bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adoptive Transfer of CMV-Specific TCR-T Cells for the Treatment of CMV Infection after Haploidentical Hematopoietic Stem Cell Transplantation
Authors: Chao Ma , Daihong Liu , Dou Liping 
Published: December 2023

Background: CMV reactivation after unmanipulated haploidentical stem cell transplantation (SCT) frequently occurs, causing life-threatening morbidities and transplantation failure. Preemptive therapy upon the detection of CMV viremia using antiviral agents is currently the standard of care but its efficacy is quite limited and associated with significant toxicity. The CMV antigen-specific cytotoxic T lymphocytes (CTLs) therapy was limited by the time-consuming manufacture process and relatively low success rate. More effective and safer approaches for the treatment of CMV reactivation after haploidentical SCT are in urgent need.
Methods: A single-arm, open-label Phase I clinical trial evaluating the safety and efficacy of CMV-targeting TCR-T (CMV-TCR-T) cell therapy as the first-line preemptive therapy for patients with CMV reactivation post haploidentical PBSCT was conducted in the Chinese PLA General Hospital (ClinicalTrials.gov Identifier: NCT05140187). Six patients with CMV reactivation after haploidentical SCT were adoptively transferred by 1 to 3 doses of SCT donors-derived CMV-TCR-T cells. This trial was a dose-escalation study with doses ranging from 1×10 3 CMV-TCR-T cells/kg body weight per dose to 5×10 5 CMV-TCR-T cells/kg per dose
Results: Except for the Grade 1 CRS observed in one patient and mild fever in 2 patients, no other adverse events were observed. Four patients had complete response within a month after CMV-TCR-T cells infusion without the administration of any antiviral agents. The other two patients who initially didn't completely respond to CMV-TCR-T cell therapy had salvage ganciclovir and foscarnet administration and then had rapid CMV clearance. The CMV-TCR-T cells displayed overall robust expansion and persistence in the peripheral blood after infusion. The CMV-TCR-T cells were firstly detected in the peripheral blood of these patients 3 to 7 days after the 1 st dose of CMV-TCR-T infusion, rapidly expanded and persisted for at least 1~4 months, providing long-term protections against CMV reactivation. In one patient, the CMV-TCR-T cells started to expand even when the anti-GvHD reagents were still being used , further indicating the proliferation potential of CMV-TCR-T cells.
Conclusions: Our study firstly showed CMV-targeting TCR-T cell as a highly feasible, safe and effective first-line preemptive treatment for CMV reactivation after haploidentical PBSCT.
The Mechanism of Setd2 in Mesenchymal Stromal Cell in MDS-Associated Ineffective Hematopoiety
Authors: Wang Roujia , Wang Lan , Chang Chunkang 
Published: December 2023

Myelodysplastic syndrome (MDS) is a heterogeneous, clonal, hematopoietic stem progenitor cell malignant disease. Bone marrow microenvironment participates in the occurrence and development of MDS and plays a key role.
Histone methyltransferase Setd2 is the only enzyme that can catalyze the methylation of mammalian histone H3K36Me3. Previous studies have confirmed that Setd2 is involved in the the transformation of MDS to leukemia, and the expression of S100A9 decreases after Setd2 knockout. This study is based on NHD13 mice with mesenchymal stem cell (MSC) targeted knockout of Setd2. Through experimental methods such as flow cytometry, peripheral blood routine, Wright staining, HE staining, and Gomori staining, it finds that Setd2 targeted knockout mice undergo faster leukemia transformation compared to NHD13 mice; Peripheral blood white blood cells increase, while hemoglobin and platelets decrease; The differentiation of hematopoietic stem progenitor cells in bone marrow is abnormal, with abnormal hematopoiesis and increased blast cells; Spleen fibrosis aggravates and megakaryocyte infiltrates; and Extramedullary hematopoiesis in liver. Through flow cytometry,β- Galactosidase staining and other experimental methods, we finds that compared with NHD13 mice, the MSC morphology of knockout mice is irregular, and the support function for hematopoietic cells is weakened. This study demonstrates that MSC targeted knockout of Setd2 has a promoting effect on the progression of MDS.
We used mass spectrometry to identify proteins that are significantly upregulated in the targeted knockout mouse bone marrow supernatant compared to the NHD13 mouse bone marrow supernatant, and and select the protein Uba5 related to hematopoiesis from them. The expression of Uba5 in the bone marrow supernatant of knockout mice is significantly higher than that in the NHD13 group through WB validation. It is speculated that the hematopoietic changes in Setd2 targeted knockout mice may be related to the downstream target protein Uba5 of Setd2.
This study takes the change of bone marrow microenvironment as an entry point to explore the possible mechanism of MDS ineffective hematopoiesis, hoping to provide microenvironment regulation therapy for some MDS patients with genes that hematopoietic stem cell transplantation that cannot cure(such as TP53, NRAS, KRAS), and provide new ideas for MDS treatment. There are still many limitations in this study, further clinical validation and vivo and vitro rescue experiments on downstream target genes and proteins are still needed in the future.
Granulocyte Colony-Stimulating Factor and Decitabine-Containing Conditioning Improves Survival in Myelodysplastic Syndrome Patients with Iron Overload Undergoing Allogeneic Hematopoietic Stem Cell Transplantation
Authors: Zhao Wenshu , Pan Danqi , Zeng Xiangzong , Xuan Li , Fan Zhiping , Huang Fen , Xu Na , Sun Jing , Liu Qifa , Dai Min 
Published: December 2023

Background: Iron overload is considered as an unfavorable prognosis in myelodysplastic syndrome (MDS) even in those undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although iron chelation therapy has improved the prognosis of these patients to some extent, the effect has not yet been satisfactory. This study aimed to investigate whether granulocyte colony-stimulating factor and decitabine (G-DAC)-containing conditioning improve the prognosis of iron-overloaded MDS patients undergoing allo-HSCT.
Study Design: One hundred and ninety-seven patients were enrolled in this retrospective study. Based on the level of serum ferritin (SF) and conditioning regimen, all patients enrolled were divided into 4 groups: SF﹤1000µg/L with G-DAC conditioning (cohort 1), SF﹤1000µg/L with non-G-DAC conditioning (cohort 2), SF ≥ 1000µg/L with G-DAC conditioning (cohort 3) and SF ≥ 1000µg/L with non-G-DAC conditioning (cohort 4). The clinical features and prognosis between cohorts were analyzed.
Results: The 2-year overall survival (OS) was 77.0%, 72.1%, 73.8% and 38.7% ( P = 0.001), and disease-free survival (DFS) was 75.7%, 62.8%, 71.7% and 35.5% ( P = 0.001), and the cumulative incidence of non-relapse mortality (NRM) was 17.3%, 25.0%, 15.2% and 53.1% ( P = 0.001), and the incidence of relapse was 8%, 13.6%, 13.1% and 12.5% ( P = 0.592), respectively, in the four groups. To be specific, cohort4 had worse OS and DFS and higher NRM than the other three groups (all P﹤0.05). Multivariate analysis revealed that SF ≥ 1000µg/L was a risk factor for OS, DFS and NRM ( P = 0.022, P = 0.025, P = 0.036), while G-DAC-containin g conditioning was a protective factor ( P = 0.009, P = 0.003, P = 0.004). Intriguingly, when cohort 1 to cohort4 were included in the multivariate analysis, only cohort4 was a risk factor for OS, DFS and NRM (all P﹤0.05), but cohort3, namely iron-overloaded patients who received G-DAC-containing conditioning had no difference in prognosis compared with patients with SF﹤1000µg/L.
Conclusions: The poor prognosis of patients with iron overload could be overcome by G-DAC-containing conditioning partly.
Corelationship of Lymphocyte Subsets and Prognosis in Juvenile Myelomonocytic Leukemia
Authors: Chen Yunlong , Zhang Jingliao , Liu Fang , Zhang Yingchi , Zhu Xiaofan , Yang Wenyu 
Published: December 2023

Objective: Juvenile Myelomonocytic Leukemia (JMML) is a rare hematopoietic stem cell malignant clonal disease in children and has the characteristics of MDS/MPN. Due to the poor response of JMML to chemotherapy, hematopoietic stem cell transplantation (HSCT) is currently the only cure method for JMML. Currently, there are some reports on the distribution of lymphocyte subsets in MDS. A significant lymphocytopenia was observed in MDS patients compared with similarly aged healthy people. Another study form Polish found patients with MDS have more CD3+T cells than healthy donors. But there are few related reports in JMML. The objective of this study is to analyze relationship between the distribution of lymphocyte subsets and prognosis in JMML patients, and explore new risk stratification indicatorsfor JMML personalized therapy.
Method: This study retrospectively collected JMML patients clinical data from Institute of Hematology & Blood Diseases Hospital of CAMS during Jan 2008 and Nov 2022. All completed complete blood count and peripheral blood classification, fetal hemoglobin (HbF), granulocyte colony-stimulating factor (GM-CSF) high sensitivity, bone marrow morphology, immune typing, molecular biology, cytogenetics and other tests. We analyzed the relationship between distribution of lymphocyte and prognosis in these patients.
Result: 96 newly diagnosed JMML patients were admitted to this study. The median age at diagnosis was 25 (2-82) months, and male to female was 3.8:1. 66 cases underwent lymphocyte subsets by flow cytometry. Median percentage of lymphocytes is 34.3%(1.8%-90.4%, normal range: 20%-40%), median percentage of CD3+T cells is 54.85%(10.3%-91%, normal range: 57%-88%), median percentage of CD3+CD4+ T cells is 30.95%(3.54%-53%, normal range: 27%-58%), median percentage of CD3+CD8+ T cells is 18.4%(2%-49.1%, normal range: 16%-45%), median percentage of NK cells is 13.95%(2.3%-50.3%, normal range: 3%-32%), and median percentage of B cells is 23.25%(4%-69.6%, normal range: 4%-18%). These cases were followed-up to Jan 2023, with a median follow-up time of 12 months (1-104 months). The overall survival (OS) rate was 52.67%, with a median OS time of 12 months (0-103 months). There were significant differences in the percentage of CD3+T cells (30.89% ± 9.46% vs 59.05% ± 10%, P&lt;0.01) and CD3+CD4+T cells (14.87% ± 5.82% vs 35% ± 5.41%, P&lt;0.01) between children with single NRAS mutation and compound mutations (≥2 classical five Ras pathway mutation ). In PTPN11 mutation patients, there is a significant positive correlation between OS and the percentage of lymphocytes (P=0.018).
Conclusion: Our study shows differences in percentage of CD3+ T cells and CD3+CD4+ T cells between patients with NRAS mutation and compound mutations. Patients with PTPN11 mutation and higher percentage of lymphocytes will have better prognosis.
Histone Methyltranferase SETD8 Stimulates Proliferation and Anti-Apoptotic Effect in Acute Myeloid Leukemia through Epigenetically Upregulation of CXCR4
Authors: Cui Zelong , Xu Man , Gao Zhenxing , Zhou Minran , Fu Yue , Feng Huimin , Zhang Lu , Chen Chunyan 
Published: December 2023

Acute myeloid leukemia (AML) is the most common type of leukemia., characterized by the rapid and uncontrolled growth of abnormal cells. Epigenetic modifications control gene expression without affecting its DNA sequence. H4K20me1 is the mono-methylation occurring on lysine 20 of histone H4, acting as an epigenetic mark that promotes gene expression. SETD8 is responsible for catalyzing the mono-methylation of lysine 20 on histone H4. While SETD8's role and mechanisms have been studied in various types of cancer, its function in AML remains unclear.
This study is based on the screening of a small molecule compound library, which identified the inhibitor UNC0379 targeting the histone methyltransferase SETD8, showing clear inhibitory effects on three non-M3 AML cell lines. Bioinformatical analysis revealed elevated SETD8 expression in AML patients compared to normal individuals and higher SETD8 expression corresponded with lower overall survival rates (OS). Further investigations using AML cell lines and normal peripheral blood mononuclear cells (PBMCs) demonstrated that SETD8 knockdown or UNC0379 intervention inhibited cell proliferation and increased apoptosis in AML cells, while PBMC cells remained unaffected. In an AML model constructed with C1498 cell injection in C57BL/6J mice, we observed that SETD8 knockdown and treatment with UNC0379 inhibited tumor formation and infiltration.
To elucidate the molecular mechanism of SETD8's impact on AML, we conducted transcriptome sequencing on SETD8 knockdown AML cells and found significantly decreased mRNA expression of CXCR4, which was further validated at the protein level. Also, BeatAML and TCGA datasets demonstrated a positive correlation between SETD8 and CXCR4 mRNA expression. These phenomena motivated us to clarify the regulation relationship between SETD8 and CXCR4.
CXCR4 is a crucial membrane receptor. After stimulation, it promotes AML cell proliferation and survival. CHIP experiments in SETD8 knockdown cells revealed reduced H4K20me1 enrichment in the CXCR4 promoter region. Subsequent rescue experiments demonstrated that overexpressing CXCR4 or using its corresponding agonist SDF-1alpha in SETD8 knockdown cells moderately restored cell proliferation and reduced apoptosis.
In summary, we discovered that SETD8 promotes CXCR4 transcriptional activation through catalyzing the H4K20me1 mark, thereby influencing AML cell proliferation and apoptosis. SETD8 presents a promising therapeutic target through genetic or pharmacological intervention.
Potential Significance of CTPS1 in Multiple Myeloma
Authors: Huang Han-Ying , Liang Yang 
Published: December 2023

Introduction
With the increasing interest to the metabolism of multiple myeloma (MM), a number of metabolic genes and their associated regulatory networks are essential for MM progression. Our team's prior work indicated that high expression of metabolism-related gene CTPS1 (CTP synthase 1) correlated with poor prognosis of MM. However, the mechanism of CTPS1 in MM is unknown.
Methods
The difference in CTPS1 expression level in patients with plasma cell malignancies, various stages, and drug-resistant recurrence was examined using publicly available data. MM cell lines were engineered to overexpress or knockdown CTPS1. RNA-seq was used to unbiased interrogate the downstream effector genes modulated by CTPS1. Severe immunodeficient NRG-SGM mice expressing humanized myeloid cytokines were used to determine the effect of CTPS1 on MM proliferation in vivo.
Results
CTPS1 expression levels varied substantially in various plasma cell malignancies. High CTPS1 expression was associated with shorter overall survival and progression-free survival in newly diagnosed MM patients. Moreover, there was a strong correlation between CTPS1 and sex, albumin, β2 microglobulin, lactate dehydrogenase, and ISS/R-ISS stage. In vitro, CTPS1 overexpressing cells proliferated more quickly than CTPS1 knockdown cells. NRG-SGM3 mouse experiments demonstrated that tumor growth was significantly accelerated in vivo in the CTPS1 overexpression group. In the pharmacological experiment, CTPS1 overexpression group's sensitivity to 20 nM bortezomib reduced, but CTPS1 down-knocked group's sensitivity to 20 nM bortezomib rose. CTPS1 was also shown to be involved in metabolism and synthesis, notably mRNA metabolism, DNA metabolism, chromosomal combination, cell cycle, spliceosome, and cellular senescence. Furthermore, Myc was found to be tightly connected to CTPS1 in both MM cell lines and newly diagnosed MM.
Conclusions
This study found that increased CTPS1 expression is associated with a poor prognosis and reduced sensitivity to bortezomib in MM. Additionally, CTPS1 upregulated Myc-signaling genes, which promoted progression of MM.
NLRP3/Caspase 1/Gsdmd Mediated Pyroptosis Exerts a Crucial Role in Blood-Brain Barrier Pathological Injury Induced By Cryptococcus Neoformans
Authors: Zhang Zile , Zou Jinhu , Luo Zifeng , Luo Jiamin , Cao Hong 
Published: December 2023

Background:
Cryptococcus neoformans (Cn) traversal of the blood-brain barrier (BBB), composed of human brain microvascular endothelial cells (HBMECs), is a critical process in central nervous system infection. Previous studies have achieved significant progress in pathogenic mechanisms how Cn penetrates BBB. However, it is still largely elusive about the cellular and molecular mechanism of the abrasive interaction between Cn and HBMECs. Here, we focus on the pathogenesis of direct damage to the HBMECs by Cn.
Methods:
The immortalized HBMECs were incubated with Cryptococcus neoformans strain B-4500FO2 in different final concentrations (1×10^6/mL, 1×10^7/mL) for 3 hours at 37℃. The control group was treated with isovolumic phosphate buffered saline. Then, total soluble proteins of HBMECs in each group were extracted with RIPA lysis buffer for immunoblotting analysis. ImageJ and SPSS software were used for grayscale and statistical analysis respectively.
Results:
We found that the expression levels of pyroptosis-related protein, including NLRP3, Caspase 1, N-terminal of GSDMD especially, were significantly upregulated in HBMECs treated with higher concentrations (1×10^7/mL) of Cn compared to control group ( P &lt; 0.05), in spite of the fact that there was no statistically significant difference in the expression of GSDMD ( P &gt; 0.05).
Conclusion:
These data support that Cn might impair BBB through facilitating pyroptosis of HBMECs via NLRP3/Caspase 1/GSDMD signaling. More intensive investigations are still required to elucidate the underlying mechanism how pyroptosis is triggered and transmitted in the interaction between Cn and HBMECs.
( Acknowledgements: Undergraduate Training Program for Innovation and Entrepreneurship of SMU, China, No. 202312121330; Corresponding author: Hong Cao, gzhcao@smu.edu.cn )
Efficacy and Safety of Olverembatinib-Based Therapies in Patients with Ph/BCR-ABL1-Positive Acute Lymphoblastic Leukemia
Authors: Zhao Yanhong , Zhai Weihua , Pang Aiming , Yang Donglin , He Yi , Wei Jialin , MA Qiaoling , Chen Xin , Han Mingzhe , Feng Sizhou , Zhang Rongli , Jiang Erlie 
Published: December 2023

Background Olverembatinib is an oral, third-generation BCR-ABL1 tyrosine kinase inhibitor (TKI) developed in China that could effectively target wild-type and mutant BCR-ABL1 kinase. It is effective and well-tolerated for patients with chronic myeloid leukemia. However, its efficacy and safety in patients with Ph/BCR-ABL1-Positive acute lymphoblastic leukemia are not entirely clear. This study aims to analyze the efficacy and safety of Olverembatinib-based therapies in patients with Ph/BCR-ABL1-Positive acute lymphoblastic leukemia.
Methods Patients with Ph/BCR-ABL1-positive acute lymphoblastic leukemia who received the Olverembatinib-based therapies in our center were included to conduct this retrospective study.
Results Totally 33 patients with Ph/BCR-ABL1-positive acute lymphoblastic leukemia were included in this study with a median age of 41 years, of whom 21 (63.63%) were female. Among them, 19 subjects who had been previously treated with standard chemotherapy and three newly diagnosed patients received Olverembatinib-based therapies, while 11 patients received Olverembatinib-based therapies after hematopoietic stem cell transplantation. After the first course of the Olverembatinib-based regimen, 33 (100%) patients experienced remission (CR/CRi/ Morphologic leukemia-free state), and 22 (66.67%) achieved the negative minimal residual disease (MRD) of BCR-ABL1/ ABL. Followed by the first course of the Olverembatinib-based regimen, five patients with negative MRD were treated with HSCT successfully and 27 received the second course of Olverembatinib-based therapy, of whom 21 (77.78%) acquired negative MRD. For six patients with negative MRD of BCR-ABL1/ ABL1, Olverembatinib-based therapies could remain the negative MRD. These patients with negative MRD will receive allogeneic hematopoietic stem cell transplantation after Olverembatinib-based therapy. Olverembatinib-based therapies were generally well tolerated. Common hematologic adverse events included leukopenia, neutropenia, anemia, and thrombocytopenia. Common nonhematologic adverse events included hepatotoxicity, nephrotoxicity, myalgia, arthralgia, rash, and edema, of which most were mild.
Conclusion Olverembatinib-based therapies are generally well tolerated and efficacious for patients with Ph/BCR-ABL1-positive acute lymphoblastic leukemia.
Immune Checkpoint Blockade Promotes Thrombosis Via T-Cell and Neutrophil Activation, and Tumor-Cell Associated Tissue Factor (TF) in a Murine Model of Colorectal Cancer
Authors: Shim Young Jun , Sharma Bal Krishan , Hisada Yohei , Mackman Nigel , Palumbo Joseph S , Diaz-Montero Claudia , Khorana Alok A. , McCrae Keith R. 
Published: December 2023

Background:
Immune checkpoint blockade (ICB) is a paradigm shift in cancer therapy. Despite clinical benefit, various adverse events are associated with ICB using immune checkpoint inhibitors (ICI). Venous thrombosis is an emerging clinical problem in cancer patients receiving ICI. Little is known about how ICI promotes thrombosis in cancer. To define the underlying mechanisms of ICI-associated thrombosis (IAT), we assessed the ICI impact on blood hypercoagulability and tumor TF expression using a mouse thrombosis model of colorectal cancer.
Methods:
We developed a CT26 mouse thrombosis model with ICI (aPD-1+aCTLA4) administration. Systemic hypercoagulability was assessed by measurement of circulating nucleosomes (Nu.Q H3.1 Immunoassay), neutrophil extracellular traps (NETs) (CitH3 immunofluorescence), circulating neutrophil-platelet aggregates (immunophenotyping), and levels of thrombin-antithrombin (TAT) complexes in ICI-treated mice. Tumor-associated TF and TF activity in circulating extracellular vesicles (EV) were assessed by western blotting and Factor Xa generation assays. To gain mechanistic insights into TF induction by ICI, tumor cytokines were also profiled (proteome profiler array). The role of cancer cell-derived TF in IAT was determined using mice implanted with CRISPR/Cas9-mediated TF knockout (TFKO) CT26 tumor cells.
Results:
ICI treated tumor-bearing mice developed larger thrombi than control mice and demonstrated elevated levels of circulating nucleosomes (114 vs 82 ng/ml), NET (15.8 vs 6.8%), platelet-neutrophil aggregates (44.8 vs 22.3%), and TAT complexes (12.6 vs 7.9 ng/ml) compared to IgG-treated mice. Tumor TF expression was increased in tumor extracts. Cytokine profiling suggested enhanced production of IFNɣ (2-fold) and TNFα (5-fold), as well as CXCL11 (6.8-fold), an effector T cell-recruiting chemokine. IFNɣ-increased TF expression in cultured CT26 cells, suggesting that T-cell derived IFNɣ may play a role in enhancing tumor TF expression in vivo. Circulating EV TF antigen and procoagulant activity were increased by ICI. CRISPR/Cas9-mediated TF KO in CT26 cells resulted in complete loss of TF expression, and mice bearing these cells showed reduced tumor and EV-associated TF (Fig. A), associated with a significant decrease in thrombus formation after ICI treatment (24.2 vs 20.5 mg, Fig. B). There was no significant difference in growth of wild-type and TF KO CT26 tumors.
Conclusions:
Our findings provide insights into cellular and molecular mechanisms associated with ICI-induced thrombosis, and suggest a role for neutrophil and granulocyte activation, and tumor-associated TF. Additional studies are ongoing to further define these pathways in the murine model and in patients.