bluuebunny/crossref_metadata_2025_split huggingface dataset:
MicroRNA Analysis in the Cerebrospinal Fluid and Blood Serum of Lymphoma Patients At Diagnosis and in Response to Therapy,
Authors: Pospisil Vit , Mocikova Heidi , Jankovska Milada , Markova Jana , Gaherova Lubica , Kozak Tomas , Stopka Tomas 
Published: October 2019

Abstract
Abstract 3663
MicroRNAs are non-coding RNAs (19–23nt) that negatively regulate expression by interference with the translation or RNA stability of up to hundreds of targets. Tumors including leukemias and lymphomas are associated with deregulated expression of microRNAs (Volinia et al. 2010). Several microRNAs including miR-17-92 cluster members and miR-155 are overexpressed in lymphomas and showed promise in predicting relapse (Fabbri M et al. 2011). Unlike longer species of RNA, microRNAs are preserved and detectable in human fluids including serum, plasma and cerebrospinal fluid (CSF) (Baraniskin A. et al. 2011). We investigated abundance of microRNAs in the CSF and sera and tested whether their levels are associated with diagnosis or treatment of lymphomas. We studied patients with primary central nervous system lymphomas (PCNSL) (N=3) and systemic lymphomas either with CNS dissemination (N=3) or without (N=11). Histological type was predominantly diffuse large B cell lymphoma - DLBCL (N=13). Sera of a control group were also used. 200 uL of CSF and sera (both cell free) were used for total RNA isolation followed by PCR and the data were adjusted to levels of control microRNAs (miR-let-7a or miR-24). Our data demonstrate that the microRNAs of the miR-17-92 cluster (miR-19a, miR-20a, miR-92a) and miR-106b-25 cluster (miR-106b, miR-25) and miR-155 are detectable (unlike miR-106a, miR-18a) in both the CSF and sera. Particularly, miR-17-92 and miR-106b-25 are increased in CSF of the PCNSL and the systemic lymphomas with CNS involvement compared with systemic lymphomas without CNS involvement. Conversely, in the sera, expression of miR-17-92 is increased in systemic lymphomas compared to PCNSL and the control group. Next, we investigated the levels of these microRNAs in a patient with systemic DLBCL with CNS dissemination during the treatment (R-CHOP alternating with RMPV) resulting in complete remission. Analysis of the CSF and also of the sera (at 9 different time points within three months) revealed a gradual decrease (∼6 fold) of the levels of miR-19a, miR-20a, miR-92a, miR-106b, and miR-25. However, the levels of these microRNAs in the sera at the stage of complete remission were still 2 fold higher compared to controls. We conclude that measurement of microRNAs in both CSF and sera represents novel sensitive tool, additional to other approaches, for tumor detection and/or for estimation of the therapy efficacy. (Grants: NT10310-3/2009, MPO FR-TI2/509, NPVII 2B06077, MSM 0021620806, LC 06044, SVV-2011-262507)

Disclosures:
No relevant conflicts of interest to declare.
CD11b Expression Is An Independent Adverse Prognostic Factor in Pediatric Acute Myeloid Leukemia Treated with Allogeneic Stem Cell Transplantation,
Authors: Albitar Maher , Alseraihy Amal , Ayas Mouhab , Al-Jefri Abdullah , Al-Ahmari Ali , Belgaumi Asim F , Walter Claudia Ulrike , Nounou Randa , Khalil Salem , Bakshi Nasir , Owaidah Tarek , El-Solh Hassan 
Published: October 2019

Abstract
Abstract 4092

Background:
The adhesion molecule CD11b, which associates with the beta2-integrin to form the Mac-1 complex, is expressed in monocytic leukemias as well as other myeloid leukemias. Its expression on the lekemic cells has been reported to correlate with more aggressive course in adult patients with AML. The clinical relevance of CD11b expression in pediatric AML is not known. More importantly, the clinical relevance of CD11b expression on pediatric AML when these patients are treated with allogeneic hematopoietic stem cell transplant (HSCT). We investigated whether HSCT can modify the expected adverse effects of CD11b expression in pediatric AML patients.

Methods:
Immunophenotype data of 62 pediatric patients with AML, all treated with allogeneic HSCT, was reviewed. The expression of the CD11b on the blast population as determined by flow cytometry was correlated with clinical data and outcome. The median age of patients was 8 years (range: 8 months–14 years) and included 47 (76%) patients transplanted in their first complete remission (CR1). Fourty six (74%) of all patients had intermediate-risk cytogenetics and the remaining (26%) had adverse-risk cytogenetic abnormalities.

Results:
Twenty five (40%) of all studied patients expressed CD11b on the surface of the blasts at diagnosis. Patients with CD11b expression had significantly shorter overall survival (P=0.038) with median survival of approximately 11 months while the median survival in the CD11b negative patients has not been reached. Similarly, CD11 positive patients had significantly shorter event free survival (EFS) (P=0.03). When we considered only patients treated with HSCT in CR1, the OS and EFS for the CD11b positive patients were both significantly shorter (P=0.04 and P=0.05, respectively). Multivariate analysis including CD11b expression and cytogenetic risk identified CD11b expression as an independent prognostic factor for survival and EFS, while cytogenetic-risk stratification became no longer a predictor.

Conclusion:
Our data suggests that the expression of CD11b in pediatric AML is a significant independent poor prognostic factor and HSCT does not change this trend. Most of the CD11b positive patients die in their first year after HSCT and the management of these patients should be modified to address the biological basis of this subset of AML that expresses CD11b adhesion molecule.

Disclosures:
No relevant conflicts of interest to declare.
Impact of High Dose Cyclophosphamide on the Outcome of Autologous Stem Cell Transplant in Patients with Newly Diagnosed Multiple Myeloma,
Authors: Bacon Wendi A. , Long Gwynn D. , Rizzieri David A. , Horwitz Mitchell E. , Chute John P. , Sullivan Keith M. , Yopp Amanda , Johns Angela , Chao Nelson J. , Gasparetto Cristina 
Published: October 2019

Abstract
Abstract 4127
In the age of novel targeted agents, autologous stem cell transplant (ASCT) remains the standard of care for younger patients with newly diagnosed multiple myeloma (MM), offering similar treatment responses and overall survivals as standard chemotherapeutic agents but with the added benefit of a prolonged treatment-free period. Nevertheless, a standard of care for stem cell mobilization for ASCT has yet to be determined. Even in the era of new mobilization agents such as Plerixafor, Cyclophosphamide (Cy) and G-CSF combination remains the preferred mobilizing approach for patients with MM. Several studies have shown that Cy improves the stem cell yield at the expense of increased toxicity, but whether the administration of this chemotherapeutic agent pre-transplant has any impact on the long-term event-free and/or overall survival of myeloma patients remains controversial.
In this study, we present a retrospective analysis of 186 patients with newly diagnosed MM who underwent ASCT with high-dose melphalan 200 mg/m2 (HDM) between December of 2000 and 2008 at our Institution. Eighty-three patients were mobilized with single agent G-CSF and 103 patients received high dose Cy (4 gm/m2) and G-CSF combination. Patient characteristics were similar between the treatment groups, including: age, gender, disease stage, and disease status prior to transplant. However, toxicity post-mobilization with Cy/G-CSF was significantly higher compared with G-SCF alone, including: febrile neutropenia (23%), hemorrhagic cystitis (8%), GI toxicity (57%), re-hospitalization due to complications and transplant delay (14%).
The overall post-transplant toxicity was similar in the 2 groups, though the treatment related mortality was slightly higher in the Cy/G-CSF arm (4% versus 2%). Post transplant responses were not significantly different in the 2 groups, with 60% of patients achieving a VGPR or better after ASCT in the G-CSF group and 49% in the Cy/G-CSF group (p = 0.33). The median event-free survivals (EFS) for the Cy/G-CSF and G-CSF cohorts were 21.6 and 22.6 months, respectively, (p = 0.62) yielding no significant difference (Figure 1). Similarly, with a median follow up for surviving patients of 34.3 and 32.7 months, the median overall survivals were 68.2 and 62.3 months (p = 0.23) for the Cy/G-CSF and G-CSF cohorts, respectively (Figure 2). This retrospective analysis confirms that the addition of high dose Cy as part of the mobilizing regimen offers no improvement on the transplant outcome for patients with newly diagnosed myeloma and should therefore only be used in cases of difficult stem cell mobilization.

Disclosures:
No relevant conflicts of interest to declare.
Hematopoietic Cell Transplantation for FA Patients with Acute Leukemia or Advanced MDS,
Authors: MacMillan Margaret L. , Defor Todd E. , Wagner John E. 
Published: October 2019

Abstract
Abstract 4146
Fanconi anemia (FA) patients with acute leukemia or advanced myelodysplastic syndrome (MDS with ≥ 5% blasts) have a poor prognosis and hematopoietic cell transplantation (HCT) experience is limited. We report on the outcomes of 21 patients with FA and acute myeloid leukemia (AML, n=16), acute lymphocytic leukemia (ALL, n=2) or advanced MDS (n=3) who underwent allogeneic HCT at the University of Minnesota from 1988–2009. Five patients had biallelic BRCA2 mutations. Only 2 of 7 patients who received chemotherapy before HCT achieved remission both of whom had biallelic BRCA-2 mutations and T-cell ALL. Median age was 15.5 (range 1.1–48.5) years. Median age for BRCA2 patients was substantially lower at 1.9 (range 1.4–6.6) years. Patients received cyclophosphamide, antithymocyte globulin and either single fraction total body irradiation (n=15) or busulfan (n=6), with (n=11) or without (n=10) fludarabine (FLU) followed by HLA-matched sibling (n=3) or alternative (n=18) donor stem cells consisting of bone marrow (BM, n=14), single (n=4) or double (n=3) umbilical cord blood (UCB). Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine A and methylprednisolone. Probability of neutrophil engraftment was 75% without FLU and 100% with FLU based regimens. Probability of acute GVHD was 19%. For the entire cohort, probability of survival at 1 year was 43%. A higher rate of survival was observed in the 11 patients who received a FLU based regimens (50%; figure) and in the 5 patients with BRCA2 mutations (60%). Probability of relapse was 21% in these 11 patients.

These data suggest that HCT with a FLU based preparative regimen offers potential curative therapy for FA patients with acute leukemia or advanced MDS. Except perhaps for BRCA-2 patients with T cell ALL, there is no clear role for chemotherapy prior to HCT in FA patients with AML or MDS.

Disclosures:
No relevant conflicts of interest to declare.
Unrelated Donor Hematopoietic Stem Cell Transplantation for the Treatment of Advanced Multiple Myeloma: The John Theurer Cancer Center Experience,
Authors: Donato Michele L. , Vesole David H. , Siegel David S. , Adler-Brecher Barbara , Chichili Eiswarya , Pecora Andrew L , McKiernan Phyllis , Baker Melissa , Nyirenda Themba , Bilotti Elizabeth , Goy Andre , Rowley Scott D 
Published: October 2019

Abstract
Abstract 4162
Allogeneic hematopoietic stem cell transplantation has been associated with a graft-versus-myeloma effect and may be the only potentially curative option for patients with advanced multiple myeloma. Unfortunately, the majority of patients (pts) do not have an HLA-identical sibling. Unrelated donor transplantation is an alternative option for these patients. We conducted a retrospective analysis of patients with advanced multiple myeloma who received an unrelated donor (URD) hematopoietic stem cell transplant at our institution between 11/1997 and 11/2010. Analyses were done using Kaplan Meier survival and Cox regression on the SAS version 9.2 software.
Between 11/1997 and 11/2010, 27 pts with multiple myeloma received an URD transplant. One patient was lost to follow-up and 26 pts are evaluable. Patient characteristics included: median age 52 years (range 37–64), male 58%, all patients had prior autologous transplant (23% of pts had 2 or more prior transplants), and 43% of pts had refractory or progressive disease at the time of transplantation. Transplant regimens were ablative (defined as melphalan dose > 140mg/m2) in 73% of pts. GVHD prophylaxis was tacrolimus-based in all. Hematopoietic cell source was peripheral blood for 92% of pts, 46% of pts received a 10/10 HLA matched URD and 54% had a least one allele mismatch.

Results:
With a median follow up of 3.4 years, 54% of patients are alive and 47% of patients are progression-free. The treatment-related mortality at day 100 was 15%. The incidence of acute graft-versus-host disease (GVHD) was 44% and chronic GVHD 25%. Univariate analysis of risk of progression or relapse and overall survival failed to show any significance for disease status at the time of transplantation, degree of donor match, transplant regimen, refractoriness to bortezomib or lenalidomide, acute or chronic GVHD.

Conclusion:
In this analysis, URD transplantation has an acceptable treatment related mortality, and leads to durable remissions even in patients with relapsed and/or refractory disease. The 3.4 year PFS is similar to the BMT CTN 0102 sibling data and compares favorably to related donor transplantation for relapsed disease. In comparison to the BMT CTN study, the preparative regimen in this current analysis was overall more intense and regimen intensity has been previously shown to be significant. Allogeneic transplantation is an effective treatment strategy in multiple myeloma. Unrelated donor transplantation should be offered to patients with advanced multiple myeloma who do not have an HLA-identical sibling.

Disclosures:
No relevant conflicts of interest to declare.
Gene Transfer to Hematopoietic Stem/Progenitor Cells As a Novel Approach for Immunotherapy Against B-Lineage Malignancies: In Vivo Xenograft Model,
Authors: De Oliveira Satiro N. , Giannoni Francesca , Hardee Cinnamon , Sahaghian Arineh , Cooper Laurence J N , Kohn Donald B. 
Published: October 2019

Abstract
Abstract 4168
Chimeric Antigen Receptors (CAR) against CD19 have been shown to direct T cells to specifically target B-lineage malignant cells in animal models and clinical trials, with efficient tumor cell lysis. But, there has been insufficient persistence of effector cells, limiting the clinical efficacy. We propose gene transfer to hematopoietic stem/progenitor cells (HSPC) as a novel approach to ensure persistent production of effector cells targeting B-lineage malignant cells, exponentially increasing the number of effectors that may be generated against tumor cells. Experiments were performed using NOD-SCID-IL2 receptor gamma chain null (NSG) mice engrafted with human CD34+ HSPCs transduced with lentiviral vectors carrying first and second generations of CD19-specific CAR. There was efficient and stable transduction with 1–2 copies of CAR/cell as determined by qPCR. Differentiation of modified HSPC in vivo was not impaired by gene transfer, as observed in vitro. Results of in vivo studies showed that CAR-transduced human HSPC successfully differentiated into all lineages, with CAR-expressing T, NK and myeloid cells populating bone marrow, spleen and peripheral blood. The human CD19+ B cell populations normally formed in the xenografted NSG mice were significantly reduced when the transplanted HSPC were transduced with the anti-CD19 CAR, demonstrating in vivo biological activity. Cells harvested from bone marrow and spleen of mice engrafted with modified HSPC lysed CD19-positive cell targets ex vivo. Leukemic challenges of engrafted mice are in progress. Our results provide evidence for the feasibility and efficacy of the modification of HSPC with CAR as a protocol for generation of effector cells for immunotherapy against B-lineage malignancies.

Disclosures:
No relevant conflicts of interest to declare.
ATP-Hydrolysing Treg in Patients with Acute Myeloid Leukemia
Authors: Schuler Patrick J , Fenton Moon , Hong Chang-Sook , Jackson Edwin K , Whiteside Theresa , Boyiadzis Michael 
Published: October 2019

Abstract
Abstract 4255
We have previously shown that the frequency and suppressor function of regulatory T cells (Treg) is increased in newly-diagnosed patients with acute myeloid leukemia (AML). Here, we show that CD4+CD39+ ATP-hydrolyzing T cells are involved in the production of immunosuppressive adenosine and that this mechanism of suppression characterizes Treg present in the blood and bone marrow of AML patients.
Peripheral blood and bone marrow samples were obtained from AML patients prior to any treatment (n=20) and healthy controls (NC, n=20). The frequency and phenotype as well as cytokine profiles of CD4+CD39+ T cells were determined using multicolor flow cytometry, real-time PCR and western blots. Adenosine production was measured by mass spectrometry. Co-cultures of CD4+CD39+ Treg with conventional T cells were tested for suppression of proliferation.
In NC and patients with AML, CD4+CD39+ Treg contained two subsets of ATP-hydrolyzing T cells in equal proportions (Fig.1): FOXP3+CD25+ and FOXP3negCD25neg.Both subsets were increased in frequency (p < 0.04) in AML relative to NC (Fig.2). In the presence of other immune cells or exosomes positive for CD73 (hydrolyzes AMP to adenosine) both subsets produced immunosuppressive adenosine. In co-cultures, CD4+CD39+FOXP3negCD25neg T cells converted to FOXP3+CD25+ T cells.
Our data suggest that Treg with the ability to produce adenosine are a major subset of suppressor cells in the blood and bone marrow of AML patients. Co-culture experiments indicate that the ability of these cells to suppress immune responses is regulated at the level of FOXP3 and CD25 expression by factors present in their microenvironment.

Disclosures:
No relevant conflicts of interest to declare.
The Natural Product Borrelidin Induces UPR and Apoptosis in AML Cell Lines
Authors: Jackson Leah , Bechler Shelby , Miller Justin , Brownell Amy , Garshott Danielle , Callaghan Michael , Fribley Andrew M 
Published: October 2019

Abstract
Abstract 4260
Acute Myelogenous Leukemia (AML) is the most common form of leukemia. Current therapies are intense and even those fortunate enough to achieve remission often relapse extending extremely poor prognoses to these patient. The most commonly used therapeutics, namely cytarabine aribinoside, the anthracyclines and etoposide, are decades old and target ubiquitous cellular processes. We have previously reported that small molecules and natural products that activate and exacerbate the unfolded protein response (UPR) can effectively and selectively induce cell death in a wide variety of solid tumor cells. We hypothesized that the UPR might be a viable new therapeutic target in AML and sought to determine whether or not the novel UPR-inducing natural product borrelidin might be used as such an agent. A luminescent proliferation assay performed with panel of four AML cell lines treated with the ER stress-inducing antibiotic tunicamycin (Tm) revealed that three of the cell lines displayed IC50 values between 0.47–2.5μ M, doses of Tm which are known to induce a low to moderate level of ER stress. We then repeated the experiment with the more general UPR-inducing natural product borrelidin, which has been shown to have potent anti-inflammatory properties in several murine assays in vivo. All four cell lines were sensitive to borrelidin, displaying IC50 values between 0.032–0.29 μ M. Time course assays performed with borrelidin revealed 4–20 fold increases in active caspase 3 and 7 indicating borrelidin-induced AML decreases in cell proliferation might be the result of apoptosis. Quantitative reverse-transcription real time PCR performed with mRNA isolated from two AML cell lines revealed an increase in the UPR-related transcripts CHOP, ATF4, and GADD34 and the cell death genes Noxa, Puma, DR5 and Bim confirming that borrelidin could induce the UPR and apoptosis in AML cells. Studies currently underway in our laboratory will determine the ability of borrelidin and other UPR-inducing agents to reduce leukemic burden in an in vivo xenograft model.

Disclosures:
No relevant conflicts of interest to declare.
MNPs-Fe3o4 Induce Leukemia Cell Apoptosis by Oxidative Stress
Authors: Wang Ying , Chen Bao-An , Xia Guo-hua , Jian Ziying , Li Zhi , Lin Yun 
Published: October 2019

Abstract
Abstract 4304
The present study evaluated on the safe concentration for normal monocyte cells whether the magnetic nanoparticles of Fe3O4 (MNPs-Fe3O4) could induce caspase3-dependent apoptosis in SKM1 cells and U937 cells by oxidative damage. The average sizes of the particles were carefully determined by transmission electron microscopy. The viability of the cells after exposed to different concentration MNPs-Fe3O4 for 12h, 24h, 48h and 72h were detected by trypan blue staining. Cell cycle after exposure for 72h was analysed by propidium iodide (PI) staining. Apoptosis of MNPs-Fe3O4 group were detected and determined by Annexin V-FITC/PI double staining, 5,5’,6,6’-tetrachloro-1, 1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) probe and Wright - Giemsa staining. ROS in SKM1 cells and U937 cells after exposure were determined by flow cytometry after dichlorofluoroescein diacetate (DCFH-DA) staining. The changes of caspase3, survivin and bcl-rambo in the cells treatment with MNPs-Fe3O4 with or wtihout trolox for 48 hours were detected by western blot analysis. The significant decreased of cell viability caused by 100 μ M MNPS-Fe3O4 exposure were observed in SKM1 cells and u937 cells, not in normal monocytes cells (p < 0.05), the exposure also induced the SKM1 cells and u937 cells G0/G1 arrest (p < 0.05). Annexin V / PI staining assay show that 100 μ M MNPs-Fe3O4 group appeared more apoptosic rate in the U937 and SKM1 cells than the control group (p < 0.05), and on the same exposed concentration the apoptotic bodies could be frequently found in the U937 and SKM1 cells, while not in normal monocyte cells by Wright - Giemsa staining. The mitochondrial membrane potential in the two kinds of cells significantly decreased after exposed 100 μ M MNPs-Fe3O4 for 24h (p < 0.05), while pretreated with antioxidants trolox, the change was allevated (p < 0.05). DCFH-DA assay show that reactive oxygen species (ROS) generation increased in 6h of exposure in SKM1 cells and U937 cells (p < 0.05). Our results also showed the particle induced caspase3-dependent apoptosis in the SKM1 cells and U937 cells, while did not in normal monocyte cells, and increasing expression of bcl-rambo and decreasing expression of survivin involve in the process. These results suggest that on the safe concentration for normal monocyte cells MNPs-Fe3O4 could induce caspase3-dependent apoptosis in SKM1 cells and U937 cells by oxidative damage, moreover, increasing expression of bcl-rambo and decreasing expression of survivin involve in the process.

Disclosures:
No relevant conflicts of interest to declare.