bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thrombosis Caused By a Novel Coagulation Factor IX Mutation (FIX Shanghai II)
Authors: WU Wenman , Wang Rui , Xu Qin , Wang Guanzheng , Jiang Shifeng , Lou Can , Wu Xi , Shao Yanyan , Lu Yeling , Liu Yu , Xu Guanquan , Ding Qiulan , Hu Xiaobo , Wang Xuefeng , Dai Jing 
Published: December 2023

Most thrombophilia patients are caused by insufficient suppression of coagulation reactions, which could be either mediated by deficiencies of protein C, protein S,antithrombin, or inhibition-resistant coagulation factor mutants, such as Factor V Leiden and prothrombin Yukuhashi. The thrombosis caused by coagulation factor mutants with increased procoagulant activity is very rare and so far only two Factor IX (FIX) variants involving residue Arg384, FIX Padua (p.Arg384Leu) and Shanghai (p.Arg384Gln), which increased FIX activity by up to 8 folds, had been reported to be associated with the incidence of thrombosis.
In a young male venous thrombosis patient we had identified a novel F9 gene missense mutation c.1018G&gt;A, resulting in p. Glu340Lys substitution (designated as FIX Shanghai II). The patient's APTT time was 28.9 seconds (normal reference 22.3~38.7s), and his PT was also within the normal reference range (11.5 seconds, normal reference 10.0~16.0s). Although the patient had the FIX: Ag comparable to the normal reference, 128.20±6.07%, the his FIX:C was around 201.69±10.38% of normal control (normal reference 50~150%). Thrombin generation assay (TGA) showed that compared with normal control, the patient had a shorter Lag time (2.67 vs 3.33 min), elevated ETP (1649.43 vs 1311.62 nM*min), and higher Peak value (387.34 vs 200.86 nM), indicating increased global coagulation activity of the patient. The activity of protein C, protein S, and antithrombin were all within the normal reference ranges and the patient was negative for antiphospholipid antibodies.
Consistent with the patient's coagulation laboratory manifestations, the purified recombinant FIX Glu340Lys mutant showed 1.82 folds higher procoagulant activity than wild type FIX as determined by FIX:C/FIX:Ag ratio. The recombinant FIX Shanghai II mutant can be converted into FIXa by activated factor XI (FXIa) in a similar fashion as FIX WT. The kinetic study of FIXa-catalyzed FX activation showed that p. Glu340Lys mutation led to accelerated FX activation by FIXa.
The p.Glu340Lys mutation also enhanced the procoagulant activity of previously reported thrombosis-causing FIX mutants, FIX Shanghai and Padua, in a similar fashion as it did on FIX WT. The FIX:C/FIX:Ag ratio of double mutants containing Glu340Lys and Arg384Gln (FIX Shanghai) or Arg384Leu (FIX Padua) was about twice of FIX Shanghai or Padua alone, 11.48- and 22.53-folds vs 6.16- and 8.13- folds higher than FIX WT respectively.
The Oncoprotein SKI As a Driver in KMT2A-MLLT3/NRasG12D-Positive Acute Myeloid Leukemia
Authors: Frech Miriam , Schaprian Janine , Ems Miriam , Teichler Sabine , Bauer Uta-Maria , Neubauer Andreas 
Published: December 2023

KMT2A-rearranged acute myeloid leukemias (AML) have an intermediate to adverse prognosis. It was shown in a mouse model that the oncogene Myb contributes to AML maintenance in the KMT2A-MLLT3/NRasG12D genetic background (Zuber et al., 2011). Interestingly, the Myb-regulated genetic network mostly overlaps with KMT2A-MLLT3 target genes, especially with leukemic stem cell-associated gene sets. However, canonical KMT2A target genes are not included. Hence, MYB seems not to act as a co-factor for KMT2A complexes. SKI is a nuclear co-regulator and an oncoprotein, which is overexpressed in AML and several solid tumors (Bonnon & Atanasoski, 2012). Overexpression (OE) of SKI is prognostic in AML and correlates with shorter overall survival. Our group showed that SKI OE in AML is partially regulated by MYB (Frech et al., 2018). As a MYB target gene, we wanted to know if SKI can mediate MYB oncogenic activity in KMT2A-MLLT3/NRasG12D-positive AML.
We adopted the Zuber mouse model and performed an shRNA-mediated Ski knockdown (KD) in murine KMT2A-MLLT3/NRasG12D-positive AML cells (RN2). Mice were transplanted with RN2 cells carrying two different anti-Ski shRNAs or an anti-Renilla shRNA as control. The expansion of RN2 cells in mice was monitored by luciferase-based bioimaging. Upon disease onset, shRNA expression was induced by doxycycline treatment. The course of the disease was further monitored by bioimaging. Bone marrow cells were isolated and analyzed via flow cytometry. The expression of KMT2A target genes was analyzed by RT-qPCR in doxycycline-induced RN2 cells.
Ski KD induced a decreased viability of the RN2 cells in vitro. Moreover, SKI KD led to the eradication of RN2 cells in the mice in vivo and was associated with longer overall survival (log-rank p&lt;0.0005). In line with the SKI OE mouse model of Singbrant et al. (2014), Ski KD led vice versa in our model to a decrease of myeloid cells and an increase of B cells. In contrast to Myb KD in the RN2 cells, Ski KD led to a lower expression of canonical KMT2A target genes, implicating that Ski may be part of KMT2A complexes in AML.
In summary, SKI seems to be crucial for KMT2A-MLLT3/NRasG12D-driven AML and may be a potential target for new therapeutic strategies.
Clinical Significance of Serum High-Density Lipoprotein Cholesterol and Its Dynamic Change in Patients with Lymphoma-Associated Hemophagocytic Lymphohistiocytosis
Authors: Shi Wenyu , Miao Yi , Zhang Jing , Li Jianyong 
Published: December 2023

Background: The lipid profile in serum is frequently dysregulated in lymphoma-associated hemophagocytic lymphohistiocytosis (HLH). We aimed to explore the clinical significance of serum lipoprotein cholesterol in lymphoma-associated HLH.
Methods: Patients with lymphoma-associated HLH were included. Serum levels of baseline low-density lipoprotein cholesterol (LDL-c) and high-density lipoprotein cholesterol (HDL-c) were collected. Serum levels of LDL-c and HDL-c after treatment (first test after treatment, time interval ≥ 14 days) were also collected.
Results: A total of 169 lymphoma-associated HLH were included. T/NK-cell lymphomas (n=88, 52.1%) account for over half of cases. Low HDL-c (&lt; 1.03 mmol/L) was observed in 161 (95.3%) patients and low LDL-c (&lt; 2.6 mmol/L) was presented in 145 (85.8%) patients. The best cutoff of 0.50 mmol/L for HDL-c was determined to predict the 60-day survival. Patients with HDL-c ≥ 0.50 mmol/L showed a significantly higher 60-day survival rate than patients with HDL-c &lt;0.50 mmol/L (70.9% vs 42.2%, p &lt; 0.001.). Patients with HDL-c ≥ 0.50 mmol/L also showed longer overall survival (OS) (median OS: 346 days vs 34 days, p &lt; 0.001). LDL-c showed no significant impacts on 60-day survival and OS. Dynamic data of HDL-c were available in 69.2% (117) of the patients. The median interval between the two tests was 18 (IQR: 15-25) days. The level of HDL-c after treatment increased in 80.3% (94/117) of the patients. The ratio of post-treatment HDL-c/baseline HDL-c was calculated and the median number was 1.67 (IQR: 1.05-2.48). The ratio was significantly lower in the patients with T/NK-cell malignancies than those with B-cell lymphomas (median 1.40 versus 1.97, p &lt; 0.001). A ratio of 0.78 was determined as the best cutoff to predict the 60-day survival. Patients with a ratio&lt;0.78 had significantly reduced 60-day survival (42.4% vs 76.6%, P=0.0035, Figure 1A) and OS (median 36 days vs 407 days, P=0.0001, Figure 1B). Then we divided these patients with available data into two groups based on baseline HDL-c and the ratio of post-treatment HDL-c/baseline HDL-c: Group 1 included patients with baseline HDL-c ≥ 0.5 mmol/L and Ratio ≥ 0.78, while others were classified as Group 2. Patients in Group 2 had significantly reduced 60-day survival (57.5% vs 87.6%, P=0.0003, Figure 1C) and OS (median 107 days vs not reached, P=0.0001, Figure 1D) than Group 1.
Conclusion: Our study demonstrated serum HDL-c and its dynamic Change were robust and clinically feasible markers for predicting the outcomes of patients with lymphoma-associated HLH.
Associations and Hospital Outcomes of Splanchnic Vein Thrombosis in Patients with Polycythemia Vera
Authors: Olusoji Rahman , Abbey Enoch , Mohamed Salih Rajaa , Chaity Monjila , Ahluwalia Meena , Omore Ibrahim 
Published: December 2023

Background: Thrombosis means the presence of a blood clot in a blood vessel. While thrombosis is a relatively common condition in deep veins of the limbs and the lungs, its incidence in splanchnic veins is at least 25 times lower compared to the former. Splanchnic Vein Thrombosis (SVT) involves portal, hepatic, splenic or mesenteric veins. Polycythemia Vera (PV) is one of the significant myeloproliferative neoplasms (MPN) that cause SVT.
Objectives: This study examines the relationship between PV and SVT and its impact on hospital outcomes, explicitly focusing on mortality rates in affected patients.
Methods: This retrospective cohort study uses the 2018 to 2020 National Inpatient Sample (NIS) database. Our inclusion criteria were for patients 18 years or older with Splanchnic Vein thrombosis and Polycythemia Vera diagnosis during hospitalization. We defined splanchnic vein thrombosis as patients with portal and hepatic vein thrombosis. We examined these patients' demographics and their relationship with hospital outcomes, such as length of hospital stay, cost of hospitalization and mortality. Using univariate and multivariate logistics regression models, we measured mortality as the primary outcome.
Result: We identified 160,570 patients with SVT, among whom 1,120 (0.69%) had PV. PV patients were primarily female and white, with a mean age of 58.9. The PV cohort exhibited a lower mortality rate of 4.5% compared to the SVT from other etiology. Being black, increasing age, and increasing comorbidity indices were associated with increasing mortality. Though not statistically significant, there was a trend of decreasing mortality in both unadjusted and adjusted models for PV [0.68, 95% CI (0.26-1.05)] and [0.55, 95% CI (0.26-1.16)], likely underpowered by the rarity of diagnosis.
Conclusion: Our study shows that the mortality of SVT from PV was 45% lower compared to other causes of SVT. PV did not pose an increased mortality risk compared to other etiologies, and it was not statistically significant. There is a need for further studies to understand better and ultimately help in management guidelines for SVT in PV compared to SVT due to other causes.
Clinical Characteristics and Prognosis of Acute Myeloid Leukemia Patients with Protein Tyrosine Phosphatase Non-Receptor Type 11 Gene Mutation
Authors: Zhao Weihua , Huang Rui , Zhang Yiting , Lin Yu , Pang Ruli , Yang Zhi 
Published: December 2023

Objective: The purpose of our study was to investigate the clinical characteristics, molecular biological characteristics and prognosis of patients with acute myeloid leukemia (AML) harboring protein tyrosine phosphatase non-receptor type 11 (PTPN11) gene mutation.
Methods: The clinical data of 30 newly diagnosed adult AML patients with PTPN11 gene mutation were analyzed retrospectively. Prognostic analysis and screening of prognostic factors were conducted using the Kaplan-Meier method and Cox proportional hazards regression model.
Results: High frequency mutation sites of PTPN11 gene were located in exon 3 of chromosome 12, which are D61 and A72（16.7%）, followed by E76 (13.3%). The median variant allele frequency (VAF) of PTPN11 mutant gene was determined to be 18.4%. The patients were divided into two groups according to PTPN11 VAF 35.3% (upper quartile). We observed that the peripheral blood leukocyte count in patients with VAF ≥35.3% was significantly higher than patients with VAF &lt; 35.3% (p=0.019).Moreover, the higher VAF was closely related to M5 subtype AML (p=0.016) and internal tandem duplication (ITD) of FMS-like tyrosine kinase 3 (FLT3) (FLT3-ITD) mutation (p= 0.048). Taking PTPN11 VAF 20% and 35.3% as the cutoff value, the patients were divided into two groups, and the overall survival(OS) and event free survival (EFS) of these two groups were not significant. Multivariate analysis using the COX proportional hazards model identified that white blood cell count and Eastern Cooperative Oncology Group (ECOG) physical status score were independent risk factors affecting the prognosis of EFS.
Conclusion: Our study suggests that PTPN11 VAF may not serve as a prognostic factor in patients with PTPN11 mut AML. However, newly diagnosed patients with high white blood cell count and poor performance status were identified as independent risk factors for EFS in PTPN11 mut AML.
Keywords: Acute myeloid leukemia, PTPN11 mutation, clinical characteristics
Increased Hydroxyurea Adherence Associated with Pediatric, Adolescent and Young Adult Sickle Cell Anemia Care at US Military Healthcare Facilities: A Cross-Sectional Study
Authors: Smith Benjamin Russell , Marshal Amanda K , Ahmed Anwar , Zanetti Richard C 
Published: December 2023

Sickle Cell Anemia (SCA) causes significant acute complications, chronic complications, and premature mortality. The foundational treatment for SCA is hydroxyurea (HU) which reduces acute and chronic complications. Several barriers prevent patient HU adherence and thus treatment benefit.
We performed a cross-sectional study of 206 pediatric, adolescent, and young adult (AYA) patients with HgbSS or HgbS-beta-thalassemia-zero in the United State Military Health System who were prescribed HU to evaluate for to factors associated with HU adherence. These patients have access to military or civilian pediatric hematology care at no or low cost.
We categorised patient's adherence dichotomously according to the American Society of Hematology quality metric for HU adherence. The adherence group, 26% of the study population, had a younger mean age, 11.7 versus 14.9 (p = 0.0003), and was more likely to receive hematology care in the MHS.
Logistic regression analysis of HU adherence, adjusted for age and gender demonstrated the patients receiving pediatric hematology care at a military healthcare facility had 2.96 (1.47-5.99, 95% CI) the odds of adherence compared to patients receiving pediatric hematology care at civilian healthcare facilities. The monthly base pay of the patient's sponsor, a strong indicator of socioeconomic status was not associated with HU adherence odds ratio 1.00 (0.99-1.01, 95% CI).
Among Pediatric and AYA patients with SCA in the MHS: HU adherence appears equitable throughout the range of sponsor rank; and hematology care at military facilities is associated with increased HU adherence.
Multiomic Single-Cell Analysis Identifies Von Willebrand Factor and TIM3-Expressing BCR-ABL1+ Chronic Myeloid Leukemia Stem Cells
Authors: Nilsson Malin S. , Komic Hana , Gustafsson Johan , Sheybani Zahra , Paul Sanchari , Rolfson Ola , Hellstrand Kristoffer , Wennström Lovisa , Martner Anna , Thorén Fredrik B. 
Published: December 2023

Characterization of the leukemic stem cell (LSC) population in chronic myeloid leukemia (CML) may identify therapeutic targets for sustained elimination of leukemic cells. High-throughput single-cell RNAseq (scRNAseq) has paved the way for detailed transcriptional assessment of CML LSC. Building on this technique, multiomic CITE-seq approaches carry additional benefit in that the paired protein expression information may enable identification of associated cell surface marker profiles, subsequent FACS-based isolation and functional characterization. However, detection of BCR-ABL1 transcripts, which is arguably the only unequivocal marker to distinguish CML LSC from healthy hematopoietic stem cells (HSC), remains challenging using current high-throughput 3' end capture-based scRNAseq methods.
For the present study, we performed single-cell CITE-seq analysis of the expression of 597 genes and 51 proteins in &gt;70,000 stem and progenitor cells from 16 chronic phase CML patients and five healthy donors. Furthermore, we developed and employed a strategy allowing parallel detection of BCR-ABL1 transcripts at the single-cell level.
In bone marrow samples from CML patients, we observed pronounced expansion of erythroid and myeloid progenitors within the CD14 -CD34 + compartment, and substantial heterogeneity within the traditionally defined CD34 +CD38 -/low LSC compartment. In-depth analysis of the latter identified a group of immature BCR-ABL1 +cells displaying a CD45RA -cKIT -CD26 + TKI resistance phenotype sitting atop a hierarchy of myeloid progenitors. Within this group of potential LSC, expression of both previously reported ( e.g. CD25 and CD26) and unreported gene and protein markers was found to distinguish these cells from their healthy counterparts. Unlike HSC, the immature LSC showed high surface expression of TIM3 and high transcription of the von Willebrand factor gene ( VWF). While overexpression of VWF within the stem cell population may be linked to the aberrant myeloid-biased hematopoiesis characterizing the disease, the cell surface upregulation of TIM3 suggests that its targeting may have merit in CML.
In conclusion, we here performed detailed multiomic characterization of CML stem and progenitor cells, paired with BCR-ABL1 detection at the single-cell level. Our findings revealed LSC-specific expression patterns that may have implications for the phenotypic definition of CML LSC. Additionally, the results identify TIM3 as a conceivable target for sustained elimination of immature LSC in CML.
Non-Viral RNA-Lipid Nanoparticles for High Efficiency Genome Editing of CD34+ Hematopoietic Stem and Progenitor Cells for Advanced Cell and Gene Therapies
Authors: Geczy Reka , Park Stella , Zhao Rita , Swaminathan Meenakshi , Jain Nikita , Thomas Anitha , Zhang Angela , Clarke Samuel 
Published: December 2023

Background and Aims: The application of CRISPR-Cas9 gene editing in hematopoietic stem (HSC) and progenitor cells (HSPCs) has the potential to revolutionize the treatment of genetic blood disorders by correcting disease-causing mutations. Previously, we demonstrated the utility of a novel lipid nanoparticle (LNP) reagent for the multiplexed engineering of gene-edited CAR T cells, showing high cell viabilities, rapid onset editing, and potent CAR-mediated killing. In this current work, we demonstrate the capability of LNPs for the efficient and gentle delivery of genetic material to CD34+ HSPCs. The benefits of a non-viral LNP-mediated approach paves the way for the development of next generation gene therapy products in the HSC field.
Methods: LNPs encapsulating S.p. Cas9 mRNA and CD33 or CD45 targeted guide RNA (sgRNA) were produced using the scalable NanoAssemblr® NxGen™ microfluidic platform technology ( Fig. 1A). The LNP composition was optimized for the gentle and efficient cargo delivery to HSC/HSPCs. Purified human CD34+ cells were cultured, stimulated, and treated by the direct addition of the RNA-LNPs. Various commercial cell sources were tested including mobilized peripheral blood and cord blood. Gene knockout and viability were assessed using flow cytometry, cell proliferation using an automated cell counter, and differentiation capacity by colony-forming unit (CFU) assays.
Results: One-step addition of LNPs to CD34+ HSPCs resulted in 84 ± 6% CD33 and 81 ± 2% CD45 knockout efficiencies (n=6 CD34+ donors, Fig. 1B). After treatment, cells maintained on average 95 ± 3% absolute cell viability, compared to 99% viability of the untreated cells, Fig. 1C. Furthermore, LNP treated HSCs maintained excellent cell proliferation, with over &gt;90% relative cell proliferation to the untreated controls. The CFU assays showed no significant change in relative lineage formation for both the RNA-LNPs and the empty LNP vehicle control ( Fig. 1D). Finally, LNP production was successfully scaled-up using microfluidics from discovery to pre-clinical scales.
Conclusions: LNP-mediated CRISPR-Cas9 mRNA delivery is a promising approach for gene editing in HSPCs. The simple and gentle nature of LNP cell treatment allows for multiple genetic engineering steps for simultaneous expression and deletion of proteins for novel gene therapies. Furthermore, LNPs can be easily manufactured using microfluidics, enabling small-scale screening of RNA libraries and rapid scale-up for clinical translation.
Predictive Value of Pre-Infusion FDG-PET/CT Parameters for Clinical Outcomes and Toxicities in Relapsed/Refractory (R/R) Large B-Cell Lymphoma (LBCL) Following CAR-T Cell Therapy
Authors: Belbachir Safia , Alexander Sweeka , Dhamija Mayank , Al-Ogaili Zeyad , Purtill Duncan , Barraclough Allison A 
Published: December 2023

Introduction:
CD19-targeting chimeric antigen receptor (CAR) T-cell therapy has demonstrated significant efficacy in patients with relapsed/refractory (R/R) non-Hodgkin lymphoma but is associated with serious adverse effects such CRS and ICANS. Pre-infusion disease burden has been investigated as a potential predictor of clinical outcome and toxicity. FDG-PET/CT derived metabolic tumor volume (MTV) and total lesion glycolysis (TLG) are quantitative indicators of this baseline tumor load. We aim to assess the predictive value of pre-CART PET metrics on outcome and toxicity in R/R large B-Cell Lymphoma (LBCL) following CAR-T cell therapy.
Method:
We conducted a retrospective single site study of 25 consecutive patients with R/R LBCL treated with CAR-T therapy in our institution between May 2020 and October 2022, All patients underwent FDG-PET/CT imaging pre-CAR-T infusion (PET-0) and at day 30 post infusion (PET-1). The images were reviewed by two expert lymphoma PET physicians. Metabolic parameters including MTV, TLG (both calculated using a relative threshold of 40%), SUVmax and Deauville score were calculated. PET-1 was classified as complete metabolic response (CMR, Deauville score 1-3) or non-CMR (Deauville score 4-5) as per the Lugano classification.
Results:
25 patients were analyzed , Based on the median MTV (74,6 mL), TLG (379,4), SUVmax (11,8) values, patients were divided into two groups “low” and “high”.
The median follow-up was 12 months. Patients with low pre-infusion MTV and TLG had significantly superior day 30 CMR rates (84,6% vs 25%, p = 0.001 & 80% vs 20%, p = 0.001 respectively). Pre-infusion SUVmax ( p = 0.29 ) and Deauville score ( p = 0.103), however, did not predict response to treatment. In our population, pre-infusion PET parameters were not associated with CRS or ICANS .
Patients with low pre-infusion MTV and TLG had significantly superior PFS (MTV: p=0.02; TLG: p=0.019) and OS (MTV: p=0.017; TLG: p=0.05), Pre-infusion SUVmax had no statistical significant impact on OS ( p = 0.61) or PFS ( p = 0.37)
Conclusion:
In our study, pre-infusion quantitative PET metrics, TLG and MTV, predict response and outcomes of CART therapy in R/R LBCL. Our results highlight the importance of adequate disease control before CAR-T administration and the value of pre-infusion FDG-PET/CT as a predictor of outcomes in R/R LBCL patients treated by CAR-T-therapy.