bluuebunny/crossref_metadata_2025_split huggingface dataset:
ABT-737 Sensitizes B Cell Tumors for Killing by CD19-Retargeted T Cells,
Authors: Karlsson Hannah S C , Lindqvist Camilla , Paul-Wetterberg Gabriella , Wiklund Helena Jernberg , Nilsson Kenneth , Simonsson Bengt , Frisk Per , Loskog Angelica 
Published: October 2019

Abstract
Abstract 4032

Introduction:
T cells expressing tumor-targeting chimeric antigen receptors are showing promise in clinical trials for patients with B cell leukemia and lymphoma. However, increased levels of anti-apoptotic proteins, a common trait among B-cell tumors, may hamper treatment efficacy. ABT-737 is a small molecule inhibitor of anti-apoptotic proteins such as BCL-2, BCL-xL, BCL-w, and MCL-1, which induces apoptosis via the intrinsic apoptosis pathway in contrast to T-cells that utilize the extrinsic pathway controlled by death receptors and their ligands. ABT-737 has been shown to efficiently promote apoptosis in B-cell tumors as exemplified in models of pre-B-ALL. Recently, ABT-737 was shown to synergize with TRAIL to induce apoptosis. This prompted us to investigate if ABT-737 could be combined with T-cell therapy to enhance tumor cell death.

Methods:
PBMCs from healthy donors and patients with pre-B-ALL was genetically engineered with a second generation chimeric antigen receptor (CAR) targeting CD19 on B-cells. The T-cells and ABT-737 were tested both individually, and in combination, for their cytotoxic capacity in in vitro assays such as flow cytometry and the Caspase-Glo® 3/7 assay. The effects were studied in a panel of B-cell tumor cell lines (Daudi, U698, Karpas422, DG75, Nall-1) since they may exhibit different apoptosis resistance profiles. The expression of anti-apoptosis molecules in these cell lines was investigated by PCR.

Results:
PCR confirmed expression of BCL family proteins in the cell lines tested. CD19-targeting T-cells specifically induced apoptosis in CD19+ tumor cells. Similarly, but less efficiently, ABT-737 as single agent increased apoptosis in the various tumor cell lines. When combining T-cell and ABT-737 therapy, the tumor cell death was significantly increased to that of single agent treatment. The effect varied from additive to synergistic effects. The tumor cell lines did not change the level of antigen presenting molecules (MHC I and II), death receptors (Fas) or adhesion or costimulatory molecules (ICAM-I, CD80, CD86) upon ABT-737 treatment. Hence, the effect did not likely represent increased killing by enhanced physical interaction between T-cells and tumors but rather simultaneous engagement of both intrinsic and extrinsic apoptosis pathways.

Conclusion:
The apoptosis inducer ABT-737 is potently enhancing CD19-targeting T-cell therapy. By triggering both intrinsic and extrinsic apoptosis pathways also resistant tumors may succumb to treatment.

Disclosures:
Simonsson: Novartis, BMS, Merck, Pfizer: Consultancy, Honoraria.
Neutropenic Fever During Stem Cell Mobilization Is Associated with Decreased CD34+ Cell Collection and Poor Survival Following Autologous Stem Cell Transplantation,
Authors: Duong Hien , Bolwell Brian J. , Rybicki Lisa , Sweetenham John William , Pohlman Brad , Dean Robert , Andresen Steven , Kalaycio Matt , Sobecks Ronald , Koo Anna , Copelan Edward A. 
Published: October 2019

Abstract
Abstract 4053
High-dose chemotherapy followed by autologous stem cell transplant can improve long-term outcome of patients with relapsed non-Hodgkin and Hodgkin lymphoma. Stem cell mobilization with chemotherapy in addition to G-CSF improves stem cell collection, however it is also associated with increased risk of neutropenic fever. We analyze whether hospitalization for neutropenic fever (NF) affects outcomes after transplantation.
From 1/1998 to 12/2008, 599 patients underwent chemotherapy priming with etoposide + G-CSF (27 patients also received rituximab). Forty five (8%) did not proceed to transplant. Of the 554 (92%) patients who were transplanted, the median age was 51 years, and 348 (63%) were male. The diagnoses were 422 non-Hodgkin lymphoma (63%) and 132 Hodgkin lymphoma (24%). A majority of these patients (92%) had received 3 or less prior chemotherapy regimens. Only 6% had been exposed to fludarabine therapy. Most patients had chemosensitive disease (88% were in CR or PR) at time of transplant. For preparative regimen: 517 (93%) received busulfan/etoposide/cyclophosphamide, 32 (5.8%) received melphalan, 5 (0.9%) received other chemotherapy.
Of the transplanted patients, 121 (21.8%) were admitted for NF during stem cell mobilization. The median duration of admission for NF was 5 days (range 1–20). Six (5%) patients had bacteremia, 1 (<1%) had parainfluenza virus pneumonia, and the remainder 114 (94%) did not have any infection source or organism identified. The patients hospitalized for NF had significantly lower CD34+ cell collection (7.49 × 10^6 CD34 cells vs. 9.36 × 10^6, P=0.029) and lower CD34+ cell dose (7.34 × 10^6 CD34 cells/kg vs. 8.60 × 10^6 CD34+ cells/kg, P=0.047) despite more apheresis days required for collection (4 days vs. 3 days, P=0.01). Following transplant hospitalization, the 30-day readmission rate was higher in NF patients (16.8% compared to 8.8%, P=0.012). Hospitalization for NF was also associated with worse survival as shown below:
In conclusion, admission for neutropenic fever during stem cell mobilization is associated with significantly increased number of apheresis days required for adequate stem cell collection, lower CD34+ cell collection, higher 30-day re-admission rates, and poorer survival.

Disclosures:
No relevant conflicts of interest to declare.
Matched Pair Analysis of Cryopreserved Versus Fresh Allogeneic Peripheral Blood Stem Cell Transplantation
Authors: Trummer Arne , Krauter Juergen , Stadler Michael , Ganser Arnold , Buchholz Stefanie 
Published: October 2019

Abstract
Abstract 4529
Published data about outcome after cryopreserved allogeneic peripheral blood stem cell transplantation are scarce and, so far, have only been compared to historic cohorts of fresh graft recipients. We have performed a matched pair analysis of 66 patients receiving either cryopreserved or fresh grafts from matched related donors at our institution between January 2005 and June 2011 with a median follow-up time of 576 days (range: 18–2080 days).
For matching patients we calculated a propensity score including patient age, sex, diagnosis, performance status, remission status before transplantation (first remission vs. other), conditioning therapy (standard vs. reduced intensity), GvHD prophylaxis (with or without methotrexate) and CD34 cell count in the graft. Consequently, there were no significant differences between both groups for these parameters: median patient and donor age were 53 years (range: 18–68 and 23–76 years, respectively) for 34 male and 32 female patients with a performance status of ECOG 0 (n=60) or ECOG 1 (n=6). Diagnoses were AML (n=47), ALL (8), lymphoma (10) and CMPN (1). 31 patients were in first remission before transplantation. Reduced intensity conditioning therapy (n=52) and GvHD prophylaxis without methotrexate (n=41) were more frequent than standard conditioning (n=14) and prophylaxis with methotrexate (n=25), respectively.
Median cell counts were also almost equal in fresh and cryopreserved grafts for CD34 cells (5.7×106/kg (range: 3.1–11.4×106/kg) vs. 5.1×106/kg (2.6–12.3×106/kg), respectively), total nuclear cells (10.0×108/kg (4.9–21.4×108/kg) vs. 9.6×108/kg (5.0–19.1×108/kg) and CD3 lymphocytes (3.8×108/kg (2.1–6.8×108/kg vs. 3.4×108/kg (0.6–4.5×108/kg)).
All patients engrafted. Median neutrophil engraftment with an ANC >0.5×109/l was reached after 16 days (range: 10–21) vs. 15 days (10–31) (p=.15 by paired t-test) and platelet engraftment >20×109/l (for 3 consecutive days without requiring transfusion) occurred after 13 days (8–33) vs. 14 days (9–45) (p=.27).
Median follow-up time was similar between both groups (566 vs. 586 days, p=.894) and mean overall survival time, as calculated by Kaplan-Meyer analysis, was 1113 days for patients receiving fresh compared to 1258 days for patients receiving cryopreserved grafts (p=.582 by log rank test). Relapse or progression occurred in 13 vs. 14 patients, giving a mean disease/progression-free survival time of 922 vs. 1114 days (p=.467).
In summary, we did not observe any relevant outcome differences between patients receiving fresh or cryopreserved peripheral stem cell grafts of matched related donors. Therefore, the use of cryopreserved grafts can be considered safe and may even allow a more flexible transplant scheduling compared to fresh allografts.

Disclosures:
No relevant conflicts of interest to declare.
Oral Loading of Tacrolimus and Sub-Therapeutic Levels on the Day of Transplant Do Not Predict the Incidence of Acute Graft Versus Disease or Survival After Allogeneic Stem Cell Transplant
Authors: MacPherson Jill , Bachier Carlos , LeMaistre Charles F. , Sowell Hannah , Garcia Luis , Curel-Sanchez Alessandra , Cain Wilma , Caga Ursula , Means Jeremy , Shaughnessy Paul 
Published: October 2019

Abstract
Abstract 4543
Tacrolimus is proven to be effective in the prophylaxis of acute graft versus host disease (aGVHD) after allogeneic stem cell transplantation (ASCT). Published studies that initiate tacrolimus loading intravenously indicate that levels should be between 5 and 20ug/L on the day of transplant. Our institution often performs ASCT in the outpatient setting and therefore we developed a process for oral loading of tacrolimus. We retrospectively reviewed our experience in 94 consecutive patients (pts) who received ASCT from 10/10 HLA matched donors, 47 were matched sibling donors (MSD) and 47 were matched unrelated donors (MUD) with a median age of 50 and 52 respectively. Preparative regimens were IV busulfan based in 68 pts (BuFlu n=44, BuCY n=24), TBI based in 19pts, and other in 7 pts. All pts received GVHD prophylaxis with tacrolimus started orally between day -3 and -1 at a dose of 0.06 mg/kg/day divided twice daily. Standard short course methotrexate (MTX) was prescribed for all pts (D1 dose 15mg/m2, D3, 6, and 11 dose 10mg/m2). Pts who had sub-therapeutic tacrolimus levels (less than 5ug/L) on the day of transplant (Day 0) were compared to pts who had therapeutic tacrolimus levels (greater than 5ug/L) for incidence of any aGVHD and overall survival. Sixty nine patients had tacrolimus levels less than 5 ug/L on day 0 and 25 patients had tacrolimus levels of 5 ug/L or greater. There was no difference in incidence of any GVHD or survival between the groups (Table 1). Thirty one pts did not receive the entire MTX dose prescribed (most commonly because of withholding day 11 MTX dose), but there was no significant difference in the incidence of aGVHD in these pts despite the level of tacrolimus on D0. With a median follow up of 441 days 54/94 pts (57%) survive; 40/69 pts (58%) with day 0 tacrolimus levels that were less than 5 ug/L, and 14/25 pts (56%) with day 0 tacrolimus levels that were greater than or equal to 5 ug/L. In conclusion, oral loading of tacrolimus prior to ASCT at our institution results in sub-therapeutic levels on the day of transplant in the majority of patients, however, this does not appear to impact the overall incidence of aGVHD or survival. This study was limited by its retrospective nature, small size, and single center experience. Prospective randomized studies will be needed to determine if oral loading of tacrolimus impacts outcomes of ASCT.
Tacro; tacrolimus, OS; overall survival, MUD; matched unrelated donor, SIB; sibling. Survival determined in mean months from date of transplant.

Disclosures:
Shaughnessy: Genzyme: Consultancy, Honoraria, Research Funding, Speakers Bureau; Millenium: Honoraria, Speakers Bureau; Otsuka: Honoraria, Speakers Bureau.
Sequential Therapy with Activated Prothrombin Complex Concentrates and Recombinant Factor VIIa in the Treatment of Unresponsive Bleeding in Patients with Hemophilia and Inhibitors
Authors: Park Young shil , Choi Yong-Mook , Park Ji Kyoung 
Published: October 2019

Abstract
Abstract 4664
The development of inhibitors to factor VIII or IX is the most common, severe, challenging and expensive complication of the treatment of patients with hemophilia. Hemophilia patients with inhibitors can develop bleeding episodes, which are refractory to monotherapy with either recombinant factor VIIa (rFVIIa) or activated prothrombin complex concentrates (APCC). Management of such bleeds is often difficult.
This report describes the results of a retrospective chart review of four hospitalized patients with severe hemophilia and inhibitors who have been treated with sequential doses of APCC and rFVIIa for refractory bleeding.
Sequential therapy was defined as the administration of both rFVIIa and APCC within 12 h. In 4 cases of patients with inhibitor, bleeding was not controlled by initial bypassing treatment. One patient had historical peak inhibitor titer increased to 1126 BU, and during peripheral inserted central catheter insertion for immune tolerance induction therapy, bleeding was not controlled. After APCC and rFVIIa were administered sequentially, bleeding was controlled. The second and third patients had bleeding problem after total knee replacement for hemophilic arthropathy, and two agents were administered sequentially. The last patient had small bowel resection because of intestinal obstruction, and post-operation bleeding was not controlled well. DIC was developed. Two agents were administered sequentially for bleeding problem and bleeding was controlled. Sequential therapy administration alternated one APCC dose to 1–2 rFVIIa doses: dosing intervals ranged between 3 and 6 h; APCC (75–100 U kg) was given every 12 h; rFVIIa (90 μg/ kg) was given every 3–6 h. Bleeding control was achieved in 12–24 h in all patients. Sequential therapy was discontinued after 2–5 days. No clinical adverse events were observed.
Sequential therapy with APCC and rFVIIa was efficacious without adverse events. A prospective clinical trial is needed to provide further evidence.

Disclosures:
No relevant conflicts of interest to declare.
Experimental Study on Construction of Human Hiwi Adenovirus Vector and Apoptosis of K562 Cells Induced by Transfected Vector
Authors: Dong Xiaoyan , Jiang Yanfang , Liu Mengmeng , Li Wei , Liu Ziling 
Published: October 2019

Abstract
Abstract 4721

Background:
Leukemia stem cells are responsible for the genesis, progression, drug resistance and relapse of leukemia. Hiwi gene is an important divisive regulatory factor, which plays a role in maintaining the resting stage and down-regulating the cell cycle of stem/progenitor cells. We acquired human overall Hiwi coding region genes and constructed an adenovirus vector carrying human Hiwi with fluorescin. This study will not only establish the foundation of the further study in function and mechanism of Hiwi induce the differentiation and apoptosis of leukemia stem cells, but also provide the theoretical basis for searching new therapeutic target and method for leukemia.

Methods:
Using the overlap extension PCR method to amplify overall Hiwi coding region genes and insert overall Hiwi coding region genes into Flag-IRES –hrGFP vector that carrying green fluorescin with Gateway clone technology to construct pDown-Hiwi-3×flag-IRES-hrGFP. After transform the vector to Stb13, we screen the positive clone with PCR method, extract the plasmid and process recombination reaction between pDown-Hiwi-3×flag-IRES-hrGFP clone vector and pAV.Des1d expression vector to abtain the pAV.Ex1d-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant. Screening the positive clone with PCR and extracting the plasmid which were digested by enzyme and then examined by sequencing consequences. At last packaged Ad-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant is obtained. Transfect the recombinant adenovirus vector to K562 cells, and then identify the function of the cells.

Results:
Successfully clone human overall Hiwi coding region genes with the technology of overlap extension PCR and construct the adenovirus vector recombinant with enzyme cutting identification and gene sequencing examination.

Conclusions:
Successfully construct Ad-Hiwi-3×flag-IRES-hrGFP adenovirus vector recombinant and establish the foundation for the further study of Hiwi gene in leukemia stem cells. The transfected recombinant adenovirus vector can induce K562 cells to apoptosis. [U1]É¾3ý£z

Disclosures:
No relevant conflicts of interest to declare.
Results of Radiation Therapy for Primary Extranodal Lymphoma of the Head and Neck: A Report of Case Series
Authors: Turaka Aruna , Tianyu Li , Cohen Adam D , Pro Barbara , Millenson Michael , Robu Valentine , Al-Saleem Tahseen I. , Smith Mitchell R 
Published: October 2019

Abstract
Abstract 4761

Purpose:
Radiation therapy is an important treatment modality for the extra-nodal lymphoma (ENL) of the head and neck. Intensity Modulated radiation therapy (IMRT) has been shown for head and neck cancers to be associated with decreased late side effects like Xerostomia compared with conventional RT techniques (Conv.RT). The purpose of the study is to determine the clinico-pathological features, treatment outcomes and late toxicities in patients (pts) with ENL of head and neck treated with different RT techniques.

Methods:
Retrospective review of records from 2007–2010 identified 14 pts with ENL of the head and neck treated with RT at Fox Chase Cancer Center. Eight were treated with IMRT and 6 with conventional RT technique. Thirteen had CD20+ non-Hodgkin lymphoma [NHL; 7 diffuse large B cell (DLBCL), 3 MALT, 3 others], and one Hodgkin's lymphoma (HL; classic). All pts underwent routine staging work-up with negative bone marrow biopsy for all. Pre and post-treatment PET scans were done in 7 pts. Initial chemotherapy (R-CHOP × 4–6 courses for DLBCL, ABVD × 4 for HL), or no chemotherapy (MALT) was followed by involved field radiation therapy (median RT dose: 36 Gy for NHL, 30 Gy HL, 150– 180 cGy per fraction in 20 fractions over 4 weeks).

Results:
The median follow up was 28 months (range: 1– 52). The median age was 60 years (range: 42–95; 7 males and 7 female). Nine had stage IEA disease, 3 had stage II and 2 stage IV. Five had oropharynx involvement, two had paranasal sinus (DLBCL) 2 nasopharynx (1 DLBCL, 1 MALT) and 1 vallecula (HL). The overall response rate after combined modality treatment was 100%. There were no FDG avid lesion noted on post-treatment PET. There were no local or neck node relapses at last follow up. The 2-year actuarial survival rate was 80%. One DLBCL patient had systemic relapse involving stomach and bilateral testes, was treated with second-line chemotherapy and radiation therapy. At the last follow up, 12/14 were alive. RTOG grade 2 acute skin changes were noted in 6 pts (3 by each RT technique), grade 2 mucositis in 5 pts (3 with Conv.RT) and grade 2 xerostomia in 6 pts. None of the pts developed grade 3 xerostomia (both acute and late). Late grade 2 Xerostomia was seen in 5 pts treated with Conv.RT but not with IMRT.

Conclusions:
RT following chemotherapy or used alone in ENL of head and neck was associated with local and distant disease control in the majority of patients, with a toxicity profile that appears favorable for IMRT compared with conventional radiotherapy techniques. Larger studies are required to confirm that IMRT is as effective as conventional involved-field RT.

Disclosures:
Pro: Celgene: Consultancy, Honoraria.
Healthy People 2020 Blood Disorders and Blood Safety: A Vision of Improved Health for Patients with Sickle Cell Disease, Thalassemias and Hemophilia
Authors: Werner Ellen M. , Brown Lorraine , Hassell Kathryn L. , DiMichele Donna 
Published: October 2019

Abstract
Abstract 4771
In December 2010, the US Department of Health and Human Services released Healthy People 2020 (HP2020) Objectives for the nation. For the first time in its thirty-year history, US health objectives include rare blood disorders and blood safety (BDBS). HP2020 is a public health program that incorporates current clinical evidence and best practices in medicine and public health to identify public health priorities. The objectives are not clinical practice guidelines. Rather, they are scientifically-based, quantified goals to improve the health of specific populations, such as children and older adults. Their development and implementation necessitate partnerships among governmental agencies, organizations, private companies, and medical and health associations at the local, state and national levels. Partners collaborate on implementing activities to reach targeted health objectives for specific populations using educational campaigns, media and other communication venues.
NHLBI led the effort with HRSA, CDC, FDA, and the Office of the Secretary to develop, vet and plan implementation of the BDBS objectives. The development process was based on qualitative research methods such as key informant interviews, semi-structured interviews and categorization of data into patterns. For example, categorization of interview results (“data”) identified specific objective topics, such as screening for complications and penicillin prophylaxis in sickle cell patients aged 4 months to 5 years. Other BDBS Objective topics are: vaccinations; patient and family referrals; care in a medical home; disease-modifying therapies; avoidable hospitalizations; high school completion; community health education; awareness of carrier status; joint health; and alloimmunization in chronic transfusion. http://healthypeople.gov/2020/topicsobjectives2020/default.aspx By sponsoring the BDBS objectives, lead governmental agencies commit to collecting, analyzing and reporting data on progress.
We adopted the default target of a 10% improvement for each objective by 2020. If targets for each objective are met, the question will be: What is the impact of reaching these targets on morbidity, mortality and quality of life for patients with sickle cell disease, thalassemias and hemophilia?

Disclosures:
Hassell: NIH: Research Funding.
Gro 3, Osteoprotegerin and Interleukin 32 Function As Hematopoietic Growth Factors
Authors: Moldenhauer Anja , Futschik Matthias , Lu Huili , Helmig Melanie , Götze Patricc , Bal Gürkan , Zenke Martin , Han Wei , Salama Abdulgabar 
Published: October 2019

Abstract
Abstract 4784
The identification of soluble factors involved in stem cell renewal is a major goal in the assessment of the BM niche. We have previously shown that human endothelial cell (EC) supernatants can induce the proliferation of hematopoietic progenitor cells (HPCs), especially after stimulation with IL-1b. To identify new potential growth factors, we compared the expression profile of IL-1b-stimulated ECs over 4, 8 and 16 h with non-stimulated Ecs using oligonucleotide microarrays covering more than 46 000 transcripts. Most significant changes were detected after 4 h. Utilization of Gene Ontology annotation for the stimulated EC transcriptome indicated multiple upregulated genes encoding extracellular proteins with a cell–cell signaling function. Using flow cytometry, delta, colony and cobblestone assays, we assessed the proliferative capacities of 11 gene products, i.e. IL-8, IL-32, FGF-18, osteoprotegerin, Gro 1–3, ENA78, GCP-2, CCL2 and CCL20, which are not known to induce HPC expansion. Notably, IL-32 and to a lesser degree osteoprotegerin and Gro 3 significantly induced the proliferation of HPCs. Furthermore, IL-32 attenuated chemotherapy-related BM cytotoxicities by increasing the number of colony forming cells in mice (7.4±0.87×103 versus 0.7±0.25×103, p=0.003). Our findings confirm that the combination of microarrays and gene annotation helps to identify new hematopoietic growth factors.

Disclosures:
No relevant conflicts of interest to declare.