bluuebunny/crossref_metadata_2025_split huggingface dataset:
Haploidentical/Mismatched Hematopoietic Stem Cell Transplantation for Nonresponders to Immunosuppressive Therapy Compared with First-Line Rabbit Antithymocyte Immunoglobulin Treatment Against Acquired Severe Aplastic Anemia
Authors: Liu Limin , Wang Xiuli , Jin Song , Hao Lin , Zhang Yanming , Zhang Xingxia , Depei Wu 
Published: October 2019

Abstract
We treated 86 patients with SAA, 31 patients who failed to respond to previous therapy underwent haploidentical hematopoietic SCT (haplo-HSCT, n=26) or mismatched unrelated donor HSCT (MMUD-HSCT, n=5). The other 55 patients were treated with immunosuppressive therapy (IST). At 6 months post-treatment, the treatment failure rates of HSCT and IST groups were 19.35% and 29.09% (P=0.320). Hematopoietic recovery time was shorter in the HSCT group than IST group (P > 0.05). The estimated OS at 3 years was 79.2% ± 7.7% in HSCT group and 89.7% ± 4.4% in the IST group (P=0.058). The estimated failure-free survival (FFS) at 3 years was 79.2% ± 7.7% in the HSCT group and 62.9% ± 8.0% in the IST group (P=0.391). The estimated FFS at 3 years in SAA that had progressed from non-SAA (NSAA) of HSCT was 71.6% ± 14.0% and 16.7% ± 13.6% of IST (P=0.021). Within the HSCT group, 38.71% of the patients developed grade II-IV acute GVHD, and 18.52% of the patients experienced moderate-severe chronic GVHD. These results suggest that haplo-HSCT/MMUD-HSCT and IST have similar treatment failure rate, OS and FFS. haplo-HSCT/MMUD-HSCT might provide a better chance of FFS than IST for SAA that had progressed from NSAA.

Disclosures
No relevant conflicts of interest to declare.
Non-Transmissible Measles Virus Vector Selectively Induces Human Primed and Naïve like Pluripotent Stem Cells from Hematopoietic Cells
Authors: Hiramoto Takafumi , Tahara Maino , Sakamoto Chika , Ono Hiroaki , Kohara Hiroshi , Takeda Makoto , Tani Kenzaburo 
Published: October 2019

Abstract
Our newly gene transfer vector could transduce multiple genes simultaneously into human hematopoietic cells with higher efficiency than general gene transfer vector. By using the vector, we could induce high quality pluripotent stem cells (naïve like pluripotent stem cells) from human cord blood derived CD34+ cells.
Measles virus (MV) which belongs to negative single strand RNA viruses has been known to have high affinity for human immune cells including monocytes, B cells and T cells. Although MV induced severe diseases, most of MV induced diseases can be controlled and curable by effective vaccination against H glycoprotein. Based on this background, we have newly developed novel measles virus vector to achieve the efficient and safe gene transfer to various cells. Our MV gene transfer vector is non-transmissible and transfer multiple genes simultaneously into human cells. Exogenous gene expressions from MV vector were controlled by changing the position in the vector genome.
The MV vector which encoded 5 genes (enhanced GFP, human OCT3/4, SOX2, KLF4, and L-MYC) (MV-ΔF-OSKL-EGFP) induced transgenes in almost all of hematopoietic cells besides natural killer cells. Quiescent T cells (CD45RA and CD197 expressed T cells), which include Naive and stem cell memory T cells were also transduced by MV-ΔF-OSKL-EGFP. For the purpose of transducing genes into hematopoietic cells safety and developing novel immunotherapy for cancer using gene modified T cells, our current results strongly suggested that the newly developed MV vector is considered to be more powerful vector than conventional gene transfer methods.
Then, we generated induced pluripotent stem cells (primed iPS cells) from terminal differentiated T cells and naive like pluripotent stem cells (NL-iPS cells) from cord blood derived CD34+ cells using MV-ΔF-OSKL-EGFP in same culture method and culture media. These primed iPS and GSL-iPS cells were demonstrated to express pluripotent markers (OCT3/4, NANOG) and differentiate into three germ line tissues. NL-iPS cells also expressed TFE3 proteins in the nuclei and SSEA1 in the cytoplasm. Interestingly, Enhanced GFP from MV-ΔF-OSKL-EGFP were lost in primed iPS cells and expressed strongly in NL-iPS cells. In the case of establishing iPS cells using this MV vector from CD34+ cells, we could establish only NL-iPS cells but not primed iPS cells, whereas we could not establish NL-iPS cells by using conventional vectors including Sendai vious vector in the same methods as MV vector.
In conclusion, our newly developed MV vector is considered to lead revolutionary advance in the research and clinical fields of gene therapy and stem cell.

Disclosures
No relevant conflicts of interest to declare.
Dinamic Evaluation of Quality of Life and Late Effects in a Cohort of Acute Myeloid Leukemia Long Term Survivors
Authors: Capelli Debora , Giostra Liviana , Maravalle Denise , Leoni Pietro , Olivieri Attilio 
Published: October 2019

Abstract
Late effects and quality of life (QoL) in Acute Myeloid Leukemia (AML) long term adult survivors represent an unexplored field of interest. We prospectively evaluated late effects and dynamic QoL in 44 and 29 cured AML patients (17 <60 years, and 12 >60 years), treated at our department between 1997 and 2010 (7 Allogeneic, 16 Autologous Transplant, 21 chemotherapy alone). We administered EORTC QLQ-C30 and FACT-AN questionnaires at two different time points with a median interval of 29 months (range: 12-34 months). We stratifyed QoL scores by age at diagnosis, performance status (PS), Sorror Index, kind of leukemia treatment, comorbidity at diagnosis.
We observed a worsening of emotional (-9.03; p= 0.04) and cognitive (-6.94; p= 0.05) EORTC scale scores, while FACTG (+2.9; p=0.03), emotional (+1.1; p= 0.04) and Functional (+2.25; p=0.001) well being FACT scores increased.
Multivariate analysis showed that older patients had worse EORTC QLQ-C30 physical and emotional scale scores and higher values of pain symptoms in comparison to younger counterpart, with RR of 20.1 (p = 0.001), 22.7 ( p <0.04) and 18.4 (p=0.03) respectively. Elderly patients also had lower Total Outcome Index and FACT-An subscale scores (RR: 11.9, p= 0.02; and 8.77, p= 0.04 respectively). Sorror index > 2 was related to lower EORTC QLQ-C30 social scale and dyspnea scores (RR: 32.5; p=0.001 and 21.7; p=0.001 respectively) and FACT-An functional well being values (RR=3.9; p=0.001).
We evaluated late effects occurring in 44 patients, since the third month after the end of treatment, with a median follow-up of 70 months (range: 12-166 months). The most frequent grade II-IV late toxicity was cardiac (3 arythmia, 9 cardiomyopathy) with 89% incidence in patients with Sorror HCT-CI score>2 at diagnosis vs 8.8% in the remaining patients and 0%, 20% and 55.5% in patients receiving respectively Daunorubicin, Idarubicin and at least two different anthracyclines. Sorror Index>2 was the only factor significantly predicting cardiotoxicity at the multivariate analysis with a RR of 82.7 (p=0.001). Twelve patients developped a transient hemochromatosis secondary to transfusions, treated with phlebotomy in 3 cases. Three patients (2 males and 1 female) had been fertile; all female patients developped menopause after Transplant. Four patients had secondary neoplasia consisting of Multiple Myeloma, breast cancer, myelodisplasia and axillary sarcoma.
Our study underlines the role of Sorror Index at diagnosis in defining patients eligibility to cardio-prophylactic therapy. The analysis of larger series of cured AML patients are strongly needed in order to define guidelines for reducing long term treatment AML toxicity.

Disclosures
No relevant conflicts of interest to declare.
JQ1, a Selective Bromodomain Inhibitor, Augment the Immunogenicity of Mantle Cell Lymphoma By Influencing the Expression of PD-L1
Authors: Wang Hongwei , Cheng Fengdong , Cheng Jie , Villagra Alejandro , Tao Jianguo , Bradner James E. , Sotomayor Eduardo M. 
Published: October 2019

Abstract
Mantle cell lymphoma (MCL) is a rare and aggressive form of non-Hodgkin's B-cell lymphoma and represents 4% to 8% of NHL. New therapeutic strategies are under development to improve the clinical outcome of patients with this disease. Recent studies have highlighted the importance of BRD/acetyl-lysine binding in orchestrating molecular interactions in chromatin biology and regulating gene transcription. The Bromodomain (BRD) is a diverse family of evolutionary conserved protein-interaction modules, which recognizes acetylated lysine residues such as those on the N-terminal tails of histones.
We have recently determined the functional consequences of inhibiting Bromodomains in MCL. First, we evaluated the effects of JQ1, a selective small-molecule bromodomain inhibitor upon MCL's immunogenicity and antigen-specific CD4+ T-cell responses. In vitro treatment of MCL cells with increasing concentrations of JQ1 resulted in a decreased expression of the tolerogenic molecule PD-L1 as compared to control cells. Second, we evaluated the capabilities of JQ1-treated MCL cells to present cognate antigen to naïve or anergic antigen-specific CD4+ T-cells. We found that treatment of MCL cells with JQ1 enhances their antigen-presenting capabilities leading to effective priming of naïve CD4+ T-cells and restoring the responsiveness of tolerant T-cells as determined by their increased production of IL-2 and IFN-gamma in response to cognate antigen. More importantly, in vivo studies clearly demonstrated a strong anti-tumor effect in JQ1-treated MCL tumor bearing mice. Mechanistically, we found a decreased STAT3 phosphorylation and c-Myc expression in JQ1-treated MCL cells, which in turn might be responsible for the diminished expression of PDL1 and augmentation of MCL's immunogenicity.
Taken together, we have found that MCL cells treated with the Bromodomain specific inhibitor JQ1 are more inflammatory, display lower expression of the immunosuppressive molecule PDL1 and, are capable of restoring the responsiveness of tolerant T-cells. More importantly, JQ1 treatment in vivo inhibits tumor growth in MCL tumor bearing animals. Our studies therefore have unveiled a previously unknown immunological effect of BRD inhibitors and have identified a novel immunotherapeutic approach in MCL.

Disclosures
No relevant conflicts of interest to declare.
RAB14 and RAB5 Gtpases Regulate Human Erythropoiesis, Potentially Via Opposing Roles in Endosomal Recycling
Authors: Kim MinJung , Kingsbury Tami J. , Cheng Wen-Chih , Tan Yee Sun , Taylor Brittany M. , Koshatwar Sampada , Civin Curt I. 
Published: October 2019

Abstract
Previously, we reported that the erythroid-expressed miRs, miR-144 and miR-451, target the RAB14 GTPase during human erythropoiesis in the human TF1 erythropoietic model cell line and in primary CD34+ hematopoietic stem-progenitor cells. In response to erythropoietin, endogenous RAB14 levels decreased during erythropoiesis, and RAB14 knockdown increased the numbers of erythroid (CD34- CD71hi CD235ahi) cells, increased b-hemoglobin expression, and decreased ETO2 expression (Kim, BJH, 2015). Taken together, our findings revealed that RAB14 functions as a physiologic inhibitor of human erythropoiesis. RAB GTPases comprise a >60 member subfamily of the large RAS oncogene family, and multiple RAB GTPases play important roles in vesicle trafficking, signal transduction and receptor recycling. RAB14 is a component of the intermediate compartment of the endosomal recycling pathway, between the RAB4/RAB5-dependent early endosome and the RAB11-dependent recycling endosome (Linford, Dev. Cell, 2012). The RAB GTPases are responsible for directing cargo proteins to the recycling vs. degradation pathway during endosome maturation. To test the role of another endosomal RAB GTPases in human erythropoiesis, we examined the consequences of knocking down RAB5 protein levels. In contrast to our findings with RAB14, knockdown of RAB5C decreased the numbers of erythroid cells generated during erythropoiesis, indicating that RAB5C enhances physiologic erythropoiesis. Consistent with this observation, mRNA and protein levels for all three RAB5 isoforms (i.e. RAB5A, B, C) increased during erythropoiesis. The opposing erythropoietic effects of RAB14 and RAB5 GTPases may be due to their contrary roles in endosomal recycling, in that internalization of cell surface receptors is dependent on RAB5 GTPase, whereas RAB14 GTPase is involved in recycling of receptors back to the cell surface.
Figure 1. Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Utility of Lung Ultrasound for Evaluating Acute Chest Syndrome in Pediatric Patients with Sickle Cell Disease
Authors: Cohen Stephanie G. , Malik Zayir M , Hagbom Robert , Alazraki Adina L , Zmitrovich April G. , Adisa Olufolake , Morris Claudia R. 
Published: October 2019

Abstract
Background: Acute chest syndrome (ACS) is a common complication in patients with sickle cell disease (SCD) and is a leading cause of morbidity and mortality. Physical assessment alone is not sensitive for diagnosing ACS and therefore Chest x-ray (CXR) is recommended because of the difficulty of diagnosing ACS on clinical grounds alone. Children with SCD are repeatedly exposed to diagnostic radiation for the evaluation of ACS. Focused chest ultrasound (US) has been used to evaluate for lung consolidation. If lung US can identify patients with ACS this application could potentially limit radiation exposure in patients with SCD at risk for ACS. We evaluated the utility of physician performed US as compared to CXR to identify patients with SCD who have ACS.
Methods: This is a prospective observational study that took place from November 2014-July 2015 in 2 urban pediatric emergency departments (EDs). The study population consisted of a convenience sample of patients with SCD from birth to 18 years of age at risk for ACS and who received a CXR for suspected ACS. Medical students and clinicians with training in lung sonography consented patients and performed a focused study to evaluate for lung consolidation. A blinded expert in point-of-care US reviewed for quality assurance and agreement. Sensitivity, specificity, and likelihood ratios were calculated for test performance characteristics of ultrasound using CXR as a reference standard. Inter-observer agreement (κ) between enrolling sonologists and reviewer was also calculated.
Results: 85 patients were enrolled for a total of 98 cases. Median age was 7 years (IQR 2-13 years) and 53% of patients were male. The prevalence of ACS by CXR was 14%. Lung US was able to detect consolidation with a sensitivity of 86% (95% CI, 56-97%), specificity of 95% (95% CI, 87%-98%), positive likelihood ratio (LR) 18 (95% CI, 7-48) and negative LR 0.2 (95% CI, 0.04-0.5). The agreement between enrolling novice sonologists' interpretation and blinded reviewer's interpretation was very good with a Cohen κ of 0.86 (95% CI, 0.7-1).
Conclusions: Focused lung US was able to identify ACS with high specificity. There was very good agreement between novice and expert sonologist interpretation. Lung US may decrease the need for CXR in patients at risk for ACS. Further studies are needed to see how this test performs within current clinical practice guidelines.

Disclosures
No relevant conflicts of interest to declare.
The Role of Receptor Interacting Protein Kinase in Myelopoiesis in Health and Disease
Authors: Silke John , Ma Chunyan , Brumatti Gabriela , Lalaoui Najoua , Ekert Paul G 
Published: October 2019

Abstract
Ripk1 -/-mice die at birth from systemic inflammation and this was not transferable in hematopoietic reconstitution experiments. One caveat to interpreting this result is our observation that Ripk1-/- progenitors failed to engraft lethally irradiated hosts properly. Competitive and serial transplantation experiments suggest that Ripk1-/- hematopoietic stem and progenitor cells (HSPC) have a cell-intrinsic defect. Blocking TNF allowed Ripk1-/- progenitors to reconstitute effectively supporting the idea that the cell-intrinsic defect is due to hyper-sensitivity to TNF induced death1. TNF can induce both caspase-8 dependent apoptosis and RIPK3/MLKL dependent necroptosis. These results prompted us to determine whether RIPK1 and TNF induced cell death had a role in leukemogenesis. We therefore generated mouse models of AML leukemia by retroviral infection of fusion MLL-ENL constructs into E14 fetal liver cells from Ripk1-/-, Ripk3-/-, Mlkl-/-, Casp8-/-Ripk3-/-, Casp8-/-Mlkl-/- and Ripk3-/-Mlkl-/- mice and transplanting them into sub-lethally irradiated wild type hosts2. Except for the Ripk1-/- MLL-ENL there was no difference in the development of leukemia from these genotypes compared to MLL-ENL wild type. Surprisingly however, given the defective reconstitution potential of Ripk1-/- HSPC, leukemia induced lethality occurred more than two times faster in Ripk1-/- MLL-ENL (20 days) compared to wild type MLL-ENL (50 days). No differences in blood profile, histology, myeloid markers or cell cycling were observed. Even more surprisingly, when we retransplanted these leukemias, Ripk1-/- MLL-ENL had a prolonged disease onset (over 80 days) compared to wild type (around 25 days). I will describe experiments to discover the mechanism behind these intriguing findings.
1. Rickard JA, O'Donnell JA, Evans JM, Lalaoui N, Poh AR, et al. RIPK1 Regulates RIPK3-MLKL-Driven Systemic Inflammation and Emergency Hematopoiesis. Cell. 2014;157:1175-1188.
2. Zuber J, Radtke I, Pardee TS, Zhao Z, Rappaport AR, et al. Mouse models of human AML accurately predict chemotherapy response. Genes Dev. 2009;23:877-889.

Disclosures
No relevant conflicts of interest to declare.
Regulation of Platelet Procoagulant Activity
Authors: Heemskerk Johan 
Published: October 2019

Abstract
Platelets play prominent and unique roles in the kinetics of the coagulation process. Modulated by the flow conditions, they control coagulation in various ways: (1) by integrin activation and ensuing clot retraction; (2) via exposure of the procoagulant phospholipid phosphatidylserine (PS), which causes binding of a specific set of coagulation factors at the platelet surface with as a result tenase activation and thrombin generation; (3) by initiating fibrin fiber formation at the platelet surface, a process specifically catalyzed by coated platelets. Recent evidence points to fibrin as an active structural network, which provides binding sides for plasma factors and platelets and, thereby, provides an important feedback mechanism for the stimulation of platelet activation. This overview will focus on novel insights into the signaling and activation processes in platelets that regulate these distinct procoagulant functions. One key mechanism is the activation and secondary closure of integrin alphaIIb-beta3. Another key mechanism concerns the calcium-dependent exposure of PS, which is accompanied by swelling of the platelets. Together, these processes control not only platelet adhesion, but also the accessibility for coagulation factors. Major distortions in these procoagulant functions are seen in platelets from a patient with the Scott syndrome, a rare bleeding disorder associated with mutations in the calcium-dependent ion channel protein anoctamin-6 (gene ANO6, previously TMEM16A). These platelets are deficient in calcium-induced PS exposure, swelling, integrin inactivation and in the formation of thrombin and fibrin at the platelet surface. These dysfunctions are precisely phenocopied in mice lacking anoctamin-6 expression. Advanced platelet proteomics analysis indicated that the deficiency in anoctamin-6 was accompanied by decreased calpain-dependent cleavage of a whole spectrum of intracellular proteins, and an increased phosphorylation state of many signaling and cytoskeletal proteins. However, calcium signaling in the Scott platelets was unchanged. In contrast, reports on patients with a gain in platelet procoagulant function, the Stormorken syndrome, indicate a link with genetic mutation in the calcium signaling protein STIM1. The possible consequences of these novel insights for our understanding of haemorrhagic and thrombotic deseases will be discussed.

Disclosures
No relevant conflicts of interest to declare.
Iron Requirement in Hypoxic Conditions
Authors: Gassmann Max 
Published: October 2019

While highlanders in the Andes and Himalayas are well adapted to live at high altitude, adequate acclimatization time to slowly adjust to hypoxic conditions is among the most important aspects for ascending mountaineers. Accordingly, most reports emphasize on mechanisms that cope with reduced oxygenation. Notably, during a journey to high altitude coverage of the elevated iron demand is equally critical. Thus, it's crucial to focus on the crosstalk between oxygen and iron homeostasis. I will discuss the role of iron in the oxygen sensing process and erythropoietin (Epo) synthesis as well as in gene expression control mediated by the hypoxia-inducible factor-2 (HIF-2). The blood hormone Epo that is abundantly expressed by the kidney under hypoxic conditions stimulates erythropoiesis in the bone marrow, a process that requires high quantities of iron. To ensure that sufficient iron is provided, hypoxia-induced soluble factors - such as the novel Epo-controlled erythroferrone that is expressed in erythroid precursor cells - reach the liver where they reduce expression of the iron hormone hepcidin. In turn, suppression of hepcidin allows both, elevated iron release from storage organs and enhanced absorption of dietary iron by enterocytes. As recently observed in sojourners at high altitude, however, iron uptake may be hampered by reduced appetite and gastrointestinal bleeding. Overall, adequate systemic iron availability is an important prerequisite to adjust not only to high-altitude induced hypoxia but to treat disease-related hypoxic conditions.

Disclosures
No relevant conflicts of interest to declare.
Pharmacomechanical Catheter Directed Thrombolysis for Pregnancy Related Proximal Lower Extremity Deep Venous Thrombosis: Effectiveness in the Prevention of Post Thrombotic Syndrome
Authors: Spectre Galia , Bloom Allan , Roth Batia , Varon David , Kalish Yosef , Da'as Nael , Tzitershpiler Tal 
Published: October 2019

Abstract
Pharmacomechanical catheter-directed thrombolysis (PCDT) reduces the incidence of post thrombotic syndrome (PTS) after proximal deep vein thrombosis (DVT); however its efficacy in PTS prevention for pregnancy related DVT is unknown. Most of the patients with DVT in pregnancy present with femoral or iliofemoral involvement and the prevalence after of PTS after pregnancy related DVT in the lower extermity is 42%. The purpose of this study is to compare PTS outcome in women with pregnancy related DVT with and without PCDT.
Methods: Women with pregnancy related proximal (femoral/iliofemoral) DVT who underwent PCDT and anticoagulation or anticoagulation alone, were evaluated for PTS using the Villalta scale and VEINES-QOL/Sym questionnaire.
Results: Eleven women with iliofemoral DVT underwent PCDT and anticoagulation, two during the first trimester and nine postpartum. Eighteen women (13 with iliofemoral DVT) were treated with anticoagulation only. There was no difference in age, number of pregnancies, trimester, duration of anticoagulation or thrombophilia between the groups. The time between DVT diagnosis and study inclusion was longer in the control group - median 50.5 (range 16-120) months , compared to study group: median 27 (range 11-64) months. None of the women in the study group developed PTS. Six of 18 patients in the control group developed PTS (33.3%) P=0.03, four of whom developed severe PTS (Villalta scale ≥15). One patient in each group developed recurrent DVT, and one patient in the study group developed a calf hematoma. A reduced frequency of lower extremity symptoms was observed in the study group by VEINES-Sym questionnaire P=0.01, but there was no difference in VEINES-QOL questionnaire (p=0.11)
Conclusion: This study suggests a reduction in PTS in women with pregnancy related proximal DVT who were treated with PCDT and anticoagulation compared to women who were treated with anticoagulation alone

Disclosures
No relevant conflicts of interest to declare.