bluuebunny/crossref_metadata_2025 huggingface dataset:
Oral Posaconazole or Intravenous-Oral Itraconazole in Preventing Invasive Fungal Diseases for Patients with Acute Leukemia
Authors: Liu Li , Pei Xiaolei , Ma Runzhi , He Yi , Zhang Rongli , Wei Jialin , Ma Qiaoling , Zhai Weihua , Jiang Erlie , Pang Aiming , Yang Donglin , Feng Sizhou , Han Ming-Zhe
Published: November 2020

Background and Objectives:

Invasive fungal diseases (IFDs) are major and lethal infectious complications for patients with neutropenia after chemotherapy for acute leukemia (AL). We aimed to compare the use of oral posaconazole vs. itraconazole in an intravenous-oral regimen for primary antifungal prophylaxis in terms of efficacy, adverse events and costs among neutropenic patients with AL after chemotherapy.

Methods:

This single-center, retrospective study enrolled patients with AL. Prophylaxis with oral posaconazole or intravenous-oral itraconazole (intravenous formulation × 2 days + oral suspension) was administered for patients recovering from neutropenia after chemotherapy. Propensity-score matching was used to assemble a cohort of patients with similar baseline characteristics.

Results:

Among 342 eligible episodes from 186 patients, 110 episodes received oral posaconazole and 110 episodes received itraconazole in sequential regimen had similar propensity scores and were included in the analyses. The incidence of breakthrough IFDs was 0.0% (0/110) and 0.0% (0/110) in the posaconazole group and itraconazole group, while the incidence of proven, probable and possible IFDs was 1.8% (2/110) and 8.2% (9/110), respectively (P=0.030). In clinical failure analysis, the failure rate of posaconazole group was lower as compared to the itraconazole group (2.7% vs. 10.9%, P=0.016). The proportion of patients who needed systemic antifungal treatment were lower in posaconazole group than that in itraconazole group (2.7% vs. 10.0%, P=0.027). Five cases (4.5%) experienced adverse events possibly associated with posaconazole and thirteen cases (11.8%) with itraconazole (P=0.049). There was no significant difference of survival between patients on posaconazole or itraconazole. The acquisition costs of posaconazole were higher than those of itraconazole (P=0.000).

Conclusions:

Both oral posaconazole and intravenous-oral itraconazole are effective in preventing IFDs for acute leukemia patients recovering from neutropenia after chemotherapy, while posaconazole is slightly better. Oral posaconazole is safer and more tolerable but costs higher than intravenous-oral itraconazole.

Disclosures

No relevant conflicts of interest to declare.
Removal of Abnormal M Protein in Patients with Multiple Myeloma By Plasma-Free Exchange
Authors: Zhou Fuling , Liu Dandan , Wu Tianzhi
Published: November 2020

ObjectiveBased on the albumin substitution fluid, we performed plasma-free exchange in the removal of abnormal M protein for patients with multiple myeloma using Fresenius COM.TEC machine

MethodsFrom February 2016 to August 2018, 70 patients with multiple myeloma, who have indication of plasma exchange because of increased M protein, were treated with plasma-free exchange. Record the relevant basic information, the clinical symptoms and related disease indicators, and use SPSS 20.0 to analyze the changes of erythrocyte sedimentation rate, blood routine, albumin, renal function, serum calcium and immunoglobulin, and report adverse reactions.

ResultsAfter treatment, the patient's erythrocyte sedimentation rate, serum calcium, serum creatinine, β2-microglobulin, uric acid, IgG, IgA were significantly reduced; hemoglobin, white blood cell level were not statistically significant. The creatinine remission rate was 97%, and the platelet decline was only 17.8%, and no adverse reactions related to thrombocytopenia such as hemorrhage and ecchymoses were observed. Patients with IgG type responded better than IgA-free treatment to plasma-free exchange. There were no serious adverse reactions after treatment, 1 case of blood pressure drop, 2 cases of low calcium reaction, all in the normal range, and recovered after the end of treatment.

ConclusionPlasma-free exchange is an effective technology reducing M protein, which can effectively improve the erythrocyte sedimentation index, renal function index and blood viscous state. Patients with IgG type or higher ISS stage have more obvious effects. Plasma-free exchange significantly get rid of the application of fresh frozen plasma.

Disclosures

No relevant conflicts of interest to declare.
Utilizing Medical Students As Patient Mentors: An Intervention to Improve Transition Readiness in Young Adults with Sickle Cell Disease
Authors: Viola Adrienne , Drachtman Richard A. , Kaveney Amanda , Sridharan Ashwin , Savage Beth , Porter Jerlym S. , Devine Katie
Published: November 2020

Background:Advances in medical care have resulted in nearly 95% of all children with sickle cell disease (SCD) living to adulthood. Standardized transition programming does not currently exist, resulting in high rates of mortality and morbidity among young adults (YA) during the transition period. Mentoring and social support have been used to impact health behavior change. Medical student mentors can serve as mentors offering specialized support for YA. This study examined the feasibility and preliminary efficacy of a medical student mentor intervention to improve transition outcomes for YAs with SCD.

Methods:24 YA with sickle cell disease and 9 medical student mentors enrolled in the intervention. Feasibility and acceptability of the intervention was assessed through enrollment rates, reasons for refusal, retention rates, engagement with the intervention, satisfaction, and reasons for drop-out. The preliminary efficacy of the intervention among patient participants was assessed using dependent t-tests to evaluate changes in transition readiness, health related quality of life, self-efficacy, SCD knowledge medication adherence, and health literacy. Among medical student mentor participants, changes in attitudes towards chronic illness and SCD knowledge were explored.

Results:Patient participants demonstrated adequate enrollment (63.2%), retention (75.0%), and adherence to the intervention (88.3%) and rated the intervention components highly. Patient participants demonstrated significant improvements in transition readiness (p= .001), self-efficacy (p= .002), medication adherence (p= .02), and health literacy (p= .05). Medical students also demonstrated significant improvements in SCD knowledge (p= .01).

Discussion:A medical student mentor intervention to facilitate transition from pediatric to adult care for young adults with SCD is both feasible and acceptable to patients and medical students. Preliminary evidence suggests such an intervention may provide dual benefit for both patients and students. A randomized controlled trial is needed to evaluate efficacy.

Disclosures

No relevant conflicts of interest to declare.
CD33 Targeting Primary CAR-NK Cells Generated By CRISPR Mediated Gene Insertion Show Enhanced Anti-AML Activity
Authors: Naeimi Kararoudi Meisam , Likhite Shibi , Elmas Ezgi , Schwartz Maura , Sorathia Kinnari , Yamamoto Kenta , Chakravarti Nitin , Moriarity Branden S , Meyer Kathrin , Lee Dean Anthony
Published: November 2020

Human peripheral blood natural killer (NK) cells have intense antitumor activity and have been used successfully in several clinical trials. Modifying NK cells with a chimeric antigen receptor (CAR) can improve their targeting and increase specificity. Recently, we described an efficient method for gene targeting in NK cells using Cas9/ribonucleoprotein complexes (PMID: 29985369 and 32603414). Here we combined this approach with single-stranded (ss) or self-complementary (sc) Adeno-associated virus (AAV)-mediated gene delivery for gene insertion into a safe-harbor locus using a wide variety of homology arms for homology repair (HR) and non-homologous directed CRISPR-assisted insertion tagging (CRISPaint) approaches. We demonstrated that expansion of NK cells on feeder cells (CSTX002) expressing membrane-bound IL21 increases expression of HDR-related genes and provides optimum biological condition for targeted gene insertion. For proof-of-concept, we successfully generated stable mCherry-expressing primary NK cells (up to 89% mCherry+) and determined that sc vectors with 300bp homology arms were optimal. Then, we generated CD33-targeting CAR NK cells with differing transmembrane and signaling domains (CD4/4-1BB+CD3ζ and NKG2D/2B4+CD3ζ), which continued to show robust expansion on CSTX002 and stably maintained their CAR expression. This resulted in CAR-NK-cells of high number and purity (mean 68% CAR+) that demonstrated enhanced antileukemic activity against acute myeloid leukemia (AML) cell lines. This efficient method for site-directed insertion of genetic materials into primary NK cells has broad potential for fundamental discovery and therapeutic applications.

Keywords: CRISPR, NK, Cas9/RNP, AAV6, CRISPaint, HR, CD33CAR-NK

Figure

Disclosures

Naeimi Kararoudi: Kiadis Pharma Netherlans B.V: Patents & Royalties. Lee:Kiadis Pharma Netherlands B.V: Consultancy, Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.
Integrin αIIbβ3 Regulates Platelet-Procoagulant Activity in the Lung
Authors: Brzoska Tomasz , Kaminski Tomasz W. , Vats Ravi , Tutuncuoglu Egemen , Gladwin Mark T , Sundd Prithu
Published: November 2020

Pulmonary thrombosis is a major complication associated with high morbidity. Despite advances in diagnosis and treatment, the pathophysiology of pulmonary thrombosis remains incompletely understood. New clinical evidence suggests that in situ platelet activation resulting in enhanced procoagulant activity may promote pulmonary thrombosis. Improved understanding of the etiological mechanism would enable the development of new therapies for pulmonary thrombosis.

Collagen and thromboplastin (TF) were administered intravascularly (IV) to C57BL/6 (WT) mice and the pulmonary microcirculation was visualized using quantitative fluorescence intravital fluorescence lung microscopy (qFILM). Fluorochrome-conjugated anti-mouse CD49b Ab and dextran was administered IV for in vivo staining of circulating platelets and visualization of blood vessels, respectively. Pulmonary thrombosis was defined as occlusion of blood vessels with platelet aggregates leading to pulmonary ischemia. Additionally, quantitative microfluidic fluorescence microscopy (qMFM) was used to study the effect of platelet αIIbβ3 inhibition on platelet procoagulant activity in human blood under vascular mimetic flow conditions.

Collagen and TF triggered dose-dependent pulmonary thrombosis in mice in vivo, which involved development of platelet-rich thrombi in the pulmonary arteriolar bottlenecks (junction of pulmonary arteriole and capillaries), resulting in a transient ischemia in the arteriole and the down-stream capillary tree. The pulmonary arteriole thrombosis triggered by IV collagen or TF was protracted, lethal and completely abrogated following IV administration of αIIbβ3 receptor inhibitor (eptifibatide). Inhibition of platelet αIIbβ3 also significantly reduced platelet procoagulant activity, fibrin formation and thrombus formation in human blood flowing through microfluidic channels ex vivo.

Our current findings suggest that αIIbβ3-dependent platelet procoagulant activity promotes pulmonary thrombosis. Both our models have potential application in investigating the molecular determinants of pulmonary thrombosis in diverse pulmonary disorders as well as evaluating efficacy of new antithrombotic drugs.

Disclosures

No relevant conflicts of interest to declare.
Changes of Peripheral Blood Lymphocyte Subsets and Cytokines in Patients with Multiple Myeloma and Their Clinical Significance
Authors: Fei Li
Published: November 2020

Objective: Accumulating evidence have indicated that immune response play an essential role in development of multiple myeloma (MM),To evaluate the immune function of patients with MM by detecting the levels of lymphocyte subsets and cytokines, and to analyze the correlation between the immune function and prognosis of patients. Methods: we examined 50 patients with newly diagnosed MM, 20 patients with Relapse / refractory MM and 30 healthy volunteers. the levels of T lymphocyte subsets, activated T cells subtypes, Treg and cytokines in peripheral blood were detected by flow cytometry (FCM).Results: ① compared with healthy control group, CD3 + CD4 + T cells of newly diagnosed MM patients were significantly decreased (P = 0.04), There was no significant difference in CD3+CD8+T and CD4/CD8 of newly diagnosed MM patients with normal controls (P=0.14).The percentage of CD16 + cd56dim NK cells of newly diagnosed MM was significantly lower than that of the normal control group (P = 0.01); There was no significant difference in frequencies of CD3+CD4+T cells, CD4/CD8, CD16+CD56dimNK (P=0.34, P=0.561, P=0.88) between newly treated MM patients and relapsed/refractory MM; HLA-DR-CD8+ activated TS cells of newly diagnosed MM was significantly higher than the normal control group (P = 0.04).② The proportion of CD4 + CD25 + CD127dim T cells(treg) in total CD4 + T cells in of newly diagnosed MM was higher than that in normal control group (P < 0.05). The CD4 + CD25 +CD127dim T cells of MM patients with relapse / refractory MM was higher than that of newly treated patients, but the difference was not statistically significant (P > 0.05). Further analysis showed that CD4 + CD25 + CD127dim T cells in mSMART stratified high-risk group were higher than those in low-risk group (P = 0.03). The number of CD4 + CD25 + CD127dim T cells in patients without PR after 2 courses of PAD was higher than that in patients with≥ PR, but the difference was not statistically significant (P > 0.05). ③ The levels of IL-6, IL-8, TNF - α of newly diagnosed MM were significantly higher than normal control group, and were positively correlated with ISS stage (r=0.61, r=0.67, r=0.59, P均<0.05).④ The levels of IL-6 and IL-8 of newly diagnosed MM with renal injury were significantly higher than those in patients with normal renal function (P < 0.05). Further studies showed that the level of IL-8 was negatively correlated with the proportion of CD16 + CD56dimNK cells (r=-0.65, P<0.05).Conclusion: the abnormal expression of lymphocyte subsets and cytokine levels in MM patients may be related to the tumor load, disease progression and prognosis.

Disclosures

No relevant conflicts of interest to declare.
Low Dose Rituximab Plus High Dose Dexamethasone As First-Line Treatment for Autoimmune Hemolytic Anemia
Authors: Rodríguez-Roque Carlos Saúl , Gomez-De Leon Andres , Morcos-Sandino Michelle , López-Flores Nelson Josafat , Galindo-Calvillo David David , Zertuche-González Elsa Laura , Rodríguez-Zuñiga Anna Cecilia , Gomez-Almaguer David
Published: November 2020

Introduction

Corticosteroids are the first line therapy for autoimmune hemolytic anemia (AIHA), but are associated with significant adverse events, dependency and frequent relapses. Rituximab is reserved for severe or steroid-resistant disease. Low-dose rituximab is also effective, but its efficacy in the first line has been poorly described. We report our results with this combination.

Methods

Adults older than 16 years newly diagnosed with warm antibody AIHA either primary or secondary were included. Patients systematically received dexamethasone 40 mg for 4 days followed by a 1 mg/kg rapid prednisone taper plus rituximab 100 mg weekly for 4 doses. Our primary outcome was response at day 28 based on the First International Consensus Meeting (complete response: normalization of Hb, no evidence of hemolysis and absence of transfusions; response: increase of Hb by >2g/dl, or normalization of biochemical resolution of hemolysis or absence of transfusion in 7 days), secondary outcome was event-free survival with an event defined as a laboratory or clinical relapse or loss of response.

Results

Sixteen patients were treated with low-dose rituximab during the study period, ten women (62.5%), six men (37.5%). The median age was 34 years (range, 17-78). Three (18.75%) were secondary to lupus erythematosus. The median follow-up was 20 months (range, 0.4-66). Most received 4 doses of rituximab (87.5%). All patients responded at day 28, (100%) 31.2% achieved a complete response (CR). Subsequently, 81.3% achieved CR. Ten (62.5%) were considered steroid-dependent, however, most discontinued treatment without loss of response (75%). The event-free survival was 63.8% to 5 years.

Conclusion

Low-dose rituximab therapy as a first-line in AIHA showed encouraging results as most patients were able to discontinue treatment without relapse.

Disclosures

Gomez-Almaguer: Amgen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Novartis: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AbbVie: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Celgene/BMS: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; AstraZeneca: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Pfizer: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Roche: Membership on an entity's Board of Directors or advisory committees, Speakers Bureau.
Tgfβ-Imprinting Decrease CD38 Expression and Lead to Metabolic Reprogramming on Primary NK Cell
Authors: Pereira Marcelo de Souza Fernandes , Sezgin Yasemin , Thakkar Aarohi , Lee Dean Anthony
Published: November 2020

Transforming growth factor-β (TGF-β) plays an essential role in regulating immune responses through its immunossupressive effect on adaptive and innate immune cells. In addition, we and others have shown that alterations in cell-intrinsic metabolism profiles can significantly impact the function of immune cells. Previously, our group demonstrated that TGFβ-imprinting (TGFβi) decreases NK cell sensitivity to TGFβ through SMAD3 suppression and enhances a pro-inflammatory phenotype with hypersecretion of IFN-γ, TNF-α, and GM-CSF, which appeared to be mediated by an epigenetic mechanism. To evaluate this process, we conducted RNA-seq and ATAC-seq to evaluate the impact of chromosomal remodeling on gene expression. In addition to confirming low SMAD3 transcription, this data showed that TGFβi NK cells have decreased CD38 expression, which we confirmed at the protein level. Since CD38 is an ectoenzyme that regulates nicotinamide adenine dinucleotide (NAD+), a critical component of OXPHOS in both T and NK cells, we examined the effect of TGFβi on NK cell metabolism. TGFβi primary NK cells were generated from peripheral blood of healthy donors as previously described, and using a mitochondrial stress test assay, we observed higher oxygen consumption rates (OCR). However, in the glycolysis stress test assay we observed comparable extracellular acidification rates (ECAR) between Standard expanded and TGFbi NK cells, resulting in higher OCR/ECAR ratios in TGFbi NK cells. Since TGFβ has been shown to induce fragmented mitochondria, and inhibition of mitochondrial fragmentation improved mitochondrial metabolism, we evaluated subcellular structure of TGFβi NK cellsby transmission electron microscopy (TEM). We found no difference in mitochondrial fragmentation between STD and TGFβi NK cells. Together, these results show that TGFβi induces epigenetic reprograming of NK cells that results in increased OXPHOS through CD38 suppression. Moreover, the decreases CD38 expression in TGFβi NK cells could be a promising non-genetic alternative to generating CD38-negative NK cells to avoid fratricide in combination with DARA

Figure

Disclosures

Lee: Kiadis Pharma Netherlands B.V: Consultancy, Current equity holder in publicly-traded company, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties.
Acute Promyelocytic Leukemia (APL): Seasonal Behavior in Colombia for the Period 2009-2019
Authors: Martínez Cordero Humberto , Ospina Idarraga Juan , Castañeda Contreras Xiomara , Rico Mendoza Alejandro , Idrobo Henry , Martinez Correa Lina , Buitrago Jenifer , Porras Ramírez Alexandra
Published: November 2020

Background

Acute promyelocytic leukemia (APL) is a malignant hematologic disease with a high cure rate, but unfortunately it produces several serious complications at the onset of the disease that explain the high early mortality. Based on clinical experience and the papers that precede this document, it has been observed that the incidence of this type of hematological cancer tends to increase at certain times of the year. The present study aims to determine the possible variations in the time intervals of the prevalence of promyelocytic leukemia in Colombia in the decade 2009 to 2019, in order to identify the behavior of the event, seasonality and cyclicality.

Methods

With data from the individual records of service provision (RIPS) of the official Colombian health system, the prevalence of APL in Colombia from 2009 to 2019 was identified. A filter was made by people attended, main diagnosis, C924 - promyelocytic leukemia acute, by year, semester and month. The analysis was performed with RStudio, R version 4.0.2 (2020-06-22); The fpp2 library was used, the analysis of the monthly and quarterly time series was carried out, the decomposition of the time series was carried out, trying through this process to evaluate the seasonal indices through additive and multiplicative types.

Results

During the entire period 2009 to 2019, 2,986 APL diagnoses were found throughout the national territory of Colombia. A progressive increase in prevalence was determined in the period studied (Graph 1). Cycles of increasing cases were observed every 10 months in the period 2009 to 2015 and in the cycle from 2016 to 2019 there was a prolongation of the cycle every 17 months, which strongly suggested that APL has cyclical behavior. Quarterly seasonality was identified by the multiplicative method with increases and decreases that behave in a similar way throughout the period studied (Graph 2).

Conclusion

APL is a highly curable disease that requires adequate support at the onset of the disease to avoid early mortality. The data from this study allow us to conclude that for the period 2009 to 2019 there is a seasonal behavior of this malignant hemopathy. The possibility of predicting during which months of the year the incidence of the disease will increase can serve to properly plan the health services in charge of treating this disease. Additional studies are required to determine what is the cause of the cyclical and seasonal behavior of this APL

Disclosures

Idrobo: Amgen:Honoraria, Speakers Bureau;Takeda:Honoraria, Speakers Bureau;Janssen:Honoraria, Speakers Bureau;Tecnofarma:Honoraria, Speakers Bureau;Abbvie:Honoraria, Speakers Bureau.
Bendamustine-Bortezomib-Dexamethasone (BVD) in Heavily Pretreated Multiple Myeloma: Old/New in NOVEL Agents' Era
Authors: Cerchione Claudio , Catalano Lucio , Nappi Davide , Rocco Stefano , Palmieri Salvatore , Pareto Anna Emanuele , Musuraca Gerardo , Lucchesi Alessandro , Pane Fabrizio , Ferrara Felicetto , Martinelli Giovanni
Published: November 2020

Introduction

Bendamustine is an old bi-functional alkylating agent which has proved to be effective in relapsed, refractory and in new diagnosed Multiple Myeloma (MM).

Aims

Thus, aiming to provide further insights in this field, also in novel agents'era, we present here a retrospective, real-life analysis of patients with relapsed/refractory MM (rrMM), who had received salvage therapy with bendamustine in combination with bortezomib and dexamethasone (BVD).

Methods.

81 patients (44 M/37 F), with rrMM, median age at diagnosis 59.4 years (r. 36-82), median age at start of treatment 63.6 years (r.37-86) treated with several lines of treatments (median 6, r. 2-11), every refractory to all the drugs previously received (also Bortezomib), received BVD (B 90 mg/sqm days 1,2; V 1.3 mg/sqm days 1,4,8,11, D 20 mg days 1,2,4,5,8,9,11,12, Pegfilgrastim day +4) every 28 days, until progression.

All patients had previously received bortezomib-based and IMIDs-based treatments, and 32% (26/81) had also received radiotherapy. 69% (56/81) had undergone single or double autologous and three (2%) allogeneic stem cell transplant. All patients were relapsed and refractory to last therapies received before BVD.

Results

Bendamustine was well tolerated, with grade 3-4 transfusion-dependent anemia in 56% (46/81) of patients, and 43% (35/81) grade 3-4 neutropenia (no ospedalization was required, no septic shocks were observed). No severe extrahematologic toxicity was observed, only grade 1 gastrointestinal side effect (nausea), treated by common antiemetic drugs.

According to IMWG, ORR was 63% (51/81: 7 CR, 18 VGPR, 15 PR, 11 MR) with 11 PD and 19 patients in SD, which can be considered as an impressive result in this subset of rrMM patients. In particular, for 11 patients, BVD was, after having achieved at least a PR, a bridge to second auSCT, and for two patients a bridge to alloSCT. Eight patients have surprisingly achieved a notable PR after failure of novel agents (i.e. Carfilzomib, Daratumumab and Pomalidomide).

Median time to response was 1.3 months (r.1-3), median OS from diagnosis was 67.3 months (r.6-151), median OS from start of Bendamustine was 9.6 months (r.2-36).

Conclusions

The triplet Bendamustine-Bortezomib-Dexamethasone has shown significant efficacy in a particularly severe setting of patients, relapsed and refractory to all available therapeutic resources, and, in particular cases, it could be considered as a bridge to a second autologous or allogenic SCT, also after failure of novel agents.

Disclosures

Lucchesi: Pfizer: Honoraria; Novartis: Honoraria; Incyte: Honoraria.
Cardiovascular Toxicities of Tyrosine Kinase Inhibitor in CML
Authors: Master Samip R , Mansour Richard Preston
Published: November 2020

Background: Cardiovascular (CV) toxicity is a known toxicity of tyrosine kinase inhibitors (TKI) used for chronic myeloid leukemia (CML). Imatinib, dasatinib, nilotinib and bosutinib are all approved for first line treatment for CML. We did a retrospective analysis on adverse effects (AE) of TKIs that has been made available to public by the FDA.

Methods: The FDA has made the data on AEs of various treatments available to general public through the FDA Adverse Events Reports System (FAERS) public dashboard. We investigated the CV AEs of various TKIs for the years 2017-2019.

Results: The percentage of CV AE compared to total AEs reported for Imatinib, Dasatinib, Bosutinib , Nilotinib and Ponatinib were 7.2%, 10.5%, 15.8%, 23.4% and 23.5% respectively. The percentage of CV AE leading to death for Imatinib, Dasatinib, Bosutinib , Nilotinib and Ponatinib were 8.3%, 9.1%, 9.1%, 13.7 % and 18.6% respectively.

Conclusions: Out of the reported cases of AEs to TKIs approved for front line CML, nilotinib appears to have more CV AE compared to imatinib, dasatinib and bolutinib. Imatinib appears to have least CV AE out of the total AEs reported

Disclosures

No relevant conflicts of interest to declare.
Liver X Receptor (LXR) Is a Novel and Reversible Regulator of Trauma-Induced Coagulopathy
Authors: Einersen Peter , Mitra Sanchayita , Selby Reid , Moore Ernest E. , Kelher Marguerite , Fragoso Miguel , Schiller Daran , Swope Megan , Nunns Geoffrey , Vigneshwar Navin , Chandler James , Ghasabayan Arsen , Desisto John , Sanford Bridget , Dzieciatkowska Monika , Johnson Linda , Cohen Mitchell J. , D'Alessandro Angelo , Peltz Erik , Banerjee Anirban , Hansen Kirk , Silliman Christopher C. , Jones Kenneth
Published: November 2020

Trauma-induced coagulopathy (TIC) manifests as a bimodal disruption of normal fibrinolysis, where at the two extremes, fibrinolytic shutdown (TICFS) places the patient at increased risk for thromboembolism and post-injury multiple organ failure, and systemic hyperfibrinolysis (TICHF) results in excessive bleeding. Plasma proteomic profiles for 65 trauma patients were used to identify changes in the patient's plasma protein profile stratifying them by risk for TIC. A strong correlation was seen between total plasma protein concentration of individual patients and the concentration of coagulation proteins within that patient, suggesting that trauma-induced coagulation may be affected through a mechanism that controls the efflux of many coagulation proteins into the plasma (Figure 1). Pathway analysis, in addition to elucidating changes in the coagulation cascade, identified several proteins that are known to be regulated by the Liver X receptor/Retinoid X receptor (LXR/RXR), suggesting that LXR/RXR-mediated activity may regulate TIC (p=9.03E-26). Validation experiments in a rat model revealed that fibrinolytic response to an LXR antagonist and/or agonist can be observed in less than 60 minutes (Figure 2). Additionally, we show through reciprocal rescue of fibrinolysis that the TIC phenotypes (TICHF and TICFS) are one continuum and are ultimately co-reversible, where the reduced LXR signaling that causes sequestration of blood factors in the liver leading to excess bleeding or induced LXR signaling that causes excess efflux leading to thrombosis can be reversed through administration of the LXR agonist or antagonist, respectively. Thus, as modulation of LXR shows co-reversibility and rapid response, induction/inhibition of the LXR pathway provides a direct and novel therapeutic intervention in severely injured patients at risk for TIC.

Disclosures

No relevant conflicts of interest to declare.
Establishment ofin VitroDrug Sensitivity Screening Assay for Chronic Lymphocytic Leukemia (CLL) Primary Patient Samples
Authors: Vishwakarma Bandana Ajay , Wesa Amy
Published: November 2020

Chronic lymphocytic leukemia (CLL) is the most common leukemia in adults. CLL is characterized by proliferation and accumulation of monoclonal, small, mature CD5+ B-cells in peripheral blood, bone marrow and secondary lymphoid organs. The current treatment regimens have improved overall survival but CLL patients will eventually experience relapse. The disease is still incurable making it a necessity to discover and screen for new drugs. Patient derived CLL specimens undergo spontaneous apoptosis when grown in culture and is a major limitation to screening for new therapeutic drugs.

We have established anin vitroculture conditions which supports growth and proliferation of patient derived primary CLL cells. Champions has a bank of primary CLL patient samples across different Rai stage, genetic mutations and relapsed and refractory cases. Using the optimizedin vitroculture conditions, we tested three CLL human primary samples (models) for proliferation . All three models proliferated 1.5 to to 2-fold in the optimized media in 6 days, as measured using CellTiter-Glo®. We also examined the phenotypic changes in the cells after growing them in the established media. Flow cytometric analysis showed that the CLL cells mostly retained the primary phenotypic characteristic CD5+CD10-Cd19+CD20+ even after being in the culture for 6 days.

Next, we screened for sensitivity of primary CLL patient samples (N=10) against known standard of care (SOC) drugs venetoclax, ibrutinib, idelasib, chlorambucil and cytarabine. Primary patient samples derived from peripheral blood mononuclear cells (PBMC) were cultured in 96 well plates in the enriched media and treated with respective drugs over a concentration range over 5 logs. Drug sensitivity was assessed using CellTiter-Glo® luminescent cell viability assay on day 3. Ourin vitroassay indicated that some, but not all patient samples were sensitive to approved standard of care drugs. A relapsed bendamustine pre-treated patient sample was sensitive to all the SOC drugs tested. In addition to drug response, whole exome sequencing and RNAseq are being conducted on these samples, to compare mutational analyses with drug responsiveness. With clinically annotated patient-derived CLL samples, WES and RNAseq plus drug response to standard of care provides a comprehensiveex vivoplatform for the preclinical testing of drug candidates, which may not only provide information on agent efficacy, but that can permit potential biomarker mining and exploration of patient selection criteria for investigational new agents in CLL.

Disclosures

No relevant conflicts of interest to declare.
Sialylated Glycans Regulate MUC13 and the Proto-Oncogenes Pim-1 and Myc to Control Hematopoietic Stem and Progenitor Cell Numbers
Authors: Rivadeneyra Leonardo , Lee-Sundlov Melissa M. , Glabere Simon , Ashwood Heather , Burns Robert , Hoffmeister Karin M.
Published: November 2020

The enzyme β-1-4 galactosyltransferase 1 (β4GalT1) plays a critical role in thrombopoiesis by modulating sialo-glycan (sialyl N-acetyl-lactosamine or LacNAc) content and function of the β1 integrin on megakaryocytes (MKs) (Nat. Commun. 2020;11(1):356). Recent data, however, point to a more complex role for β4GalT1 in hematopoiesis, as the promoter region of its conserved gene, B4galt1, is rich in enhancer sequences for transcription factors associated with cell identity and pro-oncogenic regulatory programs.

Here, we investigated the homeostasis of hematopoietic stem and progenitor cells (HSPCs) in B4galt1-/- mice. We demonstrate that lack of Sialylated LacNAc synthesis perturbs HSPCs beyond the homing deficiency associated with lack of homing sialo-glycan motifs. Flow cytometry analysis showed that B4galt1-/- mouse bone marrows have increased numbers of Long-Term HSCs (LT-HSCs, defined as LineageNeg/Sca-1Pos/c-KitPos/CD150Pos/CD48Neg). The increase in HSC numbers led us to investigate their phenotypic and functional features further. While quiescence markers in B4galt1-/- LT-HSCs were indistinguishable relative to controls, LT-HSCs expressed more of the platelet marker CD41 on their surface, supporting a highly expanded CD41+ subset of LT-HSCs. Platelet-bias of LT-HSCs has been associated with inflammation and aging. However, our data do not support an increased cytokine inflammatory profile in the bone marrow.

Instead, single-cell RNA sequencing (scRNA seq) of sorted β4galt1-/- LineageNeg/Sca-1Pos/c-KitPos (LSK) cells showed a significantly increased expression of the proto-oncogene Pim-1, its target, Myc, and the heavily O-glycosylated transmembrane receptor mucin 13 (MUC13), compared to control cells. Analysis of LT-HSC glycan expression using lectin microarray showed the expected decrease in N-glycosylation associated with B4galt1 deficiency, but also an increase in O-glycans, consistent with overexpression of MUC13, expression of which was enriched compared to other surface mucins. The data show that B4galt1 deletion leads to overexpression of the proto-oncogenes Pim-1, Myc, and MUC13 in HSPCs. The data suggest that MUC13-associated O-glycans and glyco-synthetic genes are potential therapeutic targets for hematologic malignancies since mucins have anti-inflammatory functions and alterations in mucin expression are with inflammation and cancer.

Disclosures

No relevant conflicts of interest to declare.
Developing Transient Gene Therapies to Decrease the Stability of Thrombi for Coagulopathy and Thrombosis
Authors: Strilchuk Amy W , Kastrup Christian , Palumbo Joseph S. , Cullis Pieter
Published: November 2020

Thrombotic disorders are prevalent and dangerous. Thrombosis involves the formation of unwanted blood clots inside blood vessels, which can impair blood flow and lead to severe cardiovascular events, such as heart attack, stroke, and pulmonary embolism. Current therapies for thrombosis are unsatisfactory in that they require frequent re-administration, and are associated with a significant bleeding risk, a consequence of inhibiting the coagulation cascade upstream of fibrin generation. Coagulation proteins factor XIII (FXIII) and thrombin activatable fibrinolysis inhibitor (TAFI) are enzymes that act to stabilize clots, downstream of fibrin generation and polymerization. Though this makes them ideal targets to reduce the burden of thrombosis while maintaining hemostasis, no inhibitors for either protein are currently available in the clinic. As we have recently published (Strilchuk, et al. Blood. 2020), lipid nanoparticles can be used to deliver siRNA to knock-down FXIII-B and achieve depletion of FXIII-A from circulation in mice and rabbits. FXIII-A depletion causes less antiplasmin to be crosslinked to clots, resulting in clots that are more susceptible to fibrinolysis. In the current abstract, we have expanded on preliminary data showing that while clots are weaker, bleeding is not enhanced in mice or rabbits after minor or major injury. We have also leveraged the same biological tools to knock-down TAFI for days after a single administration. In the short term, these novel RNA agents will be valuable tools in investigating the biology of thrombotic disorders. The ultimate goal is to develop these agents into precise and long-acting prophylactic therapies for thrombotic disorders, that are safer and more effective than current standards of care.

Disclosures

No relevant conflicts of interest to declare.
B-lymphoblastic leukemia arising in a patient with chronic neutrophilic leukemia
Authors: Boddy Craig S. , Tan Brent T. , Aoki Joseph
Published: November 2020

Key Points

We report the first identified case of chronic neutrophilic leukemia with transformation to B-lymphoblastic leukemia. Genetic alterations involving CSF3R, ASXL1, SRSF2, and RUNX1 contributed to the unusual progression and may drive B-cell leukemogenesis.
Anion-Regulated Synthesis of ZnO 1D Necklace-Like Nanostructures with High Photocatalytic Activity
Authors: Qin Xiaoyun , Shi Dongdong , Guo Bowen , Fu Cuicui , Zhang Jin , Xie Qingqing , Shi Xiangdong , Chen Fenghua , Qin Xiaomei , Yu Wei , Feng Xiangli , Liu Yan , Luo Dan
Published: November 2020

One-dimensional (1D) nanomaterials with specific architectures have received increasing attention for both scientific and technological interests for their applications in catalysis, sensing, and energy conversion, etc. However, the development of an operable and simple method for the fabrication of 1D nanostructures remains a challenge. In this work, we developed an “anion-regulated morphology” strategy, in which anions could regulate the dimensionally-restricted anisotropic growth of ZnO nanomaterials by adjusting the surface energy of different growth facets. ZnO 1D necklace-like nanostructures (NNS) could be prepared through a hydrothermal treatment of zinc acetate and urea mixture together with a subsequent calcination procedure at 400 °C. While replacing the acetate ions to nitrate, sulfate, and chlorion ions produced ZnO nanoflowers, nanosheets and hexagonal nanoplates, respectively. Density functional theory calculations were carried out to explain the mechanism behind the anions-regulating anisotropic crystal growth. The specified ZnO 1D NNS offered improved electron transport while the grain surface could supply enlarged specific surface area, thus providing advanced photocatalytic ability in the following photodegradation of methyl orange (MO). Among the four photocatalysts with different morphologies, ZnO 1D NNS, possessing the highest catalytic activity, degraded 57.29% MO in the photocatalytic reaction, which was 2 times, 10 times and 17 times higher than nanoflowers, nanosheets and hexagonal nanoplates, respectively. Our work provides new ideas for the construction and application of ZnO 1D nanomaterials.
A cost benefit model for high capacity transport in a comprehensive line-haul network
Authors: Lindqvist David , Salman Marwan , Bergqvist Rickard
Published: November 2020

The objective of the paper is to analyse the potential of introducing High Capacity Transport (HCT) within a comprehensive line-haul network. Barriers and enablers have been identified, and a cost benefit model has been developed. The model considers environmental performance, socio-economic costs and operational costs and has been applied to a Swedish context and case study. The findings reveal that HCT can contribute to the development of road transportation in the perspectives of energy consumption and emission releases. It can further strengthen the trade and competitiveness of Swedish hauliers, as the introduction can provide a more cost-efficient system for all actors, including the transporter. However, the potential is largest when longer vehicles are meticulously and scrupulously introduced on a designated network alone.
Differential responses in placenta and fetal thymus at 12 days post infection elucidate mechanisms of viral level and fetal compromise following PRRSV2 infection
Authors: Van Goor Angelica , Pasternak Alex , Walker Kristen , Hong Linjun , Malgarin Carolina , MacPhee Daniel J. , Harding John C. S. , Lunney Joan K.
Published: November 2020

Background

A pregnant gilt infected with porcine reproductive and respiratory syndrome virus (PRRSV) can transmit the virus to her fetuses across the maternal-fetal-interface resulting in varying disease outcomes. However, the mechanisms leading to variation in fetal outcome in response to PRRSV infection are not fully understood. Our objective was to assess targeted immune-related gene expression patterns and pathways in the placenta and fetal thymus to elucidate the molecular mechanisms involved in the resistance/tolerance and susceptibility of fetuses to PRRSV2 infection. Fetuses were grouped by preservation status and PRRS viral load (VL): mock infected control (CTRL), no virus detected (UNINF), virus detected in the placenta only with viable (PLCO-VIA) or meconium-stained fetus (PLCO-MEC), low VL with viable (LVL-VIA) or meconium-stained fetus (LVL-MEC), and high VL with viable (HVL-VIA) or meconium-stained fetus (HVL-MEC).

Results

The host immune response was initiated only in fetuses with detectable levels of PRRSV. No differentially expressed genes (DEG) in either the placenta or thymus were identified in UNINF, PLCO-VIA, and PLCO-MEC when compared to CTRL fetuses. Upon fetal infection, a set of core responsive IFN-inducible genes (CXCL10,IFIH1,IFIT1,IFIT3,ISG15, andMX1) were strongly upregulated in both tissues. Gene expression in the thymus is a better differentiator of fetal VL; the strong downregulation of several innate and adaptive immune pathways (e.g., B Cell Development) are indicative of HVL. Gene expression in the placenta may be a better differentiator of fetal demise than the thymus, based-on principle component analysis clustering, gene expression patterns, and dysregulation of the Apoptosis and Ubiquitination pathways.

Conclusion

Our data supports the concept that fetal outcome in response to PRRSV2 infection is determined by fetal, and more significantly placental response, which is initiated only after fetal infection. This conceptual model represents a significant step forward in understanding the mechanisms underpinning fetal susceptibility to the virus.
Whole genome analysis of the koa wilt pathogen (Fusarium oxysporum f. sp. koae) and the development of molecular tools for early detection and monitoring
Authors: Dobbs John T. , Kim Mee-Sook , Dudley Nicklos S. , Klopfenstein Ned B. , Yeh Aileen , Hauff Robert D. , Jones Tyler C. , Dumroese R. Kasten , Cannon Philip G. , Stewart Jane E.
Published: November 2020

Background

Development and application of DNA-based methods to distinguish highly virulent isolates of Fusarium oxysporum f. sp. koae [Fo koae; cause of koa wilt disease on Acacia koa (koa)] will help disease management through early detection, enhanced monitoring, and improved disease resistance-breeding programs.

Results

This study presents whole genome analyses of one highly virulent Fo koae isolate and one non-pathogenic F. oxysporum (Fo) isolate. These analyses allowed for the identification of putative lineage-specific DNA and predicted genes necessary for disease development on koa. Using putative chromosomes and predicted gene comparisons, Fo koae-exclusive, virulence genes were identified. The putative lineage-specific DNA included identified genes encoding products secreted in xylem (e. g., SIX1 and SIX6) that may be necessary for disease development on koa. Unique genes from Fo koae were used to develop pathogen-specific PCR primers. These diagnostic primers allowed target amplification in the characterized highly virulent Fo koae isolates but did not allow product amplification in low-virulence or non-pathogenic isolates of Fo. Thus, primers developed in this study will be useful for early detection and monitoring of highly virulent strains of Fo koae. Isolate verification is also important for disease resistance-breeding programs that require a diverse set of highly virulent Fo koae isolates for their disease-screening assays to develop disease-resistant koa.

Conclusions

These results provide the framework for understanding the pathogen genes necessary for koa wilt disease and the genetic variation of Fo koae populations across the Hawaiian Islands.
Analysis of splice variants of the human protein disulfide isomerase (P4HB) gene
Authors: Kajihara Daniela , Hon Chung-Chau , Abdullah Aimi Naim , Wosniak João , Moretti Ana Iochabel S. , Poloni Joice F. , Bonatto Diego , Hashimoto Kosuke , Carninci Piero , Laurindo Francisco R. M.
Published: November 2020

Background

Protein Disulfide Isomerases are thiol oxidoreductase chaperones from thioredoxin superfamily with crucial roles in endoplasmic reticulum proteostasis, implicated in many diseases. The family prototype PDIA1 is also involved in vascular redox cell signaling. PDIA1 is coded by theP4HBgene. While forced changes inP4HBgene expression promote physiological effects, little is known about endogenousP4HBgene regulation and, in particular, gene modulation by alternative splicing. This study addressed theP4HBsplice variant landscape.

Results

Ten protein coding sequences (Ensembl) of theP4HBgene originating from alternative splicing were characterized. Structural features suggest that except forP4HB-021, other splice variants are unlikely to exert thiol isomerase activity at the endoplasmic reticulum. Extensive analyses using FANTOM5, ENCODE Consortium and GTEx project databases as RNA-seq data sources were performed. These indicated widespread expression but significant variability in the degree of isoform expression among distinct tissues and even among distinct locations of the same cell, e.g., vascular smooth muscle cells from different origins.P4HB-02,P4HB-027 andP4HB-021 were relatively more expressed across each database, the latter particularly in vascular smooth muscle. Expression of such variants was validated by qRT-PCR in some cell types. The most consistently expressed splice variant wasP4HB-021 in human mammary artery vascular smooth muscle which, together with canonicalP4HBgene, had its expression enhanced by serum starvation.

Conclusions

Our study details the splice variant landscape of theP4HBgene, indicating their potential role to diversify the functional reach of this crucial gene.P4HB-021 splice variant deserves further investigation in vascular smooth muscle cells.