bluuebunny/crossref_metadata_2025 huggingface dataset:
Correlation between Changes in Cardiac Iron and Hepatic Iron in Pediatric Patients with Thalassemia Major
Authors: Pepe Alessia , Meloni Antonella , Filosa Aldo , Pistoia Laura , Casini Tommaso , Lisi Roberto , Putti Maria Caterina , Maggio Aurelio , Gerardi Calogera , Fina Priscilla , Positano Vincenzo , Casale Maddalena
Published: November 2019

Introduction. A prospective magnetic resonance imaging (MRI) study demonstrated a good control of myocardial iron overload (MIO) in terms of prevention and treatment in children with thalassemia major (TM).

The aim of the present study was to evaluate if changes in MIO were related to baseline hepatic iron or changes in hepatic iron overload (HIO).

Methods. We considered 68 TM patients enrolled in the MIOT (Myocardial Iron Overload in Thalassemia) project with less than 18 years at the first MRI scan and who performed a follow-up (FU) study at 18±3 months.

Myocardial and hepatic iron burdens were quantified by the T2* technique. The value of 20 ms was used as conservative normal value for the global T2* value. Liver T2* values were converted into liver iron concentration (LIC) values. A LIC<3 mg/g/dw indicated significant no HIO, between 3 and 7 mg/g/dw mild HIO, between 7 and 15 mg/g/dw moderate HIO, and ≥15 mg/g/dw severe HIO.

Results. Thirty-six patients were females and mean age at the time of the baseline MRI was 13.74±3.09 years. Baseline global heart T2* values were 29.72±11.21 ms and 16 (23.5%) patients showed significant baseline MIO.

The percentage changes in global heart T2* values per month in the whole patient population were 0.66±1.70 and they resulted significantly higher in the 16 patients with significant baseline MIO versus the patients with no baseline MIO (1.99±2.53% vs 0.25±1.09% ms; P=0.002).

Percentage changes in global heart T2* values per month were not influenced by initial MRI LIC values (R=0.048; P=0.695) (Figure 1A) and were comparable among the 4 groups of patients identified on the basis of baseline MRI LIC values (14 no HIO: 0.29±1.12% vs 21 mild HIO: 0.75±1.56% vs 15 moderate HIO: 0.82±2.03% vs 18 severe HIO: 0.71±2.00%; P=0.876).

Percentage changes in global heart T2* values per month were not associated to final MRI LIC values (R=-0.134; P=0.277) (Figure 1B).

The correlation between % changes in global heart T2* and MRI LIC values did not reach the statistical significance (R=-0.244; P=0.067) (Figure 1C).

In patients with baseline MIO no correlation was found between % changes in global heart T2* values per month and initial MRI LIC values (R=-0.325; P=0.219) or % changes in MRI LIC values per month (R=-0.353; P=0.180).

Conclusion. In pediatric TM patients changes in cardiac iron are not correlated to baseline MRI LIC values and changes in hepatic iron. So, our data seem not supporting the hypothesis for which it is necessary to clean the liver before removing iron from the heart.

Figure 1

Disclosures

Pepe: Chiesi Farmaceutici S.p.A., ApoPharma Inc., and Bayer: Other: No profit support.
Pancreatic Iron and Glucidic Metabolism in Thalassemia Major
Authors: Pepe Alessia , Pistoia Laura , Campisi Saveria , Sarli Roberto , Giovangrossi Piera , Guerrini Giulia , Mattei Roberto , Maggio Aurelio , Visceglie Domenico , Carrai Valentina , Peritore Giuseppe , Positano Vincenzo , Meloni Antonella
Published: November 2019

Background. A preliminary study involving 59 patients demonstrated that pancreatic iron assessed by magnetic resonance imaging (MRI) was the strongest overall predictor of glucose dysregulation in thalassemia major (TM) patients.

Aim. In the present multicenter study we explored systematically the link between pancreatic iron and glucidic metabolism in a large cohort of TM patients.

Methods. We considered 705 TM patients (372 F, mean age 37.00±9.95 years) enrolled in the E-MIOT (Extension-Myocardial Iron Overload in Thalassemia) project. T2* measurements were performed over pancreatic head, body and tail and global value was the mean. The pattern of disturbances of glucose metabolism was assessed by means of the oral glucose tolerance test (OGTT).

Results. According to OGTT results, 546 patients (77.4%) had normal glucose tolerance (NGT), 14 (2.0%) had isolated impaired fasting glucose (IFG), 29 (4.1%) had impaired glucose tolerance (IGT), and 116 (16.5%) had diabetes mellitus (DM).

None of the 85 patients (12.1%) without pancreatic iron overload (global pancreas T2*≥26 ms) had IGT or DM (Figure 1). The 84.6% of patients with NGT had pancreatic iron overload. The global pancreatic T2* values were significantly higher in patients with NGT than in patients with DM (13.58±11.08 ms versus 8.09±4.72 ms, P<0.0001).

Receiver operator characteristic (ROC) analysis showed that a global pancreas T2*<13.73 ms was the optimal cutoff for predicting an abnormal OGTT, with an area under the curve (AUC) of 0.62.

Global pancreas T2* values showed a weak significant correlation with insulin values (R=0.160; P=0.002) and homeostasis model assessment-insulin resistance (HOMA-IR) index (R=0.122; P=0.019).

Conclusion. A normal global pancreas T2* value has a negative predictive value of 100% for IGT and DM. The low specificity of pancreatic iron overload for glucose dysregulation seems to support the hypothesis that a latency time is need before pancreatic iron burden could give impaired glucose tolerance and overt diabetes mellitus.

Figure 1

Disclosures

Pepe: Chiesi Farmaceutici S.p.A., ApoPharma Inc., and Bayer: Other: No profit support.
Effect of Marijuana Smoking Among Adult Sickle Cell Patients on Trends in Sickle Cell Crisis Hospitalizations and Acute Chest Syndrome: A Nationwide Inpatient Assessment
Authors: Sama Shashank , Tandon Varun , Kethireddy Nikhila , Boddu Prajwal
Published: November 2019

PURPOSE

Pain for sickle cell patients occurs on a daily basis, but this becomes magnified during a sickle cell crisis (SCC). The mainstay of treatment during hospitalization is intravenous fluids and opiates however this can be a nidus for an opiate addiction long term, especially in the ongoing opiate epidemic. Marijuana has been used in a variety of chronic pain disorders including those from cancer. It is known that within the sickle cell population, smoking tobacco increases the risk of acute chest syndrome. Marijuana use or its derivatives used to treat pain has not been studied on a large scale within the sickle cell population. We propose the question of whether marijuana use affects SCC admissions as well as acute chest syndrome (ACS).

METHODS

The National Inpatient Sample (NIS) dataset was queried from 2005 to 2014 to identify the primary diagnosis of SCC with the International Classification of Disease (ICD) Code 282.42, 282.62, 282.64 and 282.69, as has been done in the literature. We then identified those with marijuana use, excluding those used in the past and not currently with the ICD code 304.30, 304.31 and 304.32. Additionally we identified those with ACS with the ICD code of 517.3. We then used multivariate analysis along with Chi-square for non-continuous variables using the statistical software SAS.

RESULTS

Between the years of 2005 and the 2014, there were a total of 798,313 hospitalizations for sickle cell crisis in the United States. Around 0.08 % of these patients had marijuana use. When stratified by race marijuana use was predominantly documented among African Americans (95.2%) followed by Hispanics (3.8%). Length of stay was statistically the same among marijuana users at 4.65 vs 5.28 days without use (p = 0.11) when compared to non-users. Total charges at the end of hospitalization were also the same at $23134.2 vs $24662.7 (p=0.60) with marijuana users and non-users respectively. Marijuana patients did that lower proportions of ACS at 5.46% vs 8.48% without (p= 0.004) and a relative risk of 0.64. Proportion of death was the same with marijuana users of 0% and non-users 0.28% (p=0.16). The age at admission was higher at 30.77 when compared to 27.25 among non-users (p=0.0009).

CONCLUSIONS:

Marijuana use was associated with lower instances of ACS with a relative risk of 0.64. Additionally patients presented on average 3 years later than non-users. The fact that mortality, length of stay and total charges were the same amongst the two groups may indicate that during a crisis, the disease process is the same, however outside of having a crisis, marijuana use may help with pain control given later presentation.

CLINICAL IMPLICATIONS:

Further research should be done to look at marijuana use as an alternative to pain control for sickle cell patients in the community setting.

Disclosures

No relevant conflicts of interest to declare.
Glycolytic Biomarkers Predictive of Transformation in Patients with Follicular Lymphoma
Authors: Monrad Ida , Madsen Charlotte , Lauridsen Kristina , Honoré Bent , Plesner Trine Lindhardt , Hamilton-Dutoit Stephen , d'Amore Francesco , Ludvigsen Maja
Published: November 2019

Introduction

Follicular lymphoma (FL) is an indolent lymphoma derived from germinal center B cells, covering approximately 20% of all lymphomas. The malignancy is generally considered an incurable condition, hence with a median survival time exceeding 10 years. A portion of patients experience early progression or histological transformation (HT) to a more aggressive lymphoma, typically diffuse large B cell lymphoma which occurs in up to 45% of FL patients, reducing the median survival after transformation to 1-2 years. Early detection of reliable predictors of HT would allow pre-emptive therapeutic intervention strategies and aim at improved survival for patients with transformed FL. In this study, we focus on the two glycolytic enzymes, Aldolase A and glyceraldehyde 3-phosphate dehydrogenase (GAPDH). Both proteins have previously been found to be upregulated at diagnosis in various solid cancers and associated with poor survival. In addition to its glycolytic effects, GAPDH has been recognized for its non-glycolytic effects including regulation of DNA replication and repair, RNA nuclear export and apoptosis.

Aim

The aim of this study was to identify biomarkers predictive of HT in patients with FL, by investigating the correlation between intratumoral aldolase A and GAPDH expression levels with the risk of transformation in pre-therapeutic tumor samples from FL patients.

Materials and Methods

Immunohistochemical aldolase A and GAPDH expression levels were quantified using digital image analysis in pre-therapeutic tumor tissue from FL patients at time of diagnosis for patients without (n=51) or with (n=41) subsequent HT, as well as in sequential samples at time of transformation (n=41). Staining intensities were assessed as area fractions (AFs) of the stained area normalized to the region of interest on whole biopsy sections.

Results

At time of initial FL diagnosis, FL patients with subsequent transformation had significantly higher levels of aldolase A and GAPDH expression compared to patients with no subsequent transformation, (p<0.001 and p=0.005). High levels of aldolase A expression (75th percentile cut-off), were found to be associated with a significantly shorter transformation free survival (TFS) (p=0.015) and progression free survival (PFS) (p=0.001). Furthermore, high levels of GAPDH expression were found to be associated with a significantly shorter TFS (p<0.001), PFS (p=0.034) as well as overall survival (OS) (p=0.018).

Conclusion

These results suggest that at time of initial FL diagnosis, aldolase A expression and GAPDH expression, maybe as indicators of high metabolic turnover, may be predictive of the patient's risk of subsequent histological transformation.

Disclosures

No relevant conflicts of interest to declare.
Increased Mutagenesis during Fetal Hematopoietic Development in Down Syndrome
Authors: Hasaart Karlijn , Manders Freek , Chuva de Sousa Lopes Susana , Van Boxtel Ruben
Published: November 2019

Children with Down syndrome are predisposed to leukemia during the first years of their life. 5-10% of newborns with Down syndrome are born with transient myeloproliferative disorder (TMD), which often spontaneously disappears. The majority of these patients achieves complete remission. However, in 20-30% of all TMD patients the disease progress in acute megakaryoblastic leukemia. In addition, they have a 20 fold higher risk of developing B-lymphocyte acute leukemia (B-ALL). Leukemic development in Down syndrome is initiated during fetal development. However, it is unclear why fetuses with a trisomy of chromosome 21 have an increased risk of developing leukemia.

Previously, we have developed a method to study somatic mutations in single cells using clonal cultures. Here, we applied this method to human fetal hematopoietic stem and progenitor cells (HSPCs) from liver and bone marrow of Down syndrome human fetuses and control fetuses with two copies of chromosome 21 (D21). In addition, we characterized somatic mutation accumulation in not affected small intestine stem cells. Subsequently, we performed in depth mutational analyses to characterize active processes using mutational signatures in fetal stem cells, which potentially can drive leukemic development during early life. Recently, we have shown that that healthy adult HSPCs gradually accumulate somatic mutations in a linear fashion with an annual mutation rate of 14.2 base substitutions per year. Whereas the somatic mutation rate is significantly higher during fetal development. Subsequently, in Down syndrome fetuses the overall somatic mutation rate of fetal stem cells is significantly increased compared to D21 fetal stem cells (P-value: 0,024). We performed phylogenetic analysis to study relatedness of the cells and observed an higher somatic mutation rate in the first cell divisions. This elevated mutation rate can be explained by increased contribution of mutational process signature 1 and 5, which are already present in fetal stem cells. Therefore, Down syndrome fetal stem cells show enhanced activity or increased sensitivity to mutational processes that are normally active during development. The same mutational signatures are present in TMD blast cells, indicating that these processes can cause cancer driver mutations and subsequently contribute to leukemic development. Interestingly, some Down syndrome fetal stem cells showed very high mutation numbers that could partly be attributed to mutational signature 18, which likely reflect oxidative-stress induced mutagenesis. These findings, show increased mutagenesis in Down syndrome during fetal development in hematopoietic stem cells and small intestine stem cells. This increased mutagenesis can potentially explain why children with Down syndrome have an increased risk of developing leukemia in early life.

Disclosures

No relevant conflicts of interest to declare.
Analysis of the Safety Profiles of Anticoagulants Using the EMA Pharmacovigilance Database
Authors: Elalamy Ismail
Published: November 2019

Introduction

Anticoagulants are widely used in the treatment of cancer-associated-thrombosis (CAT). Low molecular weight heparin (LMWH) has been the standard of care for years. However, recent data reported the use of direct oral anticoagulant (DOAC) in CAT patients with a higher rate of bleeding and more specifically a higher risk a gastrointestinal (GI) bleeding. The objective of this analysis is to investigate about the GI bleeding data in cancer and non-cancer patients.

Method

The public European database of suspected adverse drug reaction reports were checked in April 2019 for all the reports concerning DOACs and LMWHs available in Europe. These reports are submitted electronically by national medicines regulatory authorities and by pharmaceutical companies that hold marketing authorizations. A patients receiving an anticancer drug was considered as a cancer patients. All data from all patients regarding the GI haemorrhage.

Results

A total of 216 540 cases were found in the database. Gender (female: 46.5%, male: 45.6%, N/A: 7.9%). Most of the patients were > 65 years (63.8%). 32 591 cases for a LMWH and 183 949 for a DOAC. Among all, 2080 received an anticancer drug and 79 783 did not received an anticancer drug. Data about co-mediation were missing for all other cases. GI haemorrhages were common in both cancer (10.3%) and non-cancer patients (16.0%) receiving anticoagulants (Table 1).

Conclusion

GI haemorrhages were common in both cancer and non-cancer patients receiving anticoagulants, suggesting that GI haemorrhages may not only be linked to GI cancer. Furthermore, these data could suggest that patients receiving LMWH were less exposed to GI haemorrhages.

Table 1. GI bleeding in patients with anticoagulants.

Table 1

Disclosures

Elalamy: Pfizer: Honoraria, Research Funding; BMS: Honoraria, Research Funding; Boehringer-Ingelheim: Honoraria, Research Funding; Sanofi: Honoraria, Research Funding; LEO Pharma: Honoraria, Research Funding; Aspen: Honoraria, Research Funding; Bayer Healthcare: Honoraria, Research Funding.
The Vasculopathy in the Bone Marrow Microenvironment of Humanized Sickle Cell Mice Is Reversible By Blood Transfusion
Authors: Park Shin-Young , Mattè Alessandro , Jung Yookyung , Ryu Jina , Anand Wilson , Han Eun Young Anna , Liu Min , Carbone Carmine , Melisi Davide , Nagasawa Takashi , Lin Charles , Silberstein Leslie E. , De Franceschi Lucia
Published: November 2019

Sickle cell disease (SCD) is a monogenic red cell disorder with high morbidity and mortality due to significant end-organ disease. Here we investigated for the first time the impact of SCD on the bone marrow (BM) vascular niche, which is critical for hematopoiesis. In SCD mice, we find a disorganized and structurally abnormal BM vasculature with highly tortuous arterioles and fragmented sinusoidal vessels, the latter filled with aggregates of sickle erythroid and myeloid cells. By in vivo imaging, sickle and control red blood cells exhibit significantly slow intravascular flow speeds in sickle cell BM but not in control BM. These abnormalities lead to increased ROS production in expanded erythroblast populations, and to up-regulation in the BM of HIF-1a and key growth factors required to support angiogenesis, e.g. VEGF-A, Ang1, Ang2. Moreover, increased levels of VCAM-1, a marker for vascular dysfunction, are detected. SCD also impacts the BM perivascular niche, i.e. the number of CXCL12-abundant reticular cells, is markedly decreased, and is associated with mobilization of hematopoietic stem/progenitor cells as well as mature leukocytes to the periphery. Strikingly, the distorted vascular network is completely reversed by blood transfusion, which highlights the plasticity of the BM vascular niche. Thus, we provide molecular and cellular features of the pathologic BM microenvironment, which suggest that the local hypoxia-induced angiogenic network combined with inflammatory vascular dysfunction is responsible for the vasculopathy. In addition, the findings may serve to develop new biomarkers and therapeutic targets of BM end-organ damage.

Figure

Disclosures

No relevant conflicts of interest to declare.
Correction of Iron Deficiency in Hospitalized Heart Failure Patients Does Not Improve Patient Outcomes
Authors: Mistry Ronak , Kohut Andrew , Locantore-Ford Patricia
Published: November 2019

Background:

Heart failure (HF) is a chronic medical condition that affects approximately 1-2% of the world's population and greater than 10% of those age 65 and older. Among patients with HF, iron deficiency (ID), defined as ferritin <100 ng/ml or transferrin saturation (TSat) <20% with ferritin 100-300 ng/ml, has an estimated prevalence of 30-50%, often without concomitant anemia. Thus, ID in HF is often underdiagnosed unless actively sought after. ID in HF has been shown to be an independent contributor of increased mortality, hospitalization and early readmission compared to HF patients without ID or HF patients with anemia without ID. A large trial, namely IRONOUT, illustrated the superiority of intravenous iron (IVFe) over oral iron supplements for HF patients who were identified as iron deficient. The FAIR-HF and CONFIRM-HF trials showed improved functional outcomes and quality of life in HF patients treated with IVFe and that these benefits were conferred even 1 year after treatment. However, these trials focus on iron store correction in patients with chronic heart failure patients. Little data are available regarding the correction of ID in patients who are admitted to hospitals for HF exacerbations.

Methods:

We performed a retrospective analysis of hospital records of patients admitted with HF as a primary or secondary diagnosis to a city-based large academic teaching health system between April 1, 2014 and April 1, 2017. Thereafter, we identified those patients who had sufficient iron studies (hemoglobin, iron, ferritin, transferrin, TSat) during their hospitalization to diagnose ID, as per the criteria above. Patients with ID were then stratified into those that did and did not receive IVFe. Patient charts were evaluated to two primary endpoints, namely one-year readmissions for HF and patient mortality.

Results:

In total, 390 patients had sufficient iron studies performed to assess ID status, of which 279 met criteria for ID (71.5%). Eighty-four of the 279 patients received IVFe. Amongst patients that received IVFe, 38 patients were readmitted for HF (45.2%) and 6 patient deaths occurred within 1 year (7.14%). Amongst the remaining 195 patients who did not receive IVFe, 93 were readmitted for HF (47.69%) and 15 patient deaths occurred within 1 year (7.69%).

Conclusion:

These data suggest that despite clinical benefit in patient functional outcomes and overall quality of life in chronic heart failure patients treated with IVFe, treatment of ID in acute HF patients does not result in a reduction in admissions for HF or improvements in patient mortality.

Disclosures

No relevant conflicts of interest to declare.
Genotypic and Alelicas Frequencies of the ABO and Rh Systems in the State of Rio Grande Do Norte, Northeast of Brazil
Authors: Morais Linduarte Varela , Paiva Aldair Sousa , Sales Valéria SF , Cavalcanti Geraldo Barroso
Published: November 2019

Objective: To determine the immunophenotyping of a population of blood donors, intended to build a database for transfusion medicine. 2) To make available to patients with chronic diseases that are systematically dependent on blood transfusions as compatible as possible in ABH, Rh (DCcEe) and Kell 1 antigen systems. Method: A cross-sectional study was carried out on 11,664 blood donors for the ABH system typing and of these, 1255 blood donors were selected randomly for the determination of blood group antigens of the Rh system and Kell antigen. Blood centrifugation methods, centrifuge hemolysis tube test and indirect Coombs test were used for blood typing. The results obtained were compared by the Chi-square test. A level of statistical significance of p ≤ 0.05 was considered. Results: Antigenic frequencies for the ABH system found: the frequency found for the blood group O 48.8%, the frequency found for the blood group A 35.4%; the frequency found for the blood group B 10.6% and the frequency found for the blood group AB 3.2%. In the Rh-Hr system the most frequent antigens found: e 94.5%, D 88.9%; c 80.6; C 56.4%; E 26.3%. For the Kell antigen, the frequency found was 6.7%. The most frequent phenotypes found were DCcee 23.3%; ddccee 18.1%, DCCee 16.7%; Dccee 11.0%; DCcEe 10.2% and DccEe 8.8%. The lowest frequency was found: DCcEE 0.64% and ddCcEe 0.08% Conclusion: The antigenic and phenotypic frequencies found show the great importance and necessity of the immunophenotyping of these antigens of blood groups in order to transfuse them, making them as compatible as possible, thus reducing the risk of alloimmunization. Due to the great miscegenation of our population, we find different frequencies in each region, making initiatives in this regard more relevant.

Disclosures

No relevant conflicts of interest to declare.
β2-m - a Trigger for NLRP3 Inflammasome Activation within Macrophages Promoting Multiple Myeloma Cell Proliferation
Authors: Hofbauer Daniel , Mougiakakos Dimitrios , Büttner-Herold Maike , Neumann Frank , Nolting Jens , Bisht Savita , Zeiser Robert , Hamarsheh Shaima'A , Ricagno Stefano , Zaiss Mario , Eberhardt Martin , Boettcher Martin , Bitterer Katrin , Mackensen Andreas , Bruns Heiko
Published: November 2019

Multiple myeloma (MM) is closely dependent on cross-talk between malignant plasma cells and cellular components of the inflammatory bone marrow milieu, which promotes disease progression, bone destruction and immune-impairment. However, the molecular mechanisms initiating this inflammatory microenvironment remain poorly defined. Beta-2-microglobulin (β2M) accumulates during MM and represents an established prognostic biomarker, but little is known regarding its immunoregulatory potential. Here, we found that β2m triggers an enhanced caspase-1 dependent release of mature IL-1β (15.8 ± 4.5 ng/ml) and IL-18 (5.8 ± 2.1 ng/ml) by macrophages in a NLRP3 dependent way. Activation of the NLRP3 inflammasome was initiated, after phagocytosis of β2m and subsequent lysosomal rupture due to amyloid fibril formation in lysosomes, in a process which partially involved lysosomal protease cathepsin B. β2m mediated NLRP3 inflammasome activation also occurs in tumor-associated macrophages (TAMs) of MM patients leading to the release of IL-1β and IL-18 towards MM cells. Moreover, TAMs in a murine MM model (5TMM) displayed an active NLRP3 inflammasome as well. Treatment of 5TMM mice with MCC950, a selective NLRP3 inhibitor, reduced tumor progression and thus attenuated the severity of MM. Our findings shed new light on β2m as macrophagic NLRP3 inflammasome activator which contributes to MM cell progression, and open a potential novel avenue for therapies of this severe malignancy.

Figure

Disclosures

Bruns: Morphosys AG: Research Funding.
Smooth Muscle Cytochrome b5 Reductase 3 Deficiency Accelerates Pulmonary Hypertension Development in Sickle Cell Mice
Authors: Wood Katherine , Durgin Brittany , Schmidt Heidi M , Hahn Scott , Baust Jeffrey , Bachman Tim , Vitturi Dario A. , Ghosh Samit , Ofori-Acquah Solomon Fiifi , Mora Ana , Gladwin Mark T , Straub Adam
Published: November 2019

Pulmonary and systemic vasculopathy are significant risk factors for early morbidity and death in patients with sickle cell disease (SCD). An underlying mechanism of SCD vasculopathy is vascular smooth muscle (VSM) nitric oxide (NO) resistance, which is mediated by NO scavenging reactions with plasma hemoglobin and reactive oxygen species that can oxidize soluble guanylyl cyclase (sGC), the NO receptor. Prior studies show that cytochrome b5 reductase 3 (CYB5R3), known as methemoglobin reductase in erythrocytes, functions in VSM as an sGC heme iron reductase critical for reducing and sensitizing sGC to NO and generating cGMP for vasodilation. Therefore, we hypothesized that VSM CYB5R3 deficiency accelerates development of pulmonary hypertension (PH) in SCD. Bone marrow transplant was used to create SCD chimeric mice with background smooth muscle cell (SMC) specific tamoxifen-inducible Cyb5r3 knockout (SMC R3 KO) and wild-type (WT) control.Three weeks after completing tamoxifen treatment, we observed 60% knockdown of pulmonary arterial SMC CYB5R3, 5-6 mmHg elevated right ventricular maximum systolic pressure (RVmaxSP) and biventricular hypertrophy in SS chimeras with SMC R3 KO (SS/R3KD) relative to wild-type (SS/R3WT). Right ventricular (RV) contractility, heart rate, hematological parameters and cell-free hemoglobin were similar between groups. When identically generated SS/R3 chimeras were studied 12-weeks after completing tamoxifen treatment, RVmaxSP in SS/R3KD had not increased further, but RV hypertrophy relative to SS/R3WT persisted. These are the first studies to establish involvement of SMC CYB5R3 in SCD-associated development of PH, which can exist in mice by 5 weeks of SMC CYB5R3 protein deficiency.

Disclosures

Vitturi: Complexa Inc: Consultancy. Ofori-Acquah:Shire Human Genetic Therapies Inc: Other: Financial Relationship. Gladwin:Globin Solutions, Inc: Patents & Royalties: Provisional patents for the use of recombinant neuroglobin and heme-based molecules as antidotes for CO poisoning; Bayer Pharmaceuticals: Other: Co-investigator; United Therapeutics: Patents & Royalties: Co-inventor on an NIH government patent for the use of nitrite salts in cardiovascular diseases . Straub:Bayer Pharmaceuticals: Research Funding.
Reduced Packed Red Blood Cell Utilization Via Implementation of Transfusion Guidelines in a Large Health System Hospital
Authors: Bowers Reid Christian , Thompson Michael A. , Van Dreser Julie , Sanchez Federico Augusto
Published: November 2019

Background: We have previously discussed how implementation of Patient Blood Management (PBM) principles such as transfusion guidelines helped reduce packed red blood cell (PRBC) usage in cardiovascular surgery patients (Bowers et al, Reduced Blood Product Utilization via Implementation of an Anemia Clinic and Consult Service in a Large Health System Hospital https://tinyurl.com/y3ucr3sz). We now discuss the effect of implementing transfusion guidelines on PRBC transfusion practices at an institutional level at St. Luke's Medical Center, Advocate Aurora Health in Milwaukee, WI.

Methods: Beginning in 2015 Aurora Health Care made 7 g/dL the standard threshold below which packed red blood cell (PRBC) transfusion is considered appropriate. Additionally, transfusing PRBCs as single units with a hemoglobin (Hgb) measurement between each unit was made standard. Education was provided to clinicians on the new standard including best practice advice in the transfusion ordering workflow. Data was gathered on institutional PRBC use and individual clinician PRBC ordering practices. Further education was provided based on the extent to which data showed conformity to the new standard.

Results: PBM implementation 2015-2018 was associated with a drop in institutional PRBC usage from 11,490 to 9301 units. The percentage of transfusions ordered with Hgb < 7 g/dL rose from 41.6% to 71.8%. The percentage of single unit orders increased from 67.3% to 91.8%. See Graph/Table 1 & 2 for further analysis of individual practitioner PRBC ordering. See Table 3 for reporting period volume data.

Conclusions: Implementation of PRBC transfusion guidelines was associated with a greater than 2000-unit annual reduction in PRBC usage. Reduction was associated with individual clinician transfusion practices that increasingly conformed with guidelines over time.

Disclosures

Thompson: Adaptive: Membership on an entity's Board of Directors or advisory committees; AbbVie: Research Funding; UpToDate: Patents & Royalties: Myeloma reviewer; BMS: Research Funding; Celgene: Membership on an entity's Board of Directors or advisory committees; Lynx Bio: Research Funding; VIA Oncology: Membership on an entity's Board of Directors or advisory committees; GSK: Membership on an entity's Board of Directors or advisory committees; Doximity: Equity Ownership, Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees, Research Funding.
Use of Multi-Omics Approach to Produce a Robust Serum-Free Media for the Expansion of T Cells
Authors: Mansour Sarya G. , Lin Pei-Yi , Pierce Anson , Civin Curt I
Published: November 2019

Chimeric antigen receptor (CAR) T-cell therapy represents a major advancement in personalized cancer treatment. Generating robust and well characterized manufacturing processes for cell therapy has become crucial. Traditional manufacturing processes activate and expand T cells in media containing human serum, which supports cell growth and viability. However, serum batches vary significantly from lot to lot and require frequent screening for contaminants that can be detrimental to patients. T cell manufacturing processes independent of human serum will render adoptive T cell therapy less expensive, more consistent, and safer for patients. In order to create a best-in class serum-free medium that is even more robust than serum-containing media, we need to gain a better understanding of the metabolic requirements of T cells during expansion. For this purpose, we have utilized a multi-omics approach to fully characterize the proteomic and metabolomic signatures of T cells expanded in a serum free, xeno free medium at different phases of growth. We sampled cells at days 3, 5, and 7 for label-free proteomics and untargeted extra- and intracellular metabolomics analysis in order to identify metabolic patterns that could be corrected with media supplementation. We identified over 6,043 proteins and 900 metabolites from 4 donors and detected 1,200 significantly different proteins and 312 metabolites. Using an in-house bioinformatics strategy to analyze the multi-omics data, we focused on several metabolic and signaling pathways that were significantly different at days 5 and 7 compared to day 3. The "corrective" media supplementation that we added based on these results demonstrated a tremendous increase in cell growth and viability yielding a 2.6- and 3.2-fold increase in cell growth on days 5 and 7, respectively, and an increase in viability of 15-20% and 11-15% on days 5 and 7, respectively, compared to the un-supplemented prototype. In addition, cells maintained high viability throughout the whole culture and phenotype was not affected. These data demonstrate that multi-omics is a powerful tool for understanding T cell metabolism and identifying components to develop robust and reproducible serum-free media that produces high quality T cells.

Disclosures

No relevant conflicts of interest to declare.
The Diagnostic Performance of a Non-Emergency Department-Based Deep Vein Thrombosis Ambulatory Care Pathway
Authors: Sene Pape-Mamadou , Yahiaoui Denis , Koolian Maral , Tagalakis Vicky
Published: November 2019

Background

Ambulatory management of isolated acute deep vein thrombosis (DVT) is the standard of care in most patient populations, and in clinical practice is mostly initiated in the emergency department (ED). However, patients referred to the ED with suspected DVT often experience unnecessary delays to diagnosis and subsequent long stays to determine appropriate treatment and follow up care. We implemented a DVT ambulatory care pathway that does not include the ED for referred patients with suspected isolated DVT that begins with pre-test risk stratification in the community and subsequent algorithm-based diagnostic work-up, treatment, and follow-up at a tertiary care centre in Montreal Canada.

Objective

To determine the diagnostic performance of a non-ED based DVT ambulatory care pathway.

Methods

We determined the prevalence of DVT over a 46 week period between August 2018 and June 2019 among ambulatory patients with suspected isolated DVT who following community-based risk stratification using the modified Well's clinical prediction score were referred for diagnostic work up and treatment using a newly implemented non-ED based DVT ambulatory care pathway.

Results

Among 122 patients referred by community physicians, 86 (70%) met pre-defined pathway criteria for assessment of suspected DVT. In all, 42 (49%) were referred with an unlikely/low modified Well's score and 44 (51%) with a likely/high score. Overall, the prevalence of DVT was 19.8%, specifically 9.5% in the unlikely/low and 30.2% in the likely/high pretest probability groups, respectively.

Conclusion

Our results show that the diagnostic performance of a non-ED based acute DVT ambulatory care pathway is in line with literature estimates. The advantage of this pathway is that it offers clear, evidence-based guidance for community physicians to diagnosis and treat patients in an ambulatory setting without using the ED. The approach is likely to result in both healthcare and economic benefits, including increased patient satisfaction and shorter ED stays.

Disclosures

Tagalakis: Sanofi Aventis: Other: investigator initiated grant;participated on ad boards; Pfizer: Other: participated on ad boards; BMS-Pfizer: Other: participated on ad boards; Servier: Other: participated on ad boards; Bayer: Other: participated on ad boards.
Anaerobic Antibiotics and the Risk of Graft-Versus-Host Disease after Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Tanaka John , Young Rebecca , Spees Lisa , Jenkins Kirsten , Sung Anthony D. , Martin Paul L. , Heston Sarah , Thompson Jillian C. , Seed Patrick , Arshad Mehreen , Kelly Matthew
Published: November 2019

Background: The gut microbiota interacts extensively with the host immune system and thus may modify the risk of graft-versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). During the post-transplant neutropenic period, the majority of allogeneic HSCT recipients receive empirical broad-spectrum antibiotics for febrile neutropenia. We hypothesized that receipt of an antibiotic regimen with an anaerobic spectrum of activity is associated with a higher risk of grade II-IV acute GVHD than receipt of a non-anaerobic antibiotic regimens.

Methods: In this single-center retrospective cohort study, we evaluated associations between peri-transplant receipt of antibiotics with an anaerobic spectrum of activity and the risk and severity of GVHD among 877 adults who received an allogeneic HSCT between January 1, 2005 and December 31, 2016. We identified 609 patients who developed febrile neutropenia after HSCT and compared GVHD risk and mortality among patients who received anaerobic antibiotics (piperacillin-tazobactam or carbapenems; n=333) to patients who received only antibiotics with minimal activity against anaerobes (aztreonam, ceftazidime, or cefepime; n=276). Antibiotics received by patients between 7 days before and 28 days after allogeneic HSCT and GVHD diagnoses were verified via manual review of medication orders and provider notes in electronic medical records.

Results: Receipt of anaerobic antibiotics was associated with an increased risks of grade II-IV acute GVHD (hazard ratio (HR): 1.41; 95% confidence interval (CI): 1.10-1.79; P=0.01) and acute GVHD mortality (HR: 1.87; 95% CI: 1.13, 3.11; P=0.02). This hazard was primarily associated with acute GVHD of the gut or liver (HR: 1.38; 95% CI: 1.06, 1.79; P=0.02). The association remained with even short (<7 days) courses of anaerobic antibiotics. Anaerobic antibiotic exposure was not associated with acute skin GVHD (HR: 0.97; 95% CI: 0.69, 1.37; P=0.88), chronic GVHD diagnosis (HR: 0.93; 95% CI: 0.70, 1.23; P=0.43), or chronic GVHD mortality (HR: 0.89; 95% CI: 0.44, 1.81; P=0.76).

Conclusions: Receipt of anaerobic antibiotics for febrile neutropenia post-HSCT is associated with an increased risks of acute GVHD of the gut or liver and acute GVHD mortality. Limiting use of antibiotics with an anaerobic spectrum of activity after allogeneic HSCT may reduce acute GVHD incidence and mortality.

Disclosures

Sung: Novartis: Research Funding; Merck: Research Funding; Seres: Research Funding. Martin:Novartis Pharmaceuticals: Other: research support; Jazz Pharmaceuticals: Other: research support.
Excess Serine from the Microenvironment Caused the Impaired Megakaryopoiesis and Thrombocytopia in Multiple Myeloma
Authors: Kuang Chunmei , Xia Meijuan , An Gang , Liu Cuicui , Wu Dan , Su Pei , Zhang Jingyu , Xia Jiliang , Guo Jiaojiao , Wu Xuan , Jia Wei , Qiu Lugui , Zhou Wen , Zhou Jiaxi
Published: November 2019

Background: Thrombocytopenia is major complication in a subset of patients with multiple myeloma (MM). However, a lack of detailed studies about the megakaryopoiesis, thrombopoiesis as well as their connections with the survival of patients limits us to explore whether thrombocytopenia could be used as a reliable prognostic factor for MM.

Materials and Methods: In this study, 1393 newly diagnosed MM patients were selected for investigating the potential connection between PLT counts and clinical characteristics including ISS stage, overall survival as well as progression free survival. Besides, 5T33MMvt-KaLwRij mouse model were also used to examine the megakaryopoiesis and thrombopoiesis during disease progression. The proportion and function of different subpopulations of cells including megakaryocytes, megakaryocytic-erythroid progenitors (MEPs), common myeloid progenitors (CMPs) and Lin-Sca-1+c-kit+ (LSK) cells were measured both in MM patients and mouse models. Gas chromatography-time-of-flight mass spectrometry (GC-TOFMS)-based metabolomics was used to analyze the metabolites.

Results: Of the 1393 studied patients, 298 cases of MM patients are found with thrombocytopenia at the time of diagnosis. PLT counts were lower both in stage Ⅱ (P<0.01) and Ⅲ patients (P<0.001) than stageⅠpatients. Interesting, we found MM patients with thrombocytopenia had a significantly lower OS (P<0.001) and PFS (P<0.001). In mouse model, we also found PLT counts gradually decreased in peripheral blood during the disease progression (P<0.001). Further analysis demonstrated the proportion and the absolute numbers of megakaryocytes and MEPs were diminished both in mouse and MM patients with thrombocytopenia. PLT counts were negative correlated with the percentage of plasma cells or IgG2b levels (P<0.001), suggesting a potential connection of malignant cells infiltration and thrombocytopenia. In mechanism, metabolomics analysis with BM plasma identified 16 differential metabolites in MM patients with thrombopoiesis (VIP > 1.2, P < 0.05). Among them, serine was observed significantly be elevated in MM mouse and suffice to inhibit the megakaryopoieis and thrombopoiesis in vitro.

Conclusion: PLT counts might be used as a reliable prognostic factor for MM patients since the thrombocytopenia was associated with poor survival in MM patients. The thrombocytopenia in MM might attribute to, at least partially, the inhibition by serine from the microenvironment. Our findings revealed novel mechanism of MM and might eventually shed light on the treatment of MM patients.

Disclosures

No relevant conflicts of interest to declare.
Effects of Interferon-γ on Proliferation and Ability of Secretion of Human Amniotic Mesenchymal Stem Cells
Authors: Gao Ya , Xu Ying , Li Weiru , Zhang Yintian , Ping Baohong , Liu Qifa
Published: November 2019

Objective:The immunoregulatory properties and proliferation of mesenchymal stem cells (MSCs) could be affected by inflammatory factors. However, there have been few studies about human amniotic MSCs (hAMSCs). We investigated the effects of interferon (IFN)-γ on the proliferation and apoptosis of hAMSCs, and measured the level of inflammatory factors secreted by hAMSCs. Result:hAMSCs were cultured with complete medium with different concentrations of IFN-γ. We detected the proliferation of hAMSCs by Cell Counting Kit-8 assays, analysed apoptosis by flow cytometry (FCM) at 48 h, and mesasured the level of inflammatory factors such as solube HLA-G and prostaglandin E2 (PGE2) in the supernatant at 48 h by ELISA. The level of kynurenine (KYN) was measured by ultraviolet spectrophotometry. As culture time increased, the proliferation of hAMSCs with different concentrations of IFN-γ increased rapidly from day 1 to day 4, and then the growth rate slowed. FCM indicated that there was no significant apoptosis in the 100 ng/ml IFN-γ group compared with cells without IFN-γ. The level of PGE2 and soluble HLA-G in cells with IFN-γ was higher compared with those without IFN-γ. The level of KYN increased significantly in the cells with IFN-γ. Conclusion:IFN-γ did not affect the growth and proliferation of hAMSCs, and promoted secretion of PGE2 and soluble HLA-G, and enhanced activity of indoleamine 2,3-dioxygenase (IDO), providing a theoretical basis for hAMSCs to prevent and treat graft-versus-host disease.

Disclosures

No relevant conflicts of interest to declare.
Anti-BTN3A 20.1 Agonist Monoclonal Antibody Enhances Autologous Vγ9Vδ2 T Cells Cytotoxicity Against Primary Acute Myeloid Blasts
Authors: Le Floch Anne-Charlotte , Orlanducci Florence , Briantais Antoine , Le Roy Aude , Chretien Anne-Sophie , Vey Norbert , Olive Daniel
Published: November 2019

Introduction: Vγ9Vδ2 T cells immunotherapy is an emerging strategy in acute myeloid leukemia (AML). Partial effects of phosphoantigens like zoledronate, prompted us to investigate other molecules. Using Vγ9Vδ2 T cells expanded from healthy volunteers, recent works of our team demonstrate that anti-BTN3A 20.1 agonist monoclonal antibody (mAb) improves Vγ9Vδ2 T cells cytotoxicity in vitro and in vivo, including in zoledronate resistant blasts. Efficacy of anti-BTN3A 20.1 agonist mAb remains undetermined using autologous Vγ9Vδ2 T cells and mechanism of its action on AML blasts is understudied.

Material and methods: Expression of co-stimulatory molecules on surface of AML blasts and Vγ9Vδ2 T cells, from 37 PBMC of AML patients, was assessed by flow cytometry. Vγ9Vδ2 T cells expansion was realized from the same samples with zoledronate, and administrations of Il-2 and Il-15. Purity was assessed by flow cytometry at day 13. Degranulation assay was performed at day 14.

Results: 20 degranulation assays were realized on successfully expanded Vγ9Vδ2 T cells (69% of analyzable samples). Degranulation with anti-BTN3A 20.1 agonist mAb was enhanced compared to zoledronate condition (p=0.0085) or negative control (p<0.0001). We also show a positive correlation between level of degranulation and expression of DNAM-1 on Vγ9Vδ2 T cells surface, exclusively under anti-BTN3A 20.1 agonist mAb treatment (rS=0.54; p=0.0134). Conversely, under zoledronate treatment, Vγ9Vδ2 T cells degranulation was positively correlated with expression of NKG2D on Vγ9Vδ2 T cells surface (rS=0.62; p=0.0099) and was inversely associated to their expression of PD-1 (p=0.0482).

Conclusion: Efficacy of anti-BTN3A 20.1 agonist mAb on primary AML blasts is confirmed in an autologous setting. Degranulation seems to play a major role in enhancement of Vγ9Vδ2 T cells cytotoxicity under anti-BTN3A 20.1 agonist mAb treatment, where DNAM-1 could play a determinant role. An autologous strategy for Vγ9Vδ2 T cells may be studied and considered in AML patients, with potent extrapolation to γδ T cell therapy protocols in targeting strategies.

Disclosures

Vey: Novartis: Consultancy, Honoraria; Janssen: Honoraria. Olive:ImCheck Therapeutics: Consultancy, Equity Ownership, Patents & Royalties; GlaxoSmithKline: Patents & Royalties.