bluuebunny/crossref_metadata_2025_split huggingface dataset:
Plasma Concentration of Eltrombopag Correlated with the Efficacy in the Treatment of Refractory Aplastic Anemia: A Single-Center Study in China
Authors: Zuo Wei , Zhang Bo , Ruan Jing , Chen Miao , Li Jian , Han Bing 
Published: February 2021

Background and purpose: Eltrombopag (ELT) was effective in the treatment of relapse/refractory aplastic anemia (AA) patients. Response and adverse drug reaction (ADR) differed greatly among individuals even at the same dosage of ELT.
Methods: Patients diagnosed with non-severe aplastic anemia (NSAA) between January 2018 to January 2019 in Peking Union Medical Colleague Hospital who were refractory to immunosuppressive therapy were treated with ELT and followed up for at least 6 months. Plasma concentrations of ELT were detected by the high-performance liquid chromatography-mass spectrometry after at least two months of ELT treatment and staying for that dosage for at least two weeks. Dose-concentration, concentration-response and concentration- ADR relation were evaluated.
Results: Among the 72 patients treated with ELT during that period, 44 patients with complete data were enrolled. 6 (13.6%) were males and 38 were females (86.4%) with a median age of 54 years (IQR: 38.5-63). By the time of plasma concentration detected, the median dosage of ELT was 75(IQR 50-100) mg/d and median time for total ELT exposure was 3 (IQR 2.0-6.0) months, and 37 (70.5%) patients responded to ELT. The median concentration of ELT was 10.4μg/ml (IQR 3.7-24.4μg/ml). The concentration of ELT was positively correlated with the daily dose of ELT (r = 0.68, p &lt; 0.001). Multivariate logistic regression analysis showed that the risk of inefficacy of ELT with a concentration between 11.2-15.2 μg/ml was 0.028-fold (95% CI: 0.001-0.864; P=0.041) to those with a concentration between 3.2-7.2μg/ml. The cutoff value of concentration for ELT efficacy was 12.50μg/ml by receiver operation characteristic curve. A higher risk of ADR was related to a longer total exposure to ELT (P=0.012). Although not significant, odd ratio (OR) increased with ELT concentration, suggesting a possible elevated risk of ADR correlated with blood concentration.
Conclusions: ELT is effective for the treatment of NSAA with accepTable side effects. Plasma concentration of ELT correlated with the dose and the effects of ELT.

Disclosures
No relevant conflicts of interest to declare.

OffLabel Disclosure:
Eltrombopag (ELT) has been proven to induce trilinear hematopoietic responses in relapse/refractory AA patients and has been approval as a monotherapy in relapsed/refractory SAA in the USA and Europe. However, the use of ELT is only approved for idiopathic thrombocytopenic purpura (ITP) in China by the National Medical Products Administration (NMPA), for AA it is still "off-label".
Incidence and Outcomes of Acute Transfusion Reactions in Hospitalized Patients in the United States
Authors: Kumar Rohit , Malapati Sindhu , Singh Sunny R K , Mamedov Bokhodir , Shah Myra R , Bhandari Shruti 
Published: February 2021

Introduction
Acute transfusion reactions (ATRs) have a broad spectrum of presentations ranging from benign to life-threatening. Due to the rarity of these reactions, there is a paucity of data regarding their incidence and clinical outcomes. The objectives of this study were to determine the incidence of ATRs, its risk factors, and associated mortality.
Methods:
We reviewed the National Inpatient Sample (NIS) database 2014 for admissions where the patient (&gt;=18 years old) was transfused blood products. The NIS is a large publicly available all-payer inpatient healthcare database designed to produce U.S. regional and national estimates of inpatient utilization, access, charges, quality, and outcomes. ATRs were identified using ICD-9 CM codes for transfusion-associated circulatory overload (TACO), transfusion-related acute lung injury (TRALI), febrile non-hemolytic transfusion reactions (FNHTR), acute infections, anaphylaxis, and acute hemolytic reaction. Pearson's chi-square and student's t-test were used to compare categorical and continuous variables between hospitalizations with versus without ATRs, respectively. Multivariate logistic regression analysis was done to determine the risk factors for common ATRs (TACO, TRALI, and FNHTR). A multivariate cox proportional model was built to compare the mortality of two study groups. A 2-sided p-value ≤ 0.05 was considered significant.
Results:
A total of 2,134,691 hospitalizations were associated with the transfusion of blood products. ATRs were documented in 0.2% of the hospitalizations (TACO 0.08%, TRALI 0.06%, FNHTR 0.09%, others 0.003%). The group that had ATRs was slightly younger (median age 67 vs 68 years, p=0.002), had the same proportion of females (58.3% vs 55.3%, p=0.055), less comorbidity score (28.7% vs 31.7% had Charlson Comorbidity Index &gt;3, p=0.042) and more critically ill (17.8% vs 10.5% on mechanical ventilation, p&lt;0.001) compared to group without ATRs. Hospitalizations with ATRs had longer median length of stay (7 vs 6 days, p&lt;0.001) and higher median hospital cost ($64,399 vs $53,912, p&lt;0.001) compared to without ATRs. The risk factors for common ATRs (odds ratio, OR) are mentioned in the table. ATRs were not associated with increased risk of mortality (combined HR 0.89 95%CI 0.71-1.12, p=0.321).
Conclusions:
Nationally, the incidence of ATRs is low in hospitalized patients and it is not associated with increased mortality. This large database analysis gives insight into the risk factors associated with different ATRs.

Disclosures
No relevant conflicts of interest to declare.
Endothelial Microparticles Delivering Lncrna NEAT1 Are Involved in the Pathophysiology of aGVHD Following Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Zhang Ran , Cao Weijie , Li Li , Wan Dingming , Jiang Zhongxing , Sun Kai , Xia Linghui 
Published: February 2021

Endothelial microparticles (EMPs) upregulation has been observed in the pathological process of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The biological role and therapeutic potential of long non-coding RNAs (lncRNAs) carried by EMPs in aGVHD after allo-HSCT are still open questions. Herein, we investigated the functional significance of the lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) delivered by EMPs in aGVHD. To address this question, in the present study, we performed a comprehensive analysis of the lncRNA NEAT1 expression and EMPs in aGVHD by a series of in vivo and in vitro experiments. We found that expressions of lncRNA NEAT1 in MPs were remarkably elevated in aGVHD mice than in non-aGVHD mice. Furthermore, the lncRNA NEAT1 levels in the MPs were remarkably higher than those in the plasma of the aGVHD mice. We then used TNF-α (100 ng/ml) to stimulate primary mouse aortic endothelial cells (MAECs) to shed EMPs, and the expression of lncRNA NEAT1 in the EMPs and their maternal cells was compared. The lncRNA NEAT1 levels in the EMPs produced by TNF-α-stimulated MAECs were significantly higher than that in both their maternal cells and non-stimulated EMPs. Our study demonstrates that EMPs could assemble and concentrate lncRNA NEAT1. Taken together, NEAT1 delivered by EMPs could be used as a diagnostic and prognostic surrogate marker for aGVHD, and its targeting could therefore represent a promising strategy for novel therapeutic options in life-threatening aGVHD after allo-HSCT.

Disclosures
No relevant conflicts of interest to declare.
The Serine Protease Matriptase Acts As a Tumour Suppressor in Multiple Myeloma
Authors: Steiro Ida , Abdollahi Pegah , Børset Magne , Slørdahl Tobias S. 
Published: February 2021

Both in newly diagnosed multiple myeloma (MM) and during progression of the disease, malignant plasma cells are found circulating in peripheral blood as well as in the bone marrow (BM). The disseminated nature of MM is strongly dependent on the interplay between the cancer cells and the BM microenvironment, promoting myeloma cell migration in the BM. Matriptase (ST14), a type-II transmembrane serine protease primarily found in epithelial tissues, is overexpressed in a variety of human malignancies and is sufficient to induce tumour formation in mice. Frequently, a concomitant reduction in the levels of its cognate inhibitor hepatocyte growth factor activator inhibitor (HAI)-1 (SPINT1) is observed in carcinomas, while expression and function of the related inhibitor HAI-2 (SPINT2) is yet to be clarified. Dysregulated expression causing increased matriptase proteolytic activity has been associated with cancer growth, survival and metastasis. Here, we show for the first time a role of matriptase as a possible tumour suppressor in myeloma pathogenesis. Gene expression analysis of primary cells from MM patients (n=24) and human myeloma cell lines (n=8) revealed highly variable levels of matriptase, HAI-1 and HAI-2. This observation prompted us to investigate the functional role of matriptase in vitro. We showed that stable overexpression of matriptase in INA-6, a MM cell line with no endogenous ST14 expression, reduced migration by more than 50% in response to the combination of the pro-migratory cytokines stromal cell-derived factor-1 alpha (SDF-1α) and hepatocyte growth factor (HGF, Fig. 1A). Conversely, stable knockdown of matriptase in two MM cell lines with high endogenous matriptase expression (RPMI-8226 and JJN-3) significantly enhanced migration in vitro. Mechanistically, matriptase overexpression blocked activation of Src kinase (Fig. 1B), well-known as a critical player in metastasis formation promoting cancer cell motility, invasiveness and angiogenesis. In agreement with our result, previous studies have demonstrated the activation of Src family kinases (SFK) downstream SDF-1/CXCR4-signaling. Finally, we performed survival analyses in the public available MMRF CoMMpass trial database (release version IA14). Low ST14 expression was associated with significant worse overall survival (P=0.05, Fig. 1C) and progression-free survival (P=0.02, Fig. 1D). Altogether, our data are in marked contrast to the role ascribed to matriptase in epithelial and certain non-epithelial tumours as an oncogenic protein and an unfavourable prognostic marker. In conclusion, these findings suggest a novel role of matriptase as a tumour suppressor in MM pathogenesis.

Disclosures
Slørdahl: Celgene: Consultancy; Janssen and Celgene: Honoraria.
The Neurotransmitter Receptor Gabbr1 Regulates Proliferation and Function of Hematopoietic Stem and Progenitor Cells
Authors: Elujoba-Bridenstine Adedamola , Shao Lijian , Zink Katherine , Sanchez Laura , Pajcini Kostandin V. , Tamplin Owen J. 
Published: February 2021

Hematopoietic stem and progenitor cells (HSPCs) are multipotent cells which differentiate to maintain and replenish blood lineages throughout life. Due to these characteristics, HSPC transplants represent a cure for patients with a variety of hematological disorders. HSPC function and behavior is tightly regulated by various cell types and factors in the bone marrow niche. The nervous system has been shown to indirectly influence hematopoiesis by innervating the niche; however, we present a direct route of HSPC regulation via expression of neurotransmitter receptors on HSPC surface. We have identified Gamma Aminobutyric acid (GABA) receptor B subunit 1 (Gabbr1), a hitherto unknown hematopoietic player, as a regulator of HSPC function. GABBR1 is known to be expressed on human HSPCs (Steidl et al., Blood 2004), however its function in their regulation remains unknown. Based on published RNA-seq data (Nestorowa et al., Blood 2016), we discovered that Gabbr1 is expressed on a subset of HSPCs. We confirmed this expression using RT-qPCR to assay hematopoietic populations in the bone marrow (BM). Surface receptor expression analysis showed that Gabbr1 protein is expressed on a subset of BM HSPCs. To detect GABA, the ligand for Gabbr1 in the BM microenvironment, we utilized imaging mass spectrometry (IMS). We detected regionally specific GABA signal in the endosteal region of the BM. We further identified B cells as a cellular source of GABA in the BM. To understand the role of Gabbr1 in hematopoiesis, we generated CRISPR-Cas9 Gabbr1 null mutants on a C57/BL6 background suitable for hematopoietic studies and studied their hematopoietic phenotype. We discovered a decrease in the absolute number of Lin-Sca1+cKit+ (LSK) HSPCs, but the long-term hematopoietic stem cells (LT-HSCs) remain unaffected. Further analysis of peripheral blood of Gabbr1 null mutants showed decreased white blood cells due to reduced B220+ cells. This differentiation defect was confirmed in an in vitro differentiation assay where Gabbr1 null HSPCs displayed an impaired ability to produce B cells. We show that Gabbr1 null HSCs show diminished reconstitution ability when transplanted in a competitive setting. Reduced Gabbr1 null HSC reconstitution persisted in secondary transplant recipients indicating a cell autonomous role for Gabbr1 in regulating reconstitution of HSCs in transplant recipients. Our results show a crucial role for Gabbr1 in HSPC regulation and may translate to human health as a rare human SNP within the GABBR1 locus that correlates with altered leukocyte counts has been reported (Astle et al., Cell 2016). Our studies indicate an important role for Gabbr1 in HSPC reconstitution and differentiation into B cell lineages.

Disclosures
No relevant conflicts of interest to declare.
Low-Dose Triptolide Promotes MDM2 Inhibitor Nutlin3a to Induce Acute Myeloid Leukemia Cell Death Via p53-Dependent and -Independent Mechanisms
Authors: Chen Qinwei , Deng Suqi , Lin Zhijuan , Deng Manman , Ding Lihong , Zha Jie , Zhou Yong , Jiang Yuelong , Carter Bing Z , Xu Bing 
Published: February 2021

The tumor suppressor p53 is central to hematopoietic stem cell function. Loss of p53 function is associated with poor prognosis of acute myeloid leukemia (AML). P53 negative regulator MDM2 is frequently overexpressed in AML, thus it becomes an attractive therapeutic target for the treatment of AML with wild-type p53 (Wt-p53). Targeting MDM2 to restore p53 activity has been assessed in AML, but clinical outcomes are less promising. Notably, p53-mutated AML still lacks effective treatment approaches. Therefore, our study strived to explore potent therapeutics to target AML with p53 wild-type and mutations.
Triptolide (TPL), an ancient herbal medicine, has been used to treat immune disorders for centuries. Accumulating evidence including ours have proven that low dose of TPL potentiates the efficacy of chemotherapies in a broad range of tumor types. In this study, we evaluated the anti-leukemic efficacy of combining low dose TPL with Nutlin3a, a MDM2 inhibitor, against AML with p53 wild-type and mutations. We found that low-dose TPL synergized with Nutlin3a to induce mitochondrial apoptosis in AML cells with Wt-p53 bothin vitroandin vivo. Underlying mechanism for the synergy showed that Nutlin3a upregulated pro-apoptosis factors (e.g. PUMA, p21), while low-dose TPL suppressed MDM2 transcription and reduced anti-apoptosis proteins (e.g. XIAP, Mcl-1) in Wt-p53 AML cells. Interestingly, we also observed that TPL but not Nutlin3a induced cell death in p53 shRNA knockdown, p53 deficiency cell lines (e.g. THP-1, HL-60) and primary samples. Thus, the p53-independent role of TPL was consequently focused. Our RNA-Seq analysis identified 621 differential expression genes (DEGs) after TPL treatment. These genes, including MYC, ATF4 and 4EBP1, were enriched in intrinsic apoptosis pathway responding to ER stress and thus we concluded that Triptolide played its p53 independent role via MYC-ATF4 axis.
Collectively, these findings suggest that coadministration of Nutlin3a with low dose TPL would benefit for AML patients.

Disclosures
Carter: AstraZeneca:Research Funding;Syndax:Research Funding;Ascentage:Research Funding;Amgen:Research Funding.

OffLabel Disclosure:
Triptolide (TPL), an ancient herbal medicine, is applying to treat immune disorders.
LYG1 Deficiency Attenuates the Severity of Acute Graft-Versus-Host Disease Via Skewing T Cells Polarization Towards Treg Cells
Authors: Liu Huihui , Yu Zhengyu , Tang Bo , Miao Shengchao , Qin Chenchen , Li Yuan , Liang Zeyin , Zhang Yang , Ren Hanyun , Dong Yujun 
Published: February 2021

Acute graft-versus-host disease (aGVHD) is a lethal complication after allogeneic hematopoietic stem cell transplantation (allo-HSCT). As a complex immunopathology, aGVHD depends on the recognition of host antigens by donor T cells and induces augmented response of alloreactive T cells. Despite considerable achievements in the treatment of aGVHD, it remains a major clinical problem for the patients undergoing allo-HSCT. Therefore, it is necessary to further illustrate new mechanisms and develop novel therapeutic strategies of aGVHD. Previously we reported LYG1 (Lysozyme G-like 1) as a novel classical secretory protein promoted antitumor function of T cell. In this study, the role of LYG1 in aGVHD was investigated. Firstly, we examined whether LYG1 affected the alloreactivity of CD4+ T cells in vitro by MLR assay and discovered that LYG1 deficiency reduced the activation of CD4+ T cells and Th1 ratio, but increased Treg ratio. Then we confirmed these observations using a major MHC mismatched aGVHD model by transferring T cells sorting from WT B6 or Lyg1-/- mice with bone marrow cells from WT B6 mice into lethally irradiated BALB/c mice. The alloreactive CD4+ T cells and the proportions of Th1 cells decreased whereas the proportions of Treg cells increased in spleens and livers in mice receiving Lyg1-/- T cells. LYG1-deficient T cells attenuated aGVHD severity, inhibited the expression of CXCL9 and CXCL10 and restrained CD4+ T cells infiltrating in livers. Furthermore, administration of recombinant LYG1 protein intraperitoneally aggravated aGVHD by promoting IFN-γ production. More importantly, LYG1 deficiency did not affect GVT (graft-versus-tumor) effects. In summary, we demonstrate LYG1 regulates aGVHD via modulating the alloreactivity of CD4+ T cells and differentiation of Th1/Treg cells. Our study indicates that LYG1 may be a novel target in aGVHD by mitigating aGVHD without impairing GVT function. The therapeutic effect of targeting LYG1 is required in future investigations.
Funding
This study was supported by grant from The National Natural Science Foundation of China (NSFC) (Grant Number 81600144) and grant from Research Foundation of Peking University First Hospital.

Disclosures
No relevant conflicts of interest to declare.
N-glycan–mediated shielding of ADAMTS13 prevents binding of pathogenic autoantibodies in immune-mediated TTP
Authors: Ercig Bogac , Graça Nuno A. G. , Kangro Kadri , Arfman Tom , Wichapong Kanin , Hrdinová Johana , Kaijen Paul , van Alphen Floris P. J. , van den Biggelaar Maartje , Vanhoorelbeke Karen , Veyradier Agnès , Coppo Paul , Reutelingsperger Chris , Nicolaes Gerry A. F. , Männik Andres , Voorberg Jan 
Published: February 2021

Abstract
Immune-mediated thrombotic thrombocytopenic purpura (iTTP) is an autoimmune disorder caused by the development of autoantibodies targeting different domains of ADAMTS13. Profiling studies have shown that residues R568, F592, R660, Y661, and Y665 within exosite-3 of the spacer domain provide an immunodominant region of ADAMTS13 for pathogenic autoantibodies that develop in patients with iTTP. Modification of these 5 core residues with the goal of reducing autoantibody binding revealed a significant tradeoff between autoantibody resistance and proteolytic activity. Here, we employed structural bioinformatics to identify a larger epitope landscape on the ADAMTS13 spacer domain. Models of spacer-antibody complexes predicted that residues R568, L591, F592, K608, M609, R636, L637, R639, R660, Y661, Y665, and L668 contribute to an expanded epitope within the spacer domain. Based on bioinformatics-guided predictions, we designed a panel of N-glycan insertions in this expanded epitope to reduce the binding of spacer domain autoantibodies. One N-glycan variant (NGLY3-ADAMTS13, containing a K608N substitution) showed strongly reduced reactivity with TTP patient sera (28%) as compared with WT-ADAMTS13 (100%). Insertion of an N-glycan at amino acid position 608 did not interfere with processing of von Willebrand factor, positioning the resulting NGLY3-ADAMTS13 variant as a potential novel therapeutic option for treatment of iTTP.
BAFF promotes heightened BCR responsiveness and manifestations of chronic GVHD after allogeneic stem cell transplantation
Authors: Jia Wei , Poe Jonathan C. , Su Hsuan , Anand Sarah , Matsushima Glenn K. , Rathmell Jeffrey C. , Maillard Ivan , Radojcic Vedran , Imai Kazuhiro , Reyes Nancy J. , Cardona Diana M. , Li Zhiguo , Suthers Amy N. , Curry-Chisolm Itaevia M. , DiCioccio Rachel A. , Saban Daniel R. , Chen Benny J. , Chao Nelson J. , Sarantopoulos Stefanie 
Published: February 2021

Abstract
Patients with chronic graft-versus-host disease (cGVHD) have increased B cell–activating factor (BAFF) levels, but whether BAFF promotes disease after allogeneic bone marrow transplantation (allo-BMT) remains unknown. In a major histocompatibility complex–mismatched model with cGVHD-like manifestations, we first examined B-lymphopenic μMT allo-BMT recipients and found that increased BAFF levels in cGVHD mice were not merely a reflection of B-cell number. Mice that later developed cGVHD had significantly increased numbers of recipient fibroblastic reticular cells with higher BAFF transcript levels. Increased BAFF production by donor cells also likely contributed to cGVHD, because BAFF transcript in CD4+ T cells from diseased mice and patients was increased. cGVHD manifestations in mice were associated with high BAFF/B-cell ratios and persistence of B-cell receptor (BCR)–activated B cells in peripheral blood and lesional tissue. By employing BAFF transgenic (Tg) mice donor cells, we addressed whether high BAFF contributed to BCR activation in cGVHD. BAFF increased NOTCH2 expression on B cells, augmenting BCR responsiveness to surrogate antigen and NOTCH ligand. BAFF Tg B cells had significantly increased protein levels of the proximal BCR signaling molecule SYK, and high SYK protein was maintained by BAFF after in vitro BCR activation or when alloantigen was present in vivo. Using T cell–depleted (BM only) BAFF Tg donors, we found that BAFF promoted cGVHD manifestations, circulating GL7+ B cells, and alloantibody production. We demonstrate that pathologic production of BAFF promotes an altered B-cell compartment and augments BCR responsiveness. Our findings compel studies of therapeutic targeting of BAFF and BCR pathways in patients with cGVHD.