bluuebunny/crossref_metadata_2025 huggingface dataset:
Co-Administration of Hydroxyurea and a Specific AKT2 Inhibitor Has Beneficial Effects on Acute Vaso-Occlusive Events and Survival in Sickle Cell Disease Mice
Authors: Li Jing , Barazia Andrew , Kim Kyungho , Cho Jaehyung
Published: October 2019

We recently demonstrated that the basal levels of AKT phosphorylation were significantly increased in neutrophils and platelets isolated from sickle cell disease (SCD) patients compared with those cells from healthy donors and that specific inhibition of AKT2 impaired neutrophil-endothelial cell and neutrophil-platelet interactions in venules of Berkeley (SCD) mice (Li et al. J Clin Invest 2014). Although hydroxyurea (HU), an inducer of fetal hemoglobin, is the main therapy for treatment of SCD, it is unclear whether the short-term treatment with HU has immediate benefits on acute vaso-occlusive events in SCD. Using real-time fluorescence intravital microscopy, we demonstrated that co-administration of HU (100 microg/g body weight, iv injection) and Akti XII (3 microg/g body weight, iv injection), an AKT2 inhibitor, efficiently reduced neutrophil adhesion and platelet-neutrophil aggregation in cremaster muscle venules of TNF-α-challenged Berkeley mice. Importantly, compared with HU or Akti XII treatment alone, treatment with both agents significantly improved blood flow rates and survival in the mice. Further, similar results were obtained in Berkeley mice challenged with hypoxia (8% oxygen for 3 hours) and subsequent reoxygenation (room air for 3 hours). As determined by histochemistry of cremaster muscle sections following intravital microscopy of Berkeley mice, the expression of endothelial E-selectin and intercellular adhesion molecule 1 (ICAM-1) was significantly decreased by treatment with either HU or Akti XII. Leukocyte transmigration in the lung section of Berkeley mice was also inhibited by either agent, and the inhibitory effect was potentiated by co-administration of HU and Akti XII. Using the plasma and isolated cells of Berkeley mice, we found that the level of plasma nitric oxide species (NOx) was significantly elevated by treatment with HU but not Akti XII and that AKT2 phosphorylation levels in activated neutrophils and platelets were reduced by treatment with Akti XII but not HU. Taken together, these results suggest that short-term treatment of Berkeley mice with either HU or Akti XII inhibits inflammatory conditions: treatment with HU significantly increases plasma NOx levels, treatment with Akti XII decreases AKT2 phosphorylation in neutrophils and platelets without affecting plasma NOx levels, and administration of either agent reduces the surface expression of ICAM-1 and E-selectin on activated endothelial cells. Thus, our results provide evidence that co-administration of HU and a specific AKT2 inhibitor has immediate benefits on acute vaso-occlusive events and survival in SCD mice exceeding those seen for single therapy.

Disclosures

No relevant conflicts of interest to declare.
The Association Between SERPINC1 C.883G>a and VT in the Chinese Population
Authors: Liu Jingdi , Tang Liang , Zeng Wei , Hu Yu , Liu Han
Published: October 2019

Background: Antithrombin(AT) is a major anticoagulation molecule in vivo and is encoded by the gene SERPINC1. AT plays a key role as an inhibitor of physiological haemostasis by inhibiting the procoagulation factors, especially the factor Xa and thrombin.

Objectives: To explore the variations of SERPINC1 gene associated with venous thrombosis in the Chinese population.

Methods: SERPINC1 gene sequencing was carried out. A case-control study involving 1335 patients diagnosed with VT and 1315 Age- and sex-matched control individuals without a history of thrombosis were further carried out. Furthermore, plasma AT activity, AT antigen, and thrombin generation tests (TGT) were performed to evaluate the influences of the mutations.

Results: Four different missense mutations were identified in an unreported hot spot region of SERPINC1. They were c.880C>T(p.Arg294Cys), c.881G>T(p.Arg294Leu), c.881G>A(p.Arg294His) and c.883G>A(p.Val295Met). All of the affected individuals were heterozygotes. In addition, c.883G>A was found to be a predominant mutation. In the case-control study, the mutation was proved to be a strong risk factor for venous thrombosis with an OR of 10.92(p<0.01, 95%CI 1.41-84.68). Functional assays showed that both the activities and antigens of plasma AT decreased mildly.

Conclusion: A hot spot mutation region of SERPINC1 gene was discovered. The predominant mutation of SERPINC1 c.883G>A is the most frequent cause of AT deficiency and is associated with an increased risk of venous thrombosis in the Chinese population.

Disclosures

No relevant conflicts of interest to declare.
Incidence of Catheter-Related Venous Thromboembolism Events in Acute Leukemia patients; A Comparative, Retrospective Study of the Safety of Peripherally-Inserted Vs Centrally-Inserted Central Venous Catheters
Authors: Refaei Mohammad , Fernandes Bruna , Brandwein Joseph , Goodyear M. Dawn , Wu Cynthia M
Published: October 2019

1. Background

Central venous catheters (CVCs) are a leading cause of upper extremity deep vein thrombosis (UE DVT). There is little data on patients with acute leukemia (AL). Long term CVCs are required for chemotherapy in AL. Concomitant severe thrombocytopenia makes anticoagulation for CVC related thrombosis a challenge. Incidence of UE DVT has been reported to be increased in those with peripherally inserted central venous catheters (PICC lines) vs those with centrally inserted lines.

2. Aims

Our objective is to compare the incidence rate of VTE in leukemia inpatients with a PICC vs centrally-inserted CVC.

3. Methods

We reviewed 420 charts for AL inpatients requiring a PICC line admitted to Hematology at the University of Alberta Hospital between 2003-2013. Baseline patient characteristics were recorded. All venous thromboembolic events were objectively confirmed on imaging studies. Incidence of catheter associated thrombosis was calculated.

4. Results

420 patients were identified. We present the preliminary results of the 337 patients that met our inclusion criteria, and received at least one PICC line insertion. 305 (90%) had AML, 144 (43%) were smokers, 126 (37.4%) had cardiovascular risk factor, and only 14 (4.2%) had previous VTE. Overall, there were 634 PICC line insertions, with the 5FR dual lumen being the most commonly used PICC line (80%). Out of the 634 insertions, there were 65 (10%) new ipsilateral upper extremity DVTs, 54 (83%) of which developed acutely (<1month), and 44 (68%) in thrombocytopenic patients (platelet<50). 7 (1.1%) and 15 (2.4%) patients developed recurrent and concurrent VTEs, respectively. There was an incidence of 1.85 DVT per 1000 catheter days.

5. Conclusions

The incidence rate of DVT in our AL patients is higher than predicted for a general cancer patient population. This data will be compared to a similar cohort of AL inpatients presently being reviewed, who received a centrally-inserted CVC. Updated results will be included accordingly. Determining factors that are associated with a lower risk of DVT in this high bleeding risk population will be important to optimize patient care.

Disclosures

Wu: Leopharma: Membership on an entity's Board of Directors or advisory committees; Pfizer Canada: Membership on an entity's Board of Directors or advisory committees.
Outcome of BONE Marrow Transplantation in Congenital Diskeratosis: Preliminary DATA from the European Group for BONE Marrow Transplantation (EBMT)
Authors: Fioredda Francesca , Korthof Elisabeth T , Iacobelli Simona , Knol Cora , Veelken Joan H , Veys Paul , Yoshimi Ayami , Mellegren Karin , Fagioli Franca , Zecca Marco , Maschan Alexey , O'Brien Tracey , Diaz Miguel Angel , Smith Owen , Socie Gerard , Josue de la Fuente , Or Reuven , Dufour Carlo
Published: October 2019

Hematopoietic Stem cell Transplantation (HSCT) is the definitive treatment for bone marrow failure in diskeratosis congenita (DC). Organ dysfunction (mainly lung and gastrointestinal tract) are often part of the disease and may be a limiting factor for or may affect the final outcome of HSCT. Scarce data or relatively small cohort studies are available in current literature on outcome of HSCT in this disease. We analyzed the outcome of 87 patients diagnosed with DC reported in the data base of the European Society for Blood an Bone Marrow Transplantation (EBMT) who underwent HSCT from 1979 to 2014. In particular we analyzed data on type of HSCT, characteristic of donors, source of cells, incidence of acute and chronic GvHD. Males were 76%. Median age at diagnosis of DC was 5.7 years (0-33 yrs), median age at HSCT was 11.2 years (range 0.56-41 yrs). Median time from diagnosis to HSCT was 19 months (0.62-304 mo). The cell source was bone marrow (BM) in 67%, peripheral blood (PB) in 22% and cord blood (CB) 11%. Twenty six percent of patients were engrafted from a matched related and 53% from a matched unrelated donor. Twenty one percent of subjects were engrafted from a mismatched (both related and unrelated) HSCT. Engraftment was documented in 95% of subjects: 4 % had primary graft failure and 10% lost the graft. Overall 35 patients (40%) died, and 52 were alive (60%) at last follow-up. Causes of death were: infections (43%), multi-organ failure (26%), rejection/oss of graft (14%), GvDH (8.5%) undefined (8.5%). In 32% of cases death was related to transplant. Lung injury was present in 17% of subjects who died. Acute GVDH (mainly grade 2-4) and chronic GvHD occurred in 39% and 30% of cases. Five year OS was 60 %. OS was not significantly different by calendar period (before and after year 2000: 50% vs 67% respectively, p=0.424), by age at transplant (below and above age of 12 years: 64% vs 55% respectively ; p= 0.564) and by source of cells (marrow 60%, cord blood 68%, and peripheral cells 46%; p= 0.687). Conversely, OS in HSCT from mismatched donor was inferior to that of transplants from matched donor (38% vs 65% respectively; p= 0.045). The use of radiotherapy or busulfan (any dose) in the conditioning vs other regimens did not impact on 5 y-OS ( 58% vs 62% respectively; p=0.898). HSCT from HLA matched donor can be considered a treatment option in DC. Transplant related mortality looks rater high. Multi organ failure and lung injury are amongst the main causes of death thus pointing to pre-transplant organ status as important determinant of HSCT outcome.

Disclosures

Dufour: Pfizer: Consultancy.
Identification of Novel Drug Targets in T-Cell Acute Lymphoblastic Leukemia
Authors: Laukkanen Saara , Liuksiala Thomas , Nykter Matti , Heinäniemi Merja , Lohi Olli
Published: October 2019

Acute lymphoblastic leukemia (ALL) is the most common childhood cancer and one of the leading causes of death in children with tumors. Genetic changes in T-cell acute lymphoblastic leukemia (T-ALL) are relatively well known, but the underlying molecular processes driving the disease remain insufficiently understood. Better knowledge of molecular events in T-ALL would improve our understanding of the development and maintenance of the disease and could also lead to the development of targeted and more effective treatments.

We have compiled a large gene expression data-set from microarray studies of various hematological and lymphoid malignancies and healthy tissues that use a uniform technical platform (Affymetrix HG U133 Plus 2.0) (see abstract by Liuksiala et al). Data-set includes 1302 healthy samples and 4418 leukemia samples: 1713 acute myeloid leukemias, 1648 precursor B-ALLs, 801 chronic lymphocytic leukemias, 385 T-ALLs, and 215 chronic myeloid leukemias. From this data-set, we identified a number of transcription factors (TFs) that were differentially expressed in T-ALL, including high expression of NOTCH1 and BCL11B as previously reported (Weng et al Science 2004; Gutierrez et al Blood 2011). Several novel candidate TFs with specific expression in T-ALLs were also discovered, including strong expression of two poorly characterized TFs, SIX6 and PCBP3. These findings were validated using real-time quantitative PCR (RT-qPCR) in a cell line panel consisting of T-ALL and pre-B-ALL cell lines as well as healthy controls.

We next sought to identify novel drug targets in T-ALL by comparing our leukemia expression data-set with the therapeutic target database (TTD). TTD is a database providing information about the known and explored therapeutic protein and nucleic acid targets, and the corresponding drugs aimed at these targets. We identified high expression of a nicotinic acetylcholine receptor (nAChR) subunit CHRNA3 (cholinergic receptor, nicotinic, alpha 3), which is a target of nAChR inhibitor bupropion. RT-qPCR confirmed the high expression of CHRNA3 in T-ALL cell lines but not in pre-B-ALL cells or healthy controls. The effect of bupropion was tested in Jurkat cells which represent T-ALL cell line with high expression of CHRNA3. Increasing concentrations of bupropion (1-100µM) resulted in dose-dependent decrease in proliferation of Jurkat cells as measured by cell viability assay AlamarBlue. As a control, cell lines with low level of CHRNA3 expression (CCRF-CEM and REH) were treated as well but these cells did not show any changes in the rate of proliferation.

In summary, we have identified several candidate transcription factors which could have a leukemic role in T-ALL. Furthermore, we identified high expression of CHRNA3 in T-ALL, suggesting a role for the cholinergic system in T-cell leukemia, and thus a novel avenue in search of putative therapeutic options.

Disclosures

No relevant conflicts of interest to declare.
Varying Responses of PML-Rara with Different Genetic Mutations to Arsenic Trioxide
Authors: Liu Jiangying , Zhu Hong-Hu , Jiang Hao , Jiang Qian , Huang Xiao Jun
Published: October 2019

Resistance to arsenic and/or ATRA is a challenging problem in the clinical management of acute promyelocytic leukemia (APL). Acquired genetic mutations in the PML moiety of the PML-RARA fusion gene have been found to exist virtually in APL patients at relapse. While C212/C213 mutations in the PML B2 domain of PML-RARA were shown to be capable of abolishing the effect of arsenic on target proteins in vitro, A216V and L218P mutations in the PML portion of PML-RARA were detectable in two APL patients resistant to arsenic therapy. More recently, we identified a panel of point mutations (including previously reported A216V and, newly identified S214L, A216T, L217F and S220G) from nine of 13 APL patients with arsenic resistance. These findings indicated the existence of a mutational hotspot (S214-S220) within the PML-RARA oncoprotein among APL patients at relapse. Whether each of the identified point mutations plays a role and functions distinctly in the mechanism underlying the resistance to arsenic therapy is of particular interest.

Here, we performed in vitro functional analyses in both the hematopoietic irrelevant and relevant cells, and a retrospective analysis of APL patients to investigate the effect of PML-RARA mutations on mediating the resistance to arsenic trioxide. Among the five point mutations in the PML part of PML-RARA identified in patients with relapsed APL, we found that A216V, S214L and A216T mutations could attenuate the negative regulation of arsenic on PML-RARA, which resulted in the retention of oncoproteins. In contrast, L217F and S220G mutations function weakly in this context. Furthermore, we demonstrated that high concentration of As2O3could overcome the primary arsenic resistance driven by the indicated point mutations in PML-RARA. In addition to providing more evidence to reinforce the correlation of genetic mutations in PML-RARA with arsenic efficacy, we provide a novel insight into the functional difference of acquired mutations of PML-RARA both in vitro and in the clinical setting. Our findings may help predict the prognosis and select more effective strategies during APL therapy.

Disclosures

No relevant conflicts of interest to declare.
The Impact of Breastfeeding on the Childhood Acute Leukemia Risk Among Children Attending Alexandria University Children's Hospital
Authors: Massoud Mohamed NZ , Hassab Hoda MA , Ali Aida M M , Mohamed Doha N
Published: October 2019

Objective: The present study was conducted to investigate the impact of breastfeeding on the risk of development of acute leukemia among children Admitted to Alexandria University Children's Hospital.

METHODS: A case control study included 134 ALL&AML cases & 134 matched controls for age & sex from the same family relatives to evaluate socioeconomic & genetic causes of the disease. The mothers were interviewed for the completion of interview format that included: duration & patterns of breastfeeding.

RESULTS: lower proportion of acute leukemia cases 59.7% were exclusively breast-fed babies as compared to 89.6% of the control subjects. However, higher proportions of index children were either predominant breast-fed 19.4% or complementary-fed 10.4%. Nearly 10.4% were bottle-fed babies as compared to none of their controls. The differences between cases & controls as regards pattern of breast feeding were statistically significant where p<0.001. As regards duration of breastfeeding 75.4% of index children were breast-fed for more than 6 months as compared to vast majority of their controls (97.8%). The difference between cases & controls was statistically significant where p <0.001. The association also was statistically significant when 2 groups (standard & high risk) were considered as regards pattern of breastfeeding where x2=13.055, p =0.004. The multivariate logistic regression analysis indicated that bottle feeding had the odds of 7.76 of being at high risk level for acute leukemia (OR=7.76, 95% CI 1.9-33.8).

CONCLUSION: Exclusive breast-feeding and breast-feeding for one year are protective against acute childhood leukemia.

Table 1. Bivariate and multivariate analysis for pattern & duration of breastfeeding of the studied children with acute leukemia and their controls. Pattern and duration of breastfeeding Bi variant variable Multivariate variable Type of feeding during the first 6 month of life. Cases (n=134) Control (n=134) Significance OR (95% CI) Significance No. % No. % X2 = 37.6 P>0.0001* 1 Exclusive BF 80 59.7 120 89.6 Predominant BF 26 19.4 13 9.7 1.67 (1.26 - 2.2)* X2 = 9.4 P=0.002* Complementary feeding 14 10.4 1 0.7 21 (2.8 - 436.4)* X2 = 16.13 P>0.0001* Bottle feeding 14 10.4 0 0 2.5 (2.1 - 2.9)* X2 = 19.12 P>0.0001* Duration of breastfeeding 0-≤ 3 month 2 1.7 0 0 X2 =15.331 MC P=0.0001* AN- FEP=0.18 =13.06 P=0.0003* X2=0.38 P=0.537 3-≤ 6 month 17 14.2 3 2.2 7.6 (2.1 - 33.8)* 6-12 month 16 13.3 17 12.7 1.26 0.6-2.8 More than 12 month 85 70.8 114 85.1 1 - X2: Chi-Square test FEP: Fisher's Exact test -NA-: Not Applicable *significant at P≤0.0

Disclosures

No relevant conflicts of interest to declare.
Monoamine Oxidase a (MAO A) Is Expressed Selectively in Reed-Sternberg Cells of Classical Hodgkin Lymphoma
Authors: Li Pei-Chuan , Siddiqi Imran N , Loo Eric Y. , Kong Yinfei , Cozen Wendy , Wu Chieh-Hsi , Shih Jean Chen
Published: October 2019

Monoamine oxidase A (MAO A) is a mitochondrial enzyme, which catalyzes oxidative deamination of neurotransmitters or dietary amines and produces H2O2. Recent work indicates that MAO A is overexpressed in prostate cancer. Inhibiting its activity suppresses tumorigenesis and metastasis. In this study, we showed that Hodgkin lymphoma (HL)-derived L1236 cells express high MAO A activity, while other human lymphoma cell lines examined, including both Hodgkin and non-Hodgkin lymphoma, do not express MAO A. We also analyzed MAO A expression by immunohistochemistry in formalin-fixed paraffin-embedded classical Hodgkin (n=88), nodular lymphocyte predominant Hodgkin (n=8) and non-Hodgkin lymphoma (diffuse large B-cell lymphoma, n=54; follicular, n=33; Burkitt lymphoma, n= 19; mantle cell lymphoma, n= 13). We found that 60% of the classical Hodgkin lymphoma (cHL) cases demonstrated at least some MAO A expression by the neoplastic Reed-Sternberg cells and 30% of the tumors show strong/uniform expression. In contrast, no MAO A activity was seen in normal lymphoid tissue, nodular lymphocyte predominant Hodgkin lymphoma or non-Hodgkin lymphomas. The MAO inhibitor clorgyline inhibited MAO A activity (IC50 of 8.3×10-9 M) and was cytotoxic (IC50 of 30 μM) to L1236 cells. In addition, clorgyline reduced the cell proliferation, colony formation, and invasion of L1236 cells in vitro. Taken together, we have shown for the first time that MAO A is expressed selectively in Reed-Sternberg cells of cHL. Our results suggest that MAO A may be a biomarker for classical Hodgkin lymphoma Reed-Sternberg cells and may provide biological insights as well as a potential therapeutic target.

Disclosures

No relevant conflicts of interest to declare.
Smoothened (SMO) Is an Adaptor Protein That Recruits TRAF6 and Phospholipase C Gamma 2 (PLCg2) to Enhance the Activation of NF-Kb Signaling Pathway
Authors: Agarwal Nitin , Kim Chae Hwa , Kunkalla Kranthi , Yang Gloria , Tjendra Youley , Liu Yadong , Qu Changju , Vega Francisco
Published: October 2019

Background: Constitutive activation of NF-κB signaling is a hallmark of DLBCL. Activation of NF-κB is a multifactorial process resulting from oncogenic mutations (CARD11, MYD88…), chromosomal abnormalities, chronic activation of B-cell receptor signaling (BCR) as well as stimuli from the microenvironment. Chronic activation of BCR is not only the result of gene mutations (e.g.CD79B) but also is the result of stimuli generated from the lymphoma microenvironment.

We previously found that smoothened (SMO), transducer of hedgehog (Hh) signaling, enhanced NF-κB activation in lymphoma cells, independently of the presence of oncogenic mutations (Changju et al., Blood 2013). Hh ligands are provided by the lymphoma microenvironment (e.g. stromal cells) to activate SMO in the lymphoma cells. Activated SMO, recruits and activates trimeric-G-coupled proteins to activate PKCβ/CARMA1/TRAF6/NEMO axis, followed by assembling of the CARMA1/BCL10/MALT1/TRAF6 complex to SMO resulting in NF-κB activation. Now, we reveal an additional mechanism by which SMO further contributes to the activation of NF-κB in DLBCL.

Summary of results: We explored if recombinant Hh ligand can activate NF-κB signaling in the presence of the BTK inhibitor (PCI-32765). Hh stimulation resulted in phosphorylation of PLCg2 (downstream of BTK) and partially rescued the inhibitory effect of the BTK inhibitor on phosphorylation of PLCg2 suggesting a crosstalk between SMO and BCR receptor signaling. We identified that SMO forms a complex with TRAF-6 and PLCg2 using immunoprecipitation and Duolink in situ proximity ligation assays. We found that SMO stabilizes and protects TRAF-6 from proteosomal degradation. SMO-dependent TRAF6 stabilization is mediated by the ubiquitin-specific protease-8 (USP-8) by removing ubiquitin from K48-linked lysine of TRAF6.

Conclusions: Collectively, our data reveals multilayer crosstalk between Hh and BCR/NF-κB pathways and provide significant insights into how SMO contributes to chemotolerance and progression of DLBCL. We expect that our results will delineate novel molecular mechanisms involved in the pathobiology of DLBCL that may serve as therapeutic targets.

Disclosures

Vega: Seatle Genetics: Honoraria; NIH: Research Funding.
Increasing Use of Social Media at Annual ASH Meetings
Authors: Dahiya Saurabh , Kansagra Ankit J. , Ali Syed S.
Published: October 2019

Background: The use of social media is expanding in medicine. More and more healthcare professionals, patients and advocates are using social media as a common platform to enhance communication.

Methods: Each year American Society of Hematology (ASH) creates a hashtag (#) followed by year of the annual meeting (e.g. ASH 2014 had this hashtag: #ASH14) on Twitter. We conducted a retrospective study of annual meeting's hashtag use on Twitter for past 4 annual meetings using data from Symplur, LLC. Symplur is an online forum offering analytics of the global Twitter based conversations. Following hashtags were used for analysis, #ASH11, #ASH12, #ASH13 and #ASH14.

Results: Overall there is an increasing trend for use of annual meeting's hashtag (see table 1). Last year's meeting generated over 72 million impressions. Number of tweets being sent out is doubling every year. More twitter users are engaging in conversations than the year before using annual meeting's hashtag. Last year almost 4000 individual twitter accounts were used during the annual meeting. Number of tweets per participant has stayed relatively constant over past four years. @ash_hematology remains the most commonly mentioned twitter handle every year since 2011. In 2014 itself, @ash_hematology was mentioned over 2000 times.

Conclusion: Twitter is a very powerful tool that amplifies the content of scientific meetings. Use of twitter using meeting's hashtag is increasing every year at annual ASH meetings. This analysis provides a snapshot of twitter activity at the conference. Avenues for further research are: trend identification, "influencer" identification, and qualitative analysis. Interdisciplinary research should focus on evaluation methods that can assess the quality, value, and impact of tweeting.

Table 1. Year Impressions Tweets Participants Average tweet per participant 2011 12,255,646 3123 606 5 2012 18,153,786 5,094 870 6 2013 30,018,580 12,233 1942 6 2014 72,433,270 24,185 3977 6

Disclosures

No relevant conflicts of interest to declare.
Iron Deficiency Anemia Is Common in Patients Receiving Chemotherapy
Authors: Jajeh Ahmad
Published: October 2019

Background: Iron metabolism is impaired in patients with cancer and actively receiving chemotherapy. Patients with cancer on active chemotherapy suffer with decreased appetite , poor nutrition, mucositis, gastrointestinal mucosal damage, blood loss as well as anemia of chronic disease due to cytokines release such as Tumor Necrosis Factor, Interleukins and histamins.

The purpose of this abstract is to show single institution experience with patients actively receiving chemotherapy and admitted for various reasons and found to have anemia and particularly those patients that were admitted for red blood cell transfusion. Retrospective review of three hundred and fifty patients admitted in the last two years 2014-2015 for different reasons while on chemotherapy with anemia or mainly for blood transfusion. 55% females and 45% males. 77% had solid tumors and 23% with hematological malignancies. Median age 70 years. Thirty five percent 35% of solid tumors were of GI malignancies. Seventy five percenct of all patients had low iron saturation, low serum iron with normal to high total Iron Binding Capacity TIBC. Ferritin level was low in 60% of the patients. Ferritin is a phase reactive markers was elevated in 40% of patients. All patients with elevated ferritin were having low iron saturation with low serum iron and low or normal TIBC. Sixty five 65% of patients received blood transfusion. All patients with iron deficency received intravenous elemental iron support to saturate iron stores.

In conclusion: Iron deficiency anemia is very common in patient on active chemotherapy. With Intravenous elemental iron supplemet the requirement of less future red blood transfusion and improvement of hemoglobin levels were achieved in most of patients responding to chemotherapy. Need prospective analysis on patients diagnosed with malignancies and start receiving chemotherapy to prove the value of IV iron support and less requirement of red blood cell transfusion.

Disclosures

No relevant conflicts of interest to declare.
Reduced Glutamate Channel Phosphorylation in Pain-Related Brain Areas in a Transgenic Model of Sickle Cell Disease
Authors: LeBlanc Dana Marie , Towne Brook V , Paul Jinny , Gupta Kalpna , Edwards Scott
Published: October 2019

Sickle cell disease (SCD) is associated with chronic, debilitating pain beginning in childhood and persisting throughout life. SCD patients also tend to exhibit elevated rates of depression and anxiety disorders, indicating a possible connection between recurring pain episodes and affective or emotional dysregulation in this population. How SCD physiologically manifests within brain circuitry to mediate this link is currently unknown. In the current study, we utilized a transgenic mouse model of SCD to measure protein kinase A (PKA)-mediated phosphorylation of the AMPA glutamate receptor channel subunit GluR1 in specific brain regions associated with central nociception and the emotional dimension of pain, including the periaqueductal gray (PAG), prefrontal cortex (PFC), and hippocampus. Transgenic sickle mice (HbSS-BERK) expressing human sickle hemoglobin display significant mechanical and thermal hyperalgesia and exhibit reductions in GluR1 phosphorylation compared to littermate controls (HbAA-BERK) in the PAG, PFC, and hippocampus. This neuroadaptation is indicative of reduced excitatory transmission potential in these pain-associated areas. In comparison, GluR1 phosphorylation is unchanged in the dorsal striatum, a brain region not traditionally associated with pain or negative emotionality. Our data are consistent with other preclinical pain models that have linked chronic, peripheral pain conditions with compromised excitatory brain signaling. These results make a strong case for greater appreciation and investigation into mechanisms of possible affective disorder in individuals suffering from SCD.

Disclosures

No relevant conflicts of interest to declare.
Phase Ib Studies of Benegrastim in Chinese Women with Breast Cancer Receiving Myelotoxic Chemotherapy
Authors: Cao Junning , Yuan Zhongyu , Huang Cheng , Ji Dongmei , Peng Rujun , Yan Xiaoqiang , Tang Tom
Published: October 2019

Neutropenia is common in patients receiving myelotoxic chemotherapy. BenegrastimTM(F-627), a recombinant human G-CSF dimer, is an once-per-cycle therapy for prophylactic neutropenia in cancer patients after chemotherapy. Two phase I dose finding studies of benegrastim were conducted in Chinese women with stage I-IV breast cancer receiving myelotoxic chemotherapy. The aims of the studies were to evaluate the safety profile and pharmacokinetics (PK) and pharmacodynamics (PD) properties of benegrastim.

In the first study, a total of 18 patients were enrolled to receive 3 sequential dose levels of benegrastim at 80, 240 and 360 µg/kg (n=6). The patients received epirubicin/cyclophosphamide (EC) chemotherapy on day one. Benegrastim was administered to patients on day 3 by SC injection once per chemotherapy cycle for up to 4 cycles. In the second study, 15 patients were sequentially assigned to 240 µg/kg (n=7) and 320 µg/kg (n=8) of benegrastim. These patients received doxorubicin, docetaxel and cyclophosphamide (TAC) on day one, benegrastim was administered on day 2 by SC injection for up to 6 cycles. The PK/PD properties of benegrastim were evaluated in cycle one and cycle three.

Benegrastim was well tolerated in both clinical studies. The treatment emergent adverse events related to benegrastim were back pain, arthralgia, musculoskeletal pain, and rash, commonly seen in rhG-CSF therapy. The PK/PD result in patients receiving TAC in the first chemotherapy cycle is shown in Fig 1. Dose-dependent benegrastim exposure and ANC increase were demonstrated in these two clinical trials. In breast cancer patients receiving EC or TAC, benegrastim showed a non-linear pharmacokinetics. Benegrastim shortened the duration of neutropenia post chemotherapy in a dose dependent manner. In conclusion, benegrastim may provide an alternative approach to manage neutropenia, especially, severe neutropenia in patients after chemotherapy.

Figure 1. PK/PD of F-627 (benegrastim) in Chinese breast cancer patients receiving TAC in the first chemotherapy cycle. Figure 1. PK/PD of F-627 (benegrastim) in Chinese breast cancer patients receiving TAC in the first chemotherapy cycle.

Disclosures

Cao: Generon (Shanghai) Corporation Ltd.: Research Funding. Yuan:Generon (Shanghai) Corporation Ltd.: Research Funding. Huang:Generon (Shanghai) Corporation Ltd.: Employment. Ji:Generon (Shanghai) Corporation Ltd.: Research Funding. Peng:Generon (Shanghai) Corporation Ltd.: Research Funding. Yan:Generon (Shanghai) Corporation Ltd.: Employment. Tang:Generon (Shanghai) Corporation Ltd.: Employment.
DIC and Its Association with Acute Leukemia in Pre-Treatment and Post Remission Induction Phase
Authors: Naz Arshi , Farzana Tasneem , Taj Mehwish , Shamsi Tahir
Published: October 2019

Background:

Hematological malignancy like acute leukemia (AL) is associated with thromboembolic complications.DIC is a worst complication amongst subtypes of AL especially in APML which can be life threatening.

Study design:

Descriptive & cross-sectional study.

Place and duration of study:

National Institute of Blood Disease and Bone Marrow Transplantation; May 2011 to March 2012.

Patients and methods:

110 (75 males, 45 females)diagnosed cases of acute leukemia [43 cases of AML (27 newly diagnosed, 16 in remission induction), 67 cases of ALL (38 newly diagnosed; 29 in remission induction)]were included & 40 ascontrols.Mean age of patients was 25.3±13.8.Platelet count,PT, APTT, Fibrinogen levels, D-Dimer &FDP was done for scoring of DIC on day 0 and 28. SPSS version 17 was used for data analysis.

Results:

Platelet counts significantly improved on day 28 in AML and ALL. PT and APTT levels were significantly deranged. Plasma levels of fibrinogen were higher in both types of acute leukemia in pre-treatment phase, further increased at day 28 (<0.01). FDP significantly raised at day 0 reduced at day 28(AML & ALL). Markedly elevated levels of D-dimer in AML and ALL at day 0,but showed significant reduction at day 28(<0.01). DIC score of <5 was found in 15 (55.56%) patients of AML and 16 (100%) of ALL on day 0 and >5 score was recorded in 12(44.44%) patients of AML on day 28.

Conclusion:

Strong association of DIC was found in AML and ALL at day 0. Non overt DIC did not show any significant association with specific type of acute leukemia and it was equally expressed in both type of acute leukemia at day 0 and 28.

Disclosures

No relevant conflicts of interest to declare.
Clinical, Instrumental, Serological and Histological Findings Suggest That Hemophilia B May be Less Severe Than Hemophilia A
Authors: Castaman Giancarlo , Melchiorre Daniela , Linari Silvia
Published: October 2019

Background: Recent evidences suggest that patients with severe hemophilia B (HB) may have a less severe disease compared to severe hemophilia A (HA).

Objectives: to investigate clinical, radiological, laboratory and histological differences in the arthropathy of severe HA and HB.

Patients/Methods: 70 HA and 35 HB patients with at least one joint bleeding were consecutively enrolled. In all patients were assessed: joint bleedings (<10, 10-50, >50), regimen of treatment (prophylaxis/on demand), WFH, Pettersson and ultrasound (US) scores, serum sRANKL and OPG. Expression of RANK, RANKL and OPG was also evaluated in synovial tissue from 18 HA and 4 HB patients. Chi-square test, T-test, Mann-Whitney and Spearman's rank correlation coefficient were performed.

Results: The percentage of patients with either 10-50 or >50 hemarthrosis was greater in HA than in HB (p< 0.001 and p = 0.03, respectively); that with <10 hemarthrosis was higher in HB (p < 0.0001). The mean value of WFH (36.6 vs 20.2;p <0.0001) and US scores (10.9 vs 4.3;p <0.0001) were significantly higher in HA patients. Serum OPG and s RANKL were decreased in HA versus HB (p<0.0001 and p=0.006, respectively) and also in HA patients with>50 hemarthrosis. The expression of OPG was markedly reduced in synovial tissue from HA patients.

Conclusions: The reduced number of hemarthrosis, the lower WFH and US scores and higher OPG levels in serum and expression in synovial tissue in HB suggest that HB is a less severe disease than HA. OPG reduction seems to play a pivotal role in the progression of arthropathy in HA.

Disclosures

No relevant conflicts of interest to declare.
Autosomal Recessive Inherited Bleeding Disorders in Pakistan, a Cross Sectional Study from Selected Regions
Authors: Naz Arshi , Jamal Mohammad younus , Amanat Samina Tufail , Patel Humayun , Agha Akbar , Razik fazal-E- , Ahmed Nisar , Imran Ayisha , Shamsi Tahir
Published: October 2019

INTRODUCTION: Incidence of autosomal recessive disorders is rare, includes deficiencies of clotting factors I, II, V, VII, X, XI, XIII, vitamin K dependent clotting factors, combined factor V & VIII, Von Willebrand disease type 3, Glanzmann Thrombasthenia and Bernard Soulier syndrome.

OBJECTIVE: Spectrum of autosomal recessive bleeding disorders (ARBDs] among Pakistani patients.

PATIENTS AND METHODS: This cross-sectional study was carried out at Karachi, Lahore, Islamabad and Peshawar. PT,aPTT, BT, and fibrinogen levels done. Patients with prolonged APTT were tested for factors VIII and IX. If FVIII was low, von Willebrand factor: antigen (vWF: Ag) and von Willebrand factor: ristocetin cofactor (vWF: RCo) were performed. When PT and aPTT both were prolonged, FII, FV, and FX were tested. Peripheral film and platelet aggregation studies were done for platelet disorders. Urea clot solubility test was done at the end.

RESULTS: Out of429 patients, 148 were diagnosed with Hemophilia A, remaining 281 patients had ARBDs. 95 (33.8%) had VWD type 3. Fibrinogen deficiency was found in 34 (12%), Glanzmann Thrombasthenia in 27 (9.6%), factor XIII in 13 (4.6%), factor VII in 12 (4.3%), factor V in 9 (3.2%), 8 (2.8%) in vitamin K dependent clotting factors, , Bernard Soulier in 7 patient (2.5%),factor X in 2 (0.7%), factor II in 2 (0.7%), factor XI and combined factor V and VIII in 1 (0.4%)patients each. 70 patients (16.3%) remained undiagnosed.

CONCLUSION: VWD type 3 is the most common deficiency followed by fibrinogen deficiency. Glanzmann thrombasthenia was the third most common ARBD.

Disclosures

No relevant conflicts of interest to declare.
Treatment and Outcome of the Patients with Donor-Type Aplasia after Bone Marrow Transplantation in Children with Aplastic Anemia
Authors: Jeong Kumhee , Cho Joo Hyun , Cheon Kyeong Ryeol , Jang Hae In , Baek Hee Jo , Kook Hoon
Published: October 2019

Hematopoietic stem cell transplantation (HSCT) from an HLA-matched donor is the treatment of choice for children with aplastic anemia (AA). However, graft failure (GF), either primary or secondary, remains an important and life-threatening complication. Recently, donor-type aplasia, defined as bone marrow aplasia with full donor chimerism among secondary GF, has been identified after HSCT. Clinical characteristics of donor-type aplasia after HSCT and its treatment and outcome in children with AA were retrospectively reviewed. Forty-two children with AA underwent allogeneic HSCT with 10-year overall survival rate of 85.7%. While primary GF developed in 1 (2.4%), secondary GF was seen in 12 at a median of 8 months (range: 2.0-28.5 months). Among them, 11 developed a donor-type aplasia with the cumulative incidence of 26.2%. Low infused cell number (P=0.002), immunosuppressive therapy (IST) prior to HSCT (P=0.003) and preceding transfusion >40 times (P=0.008) were associated with the development of donor-type aplasia. The survival of patients with donor-type aplasia was 81.8%. Six patients were treated with stem cell rescue as follows: peripheral blood stem cell (PBSC) boost, 3; PBSC boost followed by secondary HSCT, 2; and secondary HSCT, 1. All but 1 who had a longest interval from 1st HSCT to stem cell rescue (54.4 months) showed restoration of graft function. The remaining 5 patients were managed with conservative measures including transfusions. No one showed spontaneous improvement with the median follow-up of 30.1 months (range, 4.2 to 43.0 months). Two patients died of graft failure and infection. Altogether, 4 patients still remain transfusion-dependent. Donor-type aplasia after HSCT for AA is not uncommon. Stem cell rescue, either with PBSC boost or secondary HSCT after conditioning should be attempted to obtain sustained graft function. For those who do not have availability for stem cell rescue, a novel treatment, such as thrombopoietin receptor agonist, should be considered for the future.

Disclosures

No relevant conflicts of interest to declare.
A Novel DLL1-Anti-CD19 Chimeric Protein Effectively Targets Notch Activation to B-Cell Acute Lymphoblastic Leukemia and Induces Cell Death
Authors: Wiersma Rebecca E , Kannan Sankaranarayanan , Somanchi Srinivas S. , Zeng Lizhi , Zweidler-McKay Patrick A.
Published: October 2019

Background: Acute lymphoblastic leukemia (ALL) is the most prevalent form of cancer in children, and those who relapse continue to have poor survival. Therefore, the development of improved and specifically targeted treatment options is vital. The Notch pathway has been shown to act as a tumor suppressor in B-ALL via cell type specific induction of growth-inhibiting and pro-apoptotic pathways. In this study, we aim to therapeutically activate Notch signaling in B-ALL via targeting the Notch ligand DLL1 to B cells using an anti-CD19 scFv chimeric protein.

Methods: A soluble chimeric protein composed of the extracellular domain of the Notch ligand DLL1 linked to a validated anti-CD19 scFv was isolated from HEK-293 producer cells. Human B-ALL (SB, Nalm6) and T-ALL (Jurkat) lines were treated with DLL1-anti-CD19scFv chimeric protein, and expression of the Notch target gene HES1 and effects on cell growth and survival were measured.

Results: Both B- and T-ALL lines express Notch1 and Notch2 receptors on the cell surface. Exposure of B-ALL cells to DLL1-anti-CD19scFv chimeric protein led to an increase of Notch signaling, measured via 3-14 fold increase in HES1 mRNA expression. As expected, surface expression of the Notch receptors decreased upon chimera exposure, as Notch receptors are cleaved and destroyed upon activation. Importantly, exposure of the B-ALL lines to the chimeric protein led to a maximum 60% decrease in cell counts over 3-4 days, in contrast to T-ALL, where exposure did not effect growth significantly.

Conclusions: This study demonstrates that Notch signaling can be feasibly activated in human B-ALL cells through a soluble DLL1-anti-CD19 scFv chimeric protein. Activation of Notch signaling via this method leads to growth inhibition and cell death in B-ALL, but not T-ALL cells. Based on our findings, we suggest that this soluble DLL1-anti-CD19 chimera may be a potential therapeutic approach for B-ALL, and further in vivo testing is warranted.

Disclosures

No relevant conflicts of interest to declare.
Rapamycin Induced Acute Myelogenous Leukemia U937 Cell Autophagy and Apoptosis
Authors: Xu Wenfeng , Li Chunfu , Feng Xiaoqin , He Yuelin
Published: October 2019

Objective: To investigate the autophagy and apoptosis on acute myelogenous leukemia U937 cell induced by rapamycin.

Methods: U937 cells was subcultured, and was set up blank control group (normal) and rapamycin treated groups (3 groups, 12h, 24h, 48h). The rapamycin treated groups were separately treated by 2¦ÌM concentration of rapamycin after 12h, 24h and 48h. The cell morphology of U937 cells treated by rapamycin was observed after 12h 24h and 48h. The survival rate of cells was detected by CCK-8 method. Cell apoptosis was detected by flow cytometry using Annexin V-FITC/PI double labeled. Use real-time PCR to detect the level of mRNA expression in autophagy specific protein maker LC3-¢òin different treated times by rapamycin. Rapamycin LC3 protein expression levels after treatment was detected by western blot method.

Results: Visible under inverted microscope, the rapamycin treatment groups¡¯ cell number reduced gradually after cultured 12h, 24h and 48h, smaller volume, cell rupture, increase the nucleus pycnosis and cellular debris(Figure 1). With the extension of time, U937 cells survival rate is falling, compared with control group, with statistical significance(P=0.031; P<0.05). Rapamycin treatment U937 cells after 12 h, 24 h and 48 h, U937 cell apoptosis rate increased, compared with control group, with statistical significance (P=0.027; P< 0.05, Figure2). Rapamycin treatment U937 cells after 12 h, 24 h and 48 h, LC3-¢ò mRNA expression and protein expression were cut, compared with control group, with statistical significance(P=0.029; P<0.05, Figure 3).

Conclusion: Rapamycin can induce autophagy and apoptosis of U937 cells. Autophagy protein LC3-¢ò in gene and protein expression level lowered, LC3-¢ò may play an important role in regulating the leukemia cell autophagy.

Disclosures

No relevant conflicts of interest to declare.
CLAG-Based Induction Therapy in Previously Untreated High Risk AML Patients
Authors: Seiter Karen , Germani Stephanie , Martin Julie , Raffa Rosemarie , Reilly Michele , Ahmed Nasir , Baskind Paul , Liu Delong
Published: October 2019

The CLAG regimen (G-CSF 300 mcg sc, cladribine 5 mg/m2 over 2 hours, and cytarabine 2 gm/m2 over 4 hours beginning 2 hours after cladribine, all daily times 5 days) was originally devised by Robak et al (Leuk Lymphoma 2000; 36:121-9) as induction therapy for patients with relapsed or refractory acute myeloid leukemia. Fifty percent of patients achieved a CR with a median duration of 22.5 weeks. This group subsequently added mitoxantrone to the regimen (Wrzesien-Kus A, et al. Ann Hematol 2005; 84:557-64). We treated 20 patients with previously untreated acute myeloid leukemia who were considered unsuitable for our intensive high-dose cytarabine, high-dose mitoxantrone frontline induction regimen either due to age or cardiac dysfunction with CLAG-based therapy. Patients with a cardiac ejection fraction above 50% additionally received either mitoxantrone (mito) or idarubicin (ida), 12 mg/m2 times 3 days, concurrently with CLAG. Of 20 patients treated, 5 received CLAG, 12 received CLAG-ida and 3 received CLAG-mito. The median age was 64 years (range 42-79 years). There were 13 men and 7 women. Six patients had received prior chemo and/or RT for a previous malignancy. In addition 3 patients had a prior MPD and 1 had prior MDS (total of 10 patients with secondary AML). Patients had a median of 3 comorbidities (range 0-7). Cytogenetic risk was good: 2 patients (however one was FLT3 ITD+), intermediate: 10 patients, poor: 8 patients. Only one patient was FLT3 ITD+. Responding patients (CR or PR) received 1 (5 pts), 2 (2 pts), 3 (8 pts) or 4 (2 pts) cycles of CLAG+/- ida or mito followed by allogeneic stem cell transplant (4 pts), hidac (2 pts) or decitabine maintenance (4 pts). Most patients responded to therapy. There were 13 formal CRs (65%), 1 CRp (5%), 3 PR (15%, defined as 6-10% blasts on marrow with complete hematologic recovery in the peripheral blood), 2 failures (10%) and one early death (5%). Other than the early death, treatment was well tolerated with few toxicities other than neutropenic fever and cytopenias. Estimated overall survival by Kaplan Meier analysis is 29.6 months (95% CI 20.1-39.2 months). Duration of response is 32.3 months (95% CI 21.6-43.1 months).

CLAG-based therapy is a well-tolerated, efficacious induction strategy in previously-untreated patients with high risk AML. CLAG-based regimens should be studied in a broader group of newly diagnosed AML patients.

Figure 1. Figure 1. Figure 2. Figure 2.

Disclosures

Off Label Use: Use of cladribine in AML.