bluuebunny/crossref_metadata_2025_split huggingface dataset:
CNS Prophylaxis in Diffuse Large B-Cell Lymphoma & Its Outcomes- an Institutional Review
Authors: Vaddepally Raju , Adhami Angie , Siedlecki Charles , Kuriakose Philip 
Published: November 2019

Background: Diffuse large B-cell lymphoma (DLBCL) has an approximately 5% risk of recurrence in the central nervous system, and carries a very poor prognosis. Prophylactic intrathecal chemotherapy (ITC) with agents such as Methotrexate (MTX) can mitigate the risk of recurrence and improve survival. We report a single-institution, retrospective analysis of the effect of ITC administration on outcomes in patients with DLBCL, based on CNS-IPI risk stratification.
Methods: We screened 325 DLBCL patients from our tumor registry- of these only 292 had clinical information to risk-stratify per IPI scoring- low-risk, n = 54, intermediate-risk, n = 198, and high-risk n = 40. These patients then further categorized based on the NCCN prognostic model to assess the risk of CNS disease (CNS-IPI) which includes age >60, LDH >normal, ECOG >1, Stage III or IV, Extranodal involvement >1 site, kidney or adrenal gland involvement with scores 0-1 low-risk, 2-3 intermediate-risk, and 4-6 or kidney or adrenal involvement as high-risk. The effect of ITC administration on outcomes such as overall survival (OS) at 1-year (yr) and 5-yr were calculated using univariate two-group comparisons.
Results: Demographic and clinical variables include- Age >60- 61% (n=201); mostly Caucasians- 71%(n=199); Germinal Center(GC) 25%(n=73), non-GC 74%(n=212); mean IPI score-2.8(n=196), R-CHOP-87%(n=233); and, ITC in 15%(n=45). Seven patients in low-risk patients received ITC for unclear reasons. Intermediate risk patients treated with ITC showed a trend towards 90% (n=26) 1yr OS, and 67%(n=8) 5-yr survival compared to who did not receive ITC (non-ITC) who had 84%(n=127) 1-yr, and 55%(n=52) 5-yr OS (p-value 0.57 and 0.43 respectively). High-risk patients treated with ITC showed a trend towards 100%(n=3) 1-yr, and 0% 5-yr OS compared to non-ITC who had 85%(n=23) 1-yr , and 29%(n=5) 5-yr OS (p-value 1.0 and 1.0 respectively).
Conclusions: Our results suggest that in DLBCL patients with the intermediate-risk group may benefit from intrathecal chemotherapy. Our study is limited owing to small sample size in each subset of CNS risk stratification- this limits the interpretation of the results, and should be interpreted cautiously given nonsignificant P-values. However, we believe these results can encourage further research to evaluate the effect of CNS prophylaxis in the intermediate-risk group of DLBCL patients.

Disclosures
Kuriakose: Alexion: Consultancy, Honoraria, Speakers Bureau; Bayer: Consultancy.
Astragalus Polysaccharide Promotes Hematopoiesis in an Irradiated Mouse Model and Reduces Apoptosis of Hematopoietic Cells
Authors: Xu Wan-huang , Li Liang , Yang Liuming , Xin Hongwu , Chong Beng H , Yang Mo 
Published: November 2019

Background: Huangqi (the dried root of Radix Astragali) is a notable Chinese herb that has been used to promote hematopoiesis for centuries. Astragalus polysaccharide (ASPS), a bioactive constituent extracted from Radix Astragali, may play an essential role for hematopoiesis. The objective of this study is to investigate the hematopoietic effects of ASPS in mouse models and explore underlying mechanism, which provides scientific basis for future applications.
Methods: An irradiation-induced myelosuppressive mouse model and colony-forming unit (CFU) assays were used to determine the in vivo hematopoietic effects of ASPS. The anti-apoptotic effects of ASPS were also examined by using annexin V, caspase-3, and JC-1 assays with flow cytometry in HL-60 cells.
Results: In the myelosuppressed mouse model, ASPS enhanced the recovery of platelet and red blood cell at day 21, and white blood cell at day 14. ASPS also promoted the CFU formation of all lineages (MK, GM, E, GEMM and F). Morphological study of bone marrow revealed that as compared to the control group, tri-lineage hematopoiesis was preserved in the ASPS- and TPO-treated group. Overall cellularity (MTC/area) of ASPS-treated group was similar to that of TPO. Furthermore, in-vitro study showed that 100 μg/ml of ASPS had maximum effect on CFU formation. ASPS also reduced apoptosis via inhibiting mitochondrial/Caspase-3 signal pathway.
Conclusions: ASPS may promote hematopoiesis in irradiated myelosuppressive mice by enhancing hematopoietic proliferation and inhibiting apoptosis of hematopoietic cells.

Disclosures
No relevant conflicts of interest to declare.
Fibg390-396A Limits Metastatic Potential Via a Mechanism Independent of Fibrin-Myeloid Cell Interactions, but Prospectively through the Disruption of FXIII-Mediated Fibrin Crosslinking
Authors: Francisco Brenton J , Sharma Balkrishan , Rosenfeldt Leah A. , Mureb Duaa , Palumbo Joseph S. 
Published: November 2019

Previous studies have shown that fibrin(ogen) supports metastasis. In order to better define the mechanisms coupling fibrin(ogen) to metastasis, we compared metastatic potential in immunocompetent mice carrying specific structure/function alterations in fibrinogen and controls. FibrinogenAEK mice, which have a germ-line mutation in the thrombin cleavage site that essentially "locks" fibrinogen in its soluble state, exhibited diminished metastatic potential relative to FibWT mice. Notably, previous studies have established that factor XIII is also a significant determinant of metastasis. Taken together, these studies suggest that stable fibrin polymer formation is important in the metastatic process. However, FibAEK retained significant metastatic potential relative to mice with complete fibrinogen deficiency, indicating that fibrinogen possesses prometastatic properties in the absence of polymer formation.
Fibrin(ogen) has also been shown to mediate inflammatory cell functions through direct engagement of leukocyte integrins independently of its role in platelet aggregation. Given previous studies demonstrating that myeloid cells are promote metastatic potential, we hypothesized that fibrin(ogen)-mediated leukocyte engagement represents one mechanism by which fibrinogen drives metastasis. Consistent with this view, mice expressing a mutant fibrinogen lacking the y chain binding motif for the leukocyte integrin αMβ2 (Fibγ390-396A) had diminished metastatic potential in both spontaneous and experimental metastasis assays. Furthermore, fate analyses revealed that Fibγ390-396A results in diminished survival of newly-formed metastatic foci. The early reduction in metastatic potential observed in Fibγ390-396A mice suggests that fibrin(ogen) promotes metastasis by recruiting myeloid cells to the early metastatic niche. In order to explore this hypothesis, we performed experimental metastasis assays in immunocompetent mice in which macrophages or neutrophils were specifically depleted using either clodronate or an anti-Ly6G antibody. In contrast to mice carrying the Fibγ390-396A mutation, depletion of macrophages or neutrophils had no significant impact on the early survival of metastatic foci. These studies suggest that expression Fibγ390-396A limits metastatic potential via a mechanism independent of interactions with myeloid cells.
In addition to disrupting fibrin-αMβ2 interactions, elimination of the γ chain 390-396a binding motif has also been shown to limit factor XIII binding to fibrinogen. Given the established importance of FXIII in metastasis, it is conceivable that even subtle alterations in the kinetics of fibrin cross-linking resulting from expression of Fibγ390-396A limits metastatic potential.

Disclosures
Palumbo: Ionis Pharmaceuticals: Research Funding.
Autologous Stem Cell Transplantation Outcomes in Patients with Myeloma and Renal Dysfunction
Authors: Monge Jorge , Jayabalan David S , Niesvizky Ruben , Coleman Morton , Flicker Kari , Solomon Robin S. , Rossi Adriana C. , Guo Jin 
Published: November 2019

Background
Renal impairment (RI) is a common complication and a negative prognostic factor in patients with Multiple Myeloma (MM). Patients with severe renal dysfunction may not benefit from optimal therapy as they are often excluded from autologous stem cell transplantation (ASCT). The aim of this study was to analyze the outcome of MM patients with prior RI undergoing ASCT and to evaluate the effect of transplant on renal and hematologic response.
Methods
From 147 eligible MM patients who underwent ASCT between 2002 and 2019, 131 were included in this single-center retrospective cohort study. Renal function (eGFR in ml/min/1.73m2 by MDRD) was evaluated at the time of MM diagnosis and prior to ASCT, as well as 30 and 100 days after ASCT. Patients were classified by renal function: A) eGFR <30; B) eGFR 30-59; or C) eGFR >60 prior to ASCT. ANOVA was used for comparison of renal function at diagnosis, at transplant, 30 days post ASCT, and 100 days post ASCT. Log rank testing was used for overall survival (OS) analysis of group A, B and C
Results:
Patients with renal dysfunction at baseline had lower ORR than controls to induction therapy, 71 v 78%. ASCT provided deepening responses to both groups, ORR 83 v 86% respectively. No renal responses were seen with ASCT, whereas 16% of patients with renal dysfunction at diagnosis had renal response prior to transplant. Of note, only 51% of patients with renal dysfunction received conditioning with Mel200, compared with 94% of control group. Overall survival benefit is seen for patients with eGFR>60 over any level of dysfunction (p = 0.0079).
Conclusion
Myeloma patients with renal dysfunction are a high risk group, and require special attention. Rapid and effective induction therapy remains an important step in reversing renal damage. In patients who do not recover kidney function, autologous stem cell transplantation should be considered, providing depth of response benefit. The conditioning regimen used for these patients warrants further investigation.
Figure

Disclosures
Niesvizky: Takeda, Amgen, BMS, Janssen, Celgene: Consultancy, Research Funding. Coleman:Gilead, Bayer, Celgene: Consultancy, Research Funding, Speakers Bureau; Merck: Research Funding; Pharmacyclics: Speakers Bureau; Kite Pharmaceuticals: Equity Ownership. Rossi:BMS: Research Funding; Janssen, Celgene, Amgen: Consultancy.
Absence of Damaging Effects of Stem Cell Donation in Unrelated Donors Assessed By FISH and Gene Variance Screening
Authors: Nacheva Elisabeth P. , Ahyee Trudy , Navarette C , Addada Juanah , Pawson Rachel , Syzdlo Richard , Mackinnon Stephen , Shaw Bronwen E. , Madrigal J. Alejandro 
Published: November 2019

Granulocyte-Colony-Stimulating factor (G-CSF) is currently the standard mobilizing agent for peripheral blood stem cell (PBSC) donation. While G-CSF use is associated with mild transient side-effects, concerns that it may trigger hematological malignancy were raised by a study of PBSC donors found to display epigenetic and genetic changes similar to those observed in leukemia patients. Although several studies have failed to reproduce the original data, possible long-term effects of G-CSF mobilization on chromosome integrity remain unclear. In the setting of a multi-center clinical trial we screened blood samples from PBSC donors for aberrations at cellular and gene level that are common in hematological malignancies using fluorescence in situ hybridization (FISH) and next generation sequencing (NGS) assays. Samples collected before, on the day of donation, 90 and 180 days after G-CSF admission confirmed the absence of short-term effects in 50 PBSC donors on both quiescent and dividing cells. The data obtained from interphase cells and chromosome preparations from 50 individuals tested 3 to 5 years after PBSC donation did not differ either from individuals post bone marrow (BM) donation over a similar time period or healthy persons. In an attempt to finally resolve the residual uncertainty surrounding the effects of the G-CSF administration we used next generation sequencing to search for cryptic genome aberrations that may remain undetected by FISH screening. The gene profile of samples from 47 PBSC donors assessed using a panel targeting 54 genes (TruSight Myeloid panel, Illumina) was found to be stable throughout the study period. These data conclusively demonstrate the absence of any detrimental effects on the genome integrity of lymphocytes caused by PBSC or BM donation.

Disclosures
Shaw: Therakos: Other: Speaker Engagement.
Enabling Gene-Edited, Regulatory-like, T Cells (edTreg) for Treatment of IPEX and Other Autoimmune Disorders
Authors: Honaker Yuchi , Sommer Karen , Dahl Noelle , Xiang Yufei , Lopez Christina , Scharffenberger Samuel , Christian Michelle , Torgerson Troy R. , Scharenberg Andrew M. , Rawlings David J. 
Published: November 2019

IPEX (immunedysregulation, polyendocrinopathy, enteropathy, X-linked) syndrome is a severe congenital autoimmune disorder in males resulting from hemizygous inheritance of a mutant FOXP3 allele. FOXP3 encodes a transcription factor that governs the development, maintenance, and function of regulatory T cells (Treg). We have developed a cell therapy strategy for treatment of IPEX using a gene-editing approach in which CRISPR/Cas9 RNPs are co-delivered with an AAV6 donor template designed to integrate into the FOXP3 locus an expression cassette containing the MND promoter driving expression of a functional FOXP3 cDNA and a surface LNGFR tag linked by a 2A ribosomal skip peptide. This approach enforces heterologous FOXP3 expression in IPEX CD4 effector T cells (Teff), while simultaneously eliminating expression of the endogenous FOXP3 allele. The resultant high level and stable expression of functional FOXP3 converts Teff to Treg-like cells with immunosuppressive activity. Using an optimized protocol, we obtained efficient HDR rates across multiple healthy donors. Edited cells were consistently enriched to >95% purity by a magnetic LNGFR antibody selection and expanded 50-fold in a week. Expression of FOXP3 cDNA in edited cells was sufficient to enforce Treg-like phenotypes including the up-regulation of Treg-associated markers (CD25, CTLA-4, and ICOS), and down-regulation of CD127 and inflammatory cytokines (IL2, IFNgamma, TNFalpha). Importantly, we demonstrate sustained in vivo suppressive activity of these edited Treg-like cells (edTreg) in a xeno-GvHD mouse model. edTreg (as well as expanded natural Treg) limited effector T cell expansion and tissue infiltration and significantly protected mice from xeno-GvHD induced by co-transferred autologous effector T cells. Along with preliminary data showing successful editing in CD4 T cells from IPEX patients, our data provide key pre-clinical proof-of-concept and safety data supporting use of edTreg in a clinical trial for IPEX and, potentially, for use in other autoimmune diseases.

Disclosures
Torgerson: Shire: Consultancy; CSL Behring: Consultancy; ADMA Biosciences: Consultancy; UCB: Consultancy. Scharenberg:Casebia Therapeutics LLc: Employment, Equity Ownership; Alpine Biosciences: Consultancy, Equity Ownership; Generation Bio: Equity Ownership.
Feasibility of Peripheral Blood Stem Cell Collection from Heterozygoussickle Cell Trait Donors
Authors: Foell Juergen , Morez Morad , Troeger Anja , Kleinschmidt Katharina , Pfirstinger Beatrix , Brosig Andreas , Burkhardt Ralph , Corbacioglu Selim , Ahrens Norbert 
Published: November 2019

Introduction
Allogeneic haematopoietic stem cell transplantation (HSCT) is currently the only curative treatment for sickle cell disease (SCD) offered to patients with a fully matched sibling (MSD) or matched unrelated donor (MUD). With a MSD/MUD donor availability of below 20% a haploidentical HSCT from a family donor either with post-transplant cyclophosphamide (post-cy) or with a TCRαß/CD19 depleted graft (T-haplo) is an increasingly successful alternative for the majority of patients. In contrast to post-cy where mostly bone marrow is used, T-haploHSCT requires the generation of a G-CSF stimulated peripheral stem cell grafts. Almost uniformly haploidentical relatives of SCD patients are heterozygous carriers of SC trait. For these donors with a genetically heterozygous sickle trait, G-SCF represents a major safety concern with regard to triggering a sickle cell crisis. Therefore, the use of sickle trait donors is prohibited or not recommended in several countries, and guidelines are currently not available due to lack of evidence.
Methods
Haploidentical related donors with heterozygousSCD (group S, n=13) as well as healthy donors (group O, n=9) were stimulated with G-CSF. In all donors a similar mobilizations schedule using G-CSF was applied. All donors were applied 10µg/kg body weight (Neupogen®or Granocyte®) sub cutaneous daily with first apheresis on day 5 and day 6 when necessary. The amount of mobilized CD34+ cells were assessed after mobilization. In addition, the harvested stem cell preparations were analysed for CD34 content after apheresis and patients monitored for potential serious adverse effects (AE) during and after the apheresis.
Results
Mobilization was tolerated well with only mild and typical AE related to apheresis procedure such as citrate reactions, joint pain, ostealgia and headache, observed equally in both groups. In most of the donors (n=13/13 and 8/9), we were able to collect a sufficient amount of CD34+peripheral stem cells (mean of 16.3 and 8.2 x 10^6/kg in group S and O, respectively). We observed slight increases in LDH and reticulocyte counts in few donors also without significant differences between both groups. In particular, there were no severe neurologic side effects and no problems like sickle cell crisis despite mean pre-apheresis hemoglobin concentrations of 14.1 (range 11.9-17.5) g/dL in group S.
Conclusions
Collectively, these data indicate that in the absence of a MSD or MUD donor, haploidentical family donors with heterozygoussickle cell trait provide a safe and effective alternative for collection of peripheral CD34+stem cells for a T-haplo HSCT.

Disclosures
No relevant conflicts of interest to declare.
Megakaryocyte Ablation Ameliorates Bone Marrow Fibrosis By Reducing Myofibroblast Differentiation in a Mouse Model of Primary Myelofibrosis
Authors: Chung Yutein , Jin Huiyan , Trasorras Claire , Gonzalez Francina , Min Sang Hee 
Published: November 2019

Primary Myelofibrosis (PMF) is a chronic myeloproliferative disorder characterized by excessive bone marrow (BM) fibrosis that can lead to ineffective hematopoiesis and reduced survival. PMF patients develop several megakaryocyte (MK) abnormalities including increased proliferation, abnormal morphology and upregulated expression of pro-fibrotic genes. Although several studies support the role of MK in BM fibrosis, the significance of MK in the development of BM fibrosis has not been demonstrated in vivo.
Here, we investigated the in vivo role of MK in the pathogenesis of BM fibrosis. First, we reproduced an established bone marrow transplantation (BMT) mouse model of PMF. To achieve this, lethally irradiated wild-type mice were transplanted with wild-type hematopoietic stem cells (HSC) transduced with either MPLW515L gene construct (MPLW515L BMT) or MIGR1 empty vector (MIGR1 BMT). Consistent with previous studies, starting at day 14 post-BMT, MPLW515L BMT mice gradually developed increased number of leukocytes and platelets, increased proliferation of abnormal megakaryocytes in the BM and spleen, hepatosplenomegaly with extra-medullary hematopoiesis, and reticulin fibrosis in the BM. Notably, robust fibrosis was also seen in the spleen and liver.
Next, to study the significance of MK in the development of BM fibrosis, we developed a MPLW515L BMT mouse model in which MK lineage could be selectively ablated. We accomplished this using PF4-Cre inducible diphtheria toxin receptor transgenic mice (iDTR+/-/PF4-Cre) as BMT donors. HSCs from iDTR+/-/PF4-Cre mice were transduced with MPLW515L or MIGR1 empty vector and transplanted as described above. Beginning day 14 post-BMT, recipient mice were injected with diphtheria toxin (DT) or water every 48 hours. At day 20 and day 29, DT injection significantly depleted MK in both MPLW515L BMT and MIGR1 BMT groups compared to water injection. Importantly, DT-injected MPLW515L BMT mice displayed attenuated fibrosis in the BM, spleen, and liver compared to water-injected MPLW515L BMT mice. In addition, DT treatment decreased the level of alpha smooth muscle actin in both BM and spleen of MPLW515L BMT mice, which suggests that MKs are critical for myofibroblast differentiation. MK ablation did not leukocytosis, thrombocytosis or hepatosplenomegaly. Together, our study show that successful MK ablation in vivo reduces fibrosis development in the BM, spleen, and liver of the MPLW515L mouse model of PMF.
In summary, these results support the essential role of MK in the pathogenesis of BM fibrosis in PMF. Further studies are underway to elucidate the mechanisms by which MK contribute to fibrosis in PMF.

Disclosures
No relevant conflicts of interest to declare.
Bolder guppies do not have more mating partners, yet sire more offspring
Authors: Herdegen-Radwan Magdalena 
Published: November 2019

Abstract
Background
Intra-individual stable but inter-individually variable behaviours, i.e. personalities, are commonly reported across diverse animal groups, yet the reasons for their maintenance remain controversial. Therefore, studying fitness consequences of personality traits is necessary to discriminate between alternative explanations.

Results
Here, I measured boldness, a highly repeatable personality trait, and reproductive success in male guppies, Poecilia reticulata. I found that bolder males had higher reproductive success than their shyer conspecifics and they sired offspring with females who had larger clutches.

Conclusions
This result provides direct evidence for fitness consequences of boldness in the guppy. It suggests that the effect may be driven by bolder males mating with more fecund females.