bluuebunny/crossref_metadata_2025_split huggingface dataset:
JQ1, a c-MYC Inhibitor Targets Hexokinase II and Enhances the Cytotoxic Effects of Chemotherapeutic Agents in Diffuse Large B-Cell Lymphoma (DLBCL)
Authors: Gu Juan J , Barth Matthew J. , Zhang Qunling , Mavis Cory , Hernandez-Ilizaliturri Francisco J. 
Published: October 2019

Abstract
Background: Results from thecollaborative trial in relapsed aggressive lymphoma (CORAL) study demonstrated a poor clinical among patients treated with second-line chemo-immunotherapy in the post-rituximab era, stressing the need to identify and/or optimize novel targeted agents. C-myc expression was found correlate with a poor clinical outcome in patients with newly diagnosed or relapsed/refractory DLBCL. Previously, we found that JQ1 overcomes rituximab-chemotherapy resistance in lymphoma pre-clinical models. JQ1 exhibited synergistic activity when combined with chemotherapy agents. We also demonstrated that hexokinase II was highly expressed in rituximab resistant cell lines (RRCL), and its expression was associated with a deregulation in the glucose metabolism and an increase in the apoptotic threshold leading to chemotherapy resistance. In our current work, we evaluated molecular studies dissecting the cellular pathways affected by JQ1.
Methods: A panel of rituximab-sensitive (RSCL) and RRCL cell lines were exposed to escalating doses of JQ1 (0-8μM) for 24, 48 and 72h. Changes in cell viability and cell cycle distribution were evaluated using the Presto Blue assay and flow cytometry respectively. IC50 was calculated by Graphpad Prism 6 software. Loss of mitochondrial membrane potential (∆ψm) following JQ1 exposure was assessed by DiOC6 staining. Changes in c-MYC, HKII, HKI, Voltage dependent anion channel protein (VDAC) and p21 expression levels were determined by western blotting. HKII was silencing using siRNA interference in RRCL (Raji-4RH) and changes in HKII, HKI, VDAC, c-MYC and p21 protein expression was determined. HKII knockdown or scramble Raji-4RH cells were exposed to different chemotherapy agents and JQ1; cell viability and mitochondrial membrane potential was accessed.
Result:JQ1 induced a dose-and time- dependent cell death in cell lines. In vitro exposure cells to JQ1 resulted in a loss of mitochondrial potential and induced G1 cell cycle arrest. In vitro exposure to JQ1 decreased c-MYC and HKII expression levels and induced of p21 expression. Complete silencing of HKII re-sensitized cell to chemotherapy (doxorubicin, vincristine and bortezomib). On the other hand, HKII knockout abrogated JQ1 anti-tumor activity.
Conclusion: Our data suggests that JQ1 had anti-tumor activity against rituximab-sensitive or resistant pre-clinical models. The inhibition of HKII following JQ1 exposure may contribute to lower the mitochondrial membrane potential and enhance the chemotherapeutic effect. Our finding highlights a better understanding in the molecular events triggered by JQ1 as a potential role in the treatment of B-cell malignancies.

Disclosures
No relevant conflicts of interest to declare.
PDGFR Inhibitor Imatinib May be a Potent Drug in Treating Essential Thrombocythemia
Authors: Li Xia Zhou , Mo Yang , Jie Yu Ye , Chong Beng H 
Published: October 2019

Abstract
Approximates 80% of essential thrombocythemia (ET) patients carrying JAK2, CALR, MPL mutations. However, there is still 20% of ET patients are negative for all three mutations. Our previous studies have demonstrated that platelet-derived growth factor (PDGF) and its receptor (PDGFR) has a potential effect in the regulation of hematopoiesis and megakaryopoiesis. Therefore, we speculated that PDGF/PDGFR may plays an important role in megakaryocyte/platelet related disease, such as ET. In this study, we found an increase of PDGF-BB and PDGFR expression in ET patients comparing to the normal persons. We then used the adjusted dosage of PDGF-BB (1.5ug/kg/d) to establish an ET mimic mouse model. We found that PDGF-BB can enhance platelet production in vivo significantly, while PDGFR inhibitor imatinib can block this effect. Bone marrow histology data also confirmed the megakaryopoiesis effect of PDGF, as shown by an increased number of megakaryocytes in PDGF-treated mouse. The addition of imatinib can block the effect, evident by a decrease number of megakaryocytes and an increase of apoptosis. Furthermore, we demonstrated that PDGF-BB activated theP-JAK2, the P-STAT3 and the P-AKT expression in the megakaryocytic cell line CHRF-288, while addition of imatinib can reduce the phosphorylation level of all three genes. Our results suggested that the overexpression of PDGF-BB and PDGFR may be important in the pathogenesis of ET, especially in the JAK2-mutation negative ET. The elevated PDGF-BB activates PDGFR with subsequently activation of the JAK2/ STAT3 and PI3K/AKT pathway. Imatinib may have an effect on treating ET via blocking PDGFR and the following JAK2/STAT3 and PI3K/AKT pathway. This study suggests that PDGF/PDGFR may be involved in the pathogenesis of essential thrombocythemia, and the expression of PDGF-BB and PDGFR is potential to be a diagnostic index of ET. Blockage of PDGFR by imatinib may be a new therapeutic target for ET.
Figure 1 Effect of PDGF-BB on Jak2/Stat3 and PI3K/Akt pathway of megakaryocytes. Figure 1. Effect of PDGF-BB on Jak2/Stat3 and PI3K/Akt pathway of megakaryocytes. Figure 2 The platelet numbers and the megakaryocytes of each group mice. Figure 2. The platelet numbers and the megakaryocytes of each group mice. Figure 3 Potential mechanisms for imatinib in treating essential thrombocythemia. Figure 3. Potential mechanisms for imatinib in treating essential thrombocythemia.

Disclosures
No relevant conflicts of interest to declare.
Azacitidine Use in a Low Income Country, Effectiveness of a 100mg Fixed Dose
Authors: Torres Jorge Carlos , Zapata Nidia , Cervera Eduardo , Sanchez Sergio , Aguilar Manuel 
Published: October 2019

Abstract
Azacitidine, a pyrimidine nucleoside analog of cytidine, causes hypomethilation of DNA. Currently FDA approved for treatment of low and intermediate MDS with complete responses around 50%. And Acute myeloid leukemia (AML) in the eldery
In the CALGB studies, the usual dose is 75mg/m2 in 28 day cycles, with dose modifications according to toxicity.
In low income countries such as Mexico, one course of Azacitidine is around 500 dollars, median income in Mexico is 4,910 PPP (purchasing power parity); vs 30,616 in the USA. So, azacitidine treatment is far from reach for most of the common population, particularly those who do not have insurance.
This is a retrospective observational study, of a compassionate use program of a fixed dose of Aza at 100mg. We analyzed data from patients that were treated with Aza between 2012 and 2016, and collected data in 2016. The aim of the study was to assess the effectivity of the fixed dose. For that purpose, we collected information from the physical and electronic file. We analyzed: Hemoglobin level before and after treatment, independence of transfusion, ANC recovery, number of courses, and overall survival. We conducted our research in a public institution in Mexico (Instituto Nacional de Cancerología) and a private institution (Medica Sur).
We included acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS) patients, regardless of age, previous treatment and comorbidities (we included patients with renal failure, hypertension etc.)
We included 8 patients in our study, 6 males and 2 females, with a mean age of 69.9 years (49-87). We had 2 AML and 6 MDS.
We had 2 high risk AML and according to IPSS-R: 1 very low, 2 low, 1 intermediate, 1 high and 1 very high risk MDS. As for the Karyotype we had 1 complex KT, 4 normal KT and 1 Del 7q Del 5q +8.
All patients received at least one dose of Aza, with mean number of cycles of 4. We have a mean survival of 439 days (110-1385). 6/8 patients achieved transfusion independency within 3 doses of Aza. 6/8 patients achieved ANC but lost eventually lost response. 5/8 patients are alive in follow up. 3 patients died of infectious complications. 2 patients never achieved transfusion independence or ANC.
The information recovered suggests that a fixed dose of 100mg is as feasible as a higher dose, at least when no other treatment or higher dose can be administered. We still are analyzing the survival data in order to find other bad prognosis factor within this population.

Disclosures
No relevant conflicts of interest to declare.
Ibrutinib-Based Therapy in the Treatment of Relapsed and Refractory Chronic Lymphocytic Leukemia
Authors: Kuvshinov Alexey , Voloshin Sergei , Martynkevich Irina , Martynenko Ludmila , Garifullin Andrei , Kleina Elizaveta , Shuvaev Vasily 
Published: October 2019

Abstract
Introduction . The development of Bruton's tyrosine kinase inhibitors and their introduction into clinical practice represent a major advance in the treatment of chronic lymphocytic leukemia (CLL). Several studies in CLL have determined the achievement of minimal residual disease (MRD) negativity as an independent favorable prognostic factor, associated with superior progression-free survival (PFS) and overall survival. MRD status is the single best posttreatment predictor of long-term outcomes after treatment, regardless of the treatment protocol or the presence of other pre-existing prognostic indicators.
Aim.To estimate of response rate, PFS and MRD after ibrutinib-based therapy in the treatment patients with relapsed/refractory CLL.
M ethods.21 pts with relapsed/refractory CLL were included in the analysis. Stratification of patients into groups based on therapy. Group 1 (n = 14): 2nd and subsequent lines of rituximab-based chemotherapy (RB - 10, FCR - 4) and Group 2 (n = 12): ibrutinib-based therapy (420 mg daily oral Ibrutinib ± Chemoimmunotherapy). We have used NCI-IWCLL revised guidelines for treatment initiation and assessment of response. MRD was detected by multicolor flow cytometry of bone marrow in patients achieved a complete or partial remission: Group 1 - 7 pts, Group 2 - 5 pts.
Results. The median age was 60.5 years (45-83) in Group 1 and 62.5 years (49-82) in Group 2 and median previous lines of therapy was 1 (1-4) and 2 (1-4), respectively. Patients with unfavorable chromosomal abnormalities were detected in each group: Group 1 - 2 pts (combination del(11q) with del(13q)); Group 2 - 1 pts (del(17p)).
Overall response rate (ORR) in Group 1 was 71.4% (complete remission (CR) - 1 pt, partial remission (PR) - 9 pts, stable disease (SD) - 3 pts, progression disease - 1 pt). Group 2: ORR 91.7% (CR - 3 pts, PR - 8 pts; SD - 1 pt). Statistically significant differences in the frequency of ORR between groups were not detected (p>0.05).
MRD-negative remission rate was 40% (2/5, CR - 1 pt) in Group 2 compared to 14.3% (1/7 pts, CR) in Group 1 (p>0.05). Statistically significant differences in PFS were detected between Group 1 vs. Group 2 (p=0.0006). Median PFS in Group 2 has not been reached. Median PFS in Group 1 was 16.9 month.
Conclusion. Ibrutinib is highly effective at controlling disease, but best responses are typically partial remission, and patients must remain on treatment to maintain disease control. Evaluation of response and MRD after ibrutinib-containing therapy in the treatment of patients with relapsed/refractory chronic lymphocytic leukemia require further research.

Disclosures
Shuvaev: Pfizer: Honoraria; BMS: Honoraria; Novartis pharma: Honoraria.
Alterations in Circulating Lymphoid Populations after Post-Transplant Cyclophosphamide Administration in Haploidentical Hemopoietic Cell Transplantation
Authors: Shahin Kifah , Mattar Zamil , Bradstock Kenneth 
Published: October 2019

Abstract
High dose post-transplant cyclophosphamide (PTCy) administered early after stem cell infusion has facilitated the use of haploidentical related donors for allogeneic transplantation, but the precise immunosuppressive action of PTCy remains unclear. Using flow cytometry of peripheral blood leucocytes, we investigated changes in circulating lymphoid populations immediately before and after administration of PTCy in 10 patients undergoing haploidentical allogeneic transplantation after reduced intensity conditioning therapy. Compared to baseline levels prior to the first dose of PTCy, there were significant reductions in natural killer cell and monocyte numbers by Day 7 post-transplant, but no changes in T cell numbers. However, by Day 20 there was a significant reduction in CD8+ T effector cells, with significant increases in CD4+ T regulatory cells. The activation marker CD69 was detectable on CD8+ cells, particularly T effectors, at Day 3, but decreased significantly by the third week post-transplant. These changes were confirmed on donor T cells in 2 informative cases using HLA allotype-specific antibodies. Although other factors may have contributed, the reductions in numbers and levels of activation of effector T cells, and increases in regulatory T cells, are consistent with alterations in the immune response induced by PTCy after haploidentical transplant.

Disclosures
Bradstock: DendroCyte BioTech Ltd: Other: Laboratory IP contracted via ANZAC Research Institute to DendroCyte BioTech Ltd.
The Checkpoint for Agonist Selection Precedes Conventional Selection in Human Thymus
Authors: Verstichel Greet , Vermijlen David , Martens Liesbet , Goetgeluk Glenn , Saeys Yvan , Hilde Cheroutre , Vandekerckhove Bart 
Published: October 2019

Abstract
The thymus plays a central role in self-tolerance by preventing strongly self-reactive thymocytes from accumulating as naïve T cell receptor (TCR) αβ+ T cells in the periphery. The elimination of auto-reactive T cells from the naïve pool is in part mediated by deletion during conventional negative selection. Alternatively, self-reactive thymocytes can also be positively selected in response to strong TCR signals during agonist selection and functionally differentiate to innate TCRαβ + T cells such as the CD8αα+ double negative (DN) T cells. How thymocytes discriminate between these opposite outcomes remains unclear.
We identified a novel agonist-selected PD-1+ CD8αα+ subset of mature CD8+ T cells in human thymus. Using the same markers a similar population was also identified in cord blood at about the same frequency as TCRγδ+ cells. This population expresses high levels of Helios, indicative of strong TCR engagement, and displays an effector phenotype associated with agonist selection. Indeed, PD-1+CD8αα+ T cells exhibit innate production of IFN-γ and an elevated T-bet to Eomes ratio typical of effector CD8 T cells. These cells are CD62L-, CXCR3+ and Hobit high suggesting that these cells leave the thymus and home to the tissues. Interestingly, in vitro CD3/TCR stimulation of sorted early post-β-selection thymocyte blasts uniquely gives rise to this innate subset, whereas small CD4+CD8+ double positive precursors fail to survive strong TCR signals. The generation of the innate subset seems to arise also in vivo from early post-β-selection thymocyte blasts as these two populations have an identical TCRα repertoire: ex vivo isolated PD-1+CD8αα+ thymocytes are skewed for early 3' TRAV and 5' TRAJ rearrangements compared to conventional CD8 T cells. A similar skewing was found in early post-β-selection thymocyte blasts. As TCRα rearrangements are terminated by TCR engagement of agonist selection, this is strong evidence for a precursor progeny relationship.
Together, we conclude that human CD8αα+ T cells are preferentially selected by strong TCR engagement on a subset of progenitors that express a full TCRαβ early on, leading to the generation of a post-selection T cell population with innate functional capacity and a markedly distinct TCR repertoire. These findings uncover the heterogeneity among DP precursors in their potential to survive strong selection signals and suggests that the decision making in the thymus to divert immature thymocytes to the agonist selection pathway occurs early before conventional selection of DP cells. We propose that progression through the immature thymic developmental program influences the outcome of TCR engagement with early post-β-selection thymocytes triggered by strong TCR signals preferentially giving rise to innate CD8αα+ T cells in humans.

Disclosures
No relevant conflicts of interest to declare.
Tumor Associated Myeloid Cells and their Contribution to Tumor Outcome
Authors: Merad Miriam 
Published: October 2019

My laboratory studies the mechanisms that control the development, homeostasis and function of tissue resident myeloid cells, which mostly consists of macrophages and dendritic cells (DC).
Macrophages are hematopoietic cells that perform tissue-specific functions critical for regulating and maintaining organ homeostasis. In contrast to most other hematopoietic cells, macrophages that reside in quiescent tissues originate from early hematopoietic precursors that take residence in tissues prior to birth. In this presentation, I will discuss how tissue cues control macrophage functional diversity to ensure tissue integrity. I will also discuss recent data from our laboratory showing that developmental diversity contributes to shaping macrophage functional specialization and that ontogenically distinct macrophages differently regulate tumor response to radiation therapy.
In contrast to macrophages, DC are constantly repopulated by DC restricted precursors that are recruited from the blood circulation. Our laboratory has contributed to the mapping of the regulatory network of DCs, and the identification of a lineage of DC, the CD103+ DC, which are specialized in the induction of CD8+ T-cell immunity. Here I will discuss recent data from the laboratory that revealed the critical role for tumor-associated CD103+ DC in tumor response to checkpoint blockade and argue for the need to study human CD103+ DC equivalent in clinical cancer immunotherapy.

Disclosures
No relevant conflicts of interest to declare.
Studies on Erythropoiesis
Authors: GOLDWASSER EUGENE , JACOBSON LEON O. , FRIED WALTER , PLZAK LOUIS F. 
Published: October 2019

Abstract
It has been shown that plasma from animals that have been injected with cobaltous chloride rapidly develops a high titer of erythropoietin.
The gross properities of the active material appear to be the same from cobalt-treated as from phenylhydrazine-treated animals.
Other metal ions and a complex ion have been studied as stimulants for erythropoietin formation; none was as effective as cobaltous ion with the exception of cobaltic hexamine.
Selenoid (Crescent) Bodies
Authors: CUADRA MANUEL 
Published: October 2019

Abstract
"Selenoid (Crescent) Bodies" are known in hematology as Cuerpos en Media Luna in Spanish and Corps en demi-lune in French.
An original method of staining selenoid bodies and one which is suitable for morphologic structural studies has been described. The method is also suitable for staining stromas of erythrocytes.
Selenoid bodies originate from erythrocytes and correspond to their stromas. Selenoid bodies can be produced in vitro by the spreading of blood on a slide. Two factors influence their formation: a mechanical factor by friction of the erythrocytes against the surface of the slide and a chemical essential factor represented by the lipids of the blood, increasing the fragility of the erythrocytes.
The number of selenoid bodies in any smear of blood is directly proportional to its contents in lipids, either from an exogenous source (alimentary lipemia) or from an endogenous origin.
Selenoid bodies have also been found in the dog, sheep, rabbit, guinea pig, rat and hen.
The Life Span of Red Cells in the Rat and the Mouse as Determined by Labeling with DFP32 in Vivo
Authors: VAN PUTTEN L. M. , Croon Fineke 
Published: October 2019

Abstract
The red cells of rats and mice were tagged in vivo by injection of diisopropylphosphorofluoridate (DFP), labeled with P32, and the disappearance of radioactivity from the circulating red cells was determined. From the data obtained, it is concluded that the disappearance of the red cells is linear with time and that the red cells in both rats and mice have a true life span without measurable random destruction.
The life span of the erythrocytes was found to be 60.0 ± 3.2 (S.D.) days in the rat and 40.7 ± 1.9 (S.D.) days in the mouse.
Especially in the mouse the determination of the red cell life span with DFP32 is a simple procedure.
Virological and Immunological Features of Long-Term Human Immunodeficiency Virus–Infected Individuals Who Have Remained Asymptomatic Compared With Those Who Have Progressed to Acquired Immunodeficiency Syndrome
Authors: Barker Edward , Mackewicz Carl E. , Reyes-Terán Gustavo , Sato Akihiko , Stranford Sharon A. , Fujimura Sue H. , Christopherson Cindy , Chang Sheng-Yung , Levy Jay A. 
Published: October 2019

Infection with the human immunodeficiency virus (HIV) leads to a decrease in CD4+ T cells and disease progression within a decade of seroconversion. However, a small group of infected people, despite being infected by HIV for 10 or more years, remain clinically asymptomatic and have stable CD4+ cell counts without taking antiretroviral medication. To determine why these individuals, known as long-term survivors (LTS), remain healthy, the hematological profiles, viral load and properties, HIV coreceptor genotype, and anti-HIV immune responses of these people were compared with those of individuals who have progressed to disease (Progressors) over the same time period. Unlike Progressors, LTS have a low circulating viral load and a low number of HIV-infected cells. These differences in the levels of the viral load were not associated with a dominant biologic viral phenotype, varying growth kinetics of the virus, mutation in the cellular CCR5 gene, or the presence of neutralizing antibodies. Importantly, the difference in viral load could be explained by the enhanced ability of CD8+ cells from LTS to suppress HIV replication.© 1998 by The American Society of Hematology.