bluuebunny/crossref_metadata_2025_split huggingface dataset:
Allogeneic Transplant Outcomes in Imatinib-Refractory Chronic Myeloid Leukaemia (CML) Are Similar to Transplant Outcomes in Imatinib-Responsive/Imatinib-Naïve CML and Appear To Be Predicted by the EBMT Risk Score.
Authors: Stylian Steven , Fennelly Eileen T. , Butler Jason P. , Morton James P. , Western Robyn , Hutchins Cheryl J. , Hill Geoff R. , Durrant Simon T. , Kennedy Glen A. 
Published: October 2019

Abstract
Aims: To review the outcome of allogeneic stem cell transplantation (SCT) in imatinib refractory chronic myeloid leukaemia (CML).
Methods: Outcomes of all allogeneic transplants performed after January 2001 for CML at our institution were retrospectively reviewed. Imatinib-refractory CML was defined as either lack of any cytogenetic response (CGR) after at least 6mths of imatinib, loss of CGR or progression to a more advanced disease stage (accelerated or blast phase) during imatinib therapy. Using the EBMT risk score (Lancet1998; 352: 1087), transplant outcomes for imatinib refractory CML were compared with all other CML transplants performed during the same time period. Survival analysis was performed using the Kaplan-Meier product-limit and comparison of survival data via the log-rank test.
Results: Of 31 allogeneic transplants (19M; 12F) performed for CML, 12 had been performed for imatinib refractory CML (no CGR to imatinib n=3; loss of CGR n=3; progression to AP n=3; progression to BC n=3), 5 in patients with imatinib responsive CML, and 14 in patients never exposed to imatinib. Median age at SCT was 40yrs (range 19–63yrs). Donor source included HLA-matched unrelated donors in 14 cases, HLA-identical siblings in 16 and other matched family donors in 1. Conditioning regimens included Cy/TBI (20 cases), Bu/Cy (8 cases), Flu/Mel (2 cases) and Flu/Cy (1 case); CsA + MTX was used as standard GVHD prophylaxis. EBMT risk scores were 1 (4 cases), 2 (6 cases), 3 (8 cases), 4 (5 cases), 5 (3 cases) and 6 (5 cases). At median follow-up post-SCT of 37mths (range 6–64mths), median PFS and OS are not reached; at 2yrs PFS, EFS and OS are 81%, 58% and 61% respectively. For patients with EBMT risk scores of 1–2 versus 3–4 versus 5–6, OS at 2 yrs post-SCT is 80%, 62% and 38% respectively (p=0.03). Based on EBMT risk score, no significant differences in PFS, EFS or OS were observed when comparing SCT for imatinib-refractory versus imatinib-responsive / imatinib-naïve CML.
Conclusion: Our experience suggests that survival post-SCT for imatinib-refractory CML is similar to SCT for imatinib-responsive / imatinib-naïve CML. The EBMT risk score appears to remain useful in predicting survival post-SCT in imatinib-refractory CML.
N-rasm-Transduced Fibroblasts Facilitate the Expansion and Engraftment of Umbilical Cord Blood Stem Cells.
Authors: Dolnikov Alla , Holmes Tiffany , Shen Sylvie , Knight Robert , O’Brien Tracey A. 
Published: October 2019

Abstract
Clinical trials using ex-vivo expanded umbilical cord blood (UCB) cells have thus far failed to demonstrate significant improvement in speed of hematopoietic recovery suggesting better understanding of self-renewal, proliferation, homing and engraftment capabilities of cord blood is needed. Recent progress has been made in the identification and understanding of the cellular niche for hematopoietic stem cells (HSCs) within bone marrow however attempts to identify soluble factors that regulate HSC engraftment to bone marrow have been less successful. We have demonstrated that N-rasm transduced NIH 3T3 fibroblasts release soluble factors that support the ex-vivo expansion of UCB CD34+ hematopoietic progenitor cells (HPCs). UCB CD34+ cells were cultured in combination with cytokines : TPO, SCF, IL-6 and flt3-ligand(all at 100ng/ml). Significant (7-fold) expansion of total nucleated cells was seen on day 19 in cultures complemented with medium produced by N-rasm -transduced fibroblasts (N-rasm medium), compared to control cells (GFP- transduced fibroblasts (GFP medium)). Maximum expansion of UCB CD34+ cells (2-fold) and the most primitive sub-population of CD34+ CD117+ cells (2-fold) were seen on day 4 in N-rasm medium cells compared to GFP medium cells. Furthermore, increased numbers of cycling cells were observed in UCB CD34+ cell cultures supplemented with N-rasm medium. Significant (3-fold) expansion of CD34+CXCR4+ cells enriched with the NOD/SCID mice engrafting cells, was also observed. This increased expansion of CD34+CXCR4+ cells correlated with a 3-fold increased engraftment of human UCB cells in a NOD/SCID mouse model confirming retention of engraftment capability of expanded cells. This data demonstrates that N-rasm -transduced fibroblasts secrete soluble factors that promote ex-vivo expansion and in-vivo engraftment of primitive human hematopoietic progenitor/stem cells derived from UCB. Proteins that are secreted by N-rasm -transduced fibroblasts that activate hematopoiesis are currently being identified. Our data demonstrates the potential of N-rasm-transduced fibroblasts as a reservoir for the discovery of novel secreted proteins that regulate ex-vivo expansion and engraftment of human UCB HSCs and may represent a potential novel expansion strategy allowing greater use UCB as a stem cell source for allogeneic transplantation.
Comparison of Donor Cell Homing Efficiency between Intraosseus and Intravenous Injection in Primate Allogeneic Hematopoietic Stem Cell Transplantation.
Authors: Qi Feng , Phua Cindy M.L. , Tan Peik K. , Chow Pierce K.H. , Hwang William Y.K. 
Published: October 2019

Abstract
The traditional route for the delivery of hematopoietic stem cells (HSC) for transplantation by intravenous (IV) injection results in only a small percentage of the injected cells homing to the bone marrow (BM) microenvironment - where hematopoiesis could be more optimal. As such, delivery of HSCs by intra-BM injection has been investigated using syngeneic mice or autologous primate transplants. However, there has been no animal study of allogeneic HSCT directly comparing intra-BM and IV injection in terms of the seeding of donor cells in recipient BM, peripheral blood and other organs. We present the first direct comparison of donor cell homing efficiency between intra-BM and IV injection in an non-human primate (Cynomolgus Monkey) model of allogeneic hematopoietic stem cell transplant (HSCT) model using a preparative regimen modified from existing clinical protocols. Two recipients with similar age, weight and height received pre-transplant conditioning chemotherapy comprising 30mg/m2 fludarabine from Day-5 to Day-3, and 50mg/kg cyclophosphamide from Day-5 to -2. Fresh BM cells from an unrelated donor were harvested and labeled with PKH26 cell membrane dye (labeling efficiency: 99.6%) on day 0. With the recipients under general anesthesia, 2×108 donor cells were transplanted into the conditioned recipients by IV or intra-BM injection using a modified syringe suction method. Intra-BM injection recipients were found to have significantly more donor PKH26 cells, compared with IV injection recipients, in recipient BM at 2 hours (6.9% versus 2.5%, p<0.01) and 4 hours (6.7% versus 2.2% p<0.01) after transplantation. There were somewhat fewer donor cells in the peripheral blood after intra-BM delivery (3.4% versus 4.6%) at 2 hours, though the percentages were the same 4 hours after injection (1.6% in both). Interestingly, donor cell percentages in the bone marrow remained stable between 2 to 4 hours after transplant, as opposed to donor cell percentages in the peripheral blood, which fell by 2.1 to 2.9 times between 2 to 4 hours after transplant. Furthermore, compared to IV injection, intra BM injection retained more cells within BM, with less cells trapped in other organs, like the spleen (1.4% vs. 3.1%) and liver (0.8% vs. 3.5%), as well as non-hematopoietic organs like the heart (1.1% vs. 4.9%), lung (1.0% vs. 4.0%) and kidney (1.5% vs. 3.2%). Our study showed that in a allogeneic non-human primate model of HSCT, intra-BM injection of HSC resulted in superior retention of donor cells in BM with reduced seeding onto non-hematopoietic organs - suggesting possible applications for improved donor cell engraftment and reduced effects of donor-cell-mediated graft versus host disease on non-hematopoietic organs.
Figure Figure
Dendritic Cell Adenosylhomocysteine Hydrolase-Like Protein 1 (AHCYL1) and Inositol Phospholipid Signaling Are Potential Targets for Modulating Their Function.
Authors: Kato Masato , Key Brian , Carter Adrian , Angel Nicola Z. , Cooper Benjamine J. , Hart Derek 
Published: October 2019

Abstract
Recipient and donor dendritic cells (DC) initiate acute graft versus host disease after allogeneic hematopoietic stem cell transplantation and activated DC predict for severe disease. Using a differential display technique, we isolated the cDNA for an adenosylhomocysteine hydrolase-like protein 1 (AHCYL1), a novel intracellular protein with ~50% protein identity to adenosylhomocysteine hydrolase (AHCY) and showed that it was upregulated in activated human DC (Dekker et al. Immunogenetics.2002;53:993). AHCYL1 binds to the inositol 1,4,5-trisphosphate receptor (IP3R), suggesting that AHCYL1 is involved in intracellular calcium release (Ando et al. J Biol Chem.2003; 278:10602). Given that intracellular calcium levels control DC function, we reasoned that AHCYL1 was a potential target for modulating DC function. Therefore, we sought functional data using the zebrafish model. We identified two zebrafish AHCYL1 orthologs (zAHCYL1A and B) by bioinformatics and reverse transcriptase-polymerase chain reaction (RT-PCR). Unlike the ubiquitously present AHCY genes, AHCYL1 genes were only detected in segmented animals and AHCYL1 proteins were highly conserved among species. Phylogenic analysis suggested that the AHCYL1 gene diverged early from AHCY and evolved independently. Quantitative RT-PCR showed that zAHCYL1A and B mRNA expression was regulated differently to the other AHCY-like protein zAHCYL2 and zAHCY during zebrafish embryogenesis. Injection of morpholino antisense oligos against zAHCYL1A and B into zebrafish embryos inhibited zAHCYL1A and B mRNA translation specifically and induced ventralized morphologies. Conversely, human and zebrafish AHCYL1A mRNA injection into zebrafish embryos induced dorsalized morphologies, that were similar to those obtained by depleting intracellular calcium with thapsigargin. The injection of hAHCY had little effect on the embryos. These data suggest that AHCYL1 has a different function from AHCY and plays an important role in zebrafish embryogenesis by modulating IP3R function and consequent intracellular calcium release. It also suggests that blocking AHCYL1 in DC may have a significant effect on DC function, which might, ultimately, be exploited therapeutically. We have generated AHCYL1 gene deleted mice and these will be used to explore these questions further.
Pre-B Cell Acute Lymphoblastic Leukemia Cells Carrying a Bcr/Abl Translocation Retain Sensitivity to Minor Histocompatibility Antigen Specific T Cells but Are Resistant to Natural Killer Cells: Implications for Manipulation of Graft Versus Leukemia Effects.
Authors: Mullen Craig A. , Campbell Andrew , Young Faith , Jordan Craig T. 
Published: October 2019

Abstract
Background: Some acute lymphoblastic leukemias with high risk genetic lesions such as bcr/abl are treated with allogeneic BMT. Results of allogeneic BMT generally have shown only a modest improvement over high-dose chemotherapy. The graft versus leukemia (GVL) effect associated with allogeneic BMT is generally considered to be less potent in ALL than in myeloid leukemias. One hypothesis explaining reduced sensitivity of lymphoblastic leukemia cells to GVL is that such cells may be intrinsically resistant to the effects of cytolytic T or NK cells in the posttransplant immune environment.
Methods: Primary pre-B ALL leukemias with high risk human genetic lesions were induced by incubating bone marrow from mice null at the Ink4A/Arf locus with an ecotrophic retroviral vector containing a human bcr/abl fusion oncogene as well as a GFP marker. GFP+ leukemia cells were isolated and tested for sensitivity to T cells and NK cells. Minor histocompatibility antigen (mHA) specific T cells were generated by immunizing C3H.SW mice with irradiated C57BL/6 cells. NK cells were magnetically isolated from normal spleen cells using an anti-DX5 antibody. Leukemia cell populations were measured using a flow cytometry assay. 104 GFP-expressing leukemia cells were incubated with nonfluorescent T or NK cells in 96 well plates. At 48h leukemia cell population size was measured using fluorescent microspheres for quantitative internal controls. The percent survival of leukemia cells was expressed as a fraction of the absolute number of leukemia cells recovered from wells in which no cytolytic cells were present. In vivo sensitivity of the leukemia cells was measured by performing allogeneic BMT using donors that were naive or that had been previously sensitized to mHAs. 104 leukemia cells were infused at the time of transplant and survival and leukemia cell burden.
Results/Conclusions: Pre-B ALL cells exhibited exquisite sensitivity to the cytotoxic effects of anti-mHA T cells. In contrast, the lymphoblastic leukemia cells were resistant to the effects of NK cells. In vivo survival was prolonged and leukemia burden was reduced in those animals transplanted with specific anti-mHA T cells. These results suggest that mHA genes expressed in lymphoblastic leukemia cells may be suitable targets for post-transplant immunotherapy.
Evidence for Efficacy and Safety of Lentiviral Mediated Gene Transfer in T Cells and CD34+ Cells from Wiskott-Aldrich Syndrome Patients.
Authors: Scaramuzza Samantha , Trifari Sara , Marangoni Francesco , Martino Silvana , Metin Ayse , Galy Anne , Naldini Luigi , Aiuti Alessandro , Villa Anna , Dupré Loic , Roncarolo Maria Grazia 
Published: October 2019

Abstract
Wiskott-Aldrich Syndrome (WAS) is an X-linked primary immunodeficiency characterized by eczema, recurrent infections, severe hemorrhages and lymphomas. Transplantation of hematopoietic stem cells from HLA-identical sibling donors is a resolutive treatment, but it is available only for a minority of patients. Therapy based on the transplant of genetically correct autologous stem cells could represent a valid alternative approach. We investigated the efficacy and the safety of WAS gene transfer using HIV-based lentiviral vector encoding for WAS cDNA under the control of an autologous promoter (1.6 kb). T cells obtained from WAS patients showed normal level of WAS expression after lentiviral transduction. Transduced T cells showed a correction in TCR-driven proliferation and IL-2 production. Furthermore, a selective growth advantage of transduced T cells was observed in long-term in vitro cultures. Studies in T cell clones generated from transduced WAS CD4+ T cells revealed that 1–2 vector copies were necessary and sufficient to correct T cells function. CD34+ cells, isolated from mobilized peripheral blood and bone marrow of healthy donors, were transduced using WASP or GFP-encoding lentiviral vectors. Cells were cultured in the presence of different cytokines to investigate if WAS gene transfer could have any effect on short and long-term differentiation (CFU-C, LTC-IC and B/NK assays). Transduction resulted in a comparable number of CFU-C and LTC-IC colonies and normal B and NK cells differentiation with respect to untransduced cells. Furthermore, transduction of CD34+ cells isolated from the bone marrow of a WAS patient was performed under optimized culture conditions. Lentiviral gene transfer led to restoration of WASP expression in differentiated cells with copy number ranging from 1 to 5 copies per cell. In conclusion, our data demonstrate that the WAS promoter/cDNA-containing lentiviral vector can efficiently transduce and restore WASP expression in CD34+ cells and T cells from WAS patients. Experiments in the Rag2−/−/γchain- murine model are ongoing to test the efficacy and safety of the WASP transduced CD34+ cells. Together, our studies provide a preclinical basis for the implementation of a gene therapy trial for WAS patients.
Analysis of Health-Related Quality of Life in Patients Receiving AMG 531 Using the Disease-Specific Immune Thrombocytopenic Purpura Patient Assessment Questionnaire.
Authors: George James N. , Bussel James B. , McMillan Robert , Guo Matthew , Okano Gary J. , Nichol Janet L. 
Published: October 2019

Abstract
AMG 531 is a novel thrombopoeisis-stimulating peptibody that increases platelet production by targeting the thrombopoietin receptor. The trial described here is an ongoing, open-label, extension study of patients with immune thrombocytopenic purpura (ITP) assessing the safety and efficacy of long-term AMG 531 treatment of ITP, as well as patient-reported health-related quality of life (HRQOL). Eligible patients have completed a previous AMG 531 study in ITP. Patients previously treated with AMG 531 receive a starting dose that is the same as the final dose given in the previous study; placebo-treated patients begin the extension study with an AMG 531 dose of 1 μg/kg. The dose is skipped, decreased, maintained, or increased based on platelet count. Analysis was performed on the 36 patients (25 female, 11 male; 29 white, 6 Hispanic, 1 black) for whom complete data are currently available. The mean age is 50.0 ± 13.0 (SD) years. Most patients (83%) had a splenectomy prior to this study. HRQOL is being assessed with an ITP disease-specific measure, the ITP Patient Assessment Questionnaire (ITP-PAQ). This report describes the first use of the ITP-PAQ to measure treatment effect. The ITP-PAQ has 10 scales: 4 Physical Health (PH)—Symptoms, Fatigue, Bother, and Activity; 2 Emotional Health (EH)—Psychological and Fear; 3 Quality of Life (QOL)—Overall, Social, and Work; and 1 Women’s Reproductive Health (WRH). Each scale is scored 0 (worst)—100 (best). A planned interim analysis was conducted to evaluate changes in HRQOL from week 1 (pretreatment) to week 24. Results indicated improvement in the areas of PH-Symptoms, with a 7.9-unit increase (p=0.007), PH-Fatigue, with a 7.8-unit increase (p=0.006), PH-Bother, with a 9.4-unit increase (p=0.025), PH-Activity, with a 7.4-unit increase (p=0.032), and QOL-Work, with a 7.9-unit increase (p=0.014). An analysis of minimal clinically important difference will be conducted when the sample size is large enough to determine if statistically significant values are clinically meaningful. A comparison of HRQOL in patients with a durable platelet response (doubling of the baseline count and ≥50×109/L at 6 or more weeks during weeks 17–24 in the absence of rescue medication) versus those without a durable response showed a trend for greater improvement among the responders in all 10 scales, with significant differences for PH-Symptoms (p=0.02), PH-Bother (p=0.02), PH-Activity (p=0.01), EH-Psychological (p=0.01), and QOL-Overall (p=0.03). Individualized dosing of AMG 531 provides a unique therapeutic option for managing platelet count in ITP while improving HRQOL.
Advantages wth Using the Levonorgestrel-Releasing Intrauterine System in Females with Hemostatic Disorders.
Authors: Lukes Andrea S. , Arepally Gowthami 
Published: October 2019

Abstract
BACKGROUND: The levonorgestrel-releasing intrauterine system (LNG-IUS) is a small device placed within the uterus that is used for both contraception and treatment of menorraghia. This system releases 20 micrograms of levonorgestrel every 24 hours over 5 years. We hypothesized that females with inherited or acquired bleeding disorders would have decreased menstrual bleeding with the LNG-IUS and improved quality of life.
METHODS: A small, retrospective case series of females with hemostatic disorders who used the LNG-IUS within a 2 year period and who were seen at a tertiary medical center.
RESULTS: Seven females were treated with the LNG-IUS for menorrhagia and contraception. We did not reverse anticoagulation for placement of the device and there were no bleeding complications associated with the LNG-IUS insertion. The average age was 36.7 years (range 28 – 48 years). Four females had von Willebrand’s disease (VWD). Four females were on antithrombotic therapy, 2 warfarin and 2 aspirin. One female with VWD was heterozygous for factor V Leiden and was on warfarin for treatment of deep venous thrombosis. Six females had menorrhagia and one had bleeding most of the time. The average days of menstrual bleeding prior to LNG-IUS was 8.3 days which improved to 4.5 days after placement of the LNG-IUS. Three of 7 females discontinued use of the LNG-IUS, yet one had the LNG-IUS removed because she no longer required contraception. This female became pregnant and after an uncomplicated normal spontaneous vaginal delivery elected to use the LNG-IUS again for contraception and the benefit of decreased menstrual bleeding. The other 2 females (2/7, 29%) discontinued use of the LNG-IUS because there was worsening of heavy menses or ‘failed’ therapy - one with VWD and one on anticoagulation. The female on anticoagulation in this case also had significant uterine fibroids. A ten item quality of life questionnaire developed by Kouides was administered, and showed overall improvement in quality of life.
CONCLUSION: The levonorgestrel-releasing intrauterine system is an effective method of contraception and an effective treatment for heavy menses. For females with hemostatic disorders, this device has unique advantages: it is technically easy to use in females at increased bleeding risk, offers effective control of menorrhagia, and may likely improve overall quality of life. Additional prospective large scale studies are required to validate these findings.