bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mice Deficient in Urokinase-Type Plasminogen Activator (uPA) or uPA Receptor Develop Significantly Diminished Collagen-Induced Arthritis
Authors: Thornton Sherry , Raghu Harini , Jone Alice , Cruz Carolina , Rewerts Cheryl L. , Frederick Malinda D. , Degen Jay L. , Flick Matthew J. 
Published: October 2019

Abstract
Rheumatoid arthritis (RA) is a common and debilitating autoimmune disease characterized by chronic inflammation, synovial hyperplasia, edema, cartilage and bone erosion and loss of joint function. Increasing evidence suggests that the plasminogen activation (PA) system plays a fundamental role in the mechanisms mediating inflammatory joint disease pathogenesis. However, analysis of the precise contribution of PA system components to arthritis pathogenesis has been complicated by the use of gene-targeted mice on non-susceptible genetic backgrounds or experimental models that simultaneously induce wound trauma in conjunction with arthritis induction. To rigorously define the contribution of the urokinase-type plasminogen activator system to arthritis pathogenesis, previously generated genetic deficiencies in both uPA and uPA receptor (uPAR) were inbred for 7 generations (99% inbred) to the well-characterized, collagen-induced arthritis (CIA)-susceptible strain, DBA/1J. Our results indicate a near complete amelioration of joint disease in uPA-deficient mice that was also observed in uPAR-deficient mice. Limited disease development in both uPA- and uPAR-deficient mice correlated with significantly reduced local mRNA levels of key inflammatory mediators (e.g., TNFα, IL-1β, and IL-6) in these animals. To determine if development of inflammatory joint disease in CIA-challenged mice was dependent on the expression of uPAR by non-hematopoietic- or hematopoietic-derived cells, reciprocal bone marrow transplant studies were performed. Mice in which uPAR deficiency was limited to the bone marrow compartment elicited significantly reduced macroscopic and histopathological disease in the paws and knees compared to wild-type mice or mice in which only hematopoietic-derived cells express uPAR. Our results are the first to report in the context of the highly CIA susceptible DBA/1 background that both uPA and uPAR are key determinants of inflammatory joint disease pathogenesis. Furthermore, our findings indicate a fundamental role for uPAR expression by hematopoietic cells in driving arthritis incidence and progression. Thus, these findings suggest that cell-surface associated uPA/uPAR-mediated proteolysis and/or uPAR-mediated signaling events from bone-marrow derived cells are important in promoting inflammatory joint disease, and that disrupting this key proteolytic/signaling system may provide a novel therapeutic strategy to limit clinical arthritis.

Disclosures
No relevant conflicts of interest to declare.
Lower Doses of Plerixafor Use Is Sufficient to Effectively Mobilize PBSC (Peripheral Blood Hematopoetic Stem Cells) When Compared to Standard Administration
Authors: Goker Hakan , Eliacik Eylem , Altuntas Fevzi , Tekgunduz Emre , Ozdemir Evren , Isik Ayse , Kayikci Omur , Kaya Ali Hakan , Aksu Salih , Haznedaroglu Ibrahim C. , Sayinalp Nilgun , Demiroglu Haluk , Ozcebe Osman Ilhami , Buyukasik Yahya 
Published: October 2019

Abstract
Background: Plerixafor (CXCR4 chemokine receptor antagonist), is indicated to mobilize hematopoietic stem cells (HSCs) to the peripheral blood for collection and subsequent autologous transplantation in patients who are poor mobilizers. In this retrospective analysis, we compared the efficacy of plerixafor 0.24 mg/kg by subcutaneous (SC) injection (1-2 doses) versus plerixafor (3-4 doses) 0.24 mg/kg by SC injection in terms of their effect on the hematopoietic recovery in hematopoietic cell transplantation (HCT).
Methods: We retrospectively reviewed the peripheral blood stem cell (PBSC) collections of 40 consecutive lymphoma and multiple myeloma patients who underwent autologous HCT, who had previously mobilized poorly and were given plerixafor for mobilization. Twenty-six patients received 1-2 doses of plerixafor and 14 patients received 3-4 doses. All patients gave written informed consent for the procedure. Successful engraftment time was defined as neutrophils >0.5x 109 cells/L, platelets>20 x109 cells/L for three consecutive days. SPSSincver. 20 is used for statistics.
Results: All of the 40 patients were mobilized promptly after plerixafor administration without any problem. The mean amount of autologous CD34+ PBSC was 3,8 x 10(6)/kg versus 3,2 x 10(6)/kg, for 1-2 doses of plerixafor administration ve3-4 doses, respectively(p=0,4). Median times to the neutrophil engraftment to >0,5x 109/L were 12 and 10 days for the 1-2 doses and 3-4 doses, respectively (p=0,2). Additionally, median times to the platelet engraftment to >20x 109/L were 14 and 15 days for the 1-2 doses and 3-4 doses, respectively (p=0,6).
Conclusions: These data show that the lower doses, i.e. 1-2 days of plerixafor administration evaluated in this study are sufficient to mobilize HSCs. This retrospective comparison suggest that larger prospective studies comparing two days vs. four days of plerixafor should be performed in order to further elucidate the role of two days of plerixafor use in HSC mobilization, and hence would a relatively cost effective way of mobilization in poor mobilizing patients.

Disclosures
No relevant conflicts of interest to declare.
Early Apoptotic Cells (ApoCell) As Prophylaxis of Graft-Versus-Host Disease in Myeloablative HLA-Matched Allogeneic Bone Marrow Transplantation Is Safe and Effective: 1 Year Follow-up
Authors: Mevorach Dror , Tzila Zuckerman , Nagler Arnon , Rowe Jacob M. , Or Reuven 
Published: October 2019

Abstract
Apoptotic cells infusion (ApoCell) is a novel cellular therapy that was found to be effective in a variety of mouse models of autoimmune diseases. ApoCell was tested in humans 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 that received conventional myeloablation with 35, or 70, or 140, or 210x106 cell/kg of donor ApoCell, on day -1 of transplantation (Mevorach et al. BBMT 2014).
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. We now report 1 year overall survival rate which was 62% with a non-relapse survival rate of 89% (100% for the two higher cohorts).
These results are in agreement with additional safety observations that included: ten serious adverse effects with all being not related (7) or unlikely to be related (3) to ApoCell infusion, and only three out of hundreds adverse effects, that were reported as possibly related to ApoCell infusion (with no definite or probable adverse effects).
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. GvHD clinical grade was supported by 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).
These results suggest that single-infusion high-dose donor early apoptotic cells in HLA-matched myeloablative alloBMT is a safe and effective prophylaxis for acute GVHD (clinicaltrials.gov no. NCT00524784).

Disclosures
Mevorach: Enlivex: Consultancy, Equity Ownership.
Recovery of IgG Production Following Rituximab Therapy after Allogeneic Stem Cell Transplantation
Authors: Morris Christopher L. , Morris Joan 
Published: October 2019

Rituximab is a monoclonal antibody to the CD20 antigen expressed on B lymphocytes. Several recent reports suggest that rituximab may be used to treat complications of SCT including GVHD and EBV reactivation. The long term consequences on IgG recovery aren’t well studied. This is a retrospective analysis of allogeneic pediatric SCT patients treated with rituximab at Loma Linda University from 2004 to 2013. We treated 23 patients with rituximab following allogeneic SCT (11 MUD, 7 MSD, 3 Cord) for post-transplant EBV reactivation alone (4), cGVHD alone (4), cGVHD in combination with thrombocytopenia (9), plus hemolysis (4) and EBV reactivation (2). Most patients required IVIG monthly (18) or more frequently (4) to maintain serum IgG levels >500mg/dl. We examined the length of time from end of rituximab therapy to recovery of endogenous IgG production. The effect on recovery of IgG production of cGVHD therapy, EBV infection, age, type of stem cell graft, and number of doses of rituximab were evaluated. Those treated for cGVHD plus any other indication (n=19) received a median of 11 doses of rituximab (range 4 to >30). Among these patients 11 of 14 whose cGVHD resolved and came off immune suppression recovered the ability to produce IgG completely (9) or partially (2, defined as decreasing frequency of IVIG infusions to maintain IgG>500mg/dl). Median time to recovery was 14 months (range 1-50 months) after last dose of rituximab. For patients who continued to require immune suppression for cGVHD only 1 of 5 recovered IgG production 13 months after last dose of rituximab. The remaining 4 patients required monthly or greater IVIG for >3 years after the last dose of rituximab (all still requiring IVIG). There were 4 patients treated with 2-4 doses of rituximab for EBV reactivation early after SCT, and 1 treated with 11 doses for cGVHD and EBV. Only 1 of these 5 has completely recovered IgG production. The 4 other patients remain dependent on IVIG a median of >39 months since end of rituximab therapy.
We found no relationship between age of patient or duration of rituximab therapy on the time interval between end of rituximab therapy and onset of endogenous IgG production. Recipients of MUD grafts had a non-significant increase in failure to recover endogenous IgG production, but when the impact of therapy for cGVHD was taken into account the type of stem cell graft lost significance.
Ability to recover endogenous IgG production following treatment with rituximab after SCT depends on successful treatment of cGVHD and in this group of patients requires about 1 year from end of therapy. Among those treated with rituximab whose cGVHD does not resolve and in those with early EBV reactivation, loss of IgG production persists at least several years, and this may be unrelated to rituximab therapy.

Disclosures
Off Label Use: rituximab for cGVHD.
Treatment Approaches in Acute Leukemia Patients Relapsing Following Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Topcuoglu Pervin , Gunduz Mehmet , Ozen Mehmet , Civriz Bozdag Sinem , Kocak Toprak Selami , Arslan Onder , Ozcan Muhit , Demirer Taner , Ilhan Osman , Akan Hamdi , Beksac Meral , Konuk Nahide , Gurman Gunhan 
Published: October 2019

Abstract
Aim: We would like to present our center experiences in the approaches of relapsing and/or progressive acute leukemia patients after allogeneic hematopoietic stem cell transplantation (AHSCT).
Patients: We have retrospectively evaluated 133 patients (86 Male; 47 Female) with acute leukemia who has been detected relapse and/or progression after AHSCT between June 1990 and October 2013. Median age was 30 years (16-63 years) at the transplant time. Their diagnosis were 89 AML (12 secondary AML), 41 ALL and 3 biphenotipic leukemia. Median time of relapse and/or progression was 165 days (13-4713 days) after ASCT. Relapse types were as extramedullary relapse (EMR) alone in 11, extramedullary with hematological relapse in 30 and hematological relapses (HR) alone in 92 patients.
Results: Our treatment approaches were chemotherapy and/or donor lymphocyte infusion in 109, second ASCT from same or different donor in 16 and radiotherapy alone in two patients. However, six patients were not able to receive any therapy. We have observed that the numbers of donor lymphocyte infusion or second transplantation increased after the year of 1999 (Figure 1). The probability of three-month, six-month or one-year survival after the relapse&progression were calculated as 53.7%±4.4%, 35.1 %±4.3% and 24.2%±3.9%, respectively. Survival after the relapse or progression was median 102 days (ranging between 68.7 days and 135.3 days, 95% CI). The type of relapse (EMR alone, EMR plus HR versus HR alone) and leukemic types (AML vs ALL) did not effect survival (p>0.5). When we compared second transplantation with other approaches, survival after relapse did not change (p=0.8) (Figure 2). In addition, survival was not affected by the year of relapses as 1990-1999, 2000-2009 versus 2010-2014 (p=0.7).
Conclusion: Treatment approaches in acute leukemia relapsing after transplantation vary from the cellular therapy to chemotherapy. But our experience was not able to show a difference between second transplantation and others. Moreover each center should be approached to the patients according to their experience and type of relapse until an effective treatment option will be defined.
Figure 1. Treatment approaches according to years Figure 1. Treatment approaches according to years
Figure 2. Overall survival according to treatment approaches Figure 2. Overall survival according to treatment approaches

Disclosures
No relevant conflicts of interest to declare.
Molecular Mechanisms of EML in AML1/ETO+ AML and Its Targeting Treatments
Authors: Meng Fan Yi , Jiang Ling , Zhong Qingxiu , Wang Li Chun , Yu Guopan , Zhu Xinhua 
Published: October 2019

Abstract
Our previous study has been reported that AML1/ETO positive patients with highly expressed of APP were much easier to extramedullary infiltration, and got poor prognosis£¨followed-up median 35 ( 6-96 ) months, RFS in the high expression APP group was significantly lower than the low expression group £¬40.0% vs 80.0%£¬P = 0.001). In vitro study, we constructed a cell model kasumi-1 which was consistent with AML1/ETO positive and high expressed of APP gene. The cell migration was significantly reduced after interferce of APP expression. This study was designed to investigate the molecule mechanism of extramedullary leukemia (EML) in kasumi-1 cell model and to invent a strategy for treatment in clinic.
In this study, we found p-ERK, c-Myc and MMP-2 were significantly decreased after APP expression knockdown in kasumi-1 cell. Meanwhile, we added the inhibitors to block p-ERK and c-Myc expression. The results showed that protein expression of p-ERK and c-Myc was significantly decreased after p-ERK inhibitor performance, which was proportional to the time and concentration. c-Myc and MMP-2 protein expression was significantly reduced after c-Myc inhibitor was used, but p-ERK didn't change (Fig.1B). So, we concluded that APP might regulated the AML cell migration via APP/p-ERK/c-Myc/MMP-2 pathway.
Also, we found that kasumi-1 cell was resistant to adriamycin (ADM) and Ara-C, which meant APP may be related with drug resistance. So, we detected cell surviving fraction after ADM and Ara-C performance via MTT assay. The results showed that there was no difference in control group and siAPP group. But, when compared with controls groups, cell surviving fraction in siEZH2 group was significantly decreased after ADM and Ara-C performance respectively. Furthermore, we found protein expression of EZH2 was significantly reduced after LBH589 treatment in cell culture. So, we concluded that LBH589 or SiEZH2 could increase sensitivity of kasumi-1 cell to ADM and Ara-C.
In sum, in AML1/ETO positive leukemia cells, we first report that APP gene regulates cell migration via APP/p-ERK/c-Myc/MMP-2 pathway and EZH2 gene induces drug resistantence. Interference or blocking of EZH2 and APP expression may be helpful in treating AML1/ETO positive leukemia.

Disclosures
No relevant conflicts of interest to declare.
Forewarned Is Forearmed- Comparison of Southern Network on Adverse Reactions (SONAR) Versus an FDA-Mandated Prospective Observational Registry Assessment of Gemtuzumab Ozogamycin-Associated Sinusoidal Obstructive Syndrome
Authors: Magwood Jametta , Georgantopoulos Peter , Norris LeAnn B , Bennett Charles L. 
Published: October 2019

Abstract
Gemtuzumab ozogamycin (GO) received initial approval in 2000 from the Food and Drug Administration (FDA) for patients age 60 and older suffering with relapsed acute myeloid leukemia. After 10 years of administering GO-containing combination chemotherapy regimens, the drug was voluntarily removed from the commercial market in 2010 amid reports of unexpected development of sinusoidal obstructive syndrome (SOS); which has the potential to be fatal. Four months after the initial approval for marketing in 2000, SONAR initiated an investigation of GO-associated SOS. SONAR reported an incidence rate of 38% among allogeneic stem-cell transplants and 16% among autologous stem-cell transplant (SCT) patients. Ultimately, there was an overall incidence rate of 3% when single-agent GO was administered as directed by the FDA. In 2007, as a result of the examination of the reports from clinical trials and observational studies; data mining of interim reports from the manufacturer’s Prospective Observational Registry and the FDA’s Adverse Event Reporting System was undertaken. In 2013, the pharmaceutical manufacturer reported results from its FDA-mandated prospective observational study. Major differences between the SONAR report and the mnaufacturer’s report include: timing of publication (2007 versus 2013); inclusion of information on allogeneic stem cell versus autologous stem cell transplantation prior to GO administration (in our reports only) and identification of risk factors associated with higher rates of SOS (SCT within 3 months of GO administration, off-label administration of high doses of GO, and concomitant administration of 6-TG in our analysis versus no identified risk factors in the manufacturer’s analysis). The incidence of SOS was 9.1% among 482 patients at 54 centers in the manufacturer’s registry.
Table 1: Comparison of gemtuzumab ozogamycin (GO) safety reports analyses between SONAR and manufacturer SONAR Pharmaceutical Investigation # of pts who received GO 1071 482 # of pts with SOS 99 44 DATA METHODS # of centers involved 50 centers (US and UK) 54 centers (US only) Interim analysis Not done Yes (2002 and 2005) Concomitant chemo given 67% 41% # of pts with concomitant HCT 26% 0% % with prior alloSCT 2.6% Not reported % with prior autoSCT 3.9% Not reported % with prior HSCT 6.5% 18.3% % with subsequent alloSCT 1.2% 0% % with subsequent autoSCT 2.0% 0% Publication Date* 2007 2013
* Gemtuzumab ozogamycin voluntarily removed from market in 2010 by the pharmaceutical manufacturer

Disclosures
No relevant conflicts of interest to declare.
Inhibition of ADAMTS13 Activity By Human Neutrophil Peptide 1 (HNP-1): Potential Link Between Inflammation, Onset of Thrombotic Thrombocytopenic Purpura, and Other Thrombosis
Authors: Bao Jialing , Bdeir Khalil , Siegel Don L. , Cines Douglas B. , Zheng X. Long 
Published: October 2019

Abstract
Thrombotic thrombocytopenic purpura (TTP) is a potentially fatal syndrome associated with severe deficiency of plasma ADAMTS13 activity resulting from either mutations or autoantibodies. However, patients with severe ADAMTS13 deficiency do not always develop TTP. Rather, a trigger, such as infection or inflammation, often precedes the onset of the TTP syndrome. We hypothesized that antimicrobial human neutrophil peptides 1-3 (HNPα1-3) or α-defensins, the most abundant proteins in the granules of neutrophils, which are released at site of inflammation, activate platelets, and inhibit fibrinolysis, might help to initiate TTP. This question arose because we noted that the amino acid sequences of HNPα1-3, which are nearly identical except for one residue at the N-terminus, all contain a motif (RRY) similar to exosite 3 (659RRYGEEY665) in the spacer domain of ADAMTS13 (Fig. 1) that was shown to be critical for recognition of von Willebrand factor (VWF). Here, we found that both purified and synthetic HNPα1 bind to FRETS-VWF73 and plasma-derived VWF and inhibit proteolytic cleavage of these substrates in a concentration-dependent manner. At the final concentrations of 10 micro mol/L and 150 micro mol/L, HNPα1 completely abolished the cleavage of FRETS-VWF73 (IC50=3.5 micro mol/L) (Fig. 2) and VWF (IC50=75 micro mol/L) (not shown), respectively. Such concentrations are readily attained locally after systemic infection. Deletion or alanine substitution within the RRY motif of HNPα1 completely abolished its ability to inhibit ADAMTS13 activity assessed by FRETS-VWF73 and VWF multimer analysis. This suggests that an interaction of the RRY motif in HNPα1 with the central A2 domain of VWF is required to mediate its inhibition. In support of this hypothesis, HNPα1 interacts with a human monoclonal antibody against ADAMTS13 scFv (the single chain fragment of variable region) designated 4-20, but not scFv3-1, both isolated by phage display from patients with acquired autoimmune TTP. Hydrogen-deuterium exchange mass spectrometry has shown that the binding site for scFv4-20, but not scFv3-1, contains the RRY sequence. These results suggest that HNPα1-3 released from neutrophils following infection or inflammation may inhibit residual plasma ADAMTS13 activity in vivo similar to anti-ADAMTS13 autoantibodies by interfering with its interaction with VWF, thereby triggering the onset of hereditary and acquired autoimmune TTP. Our findings suggest a potential novel link between systemic inflammation and the pathogenesis of TTP and possibility other thrombotic sequelae.
Figure 2 Figure 2.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Kit Deficiency Regulates Stable Human Hematopoietic Stem Cell Engraftment in Mice
Authors: Waskow Claudia , Rahmig Susann , Cosgun Nehir 
Published: October 2019

Abstract
Humanized mice are required for in-depth analysis of human hematopoietic stem cell (HSC) function and immunobiology. The currently available options are problematic because stable engraftment of substantial numbers of human HSCs and the continuous generation of human myeloid cell types remain difficult to achieve. We generated three novel recipient mouse strains that combine immune deficiency with a functionally impaired endogenous HSC compartment mediated by a defective Kit receptor: BALB/c Rag2- Il2rg-KitWv/Wv (BRgWv), NOD/SCID Il2rg- (NSG) KitWv/Wv (NSGWv) and NSG KitW41/W41 (NSGW41). We find that the mutant Kit receptor opens up stem cell niches across species barriers and allows for robust and sustained engraftment of human HSCs after transfer into adult mice without the necessity for irradiation conditioning prior to transplantation. Following stable engraftment in the mouse bone marrow niches, human HSCs give rise to lymphoid cells and to robust numbers of erythroid and myeloid lineage cells over extended periods of time in primary and secondary recipient mice. Particularly in NSGW41 mice, we observe improved reconstitution of human myeloid cell types compared to control irradiated NSG mice. In the bone marrow, endogenous hematopoietic progenitor cells with a defective Kit receptor are largely replaced by human Kit-proficient hematopoietic progenitor cells because progenitor cell expansion requires normal signaling by Kit. Thus, human Kit-proficient donor cells have an advantage over endogenous murine Kit-mutant cells. Increased numbers of myeloid cells are found in the bone marrow and spleen of NSGW41 transplanted mice compared with grafts established in irradiated NSG mice. We conclude that Kit-signaling regulates HSC engraftment across the human-mouse species barrier and that Kit deficient mice show great potential for the study of human HSC functions including self-renewal, differentiation and mechanisms of innate immunity.

Disclosures
No relevant conflicts of interest to declare.