bluuebunny/crossref_metadata_2025_split huggingface dataset:
Patients with Paroxysmal Nocturnal Hemoglobinuria (PNH) Treated with Pegcetacoplan: Real Life Experience
Authors: Vallejo Carlos , Bonanad Boix Santiago , Carnicero Fernando , Gaya Anna , Gonzalez Ana Pilar , Fernandez Fernando Ataulfo Gonzalez , Lavilla Esperanza , Salido Eduardo José , Sanchez Anton Piva , Vara Míriam 
Published: December 2023

Introduction: C5 inhibitors (C5i) treatment has dramatically improved the management of intravascular hemolysis (IVH) in PNH patients. However, a significant proportion of patients remain anemic due to suboptimal control of IVH and the development of extravascular hemolysis (EVH), consequence of C5i treatment. Pegcetacoplan (PEG) is the first proximal inhibitor of the complement system, through the inhibition of C3, allowing to control both IVH and EVH. PEG has recently been marketed in Spain for use in adult PNH patients who remain anemic after treatment with an C5i for at least 3 months. The objective of this work is to analyze the response of the first patients treated with PEG in real life in Spain.
Patients and methods: clinical and analytical variables were collected from a series of 10 patients diagnosed with PNH who received treatment with PEG through an early access program. PEG was self-administered subcutaneously at 1080 mg twice a week. See Table 1. Transfusion needs and the adverse effects that occurred were also collected.
Results: median (range) duration of treatment with PEG at the time of the analysis was 4 (2-8) months. Hemoglobin (Hb) values increased a median of 2.4 g/dL after PEG treatment. This improvement was rapidly observed since the beginning of the treatment (median increase at 1 week was 1.3 g/dL and at 2 weeks 2.4 g/dL). All patients reached Hb levels of &gt;8 g/dL, 80% &gt;10 g/dL and 40% &gt;12 g/dL. An increased in Hb &gt;2 g/dL was observed in 60% of the patients. Median values of parameters of hemolysis (absolute reticulocyte counts, bilirubin, LDH) became normal. See Table 2. The direct antiglobulin test turned into negative in all positive cases pre-PEG (4 out of 4). No patient suffered from acute hemolytic events, thrombosis, or required PRBC transfusion once PEG had been initiated. On a 5-point scale (0: nothing; 5: very much), subjective improvement of patient quality of life and physician satisfaction were 4 (3-5), and 5 (3-5) [median (range)]. PEG-related adverse events were just reported in one patient (10%): mild post-infusion nausea. At the end of the analysis, all patients continue on PEG.
Conclusions: the results observed in the first patients treated with PEG in real life in Spain, although with a limited follow-up, confirm those reported in phase 3 clinical trials, with significant improvement in the Hb levels and hemolysis parameters, which results in transfusion independence. PEG was also shown to act quickly from the start of treatment, to significantly improve quality of life, and to present an excellent safety profile. PEG, the first approved proximal complement inhibitor, can therefore be considered as a relevant therapeutic option in the current management of PNH.
Single-Cell RNA Sequencing Revealed the YY1/EZH2/MLH1 Axis As a Possible Therapeutic Target of Intractable Adult T-Cell Leukemia
Authors: Shimizu Takuya , Shindo Takero , Watanabe Akira , Takaori-Kondo Akifumi 
Published: December 2023

Background: Due to clinical heterogeneity of adult T-cell leukemia/lymphoma (ATL) and its diverse genetic abnormality, common therapeutic targets of ATL remains unclear. Whereas the YY1/EZH2 axis regulates global epigenetic modification of genes in ATL, its downstream target genes have not been fully elucidated. Meanwhile, DNA mismatch repair proteins may be targeted in ATL. Here we explored underlying molecular axes for ATL progression through comprehensive single-cell RNA-sequencing (sc-RNA-seq).
Methods: Peripheral blood of 42 ATL patients and asymptomatic HTLV-1 carriers in addition to two cell lines of ATL43T and ED+ were subjected to the following analyses. Molecular markers associated with their clinical phenotypes were extracted through sc-RNA-seq analysis. Expression levels of DNA mismatch repair proteins (MLH1, MSH2) and target molecules were validated by flow cytometry. Finally, their function was verified through lentiviral shRNA-dependent knockdown and molecular-targeting reagents.
Results: While the frequencies of CADM1+ cells among CD4+ T cells were generally correlated with HTLV-1 proviral loads (R = 0.76, p &lt; 0.001), substantial CADM1+ cells were detected but did not increase in some long-remitted cases; thus, CADM1+ cells are biologically heterogeneous. sc-RNA-seq of CADM1+ cells of four ATL cases identified a cluster of CD48-deficient cells. in silico promoter analysis of the cluster extracted a transcription factor YY1 as a candidate for common master regulators. EZH2 and the putative driver genes (CELF2, PTPRC, CBLB, ATXN1, IKZF2) were identified as differentially expressed genes (DEGs) in the cluster. The EZH2-overexpressed cluster was increased in acute-type ATL compared with remitted ATL and smoldering-type ATL. Interestingly, the cluster increased moderately in the other smoldering-type ATL, which required UVB irradiation due to intractable skin lesion. Given that YY1 and EZH2 were up-regulated in acute-type ATL, the cluster may represent tumor aggressiveness. Furthermore, biphasic expression of YY1/EZH2 was observed in 2 acute-type ATL, which indicates that the EZH2 high ATL cells confer malignant signature. In-house analysis on microarray dataset GSE55851 specified the MLH1 associated with YY1/EZH2. MLH1 within CADM1+ fraction was down-regulated in ATL patients with remission (n = 7) but maintained in progressive ATL (n = 6) (p &lt; 0.05). Lentiviral shYY1 knockdown resulted in down-regulation of MLH1 in both ATL43T and ED+ (p &lt; 0.01). The EZH1/2 inhibitor valemetostat down-regulated EZH2 (p = 0.018) and MLH1 (p = 0.014) in primary ATL cells. Finally, knockdown of either YY1 or MLH1 suppressed proliferation of ATL43T and ED+ (p &lt; 0.0001).
Conclusions: Our findings indicate that down-regulation of MLH1 through YY1/EZH2 inhibition plays a key role in the treatment of aggressive ATL.
Iron Uptake from Ferric Maltol in Patients with Iron Deficiency with or without Anemia: Post Hoc analysis of a Multicenter, Open-Label, Randomized, Phase 1 Clinical Study
Authors: Howaldt Stefanie , Cody Michael , Mitchell Jacqueline A 
Published: December 2023

Introduction: Iron deficiency is the most common nutritional disorder in humans, accounting for more than half of the estimated 1.76 billion cases of anemia worldwide (https://www.healthdata.org/). Anemia is a serious health problem, but, even without anemia, people with iron deficiency may experience fatigue, headaches, restless legs, and other physical and cognitive impairments, severely limiting their daily activities and quality of life. Patients may require long-term iron replacement therapy to replenish iron stores. Traditional oral iron salts are often associated with intolerable gastrointestinal toxicity and poor absorption. Ferric maltol is a European Medicines Agency- and U.S. Food and Drug Administration-approved oral iron formulation that was uniquely designed to improve gastrointestinal absorption, resulting in increased iron uptake while minimizing the risk of intestinal damage or dysbiosis. In a Phase 1 study in adults with inflammatory bowel disease and iron deficiency, ferric maltol increased iron uptake and storage over time at twice-daily doses of 30-90 mg (Bokemeyer B et al. Eur J Drug Metab Pharmacokinet 2017;42:229-238). We assessed iron measures following ferric maltol treatment in patients with iron deficiency with or without anemia.
Methods: In post hoc analyses of data from 17 of the Phase 1 study participants (4 men, 13 women), who received twice-daily ferric maltol 30 or 60 mg, we compared total iron and transferrin saturation (TSAT) maximum concentration ( Cmax) and area under the curve up to 6 hours (AUC 6h) on Days 1 and 8 in subgroups with and without anemia. Data are reported as arithmetic means (standard deviation).
Results: Four of the 17 patients (all female) had iron deficiency with anemia. Median (range) baseline hemoglobin levels were 107.5 (100-114) g/L in the subgroup with anemia and 135 (127-159) g/L in the subgroup without anemia. Total serum iron and TSAT values on Days 1 and 8 were similar between the subgroups with and without anemia (Table).
Conclusions: These data demonstrate similar iron uptake from ferric maltol in patients with iron deficiency with or without anemia. Given the adverse impact of iron deficiency, even without anemia, iron-replacement therapy should be considered in anyone with an indication of low iron availability (e.g., ferritin &lt;30 ng/mL with any TSAT value or ferritin &lt;100 ng/mL with TSAT &lt;20%), regardless of hemoglobin level. At the approved dose of 30 mg twice daily, ferric maltol is an effective, well tolerated, and convenient to take long-term iron-replacement therapy.
Sting Activation in Bone Marrow Macrophages Targets AML Blasts for Phagocytosis in an ICAM-1 Dependent Mechanism
Authors: De Silva Ravindu , Markham Matthew , Maynard Rebecca S , Hampton Katherine , Hellmich Charlotte , Fowler-Shorten Dominic , Bowles Kristian , Rushworth Stuart 
Published: December 2023

The initiation and proliferation of acute myeloid leukaemia (AML) is in part regulated by the bone marrow microenvironment. Previous work by our group has shown that the stimulator of interferon genes (STING) pathway is activated in macrophages in the AML bone marrow microenvironment (Moore et al., 2022) . We found that AML-derived mitochondrial damage-associated molecular patterns were processed by bone marrow macrophages (BMMs) via LC3-associated phagocytosis (LAP). Furthermore, activation of STING resulted in a suppression of AML growth through an LAP dependent mechanism. Here we have investigated how the activation of STING in bone marrow macrophages increases macrophage phagocytic activity and in turn supresses the growth of AML.
To understand the role of STING activation in bone marrow derived macrophages (BMDMs) in AML we used various STING agonists including DMXAA, 2'3'-cGAMP, and CpG oligonucleotides. We showed STING activation was induced through these agonists by measuring the upregulation of STING related genes ( Gbp2, Irf7 and Ifit3). We demonstrated that STING activation caused increased phagocytic clearance of live MN1 cells (mouse AML cell line) in an in vitro phagocytosis assay. In order to investigate how STING activation increases the phagocytic clearance of AML cells by BMDMs, we used three complementary methods. First, we performed a receptor tyrosine kinase array to measure the phosphorylation of mouse kinases following STING activation. Second, we investigated the transcriptomic changes resulting from STING activation with mRNA-sequencing. Third, after combining these results we identified an upregulation of key phagocytic pathways that specifically led us to focus on the adhesion molecules, in particular ICAM-1. Further investigation using an anti-ICAM-1 blocking antibody showed a reduction in the phagocytic potential of the BMDMs demonstrating the role of ICAM-1 in the phagocytic clearance of AML.
Next, we activated human macrophages (derived from peripheral blood monocytes collected from healthy donors with approval of the East of England - Cambridgeshire and Hertfordshire Research Ethics Committee and the UK Health Research Authority, IRAS Project ID 33753) through STING which depleted AML in vitro confirming the results obtained using the mouse system.
To investigate the in vivo significance of these findings, we treated C57Bl/6 animals with DMXAA and observed a significantly decreased tumour burden and increased survival following engraftment with MN1 cells. Cytotoxic T cell activity and number in the bone marrow was unchanged by STING activation in this model.
In summary, we have shown that the activation of STING in bone marrow macrophages directly supresses the growth and proliferation of AML by stimulating the phagocytic activity of the macrophages, and the clearance of the leukaemic cells.
Automated Cut&run Brings Scalable Epigenomic Profiling to Hematology
Authors: Maier Keith E , Marunde Matthew R , U. Sunitha Kumary Vishnu , Lin Carolina P , Maryanski Danielle N , Albertorio-Saez Liz , Ahimovic Dughan J , Bale Michael J , Lee Juliana J , Venters Bryan J , Keogh Michael-Christopher 
Published: December 2023

Genome-wide association studies (GWAS) and corresponding transcriptomics research have been leveraged to identify disease risk variants in hematological disorders with the hopes of developing more personalized treatment regimens. Interestingly, &gt;90% of GWAS-identified variants are found in non-coding regions of the genome, indicating they likely affect the regulatory machinery that governs chromatin structure.
Non-coding elements, such as enhancers, play important roles in supporting chromatin mechanisms, including cell-type specific gene expression, cell fates and disease states. However, these variants are difficult to study compared to protein-coding mutations and have been challenging to directly associate with phenotypic readouts. To date, efforts to understand the effects of non-coding variants on chromatin dynamics have been focused on chromatin accessibility (ATAC-seq) and DNA methylation profiling. Notably, these assays can only provide a binary, “open-or-closed,” view of chromatin and often fail to provide mechanistic insight into disease etiology.
Epigenomic features - such as histone post-translational modifications (PTMs) and chromatin-associated proteins - mark distinct genomic compartments (e.g., promoters, enhancers) and regulate chromatin structure, gene expression, and cell function. Mapping these features provides a rich context to study cell fate and has great potential for discovering new biomarkers and drug targets. Despite this, efforts to integrate epigenomics into large scale lymphatic and myeloid research have been hampered by the poor sensitivity, high background, and low throughput of traditional chromatin mapping technology (i.e., ChIP-seq).
Here, we present autoCUT&RUN, a high throughput assay for rapid, ultra-sensitive profiling of epigenomic features from FACS-isolated primary cells or tissues. This workflow generates reliable profiles from &lt;10,000 cells per reaction, and is supported by a rigorous optimization strategy, high-quality antibodies, and quantitative spike-in controls. As part of a multi-site collaboration with the Immunological Genome Consortium, we used our autoCUT&RUN platform to build a comprehensive epigenomic database of mouse immune cells - composed of &gt;1,500 epigenomic profiles from &gt;100 different FACS-isolated primary immune cell types. These studies set the stage to leverage high-throughput epigenomics for multi-site clinical studies to identify and/or characterize novel biomarkers for precision medicine applications.
Further, we will discuss our integration of CUT&RUN and Enzymatic Methylation-sequencing (CUT&RUN-EM) to deliver a true multiomic view of the co-occurrence of epigenomic features and DNA methylation, which can reveal multivalent cellular mechanisms and deconvolute sample heterogeneity.
New Suitable Eµ-Tcl1 Mouse Model for Research on BH3-Mimetics Therapy Response In Vivo
Authors: Halmer Ismini , Beckmann Laura , Knittel Gero , Kashkar Hamid , Reinhardt Hans Christian , Hallek Michael , Da Palma Guerreiro Alexandra , Frenzel Lukas Peter 
Published: December 2023

Introduction: Generation of appropriate mouse models to study clinically active compounds is inevitable. Currently, the commonly used mouse model for CLL is the TCL1 transgenic model, which shows an expression of the human TCL1 oncogene in B cells. While this model resembles many features of human CLL, it is resistant to venetoclax, a BCL2 inhibitor which represents an important treatment option for CLL and other hematological malignancies. The lack of therapeutic response to venetoclax is an important limitation of the TCL1 mouse model. As a result, the effectiveness, the mechanisms, and the development of venetoclax resistance and other BH3 mimetics cannot be investigated in the TCL1 transgenic setting.
Methods and results: To overcome the stated limitations, a novel mouse model was generated expressing human BCL2 in B cells under the control of the TCL1 promoter. We established a new mouse model (TBC) by crossbreeding Eµ-Tcl1 tg/wt mice with mice containing a B cell specific conditional Bcl-2 Rosa26/wt; Cd19Cre Cre/wt overexpression (control group Eµ-Tcl1 tg/wt; Cd19Cre Cre/wt: TC). Initial data show a strong leukocytosis ( p&lt;0,0001) and splenomegaly in TBC mice leading to a significantly shortened overall survival compared to TC mice (log-rank: p=0,0028). Immunophenotyping revealed a population of class-switched IgM - B cells within TBC mice and only a small population of Cd19 +/Cd5 dim cells. Transcriptomics followed by gene enrichment analysis showed a development of GCB-type DLBCL in TBC mice compared to TC mice.
Apoptosis assays revealed high sensitivity towards BH3 mimetics, especially BCL2 inhibitor venetoclax. In vitro experiments demonstrated that B cells from TBC mice are 10 times more sensitive towards venetoclax treatment. Based on this fact, we treated 5 TBC mice for 4 weeks with venetoclax. Even in this short treatment time frame, there was a strong loss of leukocytes in peripheral blood of about 60%, leading to normal ranges of leukocytes in TBC mice after even 14 days of treatment with venetoclax. Interestingly, we observed a massive change of Cd4 +/Cd8 + ratio towards Cd4 + cells due to venetoclax treatment, indicating that our novel model resembles human settings.
Conclusions: We were able to show a strong therapy response towards treatment with venetoclax in vivo in TBC mice leading to a loss of leukocytes, a gain of CLL typical Cd19 +/Cd5 dim cells and a strong effect on tumormicroenvironment with a significantly changed Cd4 +/Cd8 + ratio. This makes the new mouse model very suitable for testing and observing venetoclax therapy response, its side effects and outcome in vivo, to provide our patients with the best possible therapy.
HDAC Inhibitor CS014 Attenuates Thrombosis Alone and in Combination with Rivaroxaban without Increased Risk of Bleeding
Authors: Stanger Livia , Holinstat Michael , Lambert Sylviane , Yalavarthi Pooja , Bergh Niklas , Dahlof Bjorn 
Published: December 2023

Introduction: Targeting the human platelet for prevention of occlusive thrombotic events remains an active area of research due to the high incidence of myocardial infarction and stroke due to platelet-rich thrombotic clots. While, many patients taking FXa anticoagulant therapy require further inhibition of thrombosis, dual therapy including anticoagulant plus antiplatelet drugs result in an increased risk for bleeding. The current study sought to determine if a histone deacetylase (HDAC) inhibitor, either alone or in combination with the FXa inhibitor rivaroxaban, is able to effectively inhibit platelet and fibrin formation at the site of vascular injury without causing a significant increase in the bleeding risk in mice.
Methods and Results: Thrombosis and bleeding models in the mouse were assessed in the presence of the HDAC inhibitors valproic acid (VPA) or the newly developed HDAC inhibitor, CS014. Both CS014 and VPA significantly, dose-dependently inhibit platelet accumulation and fibrin formation at the site of a vascular injury using the laser-induced cremaster thrombosis assay model. Further, to determine if CS014 has the potential to be used in conditions where an anticoagulant is already on board, mice were dosed for 5 days with rivaroxaban alone or in combination with CS014 and were assessed for prevention of laser-induced thrombosis in the cremaster arteriole. Both rivaroxaban and CS014 alone cause significant attenuation of clot formation. To determine if the combination results in an increased risk for bleeding, blood from mice treated with CS014 or rivaroxaban alone, or in combination was assessed for coagulation activity and clot strength using thromboelestography (TEG). Rivaroxaban was found to significantly delay onset of clotting in TEG whereas CS014 did not show any delay in coagulation or clotting with TEG. In combination CS014 was not observed to cause any additional delay in clotting time or decrease in maximal clot strength compared to rivaroxaban alone. Finally, a tail vein bleeding assay was conducted to determine if CS014 alone or in combination with rivaroxaban result in an increased risk for bleeding. Neither CS014 alone, rivaroxaban alone, or the combination of CS014 plus rivaroxaban resulted in increased bleeding time in the resected tail experiment.
Conclusions: The data presented here suggests that while CS014 and rivaroxaban are each effective antithrombotic agents, they work well together to minimize clot formation without altering coagulation or clotting compared to rivaroxaban alone. Hence, CS014 has the potential to enrich the toolbox of anticoagulant therapies in patients with a high risk of thrombotic events.
Levels of Individual Mid and Long Chain Polyunsaturated Triacylglycerols (TAGs) Are Different between the Bone Marrow Microenvironment of Patients with Multiple Myeloma (MM) and Monoclonal Gammopathy of Unknown Significance (MGUS)
Authors: Chawla Yogesh , Anderson Emilie , Vasquez Isas Rosario , Simon Rachel A , Hitosugi Taro , Kumar Shaji Kunnathu , Gonsalves Wilson I. 
Published: December 2023

Background: It has been recently demonstrated that altered lipid metabolism involving increased free fatty acids (FFAs) uptake supports the growth and proliferation of MM cells (Panaroni et al. Blood 2022; Farell et al. Elife 2023). Whether the availability of FFAs in the bone marrow (BM) microenvironment is associated with the pathogenesis of MM from MGUS has not been previously evaluated. Thus, we evaluated if individual species of FAs or FA-containing lipids such as triacylglycerols (TAGs), diacylglycerols (DAGs), and monoacylglycerols (MAGs) are differentially present between the BM microenvironment of MGUS and MM patients.
Methods: Lipid profiles of BM plasma samples (reflective of the BM microenvironment) from patients with a spectrum of plasma cell disorders (22 MGUS and 24 MM (15 newly diagnosed and 9 relapsed/refractory)) were acquired by liquid chromatography (LC)/tandem mass spectrometry (MS), and individual lipid species were quantified based on single-point quantification using 55 isotopically labeled internal standards. Concentration-based lipid species data was analyzed by the web-based tool MetaboAnalyst 5.0. Data was normalized by quantile normalization, log transformed and scaled by Pareto scaling. p values &lt;0.05 were considered significant.
Results: A total of 993 distinct lipid species were quantified by the MS-based targeted lipidomics assessment. Of these lipid species, the levels of individual mid-chain (≤50 carbon chain length) and long-chain (≥52 carbon chain length) polyunsaturated TAGs were the most differentially present in the BM plasma of MM compared to MGUS patients (Figure 1A). Even though a principal component analysis (PCA) involving all lipid species showed moderate separation of MM from MGUS patients, five MGUS patients (categorized as MGUS-other (MGO)) who clustered more towards MM patients compared to the remaining MGUS patients had a distinct pattern of individual mid and long-chain polyunsaturated TAG levels that were comparable to MM patients (Figure 1B). Furthermore, the proliferation rate of clonal plasma cells in the BM of these MM and MGUS as measured by the S-phase% was found to be directly correlated with the level of long-chain polyunsaturated TAGs.
Conclusions: Mid and long-chain polyunsaturated TAGs are present in lower and higher levels, respectively, in the BM microenvironment of MM compared to MGUS patients. These findings continue to support the importance of FA-containing TAGs and other lipids in the pathogenesis of MM. Future confirmatory studies evaluating the biological significance of these differences are required to determine if they can be exploited for diagnostic and therapeutic purposes.