bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow Cytometric Leukemic Blasts Detection in Cerebrospinal Fluid of Children with Acute Lymphoblastic Leukemia
Authors: Levinsen Mette , Marquart Hanne Vibeke , Groth-Pedersen Line , Frandsen Thomas Leth , Albertsen Birgitte Klug , Pronk Cornelis J.H. , Heldrup Jesper , Ulvmoen Aina , Vaitkeviciene Goda , Wehner Peder Skov , Frost Britt-Marie , Abrahamsson Jonas , Harila-Saari Arja , Niinimäki Riitta , Lähteenmäki Päivi , Åsberg Ann , Lund Bendik , Gunnes Maria Winther , Norén-Nyström Ulrika , Behrendtz Mikael , Pesola Jouni , Taskinen Mervi , Schmiegelow Kjeld 
Published: October 2019

Abstract
Background: Central nervous system (CNS)-directed treatment has reduced risk of CNS relapse for childhood acute lymphoblastic leukemia (ALL), which accounts for 30-40% of initial relapses. Compared with CNS-negative patients, patients with CNS leukemia (>5 leukocytes/µL cerebrospinal fluid (CSF) and lymphoblasts) suffer from more CNS+ relapses. Conventional cytology (CC) is specific (>95%), but nonsensitive (<50%), since leukocytes in CSF decay within few hours after lumbar puncture.
Method: We prospectively assessed centralised multi-parameter flow cytometry (FCM) of fixated CSF versus local CC in Nordic/Baltic childhood ALL. Diagnostic samples from 172 children aged 0-18 years with de novo and eight children with relapsed ALL were investigated in 297 CSF samples from 180 patients. Kinetics of disappearance of leukemic cells in the CSF was evaluated until day 15. Antibody-combinations reflected the immunophenotype of leukemic blasts in bone marrow.
Result: Of 172 newly diagnosed patients, 51 (30%) had CSF involvement by FCM, while CC was positive in 16 patients (9%) (p<0.001). CSF involvement was detected by both FCM and CC in four of eight patients with relapse (50%). Among newly diagnosed patients, samples positive by FCM and CC had higher leukemic blast count compared to samples positive by FCM only (medians: 0.545 (range: 0.005-4.801) versus 0.016 (range: 0.003-1.38) leukemic blasts/µL; p<0.001). Among newly diagnosed patients with samples positive by FCM and CC, the CSF blast count was related to the CSF leukocyte count (rs=0.82; p=0.001). Compared to newly diagnosed patients who were FCM-negative, those with FCM-positivity had higher WBC (median: 37 versus 9 x 109/L; p<0.001), were younger (medians: 3 versus 5 years, p=0.04), and more often had T-cell ALL (12/51 (24%) versus (6/121 (5%), p<0.001). Five (16%) of 31 newly diagnosed patients with FCM detected blasts at diagnosis and available data at day 15 still had CSF leukemic blasts on day 15. So far the two patients who later developed CNS and/or bone marrow relapse were positive by flow (1.38 and 0.178 blasts/µL), but CNS negative by CC or had CSF leukocyte count <5/µL and lymphoblasts on CC, respectively.
Conclusion: Leukemic blasts are present in spinal fluid of one third of newly diagnosed ALL. CSF involvement is associated with other higher risk characteristics. The prognostic value of these findings awaits prospective evaluation.

Disclosures
No relevant conflicts of interest to declare.
Flamsa Reduced-Intensity Conditioning Regimen in AML Patients – Role of Re-Induction and Disease State Prior to Transplant
Authors: Holtick Udo , Shimabukuro-Vornhagen Alexander , Chakupurakal Geothy , Theurich Sebastian , Leitzke Silke , Wolf Dominik , Hallek Michael , von Bergwelt-Baildon Michael S. , Scheid Christof , Chemnitz Jens 
Published: October 2019

Abstract

Reduced intensity conditioning (RIC) regimens as compared to myeloablative protocols have demonstrated lower toxicity profiles at similar efficacy in the context of allogeneic haematopoietic stem cell transplantation (allo-SCT) for patients with acute myelogenous leukaemia (AML) in first or second remission. In addition, the FLAMSA RIC regimen, combining a cytoreductive part and a transplant-conditioning part, has been described to be efficacious in patients with refractory disease. Re-induction treatment after AML relapse or refractory disease is often accompanied by severe, in particular, infectious complications such as fungal pneumonia caused by long neutropenic phases, precluding a significant proportion of patients from completing allo-SCT. To have a closer look on the role of re-induction therapy prior to transplant, we retrospectively analysed clinical data of 118 consecutive patients with AML after allogeneic stem cell tranplantation following FLAMSA conditioning at our center. No prophylactic donor lymphocyte infusions were administered.
Median age of the cohort was 53 years (19-73 years). Donors were matched related, matched unrelated or mismatched unrelatred in 33, 49 and 18% of the transplants. Complete remission prior to transplant was detected in 29% and 21% of the patients after induction or re-induction, respectively. Twenty-five percent of the patients were transplanted with blast persistence, both in the group of patients after first induction and re-induction treatment. Median follow-up was 27 months.
The 4-yr overall survival of the whole cohort is 44% with a 4-yr relapse-free survival of 42%. Cumulative incidence of relapse was 26, 30 and 40% at one, two and four years, respectively. Cumulative incidence of non-relapse mortality (NRM) was 15, 18 and 18% at one, two and four years, respectively.
There were no significant differences regarding overall and relapse-free survival for patients transplanted in CR1, CR2 or blast persistence after induction treatment. Patients who failed remission after re-induction therapy showed significantly worse overall and relapse-free survival (p=0,049 and 0,003, repectively, log-rank test). NRM was similar in all cohorts.
FLAMSA is a highly effective conditioning regimen for AML patients even not in CR. Thus, the decision for re-induction therapy prior to allogeneic SCT has to be weighted against the potential toxicity of this approach.

Disclosures
No relevant conflicts of interest to declare.
Outcome and Prognostic Factors of 17p Deleted Multiple Myeloma Patients
Authors: Kull Miriam , Teleanu Veronica , Hayde Daniela , Wildbihler Katrin , Harsdorf Stephanie , Bunjes Donald , Liebisch Peter , Langer Christian 
Published: October 2019

Abstract
Introduction: During the last decade, the outcome of patients (pts) with symptomatic multiple myeloma (MM) has markedly improved. However, there is still a significant proportion of pts who do not achieve a longtime control of their disease. In particular, pts presenting with a deletion 17p (del17p) still have dismal prognosis. In order to better stratify this important group of MM pts we sought to investigate the prognostic impact of the following parameters in a larger cohort of del17p pts: del17p clone size, concomitant genetic abnormalities, treatment modalities and the incorporation of the novel agents lenalidomide and bortezomib.
Methods: We identified 54 MM pts diagnosed between 1998 – 2012 who had a del17p at diagnosis and were treated at the University Hospital of Ulm. The patients were screened for additional chromosomal aberrations by fluorescence in situ hybridization (FISH) performed on purified bone marrow plasma cells.
Results: The median age at MM diagnosis was 59 years and the proportion of male pts was 52%. At presentation the median del17p clone size was 83% (range: 28%-98%). In the vast majority of cases (83%) the presence of a del17p was associated with the presence of a del13q14. Other concomitant genetic abnormalities detected by FISH were t(4;14) in 17%, t(11;14) in 30% and gain at 1q21 (+1q21) in 31% of cases (figure 1). The median overall survival (OS) was poor (18.9 months) and did not change substantially over time (similar median survival in pts diagnosed before 2006 versus pts diagnosed thereafter). The del17p clone size had no impact on OS, neither the presence of a t(4;14) or a t(11;14). In patients with an additional +1q21 OS was significantly shorter (15.2 versus (v) 32.4 months; p=0,032). The incorporation of one of the novel agents into first-line treatment did not change the outcome significantly. In contrast, pts receiving at least one autologous transplantation showed a significantly longer OS (33.1 v 12.7 months; figure 2). On univariate analysis there was an improved median OS for pts undergoing an allogeneic transplantation (n=15; 32.4 v 14.4 months; p=0.025). In multivariate analysis ISS stage and the implementation of an autologous transplantation remained significant prognostic factors for OS.
Conclusions: The outcome of MM pts with a del17p remains poor, even after the introduction of lenalidomide and bortezomib into clinical practice. The development of novel therapeutic strategies therefore is urgently warranted.
Figure 1 Figure 1.
Figure 2 Figure 2.

Disclosures
No relevant conflicts of interest to declare.
Elevated Adenosine Signaling Via Adenosine A2B Receptor Induces Normal and Sickle Erythrocyte Sphingosine Kinase 1 Activity
Authors: Sun Kaiqi , Zhang Yujin , Bogdanov Mikhail , Dowhan William , Idowu Modupe , Juneja Harinder S. , Blackburn Michael R. , Kellems Rodney E. , Xia Yang 
Published: October 2019

Abstract
Sickle Cell Disease (SCD) is one of the most devastating hemolytic genetic disorders affecting millions worldwide. Erythrocytes possess high sphingosine kinase 1 (Sphk1) activity and are considered to be the major cell type for supplying plasma sphingosine-1-phosphate, a signaling lipid regulating multiple physiological and pathological functions. Recent studies revealed that erythrocyte SphK1 activity is up-regulated in sickle cell disease (SCD) and contributes to sickling and disease progression. However, how erythrocyte Sphk1 activity is regulated in SCD remains unknown. In an effort to identify specific factors and signaling pathways involved in regulation of erythrocyte SphK1 activities in SCD, we first chose to screen the effects of molecules known to induce SphK1 activities in other cell types and/or reported to be elevated in the circulation of SCD including tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), endothelin 1 (ET-1), vascular endothelial growth factor (VEGF), S1P and adenosine, on the activities of SphK1 in cultured primary mouse normal erythrocytes. Among all of those molecules tested, we found that adenosine is a previously unidentified hypoxia inducible molecule directly inducing SphK1 activity in vitro in a time and dosage-dependent manner. Next, using four adenosine receptor deficient mice and pharmacological approaches, we determined that the A2B adenosine receptor (ADORA2B) is essential for adenosine-induced SphK1 activity in cultured primary mouse normal and sickle erythrocytes. Subsequently, we provided in vivo genetic evidence that adenosine deaminase (ADA)-deficiency leads to excess plasma adenosine and elevated erythrocyte SphK1 activity. Lowering adenosine by ADA enzyme therapy or genetic deletion of ADORA2B significantly reduced excess adenosine-induced erythrocyte SphK1 activity in ADA-deficient mice. Mechanistically, we revealed that PKA functions downstream of ADORA2B mediating ERK activation and subsequently underlying adenosine-induced SphK1 activities in cultured mouse erythrocytes. Finally, we conducted human translational studies and reported that adenosine signaling via ADORA2B directly increases SphK1 activity in cultured primary human normal and sickle erythrocytes in a PKA/ERK-dependent manner. Overall, our findings reveal a novel signaling network regulating erythrocyte SphK1 and highlight innovative mechanisms to control SphK1 activity in normal and sickle setting.

Disclosures
No relevant conflicts of interest to declare.
The Tissue Factor/Thrombin/Protease-Activated Receptor 1 Pathway Enhances Double-Strand RNA Induced Immune Responses in Macrophages
Authors: Antoniak Silvio , Tatsumi Kohei , Mackman Nigel 
Published: October 2019

Abstract
Introduction: Co-regulation of the immune response and the coagulation cascade after infection is thought to be an ancient response to limit pathogen spread. Recently, we showed that activation of the thrombin receptor, protease-activated receptor 1 (PAR1), on fibroblasts enhanced the innate immune responses to RNA virus infection. Here, we investigated whether PAR1 activation by the extrinsic coagulation pathway contributes to dsRNA-induced innate immune responses in macrophages.
Methods: Activation of the type-I interferon (IFN) pathway in the murine macrophage cell line RAW264.7 and bone-marrow derived macrophages (BMDM) from WT and PAR1-/- was analyzed after dsRNA (poly I:C) and/or PAR-1 stimulation. In addition, innate immune responses in the spleen were analyzed in vivo 4 hours after poly I:C (8mg/kg) injection in mice with reduced tissue factor expression (LowTF) or global PAR1 deletion (PAR1-/-) as well as in WT mice with a thrombin inhibitor (dabigatran etexilate, 10g/kg chow) or PAR-1 inhibitor (SCH79797, 25μg/kg). Lastly, we investigated the innate immune response in the spleen of WT and PAR1-/-mice after infection with the single-stranded RNA virus coxsackievirus B3.
Results: RAW264.7 and BMDM exhibited a toll-like receptor 3 dependent induction of IFNβ and CXCL10 after poly I:C stimulation. Activation of PAR-1 with either thrombin or agonist peptide enhanced poly I:C induction of IFNβ and CXCL10. A deficiency of tissue factor levels, thrombin inhibition, PAR-1 inhibition or PAR1 deficiency resulted in reduced expression levels of type-I IFNs and IFN-response genes such as CXCL10 in the spleen and plasma in mice given poly I:C. Last, PAR1-/-mice exhibited impaired IFNβ immune response 4 days after coxsackievirus B3 infection compared to WT mice.
Conclusion: Our study indicates that the coagulation dependent activation of PAR1 on macrophages is important for anti-viral responses to dsRNA. We speculate that PAR1 inhibition may interfere with anti-viral responses in humans.

Disclosures
No relevant conflicts of interest to declare.
Very High Efficiency of ICE (Ifosfamide-Carboplatin-Etoposide) in Relapse/Refractory (R/R) Primary Central Nervous System (PCNSL) and Vitreo-Retinal (VRL) Non Hodgkin Lymphoma. a LOC Network Multicenter Retrospective Study on 58 Cases
Authors: Choquet Sylvain , Grenier Adrien , Houillier Caroline , Soussain Carole , Moles Marie Pierre , Gastinne Thomas , Cassoux Nathalie , merle Beral Helene , roos Weil Damien , Leblond Veronique , hoang Xuan Khe 
Published: October 2019

Abstract
Background: R/R PCNSL and VRL carry a very poor prognosis (i.e. 4,5 months, Jahnke, J neuro-oncol 2006)). Autologous stem-cell transplantation (ASCT) is effective in this setting but essentially if the disease is in good response (CR and PR). Since recent recommendations for first line treatment recommend high dose Mtx and AraC, new salvage chemotherapies must use other drugs. ICE regimen contain three drugs known to pass the blood-brain barrier and is validated in R/R systemic lymphoma but not in PCNSL/VRL.
Methods: From June 2010 to May 2015, all relapse/refractory PCNSL and VRL already treated by Mtx and AraC in 5 departments of haematology or neuro-oncology in France, where treated by ICE: ifosfamide (5g/m² at day 2), carboplatine (AUC 5 at day 2) and etoposide (100mg/m²/d days 1 to 3) every 3 to 4 weeks. ASCT with thiotepa-busulfan-cyclophosphamide was proposed when possible.
Results: Fifty-eight patients have been treated, 25 females and 33 males, median age was 64 [28-84]. Eight received rituximab at day 1 of each cycle. Twelve where refractory, 46 in relapse, with a mean progression free survival (PFS) of 159 days [0-1763] before ICE. Thirty patients (51%) were in second line, 12 (21%) in third, 4 in fourth. Localizations where CNS in 32 cases, CNS + spinal fluid (SF) in 11, CNS + VR in 2, VRL in 9, SF in 3 and CNS+VR+SF in 1. Patients received a median of 3 cycles and 25 needed a dose reduction. Grade 3/4 WHO toxicities where: 29% neutropenic fever, 50% anemia, 69% neutropenia, 78% thrombocytopenia and 3 neurological reversible complications. ASCT have been made in 19 patients (33%): 13 in CR, 4 in PR, 2 in progression. Response rate is 70% (48% CR and 22% PR). With a mean follow-up of 670 days, 25/41 patients in response relapsed (only 7/19 after ASCT), median PFS is 133 days (NR for ASCT), 35 patients died (30 by progression, 4 by sepsis and one by sudden heart arrest in PR). Median Overall survival (OS) was 238 days for all patients but was not reached in case of ASCT, mean follow up for surviving ASCT is 856 days [278-1628]).
Conclusion: ICE regimen is very effective in relapse/refractory PCNSL and LVR heavily treated by high dose Mtx and AraC, with a manageable toxicity. If ASCT is performed, OS is very high and relapses rare. ICE + ASCT can represent a new standard in this setting.

Disclosures
Leblond: Roche: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses, Speakers Bureau; Janssen: Consultancy, Honoraria, Speakers Bureau; GSK: Consultancy, Honoraria, Speakers Bureau; Gilead: Consultancy, Honoraria, Speakers Bureau; Mundipharma: Honoraria.
USP47 Is a New Target in Chronic Myelogenous Leukemia
Authors: Lei Hu , Ma Chunmin , Liu Zhen , Gao Shenmeng , Zhou Li , Wang Weiwei , Xu Hanzhang , Gao Fenghou , Wu Yingli , Liu Han 
Published: October 2019

Abstract
Despite of the great success of tyrosine kinase inhibitor in the therapy of chronic myelogenous leukemia (CML), CML is still not curable. Further investigation on the molecular mechanism of CML pathogenesis and identification of novel therapeutic target are required. In this study, we found that overexpression of CML characterized Bcr/Abl oncoprotein could significantly upregulate the expression of ubiquitin-specific protease Usp47. Also, increased expression of Usp47 was observed in primary CML cells and its expression was associated with the treatment response of imatinib, the first generation of tyrosine kinase inhibitor. Furthermore, Usp47 knockdown could inhibit the proliferation of K562 cells in vitro and in vivo. P22077, a Usp47 inhibitor, could inhibit the proliferation of CML cell lines, which comprise BCR/ABL mutation, including the T315I mutation. P22077 inhibits the proliferation of primary CML cells sensitive or resistant to imatinib. The colony forming activity of CD34+ from CML primary cells but not from normal cord blood cells were also inhibited by P22077. We further demonstrated that inactivation of STAT5 significantly reduce the expression of Usp47, indicating that BCR/ABL-induced upregulation of Usp47 is mediated by STAT5 activition. Interestingly, Usp47 could interact with Sirt1, another STAT5 target gene. Inactivation of Usp47 reduces the expression of Sirt1 in CML cell lines and primary CML cells. Taken together, we demonstrated that Usp47 plays an critical role in CML, revealing a novel BCR/ABL/STAT5/Usp47/Sirt1 signaling pathway in CML, providing a potential target to overcome tyrosine kinase inhibitor resistance.

Disclosures
No relevant conflicts of interest to declare.
Carfilzomib Enhances Natural Killer Cell-Mediated Lysis of Myeloma Linked with Decreasing Expression of HLA Class I
Authors: Shi Jumei , Yang Guang , Kong Yuanyuan , Gao Minjie , Tao Yi , Zhang Yiwen , Wu Xiaosong 
Published: October 2019

Abstract
Multiple myeloma (MM) is a malignant disorder characterized by uncontrolled monoclonal plasma cell proliferation. It accounts for 10% of all hematological malignancies and causes 15-20% of deaths from hematological malignancies. Although new therapies were introduced and overall survival of MM was improved in the last 10 years, MM still remains an incurable disease due to drug resistance. Natural killer (NK) cell-based treatments are promising therapies for multiple myeloma (MM). Carfilzomib (CFZ), a second-generation proteasome inhibitor, is used to treat patients with MM who are refractory or intolerant to both bortezomib and lenalidomide (or thalidomide). In this study, we determined that CFZ treatment enhanced the sensitivity of MM cells to NK cell-mediated lysis. Here, we report that CFZ decreased the expression of human leukocyte antigen (HLA) class I on MM cell lines and primary MM cells, the mean reduction was 47.7 ± 9.4% and 42.8 ± 12.4%, respectively. The down-regulation caused by CFZ occurred in a dose- and time- dependent manner. We compared the cell surface levels of HLA class I on MM cells in the presence or absence of CFZ after acid treatment. CFZ also down-regulated the expression of newly formed HLA class I on MM cells. CD107a expression levels were used to measure NK-cell degranulation. When NK cells were incubated with MM cells with CFZ treatment, the percentage of NK cells expressing CD107a on the surface greatly increased (mean ± SD: 33.6 ± 2.1%, for treated cells vs 16.7 ± 2.3%, for control cells, P < 0.05). We also showed that CFZ augmented NK-cell cytotoxity by a perforin/granzyme-mediated mechanism, because such enhancement was abolished when CMA, but not anti-TRAIL or anti-Fas-L antibodies, was added. Treatment of MM with CFZ significantly sensitized patients' MM cells to NK cell-mediated lysis (mean ± SD: 43.1 ± 6.4%, for treated cells vs 16.1 ± 4.0%, for control cells at effector/target (E/T) ratio of 10:1, n = 9, P < 0.01). Furthermore, the exogenous HLA-C binding peptides, used in the CFZ treated group rescued the down-regulation of HLA-C and reduced NK cell-mediated lysis to a similar level as in the untreated group. Blocking NKG2D, NCRs and TRAIL did not have a significant impact on NK cell lysis of myeloma cells. These implied the enhancement of NK cell-mediated lysis was mainly linked with the decreased expression of HLA class I. Our findings show a novel activity of CFZ as an immunomodulating agent and suggest a possible approach to therapeutically augment NK cell function in MM patients.

Disclosures
No relevant conflicts of interest to declare.