bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL2 and Rapamycin Enhance Persistence of Adoptively-Transferred Ex-Vivo Expanded T Regulatory Cells
Authors: Furlan Scott N , Lopez Christina , Tkachev Victor , Betz Kayla , Hunt Daniel J , Hibbard James , Blazar Bruce R. , Trapnell Cole , Kean Leslie 
Published: October 2019

Abstract

As the field of regulatory T cell (Treg) adoptive therapy develops, questions concerning how to best to deploy these cells in patients must be addressed. Prominent among these questions is which adjunctive therapies will pair most synergistically with the transferred cells. Most ideally, a therapy that could prolong persistence and stabilize function of the cells is sought.
Prior work has shown that mechanistic target of rapamycin (mTOR) inhibition can afford both stability and a degree of increased persistence to ex-vivo expanded thymic Tregs (tTregs) upon adoptive transfer. In this study, we investigated whether the addition of low-dose interleukin-2 (IL2) to the mTOR inhibitor rapamycin could impart additional persistence to ex-vivoexpanded tTregs after adoptive transfer. Using a non-human primate model of CFSE-labeled autologous tTregs, we found that the addition of IL2 to rapamycin supported a near 10-fold increase in the half-life of adoptively transferred tTregs, effectively doubling the cells in the tTreg compartment for the first month after adoptive transfer.
Using a combination of single cell approaches, we were then able to show that transferred tTregs, in the setting of IL2 and rapamycin adjunctive therapy, retain high levels of Treg-specific genes, including FOXP3, after adoptive transfer. Additionally, we found that adoptively transferred tTregs are remarkably homogenous and become more transcriptionally similar to endogenous tTregs with time in vivo. Together these preclinical data support the use of combination IL2 and rapamycin as adjunctive therapy forex-vivo expanded adoptively transferred tTregs.

Disclosures
Tkachev: Regeneron Pharmaceuticals, Inc.: Research Funding. Blazar:Kadmon Corporation, LLC: Consultancy, Research Funding. Kean:Regeneron Pharmaceuticals, Inc.: Research Funding.
SMALL Molecules Mediated Hematopoietic STEM and Progenitor CELLS Expansion for GENE Editing Application
Authors: C Abisha Crystal , Thangavel Saravanabhavan , Velayudhan Shaji Ramachandran , Srivastava Alok , Nath Aneesha , Srinivasan Saranya , Venkatesan Vigneshwaran 
Published: October 2019

Abstract
Genome editing of Hematopoietic stem Cells has revolutionized the treatment strategies for genetic disorders. Despite this, it still remains a great challenge as hematopoietic stem cells tend to lose its stem-ness during the ex vivo culture and gene editing process. The need for large dose of CD34+ HSPCs for manipulation makes it a seemingly difficult strategy. Recent works suggest that the potential effects of small molecules in expanding cord blood HSPCs ex vivo promoting self-renewal and delaying differentiation. We screened several reported small molecules to identify a condition that promotes the expansion of adult HSPCs for gene manipulation process.
The mobilized Peripheral blood HSPCs are purified and cultured with a cytokine cocktail. Along with the cytokine cocktail, we tested several small molecules and in different combinations. Expression of cell surface receptors were analysed by FACS after 12 days of ex vivo culture. Our screening identified a unique culture condition that expanded the primitive stem cell population (CD34+/CD133+/CD90+cells) along with the early progenitors (CD34+/CD133+) and the progenitors (CD34+).
Our culture conditions expanded the primitive cells by 20 folds compared to the mock treated cells. Our treatment release experiments suggested that the expansion is due to our culture conditions and are reversible.The colony forming cell (CFC) assay showed about 30 fold increase in the numbers of multilineage colony forming cell (CFU-GEMM) thereby ensuring the proliferation and differentiation capacity of expanded HSPCs. Their differentiation ability was also confirmed by ex vivo differentiation into Megakaryocytes. Our treatment conditions reduced the apoptosis rate during the ex vivo culture and improved their cell migration response towards SDF. The reduced reactive oxygen species levels and increased CXCR4 expression were observed in our expanded HSPCs and these might be the possible reasons for the low apoptosis and better cell migration respectively.

Disclosures
No relevant conflicts of interest to declare.
Improving Hematologists' Skills in Caring for Patients Transitioning to Adult Care:the Effectiveness of Online Case-Based CME
Authors: Hurst Simi , Warren Charlotte 
Published: October 2019

Abstract
BACKGROUND: Hemophilia is a lifelong condition that requires ongoing evaluation of treatment and management strategies to meet the unique needs of each patient. However, these needs, whether they are physical or social, change as a patient moves from childhood, through adolescence and into adulthood and older adulthood. The goal of this study was to use continuing medical education (CME) to improve hematologists' knowledge, competence, and confidence in transitioning young adults to an independent hemophilia treatment plan.
METHODS: To address identified practice gaps among healthcare providers caring for patients with hemophilia, an online, case-based activity was developed. Featuring an 18-year old boy with hemophilia preparing for college and living away from home for the first time, the 15-minute CME/CE-certified activity utilized a multimedia approach that combined video vignettes that portrayed physician-patient communication scenarios with detailed commentary and perspective from a recognized faculty expert. The expert commentary was complemented by key supporting evidence in text and slides, as needed. The activity launched on June 30,2018 and was made available to healthcare providers without charge.
Educational effectiveness was assessed with a repeated-pairs pre-/post-assessment study design, in which each individual served as his/her own control. Responses to three multiple-choice, knowledge questions and 1 self-efficacy confidence question were analyzed. A chi-squared test assessed changes pre- to post-assessment. P values <0.05 are statistically significant. Effect sizes were evaluated using Cramer's V (<0.05 modest; 0.06-0.15 noticeable effect; 0.16-0.26 considerable effect; >0.26 extensive effect). Data were collected through 07/30/2018.
RESULTS: Data from the subset of hematologists/oncologists (n=33; hem/oncs) who answered all pre-/post-assessment questions during the study period were analyzed. Significant improvements were observed overall (P=.0193; V=.166) and in several specific areas of assessment (Table). Following activity participation, 34% of hem/oncs indicated increased confidence with regard to helping teenagers with hemophilia transition to adult care. Lastly, the findings uncovered educational needs that are the focus of ongoing interventions.
CONCLUSIONS: Case-based learning has been recognized for its benefits in modeling real-life problems, integration of knowledge into practice, and self-evaluation and reflection. Participation in this online education significantly improved hematologists' knowledge with regard to addressing the needs of patients at different points in their journey.

Disclosures
No relevant conflicts of interest to declare.
The microenvironmental niche in classic Hodgkin lymphoma is enriched for CTLA-4- positive T-cells that are PD-1-negative
Authors: Patel Sanjay S. , Weirather Jason L. , Lipschitz Mikel , Lako Ana , Chen Pei-Hsuan , Griffin Gabriel K. , Armand Philippe , Shipp Margaret A. , Rodig Scott J. 
Published: October 2019

Key Points
CTLA-4 is enriched in the microenvironmental niche in cHL, and identifies a T-cell population largely distinct from PD-1 CD86, the natural ligand for CTLA-4, is enriched in the niche and expressed on HRS cells and a subset of TAMs that also express PD-L1
Arachidonic Acid Rescues Defects on PMA-Elicited NADPH Oxidase Activity in Rac2-Deficient Neutrophils.
Authors: Kim Chaekyun , Dinauer Mary C. 
Published: October 2019

Abstract
Rac2 is a hematopoietic-specific Rho-GTPase implicated as an important constituent of the NADPH oxidase complex. We previously showed that Rac2 plays a stimulus-specific role in regulating NADPH oxidase activation and other functional responses in neutrophils [Kim and Dinauer, JI 166, 2001]. Here we investigate the effect of arachidonic acid (AA) on Rac2-regulated NADPH oxidase activity. Superoxide production in rac2-/- neutrophils was significantly lower (~4-fold) than that of wild-type when stimulated with PMA or AA alone. However, exogenously added AA (10 μM) fully restored the defect in PMA-elicited NADPH oxidase activity in rac2−/ − neutrophils, while having no effect on FMLP-elicited superoxide production. Impaired PMA- or AA-induced F-actin polymerization in rac2−/ − neutrophils was also not restored by co-stimulation with PMA and AA. Taken together, these observations suggest that there are agonist- and pathway-specific differences in the underlying basis of functional defects in rac2−/ − neutrophils. To further investigate possible mechanisms of AA-mediated rescue of PMA-stimulated NADPH oxidase activation in rac2−/ − neutrophils, we measured protein expression and activity of cytosolic phospholipase A2 (cPLA2) and protein kinase C (PKC). The expression of cPLA2 and PMA-stimulated release of AA was similar between wild-type and rac2−/ − neutrophils, suggesting that defects in AA production by PMA-stimulated rac2−/ − neutrophils do not account for the effect of exogenous AA on oxidase activity. The neutrophil expression of PKC isoforms (α, β, δ, ζ) was also similar between genotypes. The cytosolic p47phox and p67phox components of NADPH oxidase were translocated to the plasma membrane upon stimulation with PMA in both genotypes, and no additional translocation in either wild-type or rac2−/ − neutrophils was detected upon co-stimulation with AA. The level of activated Rac1-GTP was similar between genotypes following stimulation, and was not increased by co-stimulation with PMA and AA. These studies indicate that the addition of exogenous AA reconstitutes PMA-elicited superoxide production in rac2−/ − neutrophils independent of the effects on translocation of p47phox and p67phox and activation of Rac1 GTPase. We hypothesize that the effect of AA is exerted through conformational changes of the assembled NADPH oxidase.
Transplanted Human Bone Marrow Contributes to Vascular Endothelium.
Authors: Jiang Shuguang , Walker Luke , Afentoulis Michael , Anderson Daniel A. , Jauron-Mills Linda , Corless Christopher L. , Fleming William H. 
Published: October 2019

Abstract
Accumulating evidence indicates that bone marrow is a source of endothelial progenitor cells. Recently our laboratory has shown that functional endothelial cells (EC) can be clonally derived from phenotypically defined hematopoietic stem cells. To determine the endothelial cell potential of human hematopoietic cells, blood vessels in skin and gut biopsies from sex-mismatched allogeneic transplant recipients were examined. H&E sections were initially reviewed to ensure tissue integrity and the absence of significant graft versus host disease. Coded biopsy specimens from 12 sex-mismatched transplant recipients and 6 sex-matched female transplant recipients were evaluated in a blinded fashion. Endothelial cells were identified using a combination of immunohistochemistry (IHC) and XY interphase FISH. Tissues sections were first evaluated by IHC for EC markers (CD31 or vWF) and for the pan-hematopoietic marker CD45 then assayed by dual-color interphase FISH for donor derived cells (XY). Tissue samples from each recipient were assayed at least 3 times by sequential IHC/FISH. The frequency of CD31+, CD45−, Y+ cells and vWF+, CD45−, Y+ cells in biopsies from the 18 transplant recipients was evaluated by 2 independent observers. 9 of 12 sex-mismatched recipients showed evidence of donor-derived Y chromosome positive EC. The remaining 3 recipients did not exhibit hematopoietic engraftment at the time of biopsy. Donor derived endothelial cells were detected in the skin and the gut of transplant recipients with a mean frequency of 2.3% (range 0.9–3.9). Donor derived hematopoietic cells including intraepithelial lymphocytes and tissue macrophages were excluded by CD45 expression. None of the >4,000 endothelial cells examined had >2 sex chromosomes, consistent with an absence or very low incidence of cell fusion. In addition, Y chromosome signals were not detected in sex matched female recipients, thereby excluding vertical transmission of male cells. Furthermore, none of the recipients evaluated prior to hematopoietic engraftment had detectable donor derived endothelial cells, indicating a close linkage between the recovery of hematopoiesis and endothelial cell outcomes. Our results suggest that the close association between hematopoiesis and the vascular system originally identified during early development persists into adult life. Consequently it will be important to study the potential therapeutic role of bone marrow derived endothelial cells in the setting of hematopoietic and vascular disease.
Primary Central Nervous System Lymphoma in Children: A Report of 12 Cases.
Authors: Abla Oussama , Sandlund John T. , Blaser Susan , Brock Penelope , Corbett Rob , Kirov Ivan , Griffin Tim , Sung Lillian , Weitzman Sheila 
Published: October 2019

Abstract
We report the treatment and outcome of 12 children with primary central nervous system lymphoma (PCNSL) treated at 6 pediatric oncology centers from 1995 to 2003. The main purpose is to determine whether childhood PCNSL can be cured with chemotherapy alone without cranial radiation therapy (CRT). The clinical charts of 8 immunocompetent and 4 immunodeficient children with PCNSL were retrospectively reviewed. The children were diagnosed from 4 to 17 years of age. All cases were non-Hodgkin’s lymphomas: 7 (58.3%) were phenotypically B-cell (6 mature B, one B-precursor), 3 (25%) were T-cell and 2 (16.6%) were indeterminate-cell type. The histologic subtyping (REAL classification) showed 4 diffuse large cell, 4 anaplastic large cell, 2 immunoblastic large cell, 1 Burkitts and 1 high-grade lymphoma of uncertain lineage. Two patients were t(2;5) positive. Two patients had congenital immunodeficiency and 2 were HIV+. Immunologic work-up was either normal or not done in the remaining 8. EBV was positive in 2 patients, negative in 3 and not assessed in the rest. PCNSL presented as a single lesion in 4 patients and multifocal in 8. The most frequent tumor locations were frontal and parietal lobes. Nine of 12 children received chemotherapy alone, event free survival (EFS) at 4 years was 74±16%. Three had chemo plus CRT (3900–5000 cGy), EFS was 33±27% (P=0.1). The most frequently used drugs at the 6 centers were high dose (HD) MTX (5 to 8 g/m2), HD Ara-C (2 to 3 g/m2), dexamethasone, vincristine and cyclophosphamide. Three children died, 2 of whom were HIV+; 1 died after local relapse while the other died secondary to an opportunistic infection. Two other patients relapsed, one after chemotherapy alone and one after chemotherapy plus CRT. The patient who relapsed after chemotherapy alone is in second CR at 6+ months after chemotherapy, CRT and ABMT; the other patient died from progressive disease. Nine of 12 patients (75%) are alive at a median follow-up time of 72 months (range 18–105 months) in survivors. Six of the 8 children who are in CCR received chemotherapy alone, one received chemotherapy plus ABMT and one received chemotherapy plus CRT. Two of the long-term survivors (in CCR at 88+ and 105+ mo) had congenital immunodeficiency and were treated with chemotherapy alone. It appears that immunocompetent and immunodeficient children with PCNSL may be cured with chemotherapy alone without CRT, avoiding long-term radiation toxicity. Although retrospective, involving a small cohort, it is the largest pediatric series of PCNSL reported to date. Multicentre prospective studies are clearly needed in children with this rare lymphoma, whose frequency seems to be increasing especially in immunocompetent patients.
A Phase II Study of the TNF-α Inhibitor Etanercept and Thrice Weekly Rituximab: Evidence of Clinical Activity in the Absence of del(17p13.1) Genomic Abnormalities.
Authors: Lin Thomas S. , Heerema Nyla A. , Swenor Jennifer , Lucas Margaret S. , Moran Mollie E. , Cheney Carolyn M. , Lucas David M. , Young Donn , Caligiuri Michael A. , Grever Michael R. , Byrd John C. 
Published: October 2019

Abstract
The monoclonal anti-CD20 antibody rituximab (Rituxan) induces the death of chronic lymphocytic leukemia (CLL) cells in part through apoptosis. Tumor necrosis factor (TNF)-α and other cytokines up-regulate bcl-2 and other anti-apoptotic proteins, thereby inhibiting apoptosis. In addition, TNF-α may play a role in the cytokine release syndrome and infusion toxicity associated with monoclonal antibody therapy. Therefore, antagonists of TNF-α represent potential therapeutic agents that may be able to decrease infusion toxicity and enhance rituximab-induced apoptosis. We initiated a phase II trial of etanercept in combination with rituximab in patients (pts) with relapsed CLL or small lymphocytic lymphoma (SLL), with the goals of increasing clinical activity of rituximab and reducing infusion toxicity through neutralization of TNF-α. Pts received etanercept 25 mg subcutaneously (SC) twice weekly during weeks 1–5 and rituximab 375 mg/m2 intravenously (IV) three times weekly during weeks 2–5. Stepped up dosing of rituximab was used with the first two rituximab treatments. From 12/02 to 10/03, 19 pts (14 male) with a median of 2 prior therapies (range 1–8) were enrolled. Therapy was generally well tolerated, although 5 pts developed grade 4 neutropenia and 5 pts developed grade 3–4 infection or neutropenic fever. Four of 19 pts achieved an NCI 96 partial response, with no responses in 8 pts with del(17p13.1), corresponding to loss of the p53 gene. In contrast, 4 of 11 pts without del(17p13.1) achieved a PR, including 1 pt with a complex karyotype. This study was stopped for interim analysis and has reopened, with the exclusion of patients with a del(17p13.1) based upon the response outcome above and a previous preliminary report by our group demonstrating that rituximab was ineffective for del(17p13.1) CLL (Cancer Res63:36, 2003). We conclude that the combination of rituximab and etanercept has clinical activity in heavily treated, relapsed CLL pts without del(17p13.1), irrespective of prior rituximab treatment. Accrual to this study continues, and the results will be updated at the 2004 ASH meeting.
Evidence on Chromosome 11 for a Quantitative Trait Locus Influencing Levels of Activated Protein C-Protein C Inhibitor Complex.
Authors: Vossen Carla , Hasstedt Sandra , Callas Peter , Scott Bruce , Long George , Rosendaal Frits , Bovill Edwin 
Published: October 2019

Abstract
Earlier we showed high heritability for the variation in levels of activated protein C-protein C inhibitor complex in a large kindred with type I protein C deficiency (Vossen et al. 2004). To identify genomic regions, which might harbour a gene with a mutation that accounts for a difference in the levels of activated protein C-protein C inhibitor complex, we performed a variance component linkage analysis. Family members were genotyped for 375 autosomal markers at the Marshfield Medical Research Foundation with an average marker spacing of 9.4 cM (range 0–18 cM) and an average marker heterozygosity of 75% (range 42–89%). Levels of activated protein C-protein C inhibitor complex were measured using commercial paired antibody sets from Affinity Biologicals Inc. (Ancaster, Ontario, Canada). We estimated the probability of Identity By Descent (IBD) using the multipoint IBD method in Simwalk2 (Sobel & Lange 1996), in which the proportion of alleles shared identical by descent at marker loci is used to estimate IBD sharing at arbitrary points along the chromosome for each relative pair. Then, variance component linkage analysis was performed using SOLAR (Almasy & Blangero 1998) to test whether a proportion of the genetic variance in the levels of activated protein C-protein C inhibitor complex could be attributed to specific genomic locations. The levels of activated protein C-protein C inhibitor were log-transformed to reduce skewness (from 3.0 to 0.2) and kurtosis (from 11.6 to 0.4) and were assumed to distribute as a multivariate normal density with correlation =h2K + c2H + q2B + e2I, where matrix K contains the kinship coefficients, H contains 1 for pairs from the same household and 0 otherwise, B contains the IBD probabilities, and I represents the identity matrix. The parameters for heritability (h2), household effect (c2), and heritability contributed to a specific genomic location (q2) as well as the effects of the covariates were estimated simultaneously using maximum likelihood analysis. Lod scores were computed as the log10 likelihood for q2 estimated to q2=0. A lod score of 3.3 was used as cut-off point for statistical evidence for significant linkage (as suggested by Lander and Kruglyak 1995), and a lod score above 1.9 but below 3.3 for suggestive linkage evidence. We found suggestive linkage evidence for levels of activated protein C in complex with protein C inhibitor in 121 tested family members (without a history of venous thrombosis) for a region on chromosome 11 (marker D11S969, 146 cM) with a LOD-score of 2.6, with age added to the model. As no obvious candidate genes were available under the peak, and because the linkage region is too wide to be certain of linkage, we are currently finemapping the peak by adding microsatellite markers to the genome scan to narrow the region on chromosome 11.