bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bone Health Assesment in Patients Undergoing Hematopoietic Cell Transplantation
Authors: Ramirez Pablo , Florenzano Pablo , Campusano Claudia , Jara Veronica , Bertin Pablo A , Ernst Daniel M 
Published: October 2019

Abstract
INTRODUCTION: The increased survival of hematological patients has required to widen the care of these patients, with emphasis in factors related to quality of life and late mortality. Among them, osteoporosis (OP) is a fundamental problem. Patients undergoing hematopoietic cell transplantation (HCT) are at great risk of OP, mainly due to prolonged exposure to chemotherapy, immunosuppresants and the hypogonadism frequently associated to these treatments. Despite this, there is lack of strong evidence on this matter and HCT guidelines are not clear on this problem.
OBJECTIVES: To evaluate parameters related to bone health in patients undergoing HCT. To measure the frequency and severity of the alterations in this group of patients.
PATIENTS AND METHODS: Observational and retrospective analysis of patients undergoing HCT at the Catholic University Hematology-Oncology Department. All patients undergoing HCT were recommended to have determination of 25-OH vitamin D, PTHi, Calcium, Phosphorus and bone densitometry (DXA). We included all patients with the evaluation before HCT (preHCT) and 1 year after HCT (postHCT).
RESULTS: We obtained data from 20 patients preHCT and 22 patients postHCT. Main diagnosis were acute myeloid leukemia (n=13; 32%), multiple myeloma (n=11; 26%), acute lymphoblastic leukemia (n=9; 21%) and Hodgkin's Lymphoma (n=5; 12%). Mean age was 40 years (range: 17-67) and 67% were males. In the preHCT group, the median 25-OH vitamin D levels were 13,6 ng/ml (range: 4,9-26,7 ng/ml) and 100% of the patients were in the insuficiency range leveles (<30 ng/ml). Median PTHi levels were 62,6 pg/ml (range: 24,4-223,7 pg/ml), and 46% of the patients had secondary hyperparathyroidism. In the postHCT group, median 25-OH vitamina D levels were 11,4 ng/ml (range: 4-29,4 ng/ml) and 100% had insuficiency levels. Median PTHi levels were 68 pg/ml (range: 37,4-135,8 pg/ml), and 56% of the patients had secondary hyperparathyroidism. 12 patients had DXA before HCT and 3 (25%) of them had DXA lumbar Z score less than -2.0. In 19 patients postHCT, 4 (21%) had decreased DXA Z score < 2.0. No difference could be established between autologus or allogeneic HCT in any meassurement.
CONCLUSIONS: HCT patients represent a high risk group of developing severe vitamin D deficiency, secondary hyperparathyrodism and decreased DXA levels. These data are a warning that this population of patients requiere early intervention to prevent long term complications. This report is the initial evaluation for the development and treatment of bone health in a prospective matter in HCT patients in our center.

Disclosures
No relevant conflicts of interest to declare.
Hematopoietic Stem Cell Transplantation for Patients with β-Thalassemia Major Underwent Pre-Transplant Splenectomy
Authors: He Yuelin , Wu Xuedong , Ruan Yongsheng , Peng Zhiyong , Feng Xiaoqin , Liao Jianyun , Liu Huaying , Xu Wenfeng , Li Na , Li Chunfu 
Published: October 2019

Abstract
Current study analyses the effect of splenectomy on outcomes of hematopoietic stem cell transplantation (HSCT) for patients with β-thalassemia major (TM). Twenty-two class II and III (according to Nanfang’s criteria, see Blood, 2012;120 (19): 3875-3881) patients with TM had a pre-transplant splenectomy. The outcomes of the 22 transplants were compared with 193 transplants in class II and III patients between Aug. 2008 and Dec. 2013. Patients in the splenectomy group were older (8.9±5.0 vs. 6.0±3.5 year old; p=0.001) and class III patients were more (8/22 vs. 10/193, p=0.001) than non-splenectomy. In splenectomy group 10 patients received graft from HLA matched sibling, 9 from unrelated donors and 3 from haploidentical parents plus unrelated cord blood, respectively. In non-splenectomy group 82 patients received graft from matched sibling and 121 from unrelated donors, respectively.
213 cases engrafted in 4 weeks, Splenectomized patients had a significantly shorter time to absolute neutrophil count >500/mm3 (14.5±5.2 vs 19±4.2 days p=0.001). There were no significantly different at time to platelet >2000/mm3 (16.2±5.8 vs 16.1±5.3 days, p=0.415) and hemoglobin >80g/L (15.3±4.8 vs 13.1±5.3 days p=0.135). Secondary rejection occurred in 2 patients (at 3rd month and 4th month post transplantation, respectively) in splenectomy group, but rescued successfully by second transplants from unrelated donor one year later and from haploidentical parent two years later, respectively. The patients received graft from father, one died of complications of VOD; other died of lung infection on forty-five days post transplantation. Two patients rejected their grafts in non-splenectomy group.
The mean RBC transfusion was 4u (0u-12u). The mean platelet transfusion in splenectomy group had reduced (4.4±4.5 vs 7.1±4.8 units p=0.016), with 17 patients transfused less than 6u Platelet. The incidence of acute and chronic GVHD, late infections were rare and not significantly different between the two groups. No patients died of infection due to splenectomy. Patients underwent HSCT after splenectomy had a high rate of OS (95.5% vs 92.8% p=0.001). Although TFS was lower in splenectomy group than non-splenectomy group (88.5% vs 92.3% p=0.001), this may be due to more older, class III patients and higher rate of rejection (2/22 vs 2/193).
Conclusion, our study showed pre-transplant splenectomy patients with β-thalassemia major was associated with faster engraftment and decreased require of platelet and bloodtransfusion support.

Disclosures
No relevant conflicts of interest to declare.
Encouraging Outcomes of Reduced-Intensity Allogeneic Hematopoietic Stem Cell Transplantation for Pediatric Patients with Acute Lymphoblastic Leukemia in Second Complete Remission
Authors: Higuchi Kohei , Sato Maho , Kondo Osamu , Ioi Aya , Mayumi Azusa , Shimizu Mariko , Sawada Akihisa , Yasui Masahiro , Inoue Masami 
Published: October 2019

Abstract
[Background] We have been performing reduced-intensity stem cell transplantation (RIST) to avoid preconditioning-related complications. However, the effectiveness of RIST in pediatric patients with acute lymphoblastic leukemia (ALL) remains to be clarified.
[Methods] We retrospectively reviewed 37 pediatric patients with ALL in second complete remission (CR2) who underwent first allogeneic hematopoietic stem cell transplantation (allo-SCT) between 1993 and 2012 in our institute. We compared the outcomes of RIST with those of myeloablative stem cell transplantation (MAST).
[Results] The median age at allo-SCT was 9 years (range, 1 to 18 years). There were 33 B-lineage ALL, 3 T-lineage ALL, 1 lineage unknown ALL, and none of Philadelphia chromosome-positive ALL. Sixteen patients received HLA-matched bone marrow (7 related; 9 unrelated), 12 HLA-mismatched bone marrow (11 unrelated; 1 HLA haploidentical related), 4 cord blood, and 5 CD34 positive peripheral blood stem cells (HLA haploidentical related). In all patients, the 5-year overall survival (5y-OS) rate and the 5-year event free survival (5y-EFS) rate were 75.1% and 56.5%, respectively.
Seven patients underwent RIST and 30 patients underwent MAST. The median follow-up durations of RIST and MAST groups were 3.3 years (range, 0.9 to 8.2 years) and 11.3 years (range, 0 to 21.2 years), respectively. The 5y-OS rates in RIST and MAST groups were 85.7% and 59.8%, and the 5y-EFS rates were 71.4% and 53.3%, respectively. The 5-year cumulative transplant-related mortality (TRM) rates in RIST and MAST groups were 0% and 31.0%, and the 5-year cumulative relapse rates were 28.6% and 24.3%, respectively.
[Discussion] In our series, the cumulative relapse rate in RIST group was similar with that in MAST group, and the cumulative TRM rate in RIST group was lower than that of MAST group. Therefore, both of the 5y-OS and the 5y-EFS rates in RIST group seem to be better than those in MAST group. The outcomes of RIST in our series do not seem to be poorer. Although further studies are needed because of the small size of patients and short follow-up duration, RIST can be considered as the first transplantation for pediatric patients with ALL in CR2.
Figure 1 Figure 1.
Figure 2 Figure 2.
Figure 3 Figure 3.

Disclosures
No relevant conflicts of interest to declare.
Study in the Effect of EZH2 Gene Targeting siRNA on Cell Death in HL-60 Cell Line
Authors: Luhui Lin , Huang Yiqun , Ma Xudong 
Published: October 2019

Abstract
Objective: To study small interfering RNA (siRNA) targeting EZH2 gene effect on cell proliferation, apoptosis and histone modulation in HL-60 cells line.
Methods: (1) Three siRNA segments targeting EZH2 gene were designed and transfected into HL-60 cells by LipofectamineTM 2000. The optimal segment was screened by RT-PCR. (2) Cell growth inhibited by EZH2 siRNA was determined by WST-1. Cell apoptosis was measured by Flow cytometry. (3) The expression of procaspase-3, procaspase-9, Bcl-2, and Bax was detected by Western blot. The levels of histone H3 lysine27 dimethylation and trimethylation and histone acetylatiom of H3 were evaluated by Western blot.
Results: (1) The efficiency of transfection with EZH2 siRNA for 6 hours was (82 ±2.97)%. The sequence of optimal segment of EZH2 siRNA was sense 5'- GAGGGAAAGUGUAUGAUAATT -3'，antisense 5'- UUAUCAUACACUUUC CCUCTT -3'。 (2) EZH2 siRNA suppressed the proliferation of HL-60 cells. (3) EZH2 siRNA downregulates the expression of procaspase-3, procaspase-9, Bcl-2 and upregulates the expression of Bax and induced cells apoptosis. (4) EZH2 siRNA suppressed the protein and mRNA of EZH2 in HL-60 cells. The levels of histone H3 lysine27 dimethylation and trimethylation was down regulated and histone acetylatiom of H3 was up regulated.
Conclusions: (1) LipofectamineTM2000 transfected siRNA for EZH2 gene into HL-60 cells has high efficiency. (2) EZH2 siRNA inhibits cell growth and induces cell apoptosis in HL-60 cell line. It might be a new therapeutic target in human leukemia. (3) EZH2 siRNA downregulates the levels of histone H3 lysine27 dimethylation and trimethylation and upregulates histone acetylation of H3. The mechanism of EZH2 effect on histone acetylation need to be further studied. RNAi technology is expected to become a new tool for targeted gene therapy of acute leukemia, and EZH2 might be one of the targets of gene therapy for leukemia.

Disclosures
No relevant conflicts of interest to declare.
Pathway Phenotyping of B-Cell Receptor Activation and Inhibition: Translation to Novel Clinical Biomarkers
Authors: Wehrman Tom S 
Published: October 2019

Abstract
B-cell receptor (BCR) signaling has emerged as a vital and targetable component in a number of B-cell malignancies. The recent approval and numerous ongoing clinical trials of small molecule kinase inhibitors targeting this pathway has bolstered efforts to define the signaling proteins critical to BCR activity. To this end, we first mapped BCR signaling using 93 different activation state antibodies. These results provided a high level view of BCR signaling across more than 80 intracellular proteins covering canonical and non-canonical BCR effectors. The signaling responses were then modulated with known kinase inhibitors to associate kinase activity with pathway activation. Using these data as a backdrop, three BTK inhibitors (AVL-292, CNX-774, and ibrutinib) were tested to determine which components of BCR signaling are commonly inhibited by these molecules. These studies showed that ibrutinib was the most promiscuous of the inhibitors but that all three compounds blocked activation of the S6 signal transduction pathway. Interestingly, a number of other oncogenic pathways remained intact after inhibitor treatment including the MAPK-ERK and AKT signaling pathways. After identifying the S6 pathway as a commonly blocked effector of BCR signaling, we validated the pathway readouts for their use as ex vivo pharmacodynamic assays in whole blood patient samples. Our results show that the assays yield the appropriate precision and reproducibility necessary to be used as PD markers in a clinical trial setting. Thus, the workflow of pathway phenotyping to inhibitor testing and finally to clinical biomarker development further defined the activation patterns of BCR signaling and identified novel clinical readouts for measuring BCR inhibitor activity.

Disclosures
Wehrman: Primity Bio: Employment.
MiR-146a Regulates the TRAF6/TNF-Axis in Donor T Cells during Graft-Versus-Host Disease
Authors: Stickel Natalie , Prinz Gabriele , Pfeifer Dietmar , Schmitt-Graeff Annette , Follo Marie , Finke Jürgen , Duyster Justus , Salzer Ulrich , Zeiser Robert 
Published: October 2019

Abstract

Introduction: Acute graft-versus-host disease (GvHD) arises from the attack of recipient tissues by donor allogeneic T cells and represents one of the major limitations of allogeneic hematopoietic cell transplantation (allo-HCT). In spite of many clinical trials, the standard immunosuppressive regimens for prevention of acute GvHD have improved little in the last two decades. Hence, a better understanding of the biology of acute GvHD may improve therapeutic options. MicroRNA-146a (miR-146a) was found to be increased in the sera of patients with GvHD. Therefore, we aimed to decipher the role of miR-146a in allogeneic donor T cells during GvHD by functional studies and in patients undergoing allo-HCT by single nucleotide polymorphism (SNP) analysis.
Methods: We used two different murine major MHC mismatch models for acute GvHD. Recipient mice were conditioned with irradiation before transplantation of bone marrow and either wildtype or miR-146a deficient T cells from allogeneic donor mice. Furthermore, genomic DNA from 289 patients that underwent allo-HCT and their respective hematopoietic stem cell donors was isolated in order to determine their miR-146a rs2910164genotype.
Results: We observed miR-146a upregulation in T cells of mice developing acute GvHD compared to untreated mice in a major MHC and a minor histocompatibility antigen mismatch model. Transfer of miR-146a deficient T cells caused increased GvHD severity, elevated TNF serum levels and reduced survival. Conversely, the phytochemical induction of miR-146a or its overexpression in donor T cells using a specific miR-146a mimic reduced GvHD severity. TNF receptor-associated factor 6 (TRAF6), a verified target of miR-146a, was upregulated in miR-146a-/- T cells following alloantigen stimulation. Higher TRAF6 levels translated into increased NF-κB activity and TNF production in miR-146a-/- T cells, while other pro-inflammatory cytokine levels were unaffected. The detrimental effect of miR-146a deficiency in T cells could be antagonized by TNF blockade in vivo. Moreover, in contrast to WT T cells, over expression of miR-146a in Tnf deficient T cells had no effect on their alloreactivity. In the human system, the minor genotype of the SNP rs2910164, which causes reduced miR-146a expression, was more frequent in patients developing acute GvHD grade III/IV compared to all other allo-HCT recipients (n=289).
Conclusions: Taken together we show that miR-146a functions as a negative regulator of the TRAF6/TNF-axis in allogeneic donor T cells during GvHD, leading to reduced TNF transcription. Given our observation on the predictive role of the SNP leading to decreased miR-146a expression in acute GvHD in patients and the possibility to exogenously enhance miR-146a expression, we provide a novel and targeted molecular approach to mitigate GvHD.

Disclosures
No relevant conflicts of interest to declare.
Regulation of Immune Responses during Acute GvHD Via the IL-33/ST2 Axis
Authors: Reichenbach Dawn K , Schwarze Vincent , Matta Benjamin M , Tkachev Victor , Lieberknecht Elizabeth , Liu Quan , Koehn Brent H , Pfeifer Dietmar , Taylor Patricia A , Prinz Gabriele , Dierbach Heide , Stickel Natalie , Beck Yvonne , Warncke Max , Junt Tobias , Schmitt-Graeff Annette , Nakae Susumu , Follo Marie , Wertheimer Tobias , Schwab Lukas , Devlin Jason , Watkins Simon C , Duyster Justus , Ferrara James L.M. , Turnquist Heth R. , Zeiser Robert , Blazar Bruce R. 
Published: October 2019

Abstract
The IL-1 superfamily member IL-33 is produced in barrier tissues. IL-33 binds to the receptor suppression of tumorigenicity 2 (ST2), expressed on stromal cells, regulatory T cells (Tregs), myeloid derived suppressor cells (MDSCs), and macrophages. IL-33 has both anti-inflammatory and pro-inflammatory properties. It is not known if IL-33 plays a role in acute GvHD, and if so what properties it exerts. By immunohistochemistry staining of gut tissues, IL-33 production by non-hematopoietic cells was increased in mice post-conditioning and in patients during GvHD. To determine whether IL-33 could augment GvHD via a host signaling mechanism, we compared st2-/-to wildtype (wt) hosts and observed decreased GvHD lethality (Figure 1A). Additionally, IL-33-/- versus wt hosts had a marked decrease in GvHD lethality and reduced TNFα production. Conversely, IL-33 administration during the peak inflammatory response worsened GvHD. Previous studies have shown increased levels of the soluble form of ST2 (sST2) are a biomarker for steroid-refractory GvHD (Vander Lugt, NEJM, 2013). We hypothesized that sST2 acted not only as an indicator of tissue injury and biomarker of GvHD but also as an immune modulator during GvHD. In rodents, we found that ST2 was upregulated on alloreactive T cells and sST2 increased as GvHD progressed. St2-/-versus wt donor T cells had a marked reduction in GvHD lethality (Figure 1B) without compromise of graft-vs-leukemia responses. Comparable data was seen in 2 different strain combinations. Alloantigen-induced IL-18 receptor upregulation was significantly lower in the absence of ST2, which was linked to significantly reduced IFNγ production by st2-/- vs wt CD4 and CD8 T cells during GvHD. Similarly, sST2 transgenic hosts and wt recipients given exogenous sST2-Fc fusion protein infusions (Figure 1C) to block ST2/IL-33 interaction each had significantly reduced GVHD lethality, establishing the functional role of ST2 as a decoy receptor modulating GVHD. During the peak of the GvHD inflammatory response, IL-33 signalling of either donor or host cells promoted activation of donor T cells, while the use of exogenous sST2-Fc protein to prevent IL33/ST2 engagement ameliorates disease. Together, these studies point to targeting of the IL-33/ST2 axis as a novel and potent target for GvHD therapy.

Disclosures
Warncke: Novartis Pharma AG: Employment. Junt:Novartis Pharma AG: Employment.
Polycomb Group Member Rybp Is a Functional Tumor Suppressor Repressed By Mir-9 in MLL-Rearranged AML
Authors: Price Colles , Chen Ping , Li Shenglai , Li Zejuan , Li Yuanyuan , Jiang Xi , Huang Hao , Weng Hengyou , Ulrich Bryan , Arnovitz Stephen , Neilly Mary Beth , Chen Jianjun 
Published: October 2019

Abstract
MicroRNAs (miRNAs), are small non-coding RNA molecules known to be important regulators of cancer biology. Notably, we and others have shown that miRNAs play important roles in Acute Myeloid Leukemia (AML), a heterogeneous malignancies with multiple chromosomal and molecular abnormalities. Patients with chromosomal rearrangements involving mixed lineage leukemia (MLL), the mammalian homology of trithorax gene, are associated with poor survival. Previously, we have found that MLL-rearranged AML drives aberrant expression of several miRNAs, most notably microRNA-9 (miR-9). Expression of miR-9 with MLL-AF9, a common MLL-translocation, was sufficient to promote transformation normal hematopoietic progenitor cells in vitro and leukemogenesis in vivo. We previously found that miR-9 reduces expression of several genes but we did not know which genes were critical tumor suppressors. We found that the polycomb group member RING1- and YY1-Bindin Protein (RYBP) was consistently inhibited upon miR-9 expression. To assess the regulation of RYBP we used publically available data from the Cancer Genome Atlas (TCGA) and looked at genome-wide Illumina 450K methylation data. We did not find a strong correlation with methylation and RYBP expression, suggesting that expression of RYBP is likely not regulated by the DNA methylation machinery in patients. Upon looking at copy number alterations we found that a small population of AML patients contained either homozygous or heterozygous loss of RYBP, suggesting a potential role of RYBP in leukemia pathogenesis. To assess the role of RYBP we did a series of in vitro experiments. We found that expression of RYBP was sufficient to attenuate colony-forming growth driven by MLL- AF9. Furthermore, RYBP expression was able to reduce proliferation, increase apoptosis, and significantly reduce immature cell population. To determine the role of RYBP expression in vivo, we transplanted lethally irradiated mice with progenitors retrovirally transduced with MLL-AF9 compared to MLL-AF9 and RYBP. We found that expression of RYBP was sufficient to reduce leukemia burden in vivo as well as induce differentiation as shown by flow cytometry and histological analysis. Thus, this demonstrates that RYBP is a functional tumor suppressor in MLL-rearranged AML. In conclusion, we have demonstrated that chromosomal rearrangements involving MLL, the mammalian homology of trithorax, downregulates a member of the polycomb complex through upregulation of miR-9.

Disclosures
No relevant conflicts of interest to declare.
Determinants of Low Bone Mineral Density after Childhood Cancer
Authors: den Hoed Marissa , Klap Bibi C. , Te Winkel Mariël L. , Stolk Lisette , Pieters Rob , van Waas Marjolein , Neggers Sebastian J.C.M.M. , Blijdorp Karin , van Dorp Wendy , Boot A.M. , Pluijm S.M.F. , van den Heuvel-Eibrink Marry M. 
Published: October 2019

Abstract
Purpose: Osteopenia is a potential complication of childhood cancer treatment, but the magnitude of this problem in survivors is unknown. We examined (determinants of) bone mineral density (BMD) status in long-term survivors of adult childhood cancer (CCS).
Methods: This retrospective single-center cohort study included 346 subjects with the most common types of childhood cancer. Acute leukemia or lymphoma was diagnosed in 273 (81%) of the patients. All subjects had a median age at diagnosis of 7.0 years (range: 0.1-16.8 years) and a median follow-up time of 16.7 years (range 5.6-39.9 years). Total body BMD (BMDTB) and BMD of the lumbar spine (BMDLS) were measured by dual-X-ray absorptiometry. Osteopenia was defined as BMD standardized deviation score (SDS) below -1. Twelve candidate single nucleotide polymorphisms (SNPs) in 11 genes (COL1A1, TNFSF11, TNFRSF11, TNRFSA11B, VDR, ESR1,WLS, LRP5, MTHFR, MTRR, IL6) were investigated.
Results: Survivors had a lower BMDTB and BMDLS (mean SDS: -0.55, p<0.001; and -0.30, p<0.001, respectively) as compared to healthy peers. This was similar in cancer survivors of leukemia and lymphoma (mean SDS: -0.49, p<0.001; and -0.32, p<0.001, respectively). Osteopenia (BMDTB and/or BMDLS) was present in 45% of the survivors (46% of the leukemia and lymphoma survivors). Multivariate logistic regression analyses identified age at diagnosis <12 years, age >30 years at follow-up, male gender, underweight at follow-up, carriers of the minor allele of rs2504063 (LRP5), treatment with cranial-spinal radiotherapy and prednisone as independent prognostic factors for osteopenia. In survivors of leukemia and lymphoma, we identified low BMI at follow-up, carriers the minor allele of rs2504063 (LRP5), treatment with cranial-spinal radiotherapy and cyclophosphamide use as prognostic factors for osteopenia.
Conclusions: This large cohort of childhood cancer survivors with the long follow-up identified osteopenia in 45% of CCS. This indicates that greater awareness for low BMD is warranted, especially in survivors who are older than 30 years, male, have underweight, have a genetic predisposition, and who were treated with cranial-spinal radiotherapy and/or steroids.

Disclosures
No relevant conflicts of interest to declare.