venous thromboembolism and stroke prevention in atrial fibrillation (AF)1. NOACs are considered non inferior to Vitamin K antagonists (VKAs) in terms of efficacy and clinical trial evidence has suggested lower rates of complications with the use of NOACs2. There is limited published literature on the complication rates of NOAC use in the district general hospital setting. In this retrospective study, the bleeding rates amongst patients on oral anticoagulants in a district general hospital in the United Kingdom were established.
Methods
All patients prescribed a NOAC (Rivaroxaban, dabigatran or apixaban) or a VKA at the Princess Alexandra Hospital, a district general hospital in Harlow, United Kingdom, between 01/01/2015 and 31/12/2015 were identified from hospital pharmacy records. Electronic patient hospital records of these individuals were reviewed to identify indication for anticoagulation and any subsequent bleeding complications after initiating therapy as classified according to ICD-10 up until 31st of June 2016.
Results
A total of 521 patients were prescribed a NOAC. 283 patients were prescribed Rivaroxaban, 217 patients Apixaban and 21 patients were prescribed Dabigatran. 969 patients were prescribed a VKA. From 1st of January 2015 to June 31st 2016, 5.9% of NOAC patients suffered a bleed requiring a hospital visit during this time, versus 7.1% patients on a VKA. Amongst patients prescribed a NOAC, common causes of bleeding included GI haemorrhage (34.5%), epistaxis (38.0%), haemoptysis (13.8%). Amongst patients prescribed a VKA, common causes of bleeding included gastrointestinal bleeding (57.0%), epistaxis (25.2%), haemoptysis (11.9%). Rates of cerebral haemorrhage were low in both groups (0.77% of patients prescribed a NOAC and 0.61% of patients prescribed a VKA).
Conclusion
This data illustrates real-world experience of bleeding rates requiring hospital admission of patients on NOACs and VKA. The data suggests a lower bleeding complication rate amongst a district general hospital patient population taking NOAC compared to VKA, in keeping with previously published data.
References
1. Beyer-Westendorf J, Förster K, Pannach S, Ebertz F, Gelbricht V, Thieme C, Michalski F, Köhler C, Werth S, Sahin K, Tittl L. Rates, management, and outcome of rivaroxaban bleeding in daily care: results from the Dresden NOAC registry. Blood. 2014 Aug 7;124(6):955-62.
2. Chai-Adisaksopha C, Crowther M, Isayama T, Lim W. The impact of bleeding complications in patients receiving target-specific oral anticoagulants: a systematic review and meta-analysis. Blood. 2014 Oct 9;124(15):2450-8.

Disclosures
No relevant conflicts of interest to declare.