bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Outcome of Pediatric Acquired Aplastic Anemia Underwent Immunosuppressive Therapy and Transplantation: A 13-Year Experience in Two Tertiary Care Centers , Southern China
Authors: Huang Junbin , Huang Lifeng , Liu Su , Cheng Yucai , Xue Hongman , Chen Chun , Lin Shaofen 
Published: November 2019

Objective To evaluate efficacy, complications and contributing factors for response of immunosuppressive therapy(IST) and allogenic hematopoietic stem cell transplantation(allo-HSCT) in children with acquired aplastic anemia; to explore optimal therapy for different clinical types of aplastic anemia.
Methods 230 children diagnosed with aplastic anemia accepted immunosuppressive therapy from January 1, 2006 to July 15, 2019 in Department of pediatrics, Sun Yat-sen memorial hospital and Department of pediatrics, The seventh affiliated hospital, Sun Yat-sen University, the response rate for 3, 6, 9, 12months for IST, complications,predictors of response and overall survival were analyzed and response rates were compared according to clinical types.
Results 230 children were followed with median time 47.3 months. Among all AA patients, there are 67 NSAA patients, 95 SAA patients and 68 VSAA patients. 214 patients had received IST initially, due to economic, medical insurance related factors or lack of well-matched donors at the time, and 16 patients had undergone allo-HSCT initially. And 21 patients , who had failed to had a CR/PR responds to IST, received allo-HSCT as a salvage regimen. The overall survival(OS) rate of all patients was 79.6±2.9% with a 46.2 months median follow-up.
For those who had received IST therapy overall CR rate was 27.1%(58/214), PR rate 37.4%(80/214), therefore overall response (CR+PR)rate was 64.5%(138/214). CR rate at 3,6,9,12 months were 7.9%, 15.9%, 16.4%, 27.1%, respectively; meanwhile PR rate 3,6,9,12 monthsatwere 28.0%, 34.1%, 34.1%, 37.4%, respectively. When we termed"transplantation and death"as an even, the transplantation-free survival(TFS) for NSAA was 90.7±4.0%, significantly superior to SAA(59.7±6.5%) and VSAA(50.5±7.5%). As for complications, serum sickness and infection are most commonly seen.
Among 37 patients who had recieved allo-HSCT, 33 patients had long term CR, while 2 died of infection, 1 of extensive aGVHD, 1 of thrombotic microangiopathy. 21 patients who underwent HSCT as a salvage regimen had 85.4±7.8% OS with a 55.6 months median follow-up, and 16 patients who recieved HSCT as initial treatment had 93.8±6.1% OS with a 69.1 months median follow-up.
Conclusions In children with AA, overall response rate to IST goes higher over time with a highest response rate 64.5% at 12 month. TFS for NSAA IST-group is significantly superior to SAA and VSAA IST-group. However, well-matched allo-HSCT still achieve nearly 90% OS as both initial or salvage treatment. Hence, we agure that IST is a pratical treatment for pediatric AA, especially NSAA, in conditions of economic status and poor medical insurance, and HSCT is still a cure for those who failed IST.
Figure

Disclosures
No relevant conflicts of interest to declare.
Automation in Hemopoietic Stem Cell Gene Therapy: Results of a Head-to-Head Comparison of a Manual Vs an Automated Procedure
Authors: Papanikolaou Eleni , Bissels Ute , Johnston Ian , Reinartz Sarah , Brams Dominik , Aivazidou Faidra , Krenz Dariusz , Bomhard Inga von , Knöbel Sebastian , Bosio Andreas 
Published: November 2019

The recent successes in gene therapy for the treatment of rare blood disorders such as primary immunodeficiencies and hemoglobinopathies have highlighted the need to generate robust and scalable manufacturing processes. Here we compare large scale lentiviral transductions of hemopoietic stem cells performed on the CliniMACS Prodigy® to manual processes in terms of efficacy and viability of the end product. In our first series of experiments, we used a low, non-saturating MOI of 30 and the results showed that transduction efficiency on the CliniMACS Prodigy® was significantly increased from an average of 23.3% (manual) to 50.5% (CliniMACS Prodigy®). These results were further confirmed when we employed an MOI of 100 and showed that the average transduction efficiency was 74% from the CliniMACS Prodigy® compared to 54.1% from the manual steps. Moreover, the total viability of the cells cultured on the CliniMACS Prodigy® remained unaffected after two days of cultivation with an average recovery of 105% but with significantly less variability (SD: 15.4) compared to the manual steps (SD: 35.7). Finally there was no difference to the average VCN which was 2.2 from the manual steps and 2.4 from the CliniMACS Prodigy®. Overall, our results indicate that the CliniMACS Prodigy® generates higher transduction rates, combined with high viability compared to the manual process, but most importantly, with significantly lower variability, suggesting that it represents a closed system able to automatically perform complex processes as successfully as when manual handling steps are performed, but with higher predictability, efficiency and with minimal user interaction.

Disclosures
Papanikolaou: Miltenyi Biotec: Employment. Bissels:Miltenyi Biotec: Employment. Johnston:Miltenyi Biotec: Employment. Reinartz:Miltenyi Biotec: Employment. Brams:Miltenyi Biotec: Employment. Aivazidou:Miltenyi Biotec: Employment. Krenz:Miltenyi Biotec: Employment. Bomhard:Miltenyi Biotec: Employment. Knöbel:Miltenyi Biotec: Employment. Bosio:Miltenyi Biotec: Employment.
Endogenous Retinoid X Receptor Ligands Act As Tumor Suppressors in MLL-AF9 Mouse Leukemia
Authors: Di Martino Orsola , Ferris Margaret Y , Gayla Hadwiger , Niu Haixia , Welch John S. 
Published: November 2019

Retinoid therapy transformed response and survival outcomes in acute promyelocytic leukemia (APL), but has demonstrated only modest activity in non-APL forms of acute myeloid leukemia (AML). The retinoic acid receptors (RARs) and retinoid X receptors (RXRs) are ligand-activated transcription factors that influence hematopoietic stem cell self-renewal and differentiation. In normal hematopoiesis, RARA and RXRA are dynamically regulated during myeloid maturation, with highest expression in mature neutrophils. In acute myeloid leukemia (AML) RARA and RXRA have parallel expression among AML subtypes, with the highest expression in M4/M5 myelomonocytic and monocytic subtypes. Using a murine reporter assay, we found that natural RXRA ligands, but not RARA ligands, were present in vivo in primary mouse myelomonocytic leukemia cells (derived with an MLL-AF9 retrovirus) and acted as tumor suppressors. RXR ligands were absent in erythroleukemia (derived with TLS-ERG) or T-cell leukemia (derived with activated Notch1), suggesting the presence of natural RXRA ligands specifically in myelomonocytic leukemia. Moreover, we found that deletion of Rxra and Rxrb was naturally selected in MLL-AF9, but not in Notch1 or TLS-ERG leukemias, and that loss of Rxra and Rxrb accelerated leukemic growth. This suggests that Rxrs act as tumor suppressors, but only when exposed to natural ligands. In MLL-AF9 derived leukemia cells, pharmacologic treatment with single-agent retinoid led to modest growth inhibition due to non-permissive activity of the RARA:RXR heterodimer, whereas concurrent activation of both RARA, with all-trans retinoic acid (a pan-RAR ligand), and RXR, by bexarotene (a pan-RXR ligand), enabled efficient co-repressor release and synergistic leukemic apoptosis. We observed that co-repressors release (SMRT/NCoR) from the RARA:RXRA heterodimer was specifically associated with RARA activation, whereas growth inhibition required RXR binding, and this occurred through apoptosis rather than maturation. Generating a series of RXRA mutant proteins we demonstrated the active contribution of RXRA to the activity of the RARA:RXRA heterodimer, specifically requiring the activation domains (AF1 and AF2) and the ability to recruit co-activator to the RXRA element. Finally, we observed a significant dose-dependent effect of combination ATRA and bexarotene treatment in in vivo in MLL-AF9 leukemic mice with a striking reduction in the tumor burden of treated mice compared to the control cohort. These data provide a strategy for clinical retinoid therapies in leukemias beyond acute promyelocytic leukemia and provide a mechanism for integrating retinoid therapy into future clinical trials.

Disclosures
No relevant conflicts of interest to declare.
The Platelet Millionaire: A Rare Cause of Thrombocytosis in an Adolescent
Authors: Walia Ritika , Sepulveda Theresa , Wretzel Sharon , Brandt Philip H 
Published: November 2019

Objectives: Primary myelofibrosis is rare in pediatrics, often manifesting as persistent idiopathic thrombocytosis.Transitions from pediatric to adult medical care can be complicated by workup requiring invasive procedures.
J.M., an 18-year-old healthy male, presented for excessive gingival bleeding after wisdom tooth extraction. Workup revealed persistent thrombocytosis to 1,165K, prompting a referral to hematology-oncology. A peripheral smear was notable for many platelets but normal RBC morphology. He had splenomegaly on abdominal ultrasound and a decreased von-Willebrand's activity to antigen ratio, suggesting acquired vWD. A bone marrow biopsy was advised; however, J.M. became lost to follow up for over 9 months owing to self-reported anxiety about the procedure. He remained asymptomatic in this interim until he re-presented to clinic for easy bruising, with no other evidence of bleeding at the time. The biopsy was pursued, revealing hypercellular marrow for age with left shifted granulocytic and erythroid maturation, abnormal megakaryocytes, and 3% blasts. This was consistent with primary early myelofibrosis (PMF), positive for MF-1, CALR, and TP53 mutations and negative for JAK2 and BCR-ABL. He was transitioned to adult hematology, maintained on baby aspirin, and referred for potential allogeneic hematopoietic stem cell transplant (HSCT).
PMF is characterized by marrow fibrosis due to secretion of fibroblast growth factor by clonally proliferative megakaryocytes. It is a disease of adulthood, with 67 years being the median age at diagnosis. Only 100 cases have been reported in children, most of which are secondary to AML, ALL or other malignancies.1 Most patients present with complications of extramedullary hematopoiesis or bleeding.2 Diagnosis is suggested by a leukoerythroblastic picture on peripheral smear and confirmed with a bone marrow biopsy "dry tap" revealing marrow fibrosis.3 Prognosis in pediatric PMF is difficult to predict but outcomes tend to be worse;4 TP53 mutation is rare and based on limited adult studies may portend a poorer prognosis.5 Our young patient with this rare mutation was therefore referred for HSCT evaluation. Further complicating this case was J.M.'s anxiety, which delayed definitive diagnosis by biopsy. He only agreed to it when, at the med-peds clinic, the concept of local pain management was discussed. Anticipation of upcoming procedures by primary care physicians and close follow-up is especially important for patients transitioning from pediatric to adult providers.

Disclosures
No relevant conflicts of interest to declare.
A Comparison of the Effect of Granulocyte-Colony Stimulating Factor (G-CSF) in Chemotherapy Versus Plerixafor for Hematopoietic Stem Sell Transplantation (HSCT) and Mobilization
Authors: Piazera Flavia zattar , Nonino Alexandre , Vasconcelos Rafael De Sá , Cunha Andre Luiz Soares , Pinto Jorge Vaz , Profiro Mariana 
Published: November 2019

INTRODUCTION: Peripheral Hematopoietic Stem Cells have been widely used as a source for Autologous Hematopoietic Stem Cell Transplantation (HSCT). The use of G-CSF associated with chemotherapy increases the number of circulating Hematopoietic Progenitor Cells; however, there is no harmony about the best medical strategy for the use of G-CSF associated with the mobilization conduct. The use of plerixafor for poorly mobilized patients has provided a safe and effective option for optimizing apheresis procedures despite the high cost. Thus, the evaluation of efficacy becomes essential for the incorporation of plerixafor into the HSCT routine. OBJECTIVE: To evaluate the mobilization of autologous HSCT in chemotherapy associated with G-CSF and cyclophosphamide (QT) versus plerixafor. METHODS: Retrospective evaluation of 66 patients mobilized for autologous HSCT with G-CSF associated with chemotherapy versus plerixafor (plerixafor was used in the first mobilization), also to evaluate the number of apheresis procedures, CD34-positive cells, cell viability and for the bone marrow engraftment date. Statistical analyses were performed with GraphPad Prism 5.0 (GraphPad Software, Inc., San Diego, CA), using the Mann-Whitney test for nonparametric data (p <0.05). RESULTS: A mobilization protocol was performed in 83 patients (60% male), with a median age of 55 years (17 to 68 years). Multiple myeloma (55.7%), Hodgkin's lymphoma (14.7%), Non-Hodgkin's lymphoma (16.39%), primary amyloidosis (4.9%) were the most common hematologic malignancies. There was no statistical difference between G-CSF and QT group versus plerixafor groups regarding age, sex, type of disease and previous QT. The plerixafor group had better CD34 positive cell counts compared to G-CSF and QT (median 6.2 versus 3.8 x106, p<0.004), shorter sessions of apheresis procedure (median 2 versus 4, p < 0.0002) and a better cell viability rate (98 versus 95%, p <0.0034). The bone marrow engraftment rate was 10 days for plerixafor group versus 15.05 days for G-CSF and QT group, p <0.0055 (Figure 1).CONCLUSION: Our results showed the mobilization group using plerixafor had superior data and statistical significance regarding the number of apheresis procedures, cell viability, number of CD34 positive stem cells and earlier bone marrow engraftment rates when compared to the G-CSF + QT. These findings suggest and encourage the use of plerixafor in the first mobilization, as well as it should be incorporated into the standard routine. In addition, the number of apheresis procedures will be reduced, cell viability and CD34-positive cells rate will be increased. As a result, there will be a better medullary recovery and the reducing of both hospital costs and infectious complication rates.

Disclosures
No relevant conflicts of interest to declare.
Clinical Characteristic and Survival Outcome of Peripheral T Cell Lymphoma in a Chinese Population
Authors: Li Wenyu , Huang Ling , Jiang Xinmiao , Wei Xiaojuan , Guo Hanguo , Sichu Liu , Feili Chen , Liang Zhanli 
Published: November 2019

Background: Peripheral T-cell lymphoma (PTCL) is a heterogeneous group comprising about 10-15% of non-Hodgkin lymphomas in Western populations. Only a minority (~30%) of patients with the most common PTCL subtypes achieve a durable remission and long-term survival. Data on the clinical features of PTCL in a Chinese population are limited.
Methods: We retrospectively reviewed 342 patients with pathologically diagnosed T cell lymphoma at Guangdong Provincial People's Hospital from June 2007 through March 2019. We evaluated the incidence of PTCL subtypes, clinical characteristics and survival status.
Results: Of the 342 patients, 17.3% (n=59) had PTCL-not otherwise specified(NOS), 9.4% (n=32) had angioimmunoblastic T-cell lymphoma(AITL), 19.6% (n=67) had anaplastic large cell lymphoma(ALCL) (anaplastic lymphoma kinase positive ALCL:12.0%, n=41; anaplastic lymphoma kinase negative ALCL: 9.6%, n=26), 36.2%(N=124) had NK/T cell lymphoma, 12.9% (n=44)had lymphoblastic lymphoma(LBL), and 4.6%(n=16) had other subtypes. For next data analysis, NK/T cell lymphoma and LBL were excluded. Finally, a total of 164 patients with integrally follow-up information were analyzed. The median age was 49.5 years (range 15-87) old and the median follow-up time was 15.8 months (range 0.5-124.3). Most patients were males (62.0%) with advanced stage disease (III/IV, 74.5%), and 51.0% of patients presented with B symptoms and 83.0% with no bulky disease. For the total population, the median progression-free survival(PFS) and overall survival(OS) was 6.5 months and 13.5 months, respectively. ALCL patients had better PFS and OS than PTCL-NOS and AITL (PFS: 51.2 vs. 5.8 vs.4.7 months, P = 0. 000; OS: not reached vs. 13.4 vs. 8.8 months, P = 0.000). There were no difference of PFS and OS between PTCL-NOS and AITL(P = 0. 550 and P = 0. 333, respectively).
Conclusions: ALCL has better outcome than other PTCL subtypes. However, no efficacy therapy has emerged for patients with relapsed/refractory PTCL and outcomes in this setting are still poor. Novel potential targets for PTCL, with particular focus on identifying markers of response and resistance need further investigation.

Disclosures
Li: Guangdong Province Hospital: Employment.
The Role of M2 Macrophage in Primary Immune Thrombocytopenia
Authors: Shao Xia , Wu Boting , Chen Pu , Zhan Yanxia , Li Feng , Hua Fanli , Sun Lihua , Cheng Yunfeng 
Published: November 2019

Background: Primary immune thrombocytopenia (ITP) is an acquired autoimmune hemorrhagic disorder, characterized by immune-mediated platelet destruction and impaired megakaryocyte maturation. Although impaired T cells have been implicated to participate in the pathogenesis of ITP, another immune cell signified as M2 macrophages has not been investigated properly in ITP patients. This study aimed to investigate the role of M2 macrophage subsets in primary immune thrombocytopenia (ITP).
Methods: Peripheral blood mononuclear cells (PBMC) from newly diagnosed ITP patients and healthy controls (HC) were isolated. M2-like macrophages (CD68+CD163+) and M2 macrophages (CX3CR1+CD163+) were measured by flow cytometry. The correlation between CD68+CD163+ cells and CX3CR1+CD163+ cells was also analyzed. The CX3CR1+cells were sorted by magnetic bead, CD68+CD163+ in PBMC of ITP patients and healthy controls were then isolated, and the proportion of m2-like macrophages before and after the sorting was analyzed. The PPAR gamma and arg-1 levels of mRNAs and proteins of CX3CR1+ M2 macrophages were examined by Real-time PCR and Western Blot, respectively.
Results: CX3CR1+CD163+M2 macrophages were positively correlated with CD68+ CD163+ M2-like macrophages in ITP patients (r = 0.54, p < 0.01). After magnetic bead separation, the proportion of CD68+CD163+ cells in CX3CR1+ cells was significantly increased (p = 0.02). Compared with HC, both the mRNA and protein levels of arg-1 of CX3CR1+ M2 macrophages were significantly increased in patients with ITP. The expression level of PPAR gamma protein was significantly increased in ITP than that of HC. However no statistical difference was detected at mRNA expression level, although it was numerically higher in ITP patients than in HC ( p = 0.19).
Conclusion: The peripheral CX3CR1+ M2 macrophage exercises similar phenotypes and functions of M2 macrophage. The remarkably increased expression of arg-1 at both transcription and protein levels and PPAR gamma at protein level of CX3CR1+M2 macrophages in ITP patients suggests potential immunomodulatory functions of these macrophage subsets during ITP pathogenesis. However, no significant change at mRNA level of PPAR gamma indicating that the increased PPAR gamma protein level might be caused by other mechanisms, such as after transcription abnormalities, which warrants further investigation.

Disclosures
No relevant conflicts of interest to declare.
Tumor Microenvironment Differs between Germinal Centre B-Cell and Non-Germinal Centre B-Cell like Diffuse Large B-Cell Lymphomas and Has Subtype-Specific Prognostic Impact on Survival
Authors: Autio Matias , Leivonen Suvi-Katri , Karjalainen-Lindsberg Marja-Liisa , Pellinen Teijo , Leppa Sirpa 
Published: November 2019

Introduction
Based on the cell of origin, diffuse large B-cell lymphoma (DLBCL) is divided into germinal center B-cell (GCB) and activated B-cell (ABC) like subtypes, which differ in their gene expression profiles and clinical presentation with the ABC DLBCLs showing a worse outcome in response to R-CHOP immunochemotherapy. However, composition of the tumor microenvironment (TME) of these molecular subtypes has not been characterized.
Methods
We used Hans algorithm to determine the molecular subtypes (GCB vs non-GCB) and multiplexed immunohistochemistry (mIHC) to characterize tumor infiltrating T-cell phenotypes, including cytotoxic T-cells (CTLs; CD8, Granzyme B, OX40, Ki67), T regulatory cells (Tregs; CD3, CD4, FoxP3), Th1 effector cells (CD3, CD4, TBET) and T-cell immune checkpoint (CD3, CD4, CD8, PD1, TIM3, LAG3) in 165 primary DLBCLs. The findings were correlated with the expression of human leukocyte antigens (HLA) I and II (beta-2 microglobulin (B2M), HLA-ABC and HLA-DR), and outcome of the patients treated with R-CHOP-like immunochemotherapy.
Results
In the whole cohort, 82 (50%) cases were classified as GCB and 83 (50%) as non-GCB DLBCLs. In the GCB subtype, cytotoxic T-cells were more often PD1+, and T-cells FoxP3+than in the non-GCB subtype (Figure 1A-B). Furthermore, GCB DLBCLs tended to be more commonly HLA-DR+(p=0.102). In the non-GCB DLBCLs in turn, HLA I positivity was more frequent (B2M, P=0.007; HLA-ABC, p=0.108), cytotoxic T-cells more often granzyme B+(Figure 1C), T-cells TBET+(p=0.018) and LAG3+TIM3+(p=0.033). A high proportion of granzyme B+cells (p=0.002), PD1+cells (p=0.02) and TIM3+CD4+T-cells (p=0.006) from all cells translated to adverse overall survival (OS) in the patients with non-GCB DLBCL, all independent of the IPI. In contrast, a high proportion of TIM3+cells (p=0.015), and FOXP3+TBET+T-cells (p=0.005) from all cells were associated with poor OS in the patients with GCB DLBCL, also independent of the IPI.
Conclusions
TME differs significantly between GCB and non-GCB DLBCLs and has subtype-specific prognostic impact on survival.
Figure 1

Disclosures
Leppa: Roche: Honoraria, Research Funding; Takeda: Honoraria, Membership on an entity's Board of Directors or advisory committees, Research Funding; Bayer: Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding; Janssen-Cilag: Research Funding; Novartis: Membership on an entity's Board of Directors or advisory committees.