bluuebunny/crossref_metadata_2025_split huggingface dataset:
HLA-Haploidentical Donors Seem to be Not Inferior to HLA-Identical Sibling Donors for Hematopoietic Stem Cell Transplantation of Patients with Paroxysmal Nocturnal Hemoglobinuria (PNH)?
Authors: Chen Feng , Wu Depei , Tang XiaoWen , Miao Miao , Fu Chengcheng , Qiu Huiying , Xue Shengli , Ma Xiao , Jin Zhengming 
Published: November 2019

Abstract
Background PNH is an acquired clonal disorder of the hemopoietic stem cells for which the only curative treatment is allogeneic hematopietic stem cell transplantation (allo-HSCT). But allo-HSCT is challenging for those who have no HLA-matched donors.Several recent studies have shown that haploidentical HSCT for patients with hematological malignancy can achieved comparable outcomes with HLA-identical sibling transplantation . There are very few reports on the use of haploidentical HSCT for PNH . Is haploidentical HSCT a valid alternative option for patients with PNH?
Methods 19 PNH patients received allo-HSCT between Dec 2007 and Oct 2015 at our institution. 12 donors were HLA-haploidentical and 7 were HLA-matched siblings. The patients were aged 8 to 54 years (median 28 years) . Of the 12 haploidentical donors, 6 were siblings, 2 fathers,2 mothers,1 son and 1 daughter. 12 patients with haploidentical donors received a myeloablative conditioning regimen consisting of busulfan, cyclophosphamide and ATG (anti-thymocyte globulin), 7 patients with identical siblings were given a reduced intensity conditioning. G-CSF-mobilized bone marrow and peripheral blood stem cells were transplanted as graft. Prophylaxis for graft-versus-host disease (GVHD) consisting of cyclosporine or tacrolimus + short-term methotrexate + mycophenolate mofetil was used for 12 patients with haploidentical donors, and cyclosporine + short-term methotrexate for 7 with identical siblings.
Results All 19 patients were engrafted successfully. The median time of neutrophils (ANC) reached to 0.5×109/L and platelets (PLT) reached to 20×109/L was 12 days and 15 days in haploidentical group , and that to identical group was 11 days and 13 days ,respectively. There were 2 patients developed grade Ⅱ acute GVHD in haploidentical group while 1 patients with grade Ⅳ aGVHD in identical group . Limited chronic GVHD was observed in 2/12 patients in haploidentical group and 1/7 patients in identical group. After a median follow-up time of 22.0 (range 4.0-42.0) months, the 3-year OS probability was 77.8±13.9% and 85.7±13.2% for haploidentical and identical group,respectively (P=0.03). 2 patients died of treatment-related mortality in haploidentical group, including severe pulmonary infection (n=1) and transplant-associated thrombotic microangiopathy (n=1) ,and 1 died of severe aGVHD in identical group. No patients were documented to have a recurrence of a PNH clone after HSCT in both groups.
Conclusion This report seemed that long-term outcomes of HLA- haploidentical HSCT in patients with PNH were comparable to that of HLA- matched donor at our institution . Haploidentical HSCT should be considered as a valid alternative option for PNH patients without HLA- matched donors .

Disclosures
No relevant conflicts of interest to declare.
Allogeneic Stem Cell Transplantation for Multiple Myeloma: a Single Center Experience
Authors: Figueiredo Amarilis , Atkins Harold , Kekre Natasha , Kew Andrea , McCurdy Arleigh 
Published: November 2019

Abstract
Introduction
The use of allogeneic stem cell transplantation (Allo-SCT) in patients with multiple myeloma (MM) remains controversial, but it offers prolonged disease free survival in some patients. It is unclear which patients should undergo Allo-SCT, what conditioning regimen should be used, and what the timing of the transplant should be in the course of the disease. Therefore, we sought to contribute our center's experience to the growing body of literature.
Methods
We performed a retrospective observational cohort study of all patients who underwent Allo-SCT for multiple myeloma at our center between January 1, 1992 and May 31, 2016. Categorical variables were compared using Pearson's chi-square test and the Kaplan-Meier method was used for the overall survival curves.
Results
Thirty-four patients underwent Allo-SCT for multiple myeloma and were included in this analysis. The median age was 40 years and 21 (62%) were male. Nineteen patients (56%) underwent Allo-SCT as upfront therapy, 1 (3%) underwent tandem autologous stem cell transplant (auto-SCT) followed by Allo-SCT, and 14 (41%) had salvage Allo-SCT at the time of relapse. Twenty-four (70.5%) patients had a matched related donor, 1 (3%) had a mis-matched related donor, 8 (23.5%) had matched unrelated donor and matching in 1 (3%) was not available. Myeloablative conditioning was given in 18 patients (52.9%) and non-myeloablative conditioning in 13 (38.2%) with 3 (8.8%) missing. The conditioning regimens included: 5 (15%) Flu-Mel, 7(20.6%) Flu-Bu, 13 (38%) Bu-Cy ± TBI, 6 (17.6%) MelVPTBI, and 3 (8.8%) were missing. Median overall survival (OS) for all patients was 72.5 months from diagnosis (figure 1) and 26.5 months from the time of Allo-SCT. For the 19 patients who had upfront Allo-SCT, median OS from diagnosis was 7.4 years compared to 5.3 years for those having salvage Allo-SCT (figure 2). However, in the upfront group 6 (32%) were alive 10 years post Allo-SCT and the survival curve reaches plateau, whereas in the salvage group, no patient was alive after 8 years post Allo-SCT. There was no difference in median survival between myeloablative and non myeloablative conditioning (2.9 versus 1.33 years, p=0.925). There have been 20 deaths in our cohort (59%); 5 (14.7%) from transplant related mortality within 1 year, 9 (26.5%) from disease progression, and 6 (17.6%) transplanted remotely whose cause of death is unknown.
Conclusions
Our data suggest that Allo-SCT offers prolonged disease free survival in some patients. In our small cohort, a greater proportion of patients undergoing upfront Allo-SCT achieved long term survival, raising the possibility that this group of patients may benefit more from Allo-SCT. Further prospective studies are needed to clarify the role of Allo-SCT in MM.

Disclosures
Kew: Celgene: Honoraria. McCurdy:Celgene: Honoraria.
CXCR3 Upregulation Effect of T Lymphocytic Leukemia Cell Invasion in Vitro
Authors: Feng Xiaoqin , Ding Wenjiao , Li Chunfu 
Published: November 2019

Abstract
Objective
The study was to explore chemokine receptor CXCR3 influence on invasion of T lymphocytic leukemia cell.
Methods
Mouse CXCR3 gene was amplified from T vector containing mouse CXCR3 by PCR, and constructed the mouse CXCR3 overexpressing lentivirus carrying GFP & Puromycin (puro). Jurkat cell line was infected by the constructed CXCR3 overexpression lentivirus. Puromycin was added to screene the successfully infected lymphoblastic leukemia cells. CXCR3 expression on Jurkat cells was analyzed by flow cytometre. Constructed CXCR3-LV5-Jurkat cells were added into the Transwell upper chamber covered with Matrigel. Ligand of CXCR3(CXCL10) was added to the lower chamber. Cell number in the upper chamber and lower chamber which used to calculate the invasion rate was counted by flow cytometry respectively.
Results
GFP+ Jurkat cells was observed less than <10% after lentivirus infection 96h. GFP+Jurkat cells achieved to 90% by Puromycin selection. The expression of CXCR3 on CXCR3-LV5-Jurkat cell group was 90% higher than LV5-Jurkat cell group. CXCR3-LV5-Jurkat cell line stably overexpressed CXCR3 was successfully constructed. The invasion rate of CXCR3-LV5-Jurkat cell lines was significantly higher than LV5-Jurkat control group (12.71 %± 1.03% vs 6.82% ± 0.49%, p <0.0001).
Conclusion
Overexpression of CXCR3 on T lymphocytic leukemia cell line (Jurkat cell) can enhance the invasion of leukemia cells in vitro, CXCR3/CXCL10 path way may induce the T lymphoblastic leukemic cell invading the central nervous system.

Disclosures
No relevant conflicts of interest to declare.
TCR-Ligand Dissociation-Rate, a Reliable and Determining Parameter, Identifying Tumor-Reactive CD8 T Cells of High Avidity and Functional Potency
Authors: Rufer Nathalie , Allard Mathilde , Couturaud Barbara , Carretero Laura , Schmidt Julien , Duong Minh Ngoc , Baumgaertner Petra , Gannon Philippe , Speiser Daniel , Hebeisen Michael 
Published: November 2019

Abstract
Background & Aims: Kinetics of T-cell receptor (TCR) binding to peptide-MHC (pMHC) contribute to T-cell activation and various cellular responses. However, due to technical reasons, the characterization of T-cells is usually only done with functional assays, without assessment of TCR-pMHC binding kinetics, thus limiting the information on the overall quality and clinical relevance of TCR-ligand binding properties. With the aim to broadly evaluate possible correlations between T-cell function and TCR binding kinetics in humans, we undertook a large-scale analysis of combined multiple functional readouts (i.e. CD107a degranulation, cytokine production and proliferation) and off-rate measurements by NTAmers (ref. 1) to characterize tumor- and virus-specific CD8 T-cell clones (n=400) isolated from melanoma patients and healthy donors.
Results: Our results show that, within an antigen-specific repertoire, TCR-ligand off-rates accurately predicted single-functional and poly-functional avidities (i.e. ligand potency) as well as co-receptors expression/modulation of CD8 T-cells. Importantly, the TCR-pMHC dissociation-rate represented a constant and highly reliable bio-physical parameter, contrasting to functional avidity which depended on the activation status of the T-cells following re-stimulation. Remarkably, we found that the TCR-ligand off-rate repertoire depended on the antigenic-specificity, as CD8 T-cells specific for the cancer-testis antigen NY-ESO-1157-165 displayed longer TCR-ligand interactions than T-cells specific for the tumor/differentiation antigen Melan-A26-35, and both these tumor-specific TCRs exhibited significantly lower avidities than those specific for persistent herpes virus antigens (CMV/pp65495-503, EBV/BMFL1259-267).
Conclusions: Our data from a large library of human T-cell clones demonstrate that the TCR-ligand dissociation-rate is a stable/reliable and determining parameter of T-cell responsiveness, arguing that it should be systematically assessed for meaningful monitoring of human T-cell responses. Furthermore, our TCR-ligand off-rate comparisons highlighted strong binding differences between non-self/virus- and self/tumor-specific repertoires. Our data from individual clones suggest that the tumor-specific repertoires contains rare but nevertheless promising TCRs with sufficiently slow off-rate and high functional avidity to potentially confer protective immune responses against cancer.
(ref. 1) Hebeisen et al., Cancer Res, 75(10):1983, 2015

Disclosures
No relevant conflicts of interest to declare.
Dissection of Mechanisms of Realgar Intervening Telomere Protein POT1, TRF1, TRF2 Expression to Regulate Telomere Dynamics in THP-1 Cells
Authors: Xu Rui-Rong , Song Nan-Xin , Wu Xiao-Long , Cui Si-Yuan , Wang Zhen-Zhen , Wang Yan , Wang Xin 
Published: November 2019

Abstract
Objective: To observe the mechanisms of realgar intervening telomere protein POT1, TRF1, TRF2 expression to regulate telomere dynamics in THP-1 cells, and elucidate the experimental and theoretical basis for realgar targeted threapy of AML.
Methods: 1.In vitro: (1)Cultured human acute myeloid leukemia THP-1 cells. (2), The cells were incubated in absence or presence of increasing concentration of realgar for 24/48 h, from which determined the IC50. Cell viability was tested by CCK-8 assay. (3)Apoptotic rate and cell cycle distribution were tested by FCM. (4)Each group of POT1, TRF1, TRF2 protein levels were tested by western-blot analysis. (5)Each group of POT1,TRF1,TRF2 mRNA levels were tested by RT-qPCR analysis. 2.In vivo: (1)Established THP-1 model in NOD/SCID mice, and the mouse were treated by realgar. (2)All the mouse were killed by institutionally approved method after 3 weeks. The apoptotic rate and cell cycle distribution of mice spleen cells were examined by FCM. (3)The changes of POT1, TRF1, TRF2 protein levels in organic tissue were detected by IHC.
Results: 1.In vitro: (1)CCK-8 assay showed that after treated by realgar, THP-1 cells growth were inhibited and the cell viability decreased in a dose- and time- dependent manner, IC50 was 0.023μg/mL. Therefore, we choose a medium concentration was 0.015μg/mL (lower than IC50) at 48h. (2)Our study demonstrated that after treatment for 48h, the apoptotic rate and the G1 arrest of these THP-1 cell increased, compared with control group. (3)As shown by western blot analysis, compared with controls, realgar group POT1, TRF1 protein levels significantly increaesed and TRF2 decreased(P<0.01). (4) As shown by by RT-qPCR analysis, POT1, TRF1, TRF2 mRNA levels were consistent with their protein levels. 2.In vivo: (1) The THP-1/NOD-SCID model was established, and survival curve showed that compared with controls, realgar group mouse had longer life span(P<0.01). (2) The results of FCM showed that after treatment of realgar, the rate of THP-1 cell apoptosis and G1 arrest significantly increased(P<0.01). (3) The results of IHC showed that compared with controls, POT1, TRF1 protein levels significantly increased and TRF2 decreased in realgar group mouse(P<0.01).
Conclusion: In vitro and in vivo studies results indicated that realgar could significantly prolong the life span of the THP-1/NOD-SCID mice, inhibit proliferation and induce apoptosis of THP-1 cell, and regulate telomere dynamics through intervening THP-1 cell telomere protein POT1, TRF1, TRF2 expression. Our studies results provided experimental and theoretical basis for realgar targeted threapy of AML.

Disclosures
No relevant conflicts of interest to declare.
Therapeutic Development of Complement C5 Inhibitor CoversinTM with Extended Half-Life Via PASylation®
Authors: Miles Nunn , Skerra Arne 
Published: November 2019

Abstract
The complement C5 inhibitor Coversin is in clinical development for treatment of paroxysmal nocturnal hemoglobinuria (PNH) and has been shown to be safe and effective in phase 1 single dose and multiple dose trials in healthy volunteers. A single phase 2 PNH patient, resistant to the standard of care eculizumab, has been treated with Coversin for a period of more than 7 months and responded well to treatment.
In contrast to eculizumab, which is intravenously administered twice monthly by a health care professional, Coversin will be self-administered as a once daily subcutaneous injection. While this may provide patient convenience compared to intravenous administration, even longer intervals between subcutaneous dosing would enhance that convenience.
To extend Coversin's half-life and explore the potential for subcutaneous administration at longer dosing intervals Coversin has been modified by the addition of a 600 amino acid proline/alanine/serine (PAS) N-terminal fusion tag to generate PAS-Coversin (68kDa). The unstructured and uncharged PAS polypeptide gives Coversin an apparent molecular size of approximately 720kDa, thus retarding kidney filtration and extending the half-life of PAS-Coversin by 52-fold in mice compared to unmodified Coversin. Furthermore, in vitro complement lytic and C5 binding assays indicate PAS-Coversin inhibits complement C5 as potently as unmodified Coversin.
Recent work has shown that PAS-Coversin can be administered to mice by subcutaneous injection and has very high bioavailability when administered via this route. The bioactive protein can be produced in both (commercially scaleable) bacterial and yeast expression systems. The further development of PAS-Coversin for therapeutic purposes with the potential for weekly dosing will be presented.

Disclosures
Miles: Akari Therapeutics plc: Employment, Equity Ownership. Skerra:XL-protein GmbH: Equity Ownership.
Molecular Mechanism Analysis the Apoptosis of Leukemia Cells Induced By CRM1 Selective Inhibitor KPT-330
Authors: Lu Jun , Hu Shaoyan , DU Weiwei , Pan Jian , Ren Junli , Zhiheng Li 
Published: November 2019

Abstract
Objective: To investigate the effect and mechanisms of KPT-330 on cell apoptosis of human leukemia cells. Methods: CCK-8 assay was used to quantify the growth inhibition of cells after exposure to KPT-330 for 24 hours. Fluorescence microscope was used to observe the morphological change of leukemia cell when treated with KPT330 for 24 hours. Annexin V/propidium iodide staining followed by flow cytometric analysis was used to detect apoptosis of leukemia cells. Then Western blot was used to detect the expression level of CRM1 and the activation of apoptosis related protein Caspase-9、Caspase-3 and PARP level. LncRNA microarray was used to analyze the differentially expressed genes from the cluster analyses. Results: The inhibition of cell proliferation of leukemia cells was in a dose-dependent manner and KPT-330 treatment induced expression of CRM1 down-regulated .Abnormal cells were observed under fluorescence microscopy. Cell apoptosis, cell cycle and activation of apoptosis markers demonstrates that KPT-330 induced apoptosis in leukemia cells. LncRNA microarray analysis showed differently expressed mRNAs and lncRNAs in KPT-330 treated HL-60 cells compared with control group. Molecular function analysis showed that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. Conclusion: In this study, KPT-330 treatment resulted in inhibition of cell proliferation and induction of apoptosis in leukemia cells. LncRNA microarray analysis showed differentially expressed genes and lncRNAs in KPT-330 treated HL-60 cells and we demonstrated that KPT-330 induced apoptosis in leukemia cells partially related with Glutathione metabolism and Toll-like receptor signaling pathway. These results may provide new clue about molecular mechanism of KPT-330 induced apoptosis; however, underlying details will be required to determine further. Taken together, our findings suggest that for the first time that KPT-330 may act as new candidate's drug for leukemia.

Disclosures
No relevant conflicts of interest to declare.
A Fit-for-Purpose Approach to Monitor Circulating Dendritic Cell Subpopulations for Clinical Investigation
Authors: Trampont Paul Charles , Lourenco Elaine , Czechowska-Kusio Kamila , Litwin Virginia 
Published: November 2019

Abstract
Dendritic cells (DC) are a heterogeneous population of antigen presenting cells involved in the capture, processing and surface presentation of antigens to T-cells. Their immune functions serve to connect innate and adaptive immune responses. In human peripheral blood, the two main subpopulations of dendritic cells, myeloid and the plasmacytoid, are of interest as biomarkers for the development of new therapies and clinical indications. A fit-for-purpose approach was applied to the validation of a two related flow cytometric method designed to evaluate both myeloid and the plasmacytoid dendritic cell populations. The iterative approach allowed us to tailor the assays for and the validation for the intended use of the data within the clinical trial. This presentation describes the rationale for the panel design and the technical validation approach.

Disclosures
No relevant conflicts of interest to declare.
Increasing Quality Improvement Capability in a Hemophilia Treatment Center
Authors: Thornburg Courtney , Lane Heidi , Farrow Katharine , Brooks Rosalie , Jahan Mina , Scopelliti Maria 
Published: November 2019

Abstract
Introduction
Persons with hemophilia require complex medical care by a multi-disciplinary team throughout life. There are opportunities for Hemophilia Treatment Centers (HTC) to enhance the care of persons with hemophilia through participation in quality improvement (QI) initiatives. Maintaining comprehensive care and excellent health outcomes are national priorities for Health Resources and Services Administration (HRSA) and the American Thrombosis and Hemostasis Network (ATHN), the National Hemophilia Program Coordinating Center (NHPCC). Rady Children's Hospital San Diego (RCHSD) HTC participated in the NHPCC Dartmouth Improvement Program pilot program with the aim to develop QI capabilities within the HTCs to enhance the care of persons with hemophilia.
Methods
RCHSD participated in the NHPCC Dartmouth Improvement Program starting in December 2015. A QI team was established including the HTC medical director, nurse case managers, pediatric social worker and the social worker of the collaborating adult HTC. The QI team was coached by an expert TDIMA coach and a "coach in training" from another HTC. The team learned QI methods through in person training, web-based training and weekly team meetings with the coaches. The team assessed HTC data (The 5Ps-purpose, patients, professionals, processes, and patterns) to gain system knowledge and insights to determine a QI theme, global aim, specific aims and associated PDSA cycles.
Results
The RCHSD HTC QI team established a QI theme to focus on the transfer from pediatric to adult care. The Global Aim of the team is to improve autonomous communication in RCHSD HTC. The process begins with the 12 year old comprehensive clinic visit and ends with a new patient visit at an adult HTC. By working on the process, we expect patients to have the communication skills to be able to arrange medical insurance, call the home care company to order factor and to communicate their health and personal care needs to adult health care providers. We developed four specific aims which are in various stages of testing in PDSA cycles (Table 1). For Aim 1, we found that patients have only basic knowledge of insurance information. For Aim 2, we found that we needed to adjust our clinic process to make sure that patients receive their "after visit summary" that includes the insurance information prior to leaving clinic.
Conclusions
We developed a QI program within RCHSD HTC focused on improving transfer of care. Once we have completed the specific aim PDSA cycles we will standardize the new and improved procedures. We will continue to develop the program based on internal needs assessment and family input. This training model may be adapted by other HTCs to enhance patient care.

Disclosures
Thornburg: Mast Pharmaceuticals: Research Funding; Bayer Pharmaceuticals: Research Funding; Shire: Consultancy; Biogen Idec: Other: Data Safety Monitoring Board; Bluebird inc: Other: Data Safety Monitoring Board.