bluuebunny/crossref_metadata_2025_split huggingface dataset:
Platelets Can Phagocytose Influenza Virus Which May Contribute to the Occurrence of Thrombocytopenia during Influenza Infection
Authors: Jansen A.J. Gerard , Low Hui Zhi , van den Brand Judith , van Riel Debbie , Osterhaus Albert , van der Vries Erhard 
Published: October 2019

Abstract

Thrombocytopenia is a common and sometimes life-threatening symptom in patients with influenza, which indicates that platelets may play a significant role during virus infection. However, the mechanisms underlying influenza associated thrombocytopenia are still unclear to date. In this study, the relationship between platelets and influenza infection was studied in vivo and in vitro.
In the period 2009-2012 we randomly measured viral load and platelet count in laboratory-confirmed influenza patients (A/H1N1 subtype; n=85) presented at the emergency department of Erasmus MC. In a cross-sectional study we found, after excluding patients with HIV infection, chemotherapy treatment and immediate referral to the ICU, that a high viral load was significantly correlated with a low platelet count at presentation (n=36; Spearman correlation coefficient 0.341; P=0.042). To study this relationship further we inoculated ferrets (n=120) with an H3N2 (n=30), pandemic H1N1 (n=30), or H5N1 influenza A virus (n=30) and measured platelet counts in six ferrets per time point at day 0, 1, 2, 3 and 4 after infection. Thrombocytopenia was seen at day 3 in all ferrets compared to the control animals (n=30). The decrease in platelet counts was significant in the H5N1 infected ferrets (p<0.05). For the sialidase and influenza virus inhibitor oseltamivir, we have previously shown that it functions as an inhibitor of platelet degradation. Interestingly, thrombocytopenia in ferrets could also be inhibited by oral administration of oseltamivir. However, these results were not statistically significant.
Finally, we studied the interaction between platelets and influenza viruses by electron microscopy. In a platelet suspension incubated for 1 minute with influenza virus we observed platelets with virus containing vacuoles, suggesting that the platelets had rapidly phagocytosed the viruses. Characteristics of these platelets incubated with influenza virus were measured including membrane expression of the GPIb-IX-V complex, CD62P, and surface glycans sialic acid and GlcNac using flowcytometry and functional capacity employing aggregometry using collagen, thrombin and ADP as agonists. However, no effect was seen in expression of GPIba and GPIX, P-selectin expression and surface expression of sialic acid and GlcNAc after incubation with influenza virus. Also no effect was seen on functional capacity of these platelets.
Our study shows that influenza virus is significantly correlated with a lower platelet count. Phagocytosis of influenza virus by platelets may play an important role in the occurrence of thrombocytopenia during influenza infection and may be a mechanism of virus clearance during infection.

Disclosures
Osterhaus: Viroclinics Biosciences: Other: chief scientific officer and hold certificates of shares.
NCCN-IPI Is Superior to aaIPI and IPI in Predicting Survival of DLBCL in the Rituximab Era
Authors: Wei Yongqiang , Zhang Yuankun , Hao Xiaoxiao , Wei Xiaolei , Huang Weimin , Feng Ru 
Published: October 2019

Abstract
Objective
International prognostic index (IPI) has been widely used for predicting outcome in diffuse large B cell lymphoma. Although the introduction of rituximab to chemotherapy has dramatically improved the outcome of DLBCL, it also changed the prognostic value of IPI. The National Comprehensive Cancer Network IPI (NCCN-IPI) and age-adjusted IPI (aaIPI) were used to evaluate the prognosis for DLBCL in the rituximab era. However which one of them is more powerful in predicting survival remains unknown in Chinese patients.
Patients and Methods
A total of 334 patients with de novo diffuse large B-cell lymphoma diagnosed from 2003 to 2012 were included. All patients were treated with CHOP with or without rituximab. They were divided into CHOP and R-CHOP groups. IPI, NCCN-IPI and aaIPI score were recorded. Survival was performed according to the Kaplan-Meier (K-M) curves using the log-rank test. The predictive abilities of PI, NCCN-IPI and aaIPI were investigated by Harrell's C-statistics.
Result
Compared with the IPI, the NCCN-IPI and the aaIPI had better discrimination in different scores in all patients treated with or without rituximab. Both the NCCN-IPI and aaIPI had power in discriminating low, low-intermediate and high-intermediate risk groups. In comparison, the NCCN-IPI discriminated low-intermediate and high-intermediate risk groups (5-year overall survival [OS]: 74% vs 50%) better than the aaIPI (5-year OS: 80% vs 62%) in all patients and in the R-CHOP group with the NCCN-IPI 5-year OS: 85% vs 67% while the aaIPI: 87% vs 77%. In the CHOP group, the aaIPI discriminated low-intermediate and high-intermediate risk groups better (5-year OS: 70% vs 25%) than the NCCN-IPI (5-year OS: 60% vs 24%). According to the Harrel C statistic, the NCCN-IPI showed higher discrimination of the different scores (C_index: 0.669 ) than the aaIPI (C_index: 0.663 ) in all the patients (C_index: 0.669 vs. 0.663), especially in the R-CHOP group (C_index: 0.681 vs. 0.636). In the CHOP group, the aaIPI had stronger power (C_index: 0.711 ) than the NCCN-IPI (C_index: 0.683 ) in discriminating different scores.
Conclusion
The NCCN-IPI is more powerful than the IPI for predicting survival in the rituximab era. In patients aged 60 or younger and treated with CHOP without rituximab, aaIPI is a preferable tool for evaluating prognosis.

Disclosures
No relevant conflicts of interest to declare.
Pharmacometabolomics for Predicting Variable Busulfan Exposure in Pediatric HSCT Patients
Authors: Kim Bora , Hong Kyung Taek , Lee Ji Won , Yu Kyung-Sang , Jang In-Jin , Park Kyung Duk , Shin Hee Young , Ahn Hyo Seop , Cho Joo-Youn , Kang Hyoung Jin 
Published: October 2019

Abstract
Optimal dosing for busulfan is important for minimization of systemic toxicity from overexposure and graft failure or relapse from underexposure. Herein, we investigated potential markers for predicting individual variation in the pharmacokinetics of busulfan, and suggested possible mechanism for inter-individual variability by using pharmacometabolomics.
Fifty-nine pediatric patients undergoing busulfan-based conditioning chemotherapy for hematopoietic stem cell transplantation were divided into three groups according to the area under the concentration-time curve (AUC) of busulfan; low-, medium-, and high-AUC group. Nontargeted metabolic profiling of baseline urine samples showed that deferoxamine metabolites were abundant, while 2 acylcarnitines and phenylacetylglutamine were significantly lower in high-AUC group, compared with low-AUC group. Higher level of deferoxamine, an iron-chelating agent for the patients with a high serum ferritin level during the conditioning chemotherapy, suggested pharmacokinetic interaction between serum ferritin level and busulfan exposure. Retrospective analysis of the correlation between serum ferritin level and busulfan AUC showed positive correlation in 130 pediatric patients. The optimal busulfan dose to meet the target AUC of 18,750 μg*h/L/day was calculated to be 119.7 ± 30.1 mg/m2 in patients with ferritin < 1,000 ng/mL and 106.1 ± 29.3 mg/m2in patients with ferritin ≥ 1,000 ng/mL (P=0.021).
Mechanismly, previous studies have indicated that ferritin, acylcarnitine and phenylacetylglutamine are closely associated with liver function. Increased serum ferritin is thought to be responsible for an increased production of oxygen free radicals and activation of GSH turnover, which means reduction of GSH levels in both plasma and erythrocytes and this depletion seems to be related to the decrease of busulfan metabolism. Down regulation of acylcarnitines is associated with deregulation of mitochondrial fatty acid β-oxidation by hepatic injury. PAGN, a marker of waste nitrogen scavenger, was down-regulated which indicates ammonia-related metabolism could also be involved in busulfan exposure. Hyperammonemia, a clinical condition of elevated ammonia levels, is mainly lead by liver cell damage.
Taken together, our findings demonstrate that elevated serum ferritin levels are a potential biomarker correlated with busulfan exposure and provide some evidence that busulfan metabolism seems to decrease in the patients with reduced liver function.

Disclosures
No relevant conflicts of interest to declare.
the Role of MSCs in the Development of Acute Myeloid Leukemia
Authors: Al-Matary Yahya Saleh , Botezatu Lacramioara , Fernando Lams Robert , Thivakaran Aniththa , Köster Renata , Schütte Judith , Dührsen Ulrich , Opalka Bertram , Khandanpour Cyrus 
Published: October 2019

Abstract
Acute myeloid leukemia (AML) is a malignant disease of the bone marrow. The prognosis of AML is still poor with only 25% of patients living longer than 5 years. Thus, new therapeutic approaches are warranted to improve patients' survival. The growth of AML cells not only depends on cell intrinsic factors, but is also supported by stroma cells, among them mesenchymal stem cells (MSCs). We reported here that MSCs derived from human AML patients or murine models of human AML better supported the in vitro growth of leukemic cells than MSCs from non-leukemic patients or mice and this is dependent on cell-to-cell contact. Similarly, immortalized MSCs lines derived from AML patients supported the growth of human AML cell lines better than a MSC line originating from a healthy donor. In addition, the frequency of MSCsin the BM of leukemic mice was increased. On a molecular level, the polarization of MSCs towards an AML-supporting state depended on the expression of the transcription factor Growth factor independence 1 (Gfi1). Loss of Gfi1 abrogated the tumor-supporting state of AML-associated MSCs in vitro and in vivo. In summary, MSCs from AML patients support the growth of AML cells in vitro in a Gfi1-dependent manner which could open the path to new therapeutic approaches.

Disclosures
Dührsen: Amgen: Honoraria, Research Funding; Alexion Pharmaceuticals: Honoraria, Research Funding; Roche: Honoraria, Research Funding. Khandanpour:Hospital of Essen university: Research Funding; Max-Eder: Research Funding.
Modeling Clonal Hematopoietic Disorders in Zebrafish Using Color Barcoding
Authors: Avagyan Serine , Henninger Jonathan E , Moore Jessica , Mannherz William , Zon Leonard I. 
Published: October 2019

Abstract
Myeloid neoplasms, including myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML), are clonal hematopoietic disorders that arise from abnormal hematopoietic stem and progenitor cells (HSPCs)with acquired somatic mutations. To follow the hematopoietic output of HSPC clones endogenously, we have developed a novel marking strategy in which each HSPC clone is labeled in a unique fluorescent hue. The transgenic zebrafish line Zebrabow uses a Cre/lox system where stochastic recombination of multiple transgenic insertions harboring three fluorescent proteins (dTomato, YFP and CFP) provide stable and heritable color barcodes. Tissue-specificity is ensured by tamoxifen-induced Cre activation under the draculin promoter which is highly expressed in zebrafish HSPCs throughout development. The result of Zebrabow labeling is a collection of colored blood cells, which are grouped by their hue, and largely cluster in various groups, each representing a clonal population of cells that arise from a specific HSPC. To induce clonal hematopoietic states in zebrafish assessed by color dominance in hematopoietic cells, we introduced combinations of known genetic mutations founds in MDS and AML into Zebrabow embryos. Embryos at the 1-cell stage were injected with combinations of mutations and HSPC color labeling was performed at 36 to 48 hours post fertilization, during peak emergence of definitive blood stem cells from the dorsal aorta. The injected mutations were grouped by known co-occurence in patients with MDS and AML. The transgenic overexpression of Jak2V617F, FLT3ITD, IDH2R140Q, HOXA9 or HOXB4 under the zebrafish blood-specific promoter draculin, was combined with somatic mutagenesis of endogenous tet2, asxl1, dnmt3a, ezh2, ptena/b and tp53 via the CRISPR/Cas9 system. Mosaic combinations of these mutations resulted in the expansion of single color clones, contributing to ≥30% of granulocytes, with concomittant myeloid expansion in a third of these zebrafish. The majority of zebrafish exhibiting clonal expansion harbored frame-shift mutations in asxl1 with >25% variant allelic frequency identified by next-generation sequencing. Similarly, combinations including Jak2V617F, tet2, or asxl1 mutations lead to single color expansion in kidney marrow cells in 30% of injected zebrafish, with the occasional presence of a blast-like population of cells in the marrow and spleen. We have successfully modeled early stages of clonal hematopoietic expansion with myeloproliferation in the presence of Jak2V617F and asxl1 deletion using the unique color barcoding system in zebrafish.

Disclosures
Zon: Marauder Therapeutics: Equity Ownership, Other: Founder; Scholar Rock: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Other: Founder; Fate, Inc.: Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Other: Founder.
Deferasirox Associated to Liver Failure and Death in a Sickle Cell Anemia Patient Homozygous for the -1774delG Polymorphism in the ABCC2 Gene Encoding Multidrug Resistance Protein 2 (MRP2)
Authors: Braga Caroline Cordeiro Barroso , Benites Bruno Deltreggia , de Albuquerque Dulcineia M , Alvarez Marisa Claudia , Costa Fernando F , Gilli Simone C O , Saad Sara T.O. 
Published: October 2019

Abstract

Introduction: Regular blood transfusions may result in hemochromatosis, leading to damage of various organs. Deferasirox exhibits good efficacy as iron chelation therapy, however changes in liver parameters may occur, as well as, despite being rare, more serious complications such as liver failure. Multidrug resistance protein 2 (MRP2) is a member of the ATP-binding cassette (ABC) transporter family of membrane proteins encoded by the ABCC2 gene and is expressed in the canalicular part of the hepatocyte. MRP2 acts in the biliary transport of several endogenous compounds and many drugs, including Deferasirox. Thus, genetic polymorphisms related to the ABCC2 gene, which encodes the MRP2 protein, may influence individual susceptibility to hepatotoxicity related to Deferasirox.
Methods: Two functional genetic variants, rs369192412 (-1774delG) and rs717620 (-24C> T), of the ABCC2 gene, were studied using the DNA sequencing technique in 31 genomic DNA samples from patients with iron overload who were currently using or had previously used Deferasirox. The sequencing results were compared to clinical data related to the presence or absence of hepatotoxicity during the use of this drug.
Results: Of all patients studied, 9 (29%) of them developed hepatotoxicity based on elevation of AST and/or ALT >5 X ULN or elevation of bilirubins >3 X ULN, which occurred in an average of 239 (14-983) days after initiation of treatment. From the 9 (29%) patients with hepatotoxicity, only 2 (22.2%) of them had the polymorphism -24C> T in only one allele, and the remaining 7 (77.8%) patients did not (OR = 0.77, 95% CI = 0.06 to 5.91, p = 1). Despite no association was detected between the -17774delG polymorphism and hepatotoxicity (OR = lower, 95% CI = 0.06 to bottom, p = 0.29), this polymorphism was present in homozygosis in 1 patient with sickle cell anemia, who, coincidently, had fulminant hepatitis leading to death, with no other causes for liver toxicity and despite all introduced therapies.
Conclusions: Our study showed no correlation between ABCC2 polymorphisms (-24C> T, -1774delG) and hepatotoxicity to Deferasirox, however, the rs369192412 (-1774delG) polymorphism was present in homozygosis in the only patient who died due to fulminant hepatitis secondary to the drug. This data may indicate a possible influence of the presence of this polymorphism in the severity of liver damage related to Deferasirox.

Disclosures
No relevant conflicts of interest to declare.
Cell-Free DNA Is Elevated after Acute Arterial Injury in Infants
Authors: Kim Jina , Naqvi Ibtehaj , Nag Uttara P. , Leraas Harold J. , Otto James C. , Borst Alexandra J. , Hill Kevin D. , Fleming Gregory A. , Sullenger Bruce A. , Tracy Elisabeth T. 
Published: October 2019

Abstract
Purpose: Circulating cell-free DNA in blood may originate from cell necrosis, apoptosis, or NETosis. NETosis is a recently described inflammatory process in which neutrophils release a meshwork of DNA fibers called neutrophil extracellular traps (NETs) to attack microorganisms. Higher levels of cell-free DNA have been observed in patients with sepsis and malignancy, and it may be linked to arterial thrombosis as well. We examined changes in cell-free DNA in infants who experienced acute, iatrogenic thrombosis of the femoral artery after arterial catheterization to better understand the role of cell-free DNA in clot formation.
Methods: Plasma samples were collected from infants < 1 year of age with congenital heart disease during cardiac catheterization. Post-procedurally, femoral artery thrombus formation at the site of sheath insertion was detected by ultrasound. Cell-free DNA was quantified at two time points (before and after arterial catheterization) in three infants who developed femoral artery thrombus after catheterization and three age-matched controls. Cell-free DNA was also quantified from plasma of normal adults as a comparison.
Results: At baseline,cell-free DNA level for the infant cohort was similar to that in normal adults (13.9 ng/mL vs. 10.3 ng/mL, p = 0.20, Figure). After arterial catheterization, cell-free DNA was increased from baseline in all infants (51.0 ng/mL vs. 13.9 ng/ml, p = 0.004). However, infants who developed femoral artery thrombus after catheterization did not demonstrate higher cell-free DNA after arterial injury, compared to the control group (47.1 ng/mL vs. 54.9 ng/ml, p = 0.83).
Conclusions: Cell-free DNA is significantly and consistently elevated in infants after arterial catheterization. As part of NETosis, cell-free DNA may serve as an inflammatory mediator in infants who experience arterial injury. We plan to confirm the findings of this pilot study in a larger human infant cohort as well as in a piglet model of vascular injury.
Figure Cell-free DNA levels in infants, before and after arterial catheterization, in comparison to normal adults. Figure. Cell-free DNA levels in infants, before and after arterial catheterization, in comparison to normal adults.

Disclosures
No relevant conflicts of interest to declare.
Intravital Microscopy Reveals Fundamental Differences in the Interaction of Stem Cells and T Acute Lymphoblastic Leukaemia with the Bone Marrow Microenvironment
Authors: Lo Celso Cristina , Hawkins Edwin D , Duarte Delfim , Akinduro Olufolake , Khorshed Reema A , Passaro Diana , Nowicka Malgorzata , Straszkowski Lenny , Scott Mark K , Rothery Steve , Ruivo Nicola , Foster Katie , Waibel Michaela , Johnstone Ricky W , Harrison Simon J. , Westerman David A , Quach Hang , Gribben John , Robinson Mark D , Purton Louise E , Bonnet Dominique 
Published: October 2019

Abstract
Haematopoiesis is the complex developmental process maintaining the turnover of all blood cell lineages, i.e. erythrocytes, immune (white) cells and platelets. It takes place primarily in the bone marrow, where it critically depends on the correct functioning of rare, quiescent haematopoietic stem cells (HSCs) and more numerous HSC-derived, highly proliferative and differentiating haematopoietic progenitor cells (HPCs). This is regulated by complex interactions between stem cells and the bone marrow microenvironment, and it was suggested to change in settings involving stress. Indeed, using intravital microscopy, we previously showed that HSCs injected in irradiated recipients steadily engage with endosteal niches. Moreover, if phenotypically identical HSCs are harvested from mice harbouring a natural infection, such as that caused by T.spiralis, they show a unique 'stop and go' migratory behaviour, engage larger niches, yet remain in the proximity of osteoblastic cells.
Based on these and other studies, it has been hypothesised that also malignant haematopoietic cells rely on interactions with specific bone marrow microenvironments to grow, and especially to develop chemoresistance. To investigate this further, we performed longitudinal imaging of mice injected with murine and human T acute lymphoblastic leukaemia (T-ALL) cells over the course of hours or days, including during response to multiple chemotherapy treatments. Strikingly, we observed a very different behaviour from that of HSCs, with T-ALL cells showing a highly migratory behaviour throughout all stages of disease, from initial bone marrow infiltration to response to chemotherapy. Interestingly, in untreated mice and in patients trephine biopsies we observed rapid loss of osteoblastic cells by apoptosis, an event that likely contributes to the dramatic loss of healthy haematopoiesis observed in mice and patients. Together, these finds indicate that novel therapeutic strategies should target the ability of T-ALL cells to entertain short-lived and promiscuous interactions with any neighbouring cells, and aim to strengthen the bone marrow microenvironment to improve support for residual healthy haematopoiesis.

Disclosures
Harrison: AbbVie: Research Funding; Janssen Cilag: Research Funding, Speakers Bureau. Quach:Amgen: Membership on an entity's Board of Directors or advisory committees; Janssen Cilag: Membership on an entity's Board of Directors or advisory committees; Celgene: Membership on an entity's Board of Directors or advisory committees.
Decitabine Combined with Haag Regimen Is an Effective Salvage Treatment for Advanced Acute Myeloid Leukemia
Authors: Tang XiaoWen , Cui Wei , Jin Zhengming , Zhu Xiaming , Sun Aining , Qiu Huiying , Miao Miao , Cao Jing , Guo Dandan , Wu Depei 
Published: October 2019

Abstract
Objective To evaluate the clinical efficacy and safety of decitabine (DAC) in combination with HAAG regimen [homoharringtonine (HHT), cytarabine (Ara-C), doxorubicin (Acla) and recombinant human granulocyte colony stimulating factor (G-CSF)] for advanced patients with acute myeloid leukemia (AML). Methods Thirty-six patients with advanced AML receiving DAC combined with HAAG chemotherapy in our center from December 2012 to August 2015 were enrolled in this study. Eighteen of them were refractory or relapsed AML, and another 18 patients were those who didn't achieve complete remission (CR) after a course of induction chemotherapy. The therapeutic responses, side effects and long-time survival were retrospectively analyzed. Results After a course of treatment, the rate of CR and partial response (PR) was 58.3% (21/36) and 22.2% (8/36) respectively, while the overall response rate (ORR) was 80.6% (29/36) in the cohort. For the patients with refractory or relapse AML, CR was 61.0% (11/18), PR was 22.2% (4/18), and ORR was 83.3% (15/18). While for the other not getting CR after a course of induction chemotherapy, CR was 55.6% (10/18), PR was 22.2% (4/18), and ORR was 77.8% (14/18). Grade 4 hematological toxicities were observed in all patients, and 72.2% cases experienced infection. And all non hematological side effects were mild and well-tolerated. With a median follow-up of 7.5 (0.5~33.3) months, the 1-year overall survival (OS) rate was 43.3%, 24.2% for the refractory or relapsed AML patients, and 61.6% for those not achieving CR after a course of induction chemotherapy. The difference was significantly (P=0.01). Conclusion DAC combined with HAAG regimen is safe and effective salvage treatment for advanced stage AML patients.

Disclosures
No relevant conflicts of interest to declare.