bluuebunny/crossref_metadata_2025_split huggingface dataset:
Establishment of a Nomogram for Patients with Primary Mediastinal Large B-Cell Lymphoma Based on the SEER Database and Validation of Real-World Data
Authors: Wang Hua , Chen Guanjun , Fu Bibo 
Published: November 2021

Abstract
Background
Primary mediastinal large B-cell lymphoma (PMBCL) is a rare disease with the majority of patients being rapidly progressive anterior mediastinal large tumors. Due to the rarity of PMBCL, information on the incidence, clinical features, prognostic factors and models of PMBCL is limited. The present study is one of the largest studies on the incidence and prognostic factors of PMBCL and is the first to establish a nomogram model of PMBCL and validate it with real-world data. We also compared the newly established Nomogram with the existing IPI prognostic model. We believe that our findings can help clinicians to quickly and accurately assess the predicted survival of patients and help them to perform individualized risk stratification of patients.
Methods
Based on data from the Surveillance, Epidemiology and End Results (SEER) database, 797 patients diagnosed with PMBCL were enrolled in this study. The 797 patients were randomly divided into training and internal validation groups in a 7:3 ratio, and 116 patients diagnosed with PMBCL were included in the external validation group based on data from the Sun Yat-sen University Cancer Center and the First Affiliated Hospital of Guangzhou Medical University. Independent prognostic factors were screened by Cox regression analysis. R-coding was used to construct nomograms predicting overall survival (OS). Discriminations and corrections of the new model were assessed using the consistency index (C-index), subject operating characteristic curves (ROC) and calibration curves, and compared with the conventional international prognostic index (IPI) using decision curve analysis (DCA) to assess its accuracy and benefit.
Results
From 2001 to 2016, the incidence of primary mediastinal large B-cell lymphoma showed a relatively stable increasing trend with an APC of 11.8% (95% confidence interval 8.8-14.0, P&lt;0.05), and this trend was more pronounced in the female population. Multivariate models showed that age and Ann arbor staging were significantly associated with OS, while the variable of extra-nodal invasion was included in the modeling based on clinical experience. In the training cohort, the C-index of the nomogram for OS was 0.712. the C-index for the internal and external validation cohorts was 0.667 and 0.690, respectively. The calibration curve also showed high predictive accuracy. The C-index and ROC curves of nomogram showed better results compared to IPI scores, and also yielded better net gains in decision curve analysis.
Conclusion
In summary, we successfully established a validated nomogram for predicting OS in patients with PMBCL, which can help clinicians to select appropriate individualized treatment for their patients.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Use of Emicizumab in Patients with Hemophilia a with and without Inhibitors: A Single Center Experience
Authors: Giuffrida Gaetano , Nicolosi Daniela , Condorelli Annalisa , Markovic Uros , Di Raimondo Francesco 
Published: November 2021

Abstract
Emicizumab is a humanized bispecific recombinant monoclonal antibody that binds the enzyme factor IXa and the substrate factor X and mimics the function of FVIII. It solved some unmet needs of haemophilia A (HA) treatment, such as regimen (once weekly up to once monthly infusion) and route of administration (subcutaneous). Most importantly, emicizumab reduce bleeding episodes and improved the quality of life of these patients. Emicizumab received FDA approval for prophylaxis in HA patients with inhibitors in November 2017 and for patients without inhibitors in October 2018.The aim of this study is to provide information on treatment with emicizumab in patients with severe haemophilia A with and without inhibitors in terms of efficacy, safety and quality of life. Here we report our experience with 9 patients with severe haemophilia A (5 with inhibitors and 4 without inhibitors) that were switched from replacement therapy to emicizumab prophylaxis. Median age at initiation of treatment was 26.8 years for patients with inhibitors and 18.7 years for those without inhibitors. Median duration of therapy was 12 months for patients with inhibitors and 8 months for those without inhibitors. All patients were started on emicizumab with a loading dose of 3 mg/kg once weekly for 4 weeks followed by a maintenance dose of 1.5 mg/kg once weekly or every two weeks. The mean annual bleeding rates (ABR) in patients without inhibitor prior to emicizumab was 1.5 compared to 0 while on emicizumab prophylaxis. In subjects with inhibitors ABR was 1.8 compared to 0.4 while on emicizumab prophylaxis. Furthermore, no adverse events including thrombotic events occurred in course of emicizumab prophylaxis. Bleeding events in course of emicizumab prophylaxis did not request any treatment with replacement therapy or bypassing agents. The questionnaire evaluating quality of life (HAL v 2.0 - 2015 ITA; pedHAL v 2.0 - 2015 ITA) was administered to the patients before switch to emicizumab and after a follow up of six months. The scores showed a significant improvement in terms of quality of life in course of emicizumab prophylaxis. In conclusion, our experience suggests that emicizumab prophylaxis is safe and improved protection against bleeding and quality of life compared to replacement therapy in patients with severe A haemophilia.

Disclosures
Di Raimondo: Amgen: Honoraria; Jazz Pharmaceutical: Honoraria; Pfizer: Honoraria; Janssen Pharmaceuticals: Honoraria; Bristol Myers Squibb: Honoraria; AbbVie: Honoraria.
Neutrophil Extracellular Traps (NETs) Contribute to the Formation of Microvascular Thrombosis in Immune Thrombotic Thrombocytopenic Purpura
Authors: Yada Noritaka , Sui Jingrui , Zheng Liang , Zheng X. Long 
Published: November 2021

Abstract
Introduction. Immune thrombotic thrombocytopenic purpura (iTTP), a potentially fatal blood disorder, is primarily caused by severe deficiency of plasma ADAMTS13 activity resulting from immunoglobulin (Ig) G-mediated inhibition of plasma ADAMTS13 activity. However, severe ADAMTS13 deficiency is necessary but not sufficient to cause acute iTTP. An environmental factor such as infection or acute inflammation may be necessary to trigger the acute onset of the disease. We and others have previously reported that plasma markers of neutrophil activation and neutrophil extracellular traps (NETs) formation are significantly elevated in patients with acute iTTP, which returns to normal during remission. However, the pathogenetic role of NETs in acute iTTP is not fully understood.
Methods and results. Using flow cytometry, microfluidic shear-based assay, and confocal imaging analysis, we determined the in vivo NETosis in blood samples obtained from patients with acute episode of iTTP and ex vivo NETs formation, as well as the therapeutic efficacy of DNase I on thrombus formation under flow. We showed that by flow cytometry that only very few CitH3+/MPO+ positive neutrophils were present in the healthy donor blood. This population of cells dramatically increased after being stimulated with a bacterial toxin (i.e., Shigatoxin-2) at ~100 ng/mL for 15 min. Importantly, the number of CitH3+/MPO+ positive neutrophils in the sample obtained from a patient with acute iTTP was ~1,000 times higher than that in the healthy controls (Fig. 1), suggesting a massive NETosis in patients with acute iTTP. Microfluidic shear-based assay and confocal imaging analysis further confirmed a dramatic increase in adhesion and aggregation of murine platelets (stained with Alexa647 anti-CD41) and neutrophil (stained with Hoechst), as well as formation of NETs (stained with Syto green) following a perfusion of an Adamts13 -/- murine whole blood (anti-coagulated with thrombin inhibitor, PPACK) under arterial shear (15 dyne/cm 2) over a stimulated murine endothelial surface. Interestingly, an addition of DNase I (100 U/mL) significantly reduced the overall surface coverage of platelets and neutrophils on the murine endothelial surface under the same conditions (Fig. 2).
Conclusions. These results demonstrate for the first time NETosis and NETs formation are common in patients with acute iTTP and in Adamts13 -/- mice after being stimulated with shigatoxin; DNase I appears to be highly efficacious eliminating the NETs and platelet/neutrophil-dominant thrombosis under arterial flow. Our findings support the pathogenetic role of NETs in the onset and progression of iTTP, and the therapeutic potential of DNase I in such a fatal disease.
Figure 1 Figure 1.

Disclosures
Zheng: Alexion: Speakers Bureau; Sanofi-Genzyme: Honoraria, Speakers Bureau; Takeda: Consultancy, Honoraria; Clotsolution: Other: Co-founder; AJMC: Honoraria.
SATB1 Regulates Chromatin Organization and HSP70 Expression in Early Erythropoiesis and Is Downregulated in Models of Diamond Blackfan Anemia
Authors: Wilkes Mark C , Shibuya Aya , Scanlon Vanessa M , Chae Hee-Don , Narla Anupama , Sakamoto Kathleen M. 
Published: November 2021

Abstract
Diamond Blackfan Anemia (DBA) is a rare genetic disease predominantly caused by mutations carried within one of at least 20 ribosomal genes. DBA is characterized by red blood cell aplasia and normal myeloid and megakaryocyte progenitors, indicating that early uncommitted progenitors are relatively unaffected by the mutations. In DBA, the formation of BFU-E colonies and subsequent erythroblasts are severely restricted and indicate a defect in one of the earliest stages of erythroid expansion.
To identify critical molecular mechanisms that may regulate early erythropoiesis, we used shRNAs against the ribosomal protein RPS19 (the most commonly mutated gene in DBA) in cord blood derived CD34+ hematopoietic stem and progenitor cells (HSPCs) and performed bulk RNA-seq. After 3 days in an erythroid culture media, the transcriptomes in CD71+ erythroid progenitors were examined. We found that the special AT binding protein 1 (SATB1) was downregulated in RPS19-insufficient HSPCs compared to healthy cord blood HSPCs.
SATB1 is modestly expressed in hematopoietic stem cells but is induced during lymphoid expansion and has been previously reported to suppress myeloid/erythroid progenitor (MEP) expansion. Our results showed that maintaining SATB1 expression is required for optimal expansion of MEP progenitors and that the premature loss of SATB1 in DBA contributes to the anemia phenotype.
SATB1 binds to 3 specific regions upstream of the 5'UTR of the HSP70 genes and induces the formation of 2 chromatin loops. An enhancer element associates with the proximal promoters of the two HSP70 genes and facilitates the induction of HSP70. In DBA, HSP70 is not induced and contributes to DBA pathogenesis.
HSPA1A is induced 4.3-fold while HSPA1B is induced 3.1-fold. Increased expression of the master erythroid transcription factor GATA1 during erythropoiesis occurs in two phases. The first induction precedes a more dramatic induction that accompanies later stages of erythroid differentiation. The absence of SATB1 or HSP70 reduced the earlier GATA1 induction that accompany MEP expansion by 46.1% and 49.3% respectively. The number of MEPs in SATB1 knockdown HSPCs was reduced, resulting in a 24.5% reduction in CD235+ erythroid and 20.8% reduction in CD41+ megakaryocytes.
While SATB1-independent effects of RPS19-insufficiency contribute more significantly to erythroid defects in DBA, we have uncovered that SATB1 contributes to regulation of the earliest stages of erythropoiesis by facilitating the induction of HSP70 and subsequent stabilization of an early induction of GATA1.

Disclosures
No relevant conflicts of interest to declare.
Clinical Outcomes of COVID-19 Infection Among Children with Cancer: A Single-Center Experience
Authors: Akkoyun Esra , Sotelo Jesus , Cherry Giselle , Bisceglia Maria , Slone Tamra , Aquino Victor , Sue Paul 
Published: November 2021

Abstract

Background
Outcomes of COVID-19 infection among children with underlying malignancy remain unclear, with limited data available to date. Here we report clinical characteristics and outcomes following COVID-19 infection among children with underlying malignancy or a history of hematopoietic stem cell transplant (HSCT) during the first year of the global COVID-19 pandemic, in Dallas, TX.
Methods:
We retrospectively reviewed children with a history of malignancy or prior HSCT and positive SARS-CoV-2 polymerase chain reaction (PCR) result between March 1, 2020 - March 31, 2021. Demographic, clinical, and outcome data were reviewed, and analyzed according to underlying condition and disease severity.
Results:
Forty-six patients with underlying malignancy or HSCT met inclusion criteria. Median age was 9 years, with twenty (43%) female, and twenty-seven (59%) of Hispanic ethnicity. Underlying conditions included Acute Lymphoblastic Leukemia (67%), Acute Myelogenous Leukemia (9%), HSCT (11%), sarcoma (9%), solid tumors (9%), and Hodgkin's lymphoma (2%). Fifteen subjects (33%) were asymptomatic, with twenty-four children (52%) classified as having mild, five (11%) with severe, and two (4%) with critical COVID-19 disease. The most common symptoms at presentation were fever (46%), cough (39%), and gastroenteritis (19%).
Thirty-four children (74%) were hospitalized, with seven (15%) requiring intensive care unit (ICU) care. 17% of patients were admitted for treatment of COVID-19 alone, with other indications including neutropenic fever, relapse, and chemotherapy. Six (13%) patients required non-invasive ventilation, and two patients (4%) required invasive mechanical ventilation. Median length of stay was 9 days (IQR 4.5-24.5), with median ICU stay of 3.5 days (IQR 2-26.7).
Seven children (15%) received COVID-19 targeted therapy including steroids, remdesivir or convalescent plasma. Among severe and critically ill children, six (86%) had underlying hematologic malignancies (5 HR ALL, 1 AML), and two (28%) a prior history of HSCT. Two children (4%) died.
Thirty-six patients (78%) achieved documented clearance of SARS-CoV-2 by PCR, at median of 43.5 days (IQR 28.7-65.2).
Conclusion:
Among children with underlying malignancy or a history of HSCT, COVID-19 results in a spectrum of illness ranging from asymptomatic disease to death. Rates of hospitalization are high compared to the general pediatric population, and illness may be complicated by additional factors including co-infection, neutropenic fever, relapse of malignancy and need for chemotherapy. Children with hematologic malignancy or a history of HSCT may be at increased risk for severe disease. Additional studies are urgently needed to elucidate risk factors for severe/critical COVID-19 disease among children with underlying malignancy.

Disclosures
No relevant conflicts of interest to declare.
Engineering Human Invariant Natural Killer T (iNKT) Cells to Overexpress Immunomodulatory Cytokines
Authors: Mavers Melissa , Hollingsworth Derek , Boonchalermvichian Chaiyaporn , Liu Jeffrey , Baker Jeanette , Lopes Ramos Teresa , Lohmeyer Juliane K. , Lin Po-Yu , Roncarolo Maria Grazia , Negrin Robert S. 
Published: November 2021

Abstract
Many studies have shown the important role that invariant natural killer T (iNKT) cells play in suppression of graft-versus-host disease (GVHD). Murine studies have also demonstrated multiple subsets of iNKT cells, including those with pro-inflammatory and immunosuppressive properties. The prevention of murine acute GVHD by the adoptive transfer of immunomodulatory iNKT cells suggests that this approach would be ideal for clinical translation. However, the heterogeneity of human iNKT cells remains poorly understood, and the uncertain ability to identify immunosuppressive human iNKT cells and expand them to clinically relevant numbers remains a limitation in clinical translation. Therefore, engineering bulk human iNKT cells to enhance immunosuppressive capacity represents a novel approach to ensuring sufficient cell numbers of immunoregulatory cells for adoptive transfer. Herein, we describe the lentiviral transduction of human iNKT cells to overexpress immunomodulatory cytokines. We focused on IL-4, because murine iNKT cell-mediated GVHD suppression is IL-4 dependent, and IL-10, because an immunosuppressive regulatory T cell-like subset of murine iNKT cells highly produces IL-10. Human iNKT cells were transduced with a lentiviral construct expressing IL-4 or IL-10 (or green fluorescent protein (GFP) as a control) and an inert truncated nerve growth factor receptor (deltaNGFR) to facilitate sorting of transduced cells. Enzyme linked immunosorbent assay revealed significantly increased expression of IL-4 or IL-10 by cells transduced with the corresponding lentivirus, as compared to untransduced, mock transduced, or GFP transduced cells, replicated with 3 iNKT donors (p&lt;0.03) (Figure 1). This proof-of-concept data demonstrates the ability to effectively transduce human iNKT cells to overexpress immunomodulatory cytokines. Future studies will focus on evaluating the immunosuppressive capacity of these engineered cells both in vitro and in vivo (using the xenograft GVHD model). Successful suppression of GVHD by engineered iNKT cells will provide a critical step forward in identifying novel approaches to bring these promising cells to the clinic.
Figure 1 Figure 1.

Disclosures
No relevant conflicts of interest to declare.
Real World Experience of the BTK Inhibitors. Comparing Cardiac, Vascular and Infectious Toxicities
Authors: Master Samip R , Ramadas Poornima , Mansour Richard Preston 
Published: November 2021

Abstract
Background: Cardiac and vascular toxicities are known complications on Bruton Tyrosine kinase inhibitor (BTKi)drugs. Ibrutinib, acalibrutinib and zanubrutinib are all approved BTKi drugs. We did a retrospective analysis on adverse effects (AE) of BTKi's 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 infectious, cardiac and vascular AEs of various BTKi for the years 2019-2021.
Results: The percentage of cardiac AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 12%, 7.3% and 6.2% respectively. The most common cardiac toxicity was atrial fibrillation.
The percentage of vascular AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 8.2 %, 5.2 % and 3.9 % respectively. The most common vascular toxicity was hemorrhage.
The percentage of infectious AE compared to total AEs reported for ibrutinib, acalibrutinib and zanubrutinib were 18.3%, 13.6% and 35.5% respectively. The most common vascular toxicity was pneumonia.
Conclusions: Out of the reported cases of AEs to BTKis approved, ibrutinib have most and acalibrutinib have least cardiac AEs. Zanubrutinib has least vascular AEs. Out of the reported cases of AEs to BTKis approved, zanubrutinib have most infectious AEs.

Disclosures
Master: Blue Bird Bio: Current holder of individual stocks in a privately-held company.
The Loss of Liver Kinase B1 Accelerates Maturation Time in the Developing Erythrocyte
Authors: White III Zollie , Freeman Katie , Delehanty Lorrie L , Goldfarb Adam N 
Published: November 2021

Abstract
In the US, iron-restricted anemia contributes greatly to morbidity. The erythroid iron deprivation response, characterized by a pathway in which erythropoiesis is suppressed during iron restriction, underlies this anemia. In preliminary studies, liver kinase B1 (LKB1) was implicated as a potential key component in the erythroid iron deprivation response. Normal human CD34+ hematopoietic progenitor cells cultured for 3 days in erythroid medium with 100% or 10% transferrin saturations underwent immunoblot of whole cell lysates. Reproducibly, the levels of LKB1 did not change based on the transferrin saturation; however, immunofluorescence imaging showed a shift in subcellular localization when cells were subjected to low iron conditions.
To assess the effects of LKB1 loss in the erythroid compartment, we used control and LKB1 conditional knockout (STK11 F/F; EpoR-Cre+) mice. In steady-state conditions, the loss of LKB1 does not confer a change in RBC count, though, there is a baseline increase in the number of reticulocytes, and a large increase in the level of serum Erythropoietin (Epo). In a model to precipitate anemia, these mice were challenged with intraperitoneal injection of phenylhydrazine (PHZ). It was found that LKB1 is dispensable for an appropriate response to this type of stress erythropoiesis.
To assess the impact of LKB1 on maturation, we performed flow cytometric analysis using an ex vivo culture system of splenic erythroblasts. LKB1-deficient erythroid progenitors show increased percentages of more advanced cells as evidenced by the surface markers CD71 and Ter119 (the mouse analogue of human glycophorin A). Current studies are underway to assess if this change is due to signaling through the AMPK pathway. These studies will provide mechanistic detail of LKB1 function and activity on erythropoiesis, improving our understanding of programs involved in maturation of differentiation and lineage selection which may ultimately help to improve health outcomes and advance treatment for various types of anemias.

Disclosures
No relevant conflicts of interest to declare.