bluuebunny/crossref_metadata_2025_split huggingface dataset:
Manufacture of Clinical-Grade Universal Antigen Presenting Cells (UAPC) for the Ex Vivo Expansion and Activation of Natural Killer (NK) Cells
Authors: Liu Enli , Ang Sonny , Tong Yijiu , Li Li , Basar Rafet , Kerbauy Lucila , Daher May , Banerjee Pinaki P , Lim Francesca Lorraine Wei Inng , Kaur Indreshpal , Champlin Richard E , Shpall Elizabeth J. , Rezvani Katy 
Published: October 2019

Abstract
NK cells are potent cellular immunotherapeutic agents against a wide array of human malignancies. Ex vivo expansion of NK cells to achieve clinically relevant numbers must overcome poor in vitro growth kinetics, low starting percentages within the mononuclear cell fraction (especially from autologous donors with active disease), and limited in vivo life span.
We targeted three universally critical NK signaling pathways, namely IL-21, 4-1BB, and SLAM family member 4 (SLAMF4), to increase NK cell proliferation and enhance survival. We genetically-engineered HLA-A-ve and -B-ve K562 cells to enforce expression of membrane-bound IL-21 (mbIL21), 4-1BB-L, and CD48, forming a universal antigen presenting cell (UAPC) to generate highly potent clinical-grade umbilical cord blood (CB-NK) or peripheral blood NK cells (PB-NK). While the mbIL21 and 4-1BB signaling nexuses have been utilized previously, we highlight here SLAMF-mediated immunological sculpting of NK cells for clinical applications.
SLAMF triggering of co-receptors modulate NK cell activation, in particular through high-avidity interactions between SLAMF4 (2B4/CD244) and its heterophilic, robust affinity, and physiological ligand CD48, a glycosyl-phosphatidyl-inositol (GPI)-anchored cell surface protein. Upon ligand binding and receptor phosphorylation, SLAMF4 recruits PTPN11/SHP-2 and SH2D1A/SAP for downstream signaling, including significant increases in NK cell-mediated cytotoxicity, granule exocytosis, and production of IFN-γ and IL-2. Other than a subset of low surface density expressers from aging subjects consistently associated with inefficient and impaired activating signal transduction, the majority of NK cells express SLAMF4. The functional prominence of SLAMF4, presently recognized as an activating co-receptor, is also evidenced by loss-of-function mutations associated with X-linked lymphoproliferative (XLP) disease.
We achieved log-scale expansion of NK cells with UAPC (>1000 fold in 2 weeks), with excellent purity (>99% CD56+ve/CD3−ve and < 1% CD3+ve cells), without indications of senescence/exhaustion, even after 4 weeks of culture. Surface molecular phenotypes of UAPC-expanded CB-NK cells exhibited a phenotype similar to CB-NK cells expanded without SLAMF4. Synergistic signals from IL-21/STAT3, 4-1BB/4-1BBL, and SLAMF4 drive tonic NK propagation, supporting their clinical application. Our novel and clinically accessible platform technology for generation of high purity CB-NKs, a promising source of fresh and cryopreserved allogeneic NK cells, is well-suited for adoptive cancer immunotherapy.

Disclosures
Champlin: Sanofi: Research Funding; Otsuka: Research Funding. Shpall:Affirmed GmbH: Research Funding. Rezvani:Affirmed GmbH: Research Funding.
FANCI-FANCD2 Binds RNA, Which Stimulates Its Monoubiquitination
Authors: Liang Zhuobin , Teng Yaqun , Liu Jingchun , Longerich Simonne , Chen Xiaoyong , Green Allison M. , Collins Natalie , Lan Li , Sung Patrick , Kupfer Gary M. 
Published: October 2019

Abstract
Fanconi anemia (FA) is characterized by developmental abnormalities, bone marrow failure, and a strong cancer predisposition. FA cells are hypersensitive to DNA replicative stress, and accumulate co-transcriptional R-loops. Previous work has demonstrated that BRCA2 binds to R loops, and increased R loops are noted in FA-D2 mutant cells. Additionally, it is understood that at least one FA protein, FANCA, binds RNA.
The goal of this study was to understand the relationship between FANCD2 and RNA, especially with regard to manifestation of R loops as a part of the pathophysiology of FA. First, we confirmed the increased presence of R loops in FA mutant cells using the S9.6 monoclonal antibody immunofluorescence microscopy. RNAseH overexpression removes R loop signal and increases cell survival upon mitomycin C treatment. We also showed the presence of increased R loops in an actively transcribed region of the actin gene by bisulfite DNA sequencing. We used the Damage At RNA Transcription (DART) assay, which is designed to combine oxidative DNA damage and the genomic insertion of a hyper transcription site (Fig A). Coactivation of transcription and DNA damage results in colocalization of FANCD2 and S9.6/R loop signal at the transcriptional site (Fig B and C). Consistent with the S9.6 IF, wild type RNAseH overexpression resulted in the abrogation of FANCD2 colocalization. We then asked if FANCD2 binds RNA. FANCD2 in cell lysate bound to biotinylated RNA species, preferring GC rich RNAs. Using recombinant FANCI-FANCD2 (ID2) protein (Fig D), we found that ID2 binds preferably to single stranded RNA in a more robust manner than DNA (Fig E and F). Interestingly, an ID2 complex with a known DNA binding mutation in FANCI also was defective for RNA binding. Furthermore, ID2 bound to R loops but was mediated via the single stranded DNA component of the structure. Importantly, an in vitro monoubiquitination reconstitution system using FANCL as the E3 ligase demonstrated that monoubiquitination of ID2 was stimulated to an equal or greater degree by RNA versus DNA, with greater signal in presence of GC-rich, single-stranded RNA as well as R loops (Fig G and H and data not shown).
Collectively, our results support a novel mechanism the ID2 complex suppresses the formation of pathogenic R-loops by binding RNA species, thereby activating the FA pathway (Fig I).
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
High Incidence but Very Low Mortality of Epstein Barr Virus Related Post-Transplantation Lymphoproliferative Disorder after T-Cell Replete Haploidentical Peripheral Blood Stem Cell Transplantation
Authors: Salas Gay Queralt , Al-Shaibani Zeyad , Lam Wilson , Michelis Fotios V. , Thyagu Santhosh , Kim Dennis Dong Hwan , Lipton Jeff H. , Kumar Rajat , Viswabandya Auro , Law Arjun 
Published: October 2019

Abstract
BACKGROUND
The use of immunosuppressive agents such as antithymocyte globulin (ATG) as graft-versus host disease (GVHD) prophylaxis may increase the risk of Epstein Barr Virus (EBV) related post-transplant lymphoproliferative disorder (PTLD). The aim of the present study is to report the incidence, risk factors, and outcome of PTLD in the setting of haploidentical stem cell transplant (haploSCT) combining PTCy and ATG as GVHD prophylaxis.
METHODS
From August 2016 to May 2018, 55 adult patients diagnosed with hematological malignancies underwent T-cell replete haploSCT. All patients received a reduced intensity conditioning regimen (RIC) with fludarabine, busulfan, and 200cGy of total body irradiation (TBI), combined with rabbit ATG, PTCy and cyclosporine (Cy).
EBV titre was monitored by quantitative PCR in plasma samples. The cut-off value for test positivity was >600 copies of EBV DNA per milliliter of plasma. Testing was performed weekly from engraftment to day 100 and beyond in case the patients were on immunosuppression.
Data was collected through retrospective chart review. Last follow up was updated in June 31, 2018. Median follow-up was 9.5 months (range 1-21).
RESULTS
Patient and donor characteristics are summarized in Table 1. Median age was 57 (22-73) years, with 23 (43%) patients ≥60 years. Overall survival of the 55 patients at 6 months was 70% (95% CI 57-83), and 51% (95% CI 46-66) at 1 year.
EBV reactivation was documented in 31 (57%) patients. Median time to EBV reactivation was 54 (20-326) days. Three (5.5%) patients developed presumed PTLD with EBV-viremia higher than 1x10E6 copies and corroborative imaging. Three (5.5%) had biopsy-proven PTLD (Table 2). Median time since EBV reactivation to presumed/proven (P/P)-PTLD was 25 (3-36) days. All cases occurred early post-transplant within the first 100 days. Four patients were on therapeutic doses of cyclosporine. All 6 (11%) patients received treatment with weekly Rituximab 375 mg/m+. Five (83%) patients achieved complete clinical responses with PCR negativity with reduction of immunosuppression and single agent rituximab. One patient died of complications of viral encephalitis and bacterial pneumonia.
CONCLUSION
ATG based conditioning can be associated with increased viral reactivations. Frequent EBV monitoring and pre-emptive treatment may lead to rapid disease control. While no guidelines exist currently, further investigation is needed to define optimal monitoring strategies.

Disclosures
Lipton: BMS: Consultancy, Honoraria, Research Funding; Novartis: Consultancy, Honoraria, Research Funding; Pfizer: Consultancy, Honoraria, Research Funding; Takeda: Consultancy, Honoraria, Research Funding.
Symptomatic Hemoglobin New Mexico and a Case of Therapeutic Benefit with Acetazolamide
Authors: Ayad Ahmed , Nofal Sara , Jobson Crystal , Raizen Yuval , Alattar Mona Lisa 
Published: October 2019

Abstract
INTRODUCTION
Inherited disorders due to rare structural Hb variants are not included in national, neonatal screening tests and therefore likely underdiagnosed. Hemoglobin New Mexico is one such rare variant with only one case ever reported in 1985 in conjunction heterozygosity for Hb S.1 This Hb variant involves a missense mutation of the β chain substituting the hydrophobic Proline β100 with the hydrophilic Arginine.1 We present here the second case of Hb New Mexico variant ever reported to our knowledge.
CASE PRESENTATION
An 18-year-old athletic, Latin American female initially presented with episodic chest pain, palpitations, fatigue, headaches, and dyspnea on exertion to the point that she could no longer exercise for one year. Work up revealed erythrocytosis persistent for >1 year with hemoglobin level of 17.3 g/dL, and hematocrit of 47.2 % (Hb went as high as 19g/dL) with normal erythropoietin level, hemoglobin electrophoresis, and JAK2 V617 with reflex to JAK3 exon 12-15 negative. She had extensive evaluation to rule out acquired causes including cardiac, pulmonary, and neurologic evaluation which was all normal. She underwent therapeutic phlebotomies with no clinical improvement over 3 months and symptoms worsened as well as new iron deficiency from phlebotomies so this was discontinued. Evaluation with p50 oxygen dissociation curve revealed high oxygen affinity. Further gene sequencing revealed heterozygous inheritance of Hb New Mexico with HbA: 54.5%, HbA2: 3.6%, and Hb New Mexico: 41.9%. Beta Globin Gene sequencing identified the following mutation: Beta 100, CCT>CGT, Pro>Arg. Her symptoms worsened as well as an episode of syncope but no new neurologic, cardiac, or pulmonary etiologies were identified and at the time Hg 18g/dL. A trial of low dose acetazolamide 125mg daily was given to the patient and her symptoms improved by a 6 week follow up with notable reduction of headache, fatigue, dyspnea, and chest pain.
CONCLUSIONS
Given the rarity of this hemoglobin variant in a patient with severe symptoms, data is limited. Acetazolamide, a carbonic anhydrase inhibitor often used for migraines, is known to illicit mild acidosis and therefore potentially reduce hemoglobin-oxygen affinity and perhaps allow for relief of our patient's symptoms. Further therapeutic evaluation is warranted but likely challenging given rarity of the condition.

Disclosures
No relevant conflicts of interest to declare.
CAN WE Predict Risk Factors of Venous Thromboembolism Among Cancer Inpatients?
Authors: Shrotriya Shiva , Dhakal Prajwal , Sharma Mukta , Gardiner Joseph , Al-Janadi Anas , Lupi Alex , Delshad Ahmad , Rayamajhi Supratik 
Published: October 2019

Abstract

Introduction
Increased risk of venous thromboembolism (VTE) has been noted among cancer patients as compared to non-cancer. VTE identified as leading cause of death among those with cancer. Cancer associated thrombosis caused increased hospitalizations, increased inpatient/outpatient medical and prescription claims, and increased total health care costs per patient. Our objective was to study demographic, clinical and laboratory risk factors for venous thromboembolism (VTE) among hospitalized cancer patients and built a predictive model for VTE risk.
Methods
Ours was a retrospective cohort study focused on patients with VTE and cancer from January 2013 - September 2015. Univariate and multivariate logistic regression analysis using stepwise approach was performed. A final predictive model was derived using receiver-operating characteristics (ROC) curves and concordance indices (c-statistics).
Results
N=3948 cancer inpatients were identified which was split into a derivation cohort and a validation cohort, each with 1957. Mean age 65.9±13.8 years; 52.6% were male; 85.6% Caucasian, 7% African Americans; 15.5% were obese; common comorbidities were hypertension (46%), pulmonary disease (34.5%), diabetes (22.9%), renal disease (20.9%) and congestive heart failure (10.4%). Overall, there was 152 (3.9%) events of VTE with 77 (3.9%) in derivation and 75 (3.8%) in validation cohort. On univariate analysis, comorbidities such as infection and renal diseases, laboratory findings such as low hemoglobin and low albumin was associated with high VTE risk. The derivation set had a c-statistic or AUC of 0.668 while the validation set had an AUC of 0.65.
Conclusions
Infection, renal disease (comorbidity) and low albumin levels were associated with a higher risk of VTE. Digestive and respiratory cancers were associated with higher VTE risk. We identified three clinical and laboratory variable that was associated with increased risk of VTE in addition to the cancer group. Future research could use this analysis as a basis for forming a risk score that could be used by clinicians to identify those cancer patients at risk for VTE.

Disclosures
No relevant conflicts of interest to declare.
Co-Transplantation of Ucbmn with UC-MSC Enhances Engraftment of Ucbmn in Iron Overload NOD/SCID Mice
Authors: Huang Zhi , Xiao Yuhua , Chen Xiaomin , Li Chunfu , Feng Xiaoqin 
Published: October 2019

Abstract
Background: Umbilical cord blood (UCB)-derived hematopoietic stem cells (HSC) are ideal for allogeneic HSCT when a compatible donor is unavailable. However, the insufficient number of HSCs per UCB unit delays engraftment and may lead to graft failure and/or mortality. Besides, transfusion-induced iron overload has been reported to be involved in the delayed engraftment of HSC. Previously mesenchymal stem cells (MSC) have been implicated in playing a significant role in HSC engraftment. This study aimed to determine the effect of co-transplantation of umbilical cord mesenchymal stem cells (UC-MSC) on engraftment and hematopoietic recovery.
Methods: The iron overload NOD/SCID mice were induced by dose-escalation injection of iron dextran. And the sublethally irradiated iron overload NOD/SCID mice were divided into four groups: (1) intravenous injection of normal saline (control group, n=8), (2) intravenous injection of umbilical cord mesenchymal MN) alone (n=10), (3) intravenous injection of UCBMN and UC-MSC (n=10), and (4) intrafemoral injection of UCBMN and UC-MSC (n=12). Six weeks after injection of UC, the bone marrow (BM) was harvested and analyzed. The semi-quantitative analysis of cytokines including SDF-1a, OPN, and VEGF-α were examined by immunohistochemistry.
Results: As a result, iron deposits in bone liver, BM and spleen. In the BM of control group, no human CD45+ cells were found. Via the intra-bone marrow (iBM) injection, the percentage of human CD45+ cells was 9.86% ±1.86% (n=10) in the presence and 3.18% ±0.98% (n=6) in the absence of MSC, respectively, via the tail vein, the percentage of human CD45+ cells was 3.5%±0.94%. In additional, the levels of SDF-1a, OPN, and VEGF-a were all significantly higher in The mice that received MSC showed significantly enhanced engraftment in all three experiments (P=0.000).
Conclusion: Co-transplantation of UC-MSC is effective for improving engraftment in iron overload NOD/SCID mice. The point underlying the positive outcome may relate to cytokines including SDF-1a, OPN, and VEGF-a secreted by UC-MSC, which has been indicated to play a crucial part in homing and engraftment.

Disclosures
No relevant conflicts of interest to declare.
Pegfilgrastim Versus Filgrastim in the Management of Therapy-Related Neutropenia in Relapsed and Refractory Multiple Myeloma in Treatment with Pomalidomide-Dexamethasone
Authors: Cerchione Claudio , Nappi Davide , Pane Fabrizio , Catalano Lucio 
Published: October 2019

Abstract
Aims:
The objective of this study was to compare the efficacy and safety of pegfilgrastim in patients affected by heavily pretreated MM, treated with pomalidomide-dexamethasone, in order to determine whether a single subcutaneous injection of pegfilgrastim is as effective as daily injections of standard filgrastim, in terms of haematological toxicity, febrile neutropenic episodes, antibiotic usage and hospedalization duration.
Methods:
We enrolled 33 patients (19 male and 14 female) median age at diagnosis 69 years (r. 52-84), and median age at start of treatment 76 years (r.56-90) treated with several lines of treatments (median 7, r. 2-11), every refractory to all the drugs previously received, received Pomalidomide-Dexamethasone (Pomalidomide 4 mg for 21 days, Dexamethasone 40 mg days 1,8,15,22, pegfilgrastim day +8) every 28 days, until progression.
Results:
Since first course, received in domestic setting, with a very good compliance, patients performed blood counts once weekly and received, from day +8 to day +19 ("day + 1" when the protocol starts), prophylactic oral chinolonic antibiotics and anti-fungal drugs. During neutropenia after first cycle, Filgrastim (5 μgr/kg/day for 3 days) was given if neutrophils count was <1500 x 10^9 cells/L. Median number of filgrastim administrations was 4.8 (r. 3-6); nadir neutropenia was registered after a median of 10.7 days (r. 7-14); median of nadir neutrophil count was 1.17 x 10^9 cells/L (range 0.3 - 1.5 x 10^9 cells/L), with maximum duration of 14 days. From the second course, all patients switched to prophylaxis with pegfilgrastim (6 mg), injected subcutaneously with a single administration on day +3 independently from the neutrophil count at that time. During pegfilgrastim, neutropenia was never longer than 8 days, with a consequent reduction of neutropenia-related infections. Median nadir neutrophil count, evaluated for every patients for at least three courses of therapy (r. 3-6) registered at day +11, was 1.39 (range 0.9-2.2 x 10^9 cells/L). Only 4 patients needed a supplement of 3 administrations of filgrastim. Pegfilgrastim was well tolerated in all patients: main side effects in our patients were mild fever and bone pain (21.2%).
Conclusions:
In conclusions, in patients affected by heavily pretreated MM treated with pomalidomide-dexamethasone, pegfilgrastim seems to reduce the incidence of severe neutropenia and infections and may increase the possibility to maintain the scheduled time of treatment.

Disclosures
No relevant conflicts of interest to declare.
Secondary Skin Involvement in Non-Hodgkin's Lymphoma Does Not Impact Outcomes: SEER Database Analysis
Authors: Khurana Arushi , Al-Juhaishi Taha , Shafer Danielle 
Published: October 2019

Abstract
Background:
Non-Hodgkin's Lymphoma (NHL) comprises a diverse group of malignancies with varied presentations and clinical behavior. Different prognostic factors have been defined including age, LDH, gender, CNS involvement and molecular/genetic characteristics. We aim to study the incidence and impact of skin involvement as a secondary site in patients with mature B and T-cell NHLs.
Methods:
The Surveillance, Epidemiology and End Results (SEER) database was used to identify patients (≥18 years) diagnosed with mature B/T/NK-cell NHL with skin as secondary site of involvement between 1973 and 2014. B-cell and T-cell lymphomas of all stages were included. Overall survival (OS) was estimated using the Kaplan-Meier method, and compared using Log-Rank test.
Results:
Total of 6429 patients were included with secondary skin involvement. B-cell NHL was more common with 76%. Median age for B-cell and T-cell NHL was 65 years (48-82), and 64 years (46-82), respectively. Slight male preponderance was noted in both subtypes (B-cell-56.7%% and T-cell-61.7%). Diffuse large B cell lymphoma (DLBCL) was the most common histology among B-cell NHLs, (DLBCL, N=1920 39.6%), followed by follicular (FL, N=1381, 28.5%) and extra nodal marginal zone (EN MZL, N=1314, 27.1%) Among T cell NHL histologies, peripheral T-cell NOS was the most common (PTCL, N=819, 51.6%) followed by anaplastic large cell ALK+ (ALCL+, N=341, 21.5%), and Sezary syndrome (N=293, 18.5%) Median overall survival was 6.1 [5.2 - 7.2.; 95% CI] years in T-cell population compared to B-cell population which was 13.3 [12.6-14.0; 95% CI] years. Survival was different by histologic subtype and correlated with SEER database relative survival for the overall population described in the SEER Cancer Statistics Review 1973-2011.
Conclusion:-
Skin involvement is relatively common in both B and T-cell NHLs. DLBCL, FL, MZL, PTCL-NOS and ALCL+ are the more commonly involved subtypes, correlating with their relative incidences. Outcomes in patients with secondary skin involvement tends to mimic known survival patterns in different histological subtypes conferring no additional prognostic significance. Skin as an extra nodal site of involvement should not change management for these patients.
Figure. Figure.

Disclosures
No relevant conflicts of interest to declare.
A role for macrophages in hematopoiesis in the embryonic head
Authors: Li Zhuan , Mariani Samanta , Rodriguez-Seoane Carmen , He Wenyan , Ning Xiaowei , Liu Bing , Vink Chris S. , Dzierzak Elaine A 
Published: October 2019

Key Points
Macrophages in the hindbrain-branchial arches region of the mouse embryo play a role HS/PC expansion and/or maturation. Macrophages from embryonic head hematopoietic niches produce pro-inflammatory factor TNF-&alpha; and enhance HS/PC expansion and/or maturation
Description of a knock-in mouse model of JAK2V617F MPN emerging from a minority of mutated hematopoietic stem cells
Authors: Mansier Olivier , Kilani Badr , Guitart Amélie V. , Guy Alexandre , Gourdou-Latyszenok Virginie , Marty Caroline , Parrens Marie , Plo Isabelle , Vainchenker William , James Chloé 
Published: October 2019

Key Points
PF4iCre;JAK2 V617F/WT mice develop a full MPN that mimics polycythemia vera. The PF4iCre system induces JAK2V617F mutation in a small subset of HSC.