bluuebunny/crossref_metadata_2025 huggingface dataset:
Discovery of a Novel, First-in-Class Bfl-1 BH3 Mimetic with Pro-Apoptotic Activity
Authors: Derks Maarten , Bryan Marian C. , Rockx Cindy , Schults Tamara , Verhulst Tinne , Pietrak Beth , Maehigashi Tatsuya , Robertson James C. , DeMaria Andrew H. , Holt Kimberly , Ji Shaofei , Wong Victoria , Bueters Ruud , Koo Seong Joo , Masure Stefan , Bhogal Balpreet , Kelly Christopher B. , Mitra Kaushik , Luistro Leopoldo , Shaffer Paul L. , Szewczuk Lawrence , Elsayed Yusri , Philippar Ulrike
Published: December 2023

Background: Intrinsic apoptosis is regulated by the B Cell lymphoma 2 (BCL-2) protein family. Anti-apoptotic family members, such as BCL-2, MCL-1 and BFL-1, sequester their pro-apoptotic counterparts through a highly conserved BH3 binding groove. Disruption of this balance leads to mitochondrial pore formation and subsequent cell death. To evade apoptosis, tumors often upregulate the expression of one or more anti-apoptotic family members and as such they have become attractive targets for anti-cancer drug development. BFL-1 (‘Bcl-2 related gene expressed in fetal liver’, gene name BCL2A1) is a lesser-known anti-apoptotic family member, physiologically mainly expressed in the hematopoietic system.

BCL2A1 overexpression has been reported in many types of B cell lymphoma. Diffuse Large B Cell Lymphoma (DLBCL) patient sample analysis showed that BCL2A1 expression levels are second only to MCL1 within the anti-apoptotic family. Expression of BFL-1 was shown to be upregulated in MYC+/BCL2+ double hit lymphoma cell lines treated with the BCL-2 inhibitor, Venetoclax, in vivo. Interestingly, BFL-1 positive lymphomas are less sensitive to inhibition of BCL-2 and MCL-1. Hence, targeting BFL-1 is a rational and attractive strategy in DLBCL to improve clinical outcome.

Methods: Using hit identification screens followed by iterative structure-activity relationship (SAR) analyses and structure-based design, compound A, a first-in-class, selective BFL-1 BH3 mimetic, was identified. Compound A was evaluated using biochemical, cellular in vitro and in vivo tumor efficacy models.

Results: Compound A is a highly potent and selective compound in biochemical assays. In BFL-1 sensitive lymphoma cell lines characterized by high BFL-1 protein expression, compound A was shown to displace BAK and BIM from BFL-1, leading to downstream cleavage of caspase 3 and cell death. Interestingly, these cellular models were resistant to Venetoclax. Compound A exhibited tumor growth inhibition in an engineered BFL-1 overexpressing, MCL-1 KO human multiple myeloma xenograft model. However, in the high BFL-1 expressing SU-DHL-2 DLBCL human xenograft model, compound A did not induce significant PD (cleaved caspase 3) and efficacy in vivo.

Conclusion: First-in-class selective BFL-1 BH3 mimetics are shown to specifically inhibit cell growth of lymphoma cell lines in vitro and can induce efficacy in vivo in a BFL-1 overexpressing model. Further studies may be useful to determine whether compound A elicits anti-tumor efficacy in lymphoma alone or in combination with other anti-apoptotic agents.
Epidemiological Characteristics and Drug-Resistance Genotype of CRE in Patients with Hematological Diseases in a Single Center of China
Authors: Jiang Erlie , Feng Sizhou
Published: December 2023

Patients with blood diseases have a significantly higher risk of infections and mortality rates compared with other type of patients. Referring to the 2019 CDC criteria for CRE, we examined the carbapenemase producing genotype and resistance characteristics of CRE strains (760 strains) in patients with hematological diseases. Among these, carbapenemase producing CRE (CP-CRE) accounts for 60.39% (459/760), non-carbapenemase producing CRE (non-CP-CRE) accounts for 39.61% (301/760). The data indicated that the most common bacteria in patients with hematologic disorders were Escherichia coli, Klebsiella pneumoniae and Enterbater cloacae, which primarily produced NDM-type metallo-β-lactamases. The rates of drug resistance in CP-CRE strains with MIC≥16 to Meropenem and Imipenem were 92.51% (420/454) and 86.12% (391/454), respectively. The drug resistance rates of non-CP-CRE strain with MIC≥16 to Meropenem and Imipenem were only 22.22% and 15.34%, respectively. Furthermore, we discovered that 36.88% (111/301) of non-CP-CRE strains were resistant to Ertapenem alone but still susceptible to Meropenem and Imipenem. The therapy of such CRE may differ significantly from that of other CRE strains, ETP-CRE may not be a typical CRE. In conclusion, Our data determined that the CRE colonized or infected in patients with hematological diseases was dominated by Escherichia coli and metallo-carbapease-producing types, among which the NDM genotypes was predominant. This is quite different from the domestic statistics, indicating that the CRE types infected by hematological disease patients are special and not consistent with other patients. We discovered that some patients with hematological diseases carried or were infected with ETP-CRE, which was sensitive to Meropenem and Imipenem. In terms of therapy, ETP-CRE strains may differ from other CRE in patients with hematological diseases，it may not be a typical CRE.
Evaluation of CD70 Chimeric Antigen Receptor T (CAR-T) Cells As a Potential Treatment for Non-Hodgkin's Lymphoma: A Preclinical Study
Authors: Jiang Pengfei , Xiong Qinghui , Yuan Xiujie , Zhang Haoyu , Shen Qiushuang , Wang Yang , Gong Yiwei
Published: December 2023

Background ：CD70 is a member of the tumor necrosis factor (TNF) superfamily. While its expression is limited in healthy tissues, CD70 highly constitutive expressed on cell surface of various malignancies. This expression profile make CD70 a promising target for cancer immunotherapy. We described a CD70 CAR-T based on a potent VHH nanobody named 11C9 in treatment of renal cell carcinoma (RCC). Recently, we received approval to initiate a phase I clinical study by Center for Drug Evaluation, National Medical Products Administration (NMPA) in China. Motivated by promising evidence that CD70 aberrantly expressed on Non-Hodgkin's lymphoma (NHL), including diffuse large B-cell lymphomas, follicular lymphomas, Burkitt and mantle cell lymphomas, we conducted a preclinical evaluation of CD70 CAR-T cells as a potential treatment by preclinical model.

Method: We successfully generated CD70 CAR-T cells based on a VHH nanobody (11C9) and CD19 CAR-T cells based on murine scFv (FMC63) following the exact manufacturing protocols used for phase I and phase II clinical trial products respectively. Then, we evaluated CAR-T cell cytotoxicity, phenotype, proliferation, and cytokine production in vitro. Furthermore, we established NHL tumor cell-xenograft NOG mice models to compare their anti-tumor activity in vivo.

Result: In vitro test demonstrated two types of CAR-T cells have comparable proliferation, and cytokine production. We observed more potency for CD70 CAR-T to kill CD70 and CD19 double positive Raji cells than CD19 CAR-T cells. In vivo test showed that both CD70 CAR-T cells and CD19 CAR-T cells eradiated Raji cells xenografted NOG mice models at high dose. However, after treated at lower dose CD19 CAR-T, tumor relapsed in one week when the same lower dose CD70 CAR-T treated mice keeping tumor free statue.

Conclusion: Our results indicate that CD70 CAR-T cell is potential treatment for NHL. We will conduct an investigation of single and dual CD70 CAR-T on treatment of lymphoma especially for CD19 negative NHL.
Galician Experience with Avatrombopag: Galician Thrombosis and Hemostasis Group
Authors: Calvino-Suarez Michael , Lopez Ansoar Elsa , Lorenzo Vizcaya Alvaro , Guzman Fernandez Maria Rebeca , Dorado Lopez Andrea , Salido Toimil Fatima , Rodriguez Ruiz Matilde , Iglesias Varela Raquel , Vasco Varela Beatriz , Blanquer Blanquer Miguel , Rodriguez Lopez Manuel
Published: December 2023

Avatrombopag, an orally administered 2nd-generation thrombopoietin receptor agonist (TPO-RA), in Spain is indicated for the treatment of primary chronic (>12 months) immune thrombocytopenia (ITP) in adults with inadecuate response or intolerance to other therapies, and has been the first agonist approved for adults with chronic liver disease (CLD) scheduled for surgery. It presents some advantages in relation to the present TPO-RA that make it a very attractive drug such as oral administration, non-interference of divalent cations, no need to monitor liver function or rapidity of response.

To assess the effectiveness of Avatrombopag in patients with chronic ITP in real life, we carried out a retrospective observational study in which patients with chronic ITP and patients with secondary thrombocytopenia were included. Data are shown as percentage, median, interquartile range (IQR), and mean ± standard deviation (SD), as appropriate.

Twenty-four patients were included (mean age of 60.04 years (SD 20.38), 58.33% female). The median follow-up was 135 days (IQR 96-180). 83.33% of patients had primary ITP and in 37.5% Avatrombopag was used in second-line. The rest of the patients (62.5%) had received at least two therapies previously, a median of 3 (IQR 2-4). 37.5% was in response under other agonist when started Avatrombopag. Twenty-three patients (95.83%) started Avatrombopag at 140 mg/week, one patient at a dose of 280 mg/week. 95.83% of patients reached or maintained response (>30,000 platelets/m3) with a median number of days to reach it of 7 (IQR 6-14). At the last visit, they presented a median platelet response of 102,000/mm3 (IQR 62,000 - 147,000) and marked weekly dose variability. One patient had previously received 12 lines of treatment and only one was primary refractory. Although drug tolerance was excellent, serious adverse effects include one patient with grade 2 medullary fibrosis and one pulmonary thromboembolism .

Although the sample is small and the follow-up short due to the recent approval of the drug in Spain, Avatrombopag offers an effective alternative as a TPO-RA agonist for patients on second-line treatment, as well as for subsequent lines in case of inadecuate response or intolerance to other agonists.
A Deep Learning Framework for Classification of Immature White Blood Cells in Acute Myeloid Leukemia Using Convolutional Autoencoder and Transfer-Learning Ensemble
Authors: Elhassan Tusneem Ahmed M , Mohd Rahim Mohd Shafry , Osman Ahmed Hamza , Chaudhri Naeem A. , Aljurf Mahmoud
Published: December 2023

Acute myeloid leukemia (AML) is a type of acute leukemia that affects myeloid cells. It is characterized by uncontrolled proliferation of immature WBCs causing an accumulation of immature WBCs in the bone marrow and peripheral circulation. AML is an aggressive cancer that can be life-threatening if not treated promptly, therefore, early detection is crucial. Identification of immature WBCs and their subtypes is the first step towards AML diagnosis. Microscopic blood smear image analysis is commonly used to diagnose AML as it is less expensive non-invasive diagnostic tool compared to bone marrow biopsy and immunophenotype. Nonetheless, classifying immature WBCs presents challenges due to their high similarity and minimal interclass variation, especially for intermediate stages of myelopoiesis which make is susceptible to misclassification. The current diagnostic methods of AML are based on manual identification of immature WBCs using peripheral blood smear. These methos are time-consuming, prone to errors, and subject to inter-observer variation. Therefore, this study aims to develop a computer-aided diagnostic framework to accurately identify immature WBCs and classify them into various subtypes.

The proposed framework consists of two main components: a binary classification system to classify cells into mature versus immature cells and a multiclassification system to further classify immature cells into four subtypes including myeloblast, monoblast, and promyelocytes. The proposed framework was designed to undergo four distinct phases: the first phase involves preprocessing, which includes data augmentation techniques aimed at addressing the imbalance distribution of WBCs. Augmentation encompasses geometric transformation and generation of synthetic images using convolutional Autoencoder (CAE). Feature extraction phase involves using image embedding and transfer-learning. Image embedding representation was obtained using CAE and then utilized by two pre-trained models: VGG19 and Resnet50. Classification phase was carried out using weighted ensemble of the VGG19 and Resnet50 with the optimal weights determined through Grid search. The validation phase evaluated the model performance using overall accuracy, precision, and sensitivity. The area under the ROC curve (AUC) was used to assess model discrimination ability.

The proposed framework demonstrated excellent results, achieving an average accuracy of 99.5%, a sensitivity of 97.97%, and a precision of 94.12%. The model exhibited an excellent discriminatory capability, with an AUC of 99.9%. Furthermore, the proposed framework outperformed previous methods, showcasing improved performance.

Key word:

Acute myeloid leukemia, peripheral blood smear, immature white blood cells, deep learning, feature extraction, computer-aided diagnosis.
Real Word Data on Levels of Emicizumab and Their Correspondence with the Global Hemostatic Capacity and the Spontaneous Joint Bleeding in Patients on Emicizumab Prophylaxis
Authors: López Jaime Francisco José , Marco Rico Ana , Palomo Bravo Ángeles , Fernández-Mosteirín Núria , Nieto María del Mar , Blanquer Blanquer Miguel , Marco Vera Pascual , Fernández-Bello Ihosvany
Published: December 2023

Introduction: Emicizumab has improved the management of patients with hemophilia A. However, few real-world data are available on drug plasma levels and their correspondence with global hemostatic capacity (GHC) and annualized spontaneous joint bleeding rate (ASJBR).

Aims: Evaluation of the correlation between emicizumab plasma levels, GHC and ASJBR in patients on emicizumab prophylaxis.

Methods: Multicenter, observational and cross-sectional study was performed in 4 Spanish hospitals. Emicizumab plasma levels and thrombin generation capacity (TGT, Genesia ®, Stago ®) of patients on emicizumab prophylaxis were determined in a central laboratory. Kinetics of clot formation was evaluated by rotational thromboelastometry (ROTEM ®, naTEM ® test, Werfen ®) in one center. Time on emicizumab prophylaxis and the number of spontaneous joint bleeds (SJB) was collected retrospectively using the clinical history. Results are expressed as median (25 th-75 th percentiles) or mean ± standard deviation depending on data distribution. A p-value < 0.05 was considered statistically significant.

Results: Twenty-eight patients were included: 18 aged ≥12 years (age= 20.1 [14.7-38.9] years old) (group 1) and 10 patients aged <12 years (age= 6.1 [3.2-8.9] years old) (group 2) (p< 0.001). Globally, emicizumab levels were 46.3 ± 11.9 mcg/mL and similar between both groups: 45.2 ± 13.1 mcg/mL (group 1) and 45.9 ± 7.3 mcg/mL (group 2) (p=0.870) (Figure 1). No correlation was detected between emicizumab levels and TGT or ROTEM ®. Compared with healthy controls, patients presented lower values of thrombin generation: Peak (% compared to controls) = 41.4 (33.3-53.0) % and endogenous thrombin potential (ETP%) = 54.9 (38.9-73.4) %. However, on the contrary, ROTEM ® showed a similar procoagulant profile (example: CT and CT+CFT; p>0.05) or even higher (example: lower CFT values in patients; p= 0.023) compared to healthy subjects (Table 1). Throughout a follow-up period of 57.0 (32.3-102.3) weeks, none of the patients presented SJB (ASJBR =0).

Conclusions: Emicizumab treatment provided excellent protection to patients throughout the study period. The wide interpatient variation on emicizumab plasma levels does not appear to have an impact on the development of SJB. TGT and ROTEM ® showed contradictory results: TGT describes lower thrombin generation capacity in patients compared to healthy controls, while ROTEM ® showed a similar or even higher procoagulant function in patients. Additional studies are required to explain these findings and their clinical significance.
Research on the Relationship between Nurses' Compliance with Standard Precautions, Servant Leadership and Emotional Exhaustion:a Cross-Sectional Survey in China
Authors: Du Xin , Wang Huan , Chen Wen , Lei Wang , Feng Xue , Zhang Xi , Song Caiping
Published: December 2023

Background: To prevent and control nosocomial infections, it is important to improve nurses' compliance with standard prevention techniques. Nurses' emotional exhaustion has increased in the context of the COVID-19 pandemic. It is crucial to pay attention to the impact of emotional exhaustion on compliance with standard prevention compliance from the perspective of organizational leadership.

Objective: This study aimed to analyze the effects of service-oriented leadership on nurses' emotional failure and compliance with standard prevention and to explore the moderating effect of individual resilience.

Methods: A convenience sampling method was used to collect questionnaire data from 924 clinical nurses in a third-class general hospital in Chongqing, China. A linear mixed model in SPSS 25.0 was used for multilevel regression analysis of the mediating role of emotional exhaustion on servant leadership and standard preventive compliance. PROCESS was used to further test the moderating role of individual resilience.

Results: The emotional exhaustion score (27.42±11.77) was at the general level, and standard preventive behavior compliance (11.41±5.01) was at the general level. Servant leadership mediated by emotional exhaustion had a significant positive predictive effect on standard prevention compliance, and the indirect effect accounted for 23.8% of the total effect. Personal resilience played a negative moderating role in the relationship between servant leadership and emotional exhaustion. For nurses with low resilience, servant leadership had a greater impact on emotional exhaustion. Conclusion: The current standard prevention compliance for clinical nurses is not high due to emotional exhaustion. The level of servant leadership can alleviate nurses' emotional exhaustion and improve compliance with standard prevention. Especially for nurses with low personal resilience, the care and support of department leaders are needed.

Key Words: Compliance with standard precautions by nurses, Servant leadership, Emotional exhaustion, Resilience
Clinical Outcome of Covid-19 in Haematologic Malignancy Patients with a Fourth Dose of Anti-Sars-Cov-2 Vaccine: A Final Epicovideha Report
Authors: Karthaus Meinolf , Salmanton-García Jon , Marchesi Francesco , Glenthoej Andreas , Bilgin Yavuz M , Van Praet Jens , Rahimli Laman , Cornely Oliver , Pagano Livio
Published: December 2023

Background: There is a general scepticism on anti-SARS-CoV-2 fourth dose efficacy due to lack of data. In pts with haematological malignancies (HM), this results that only few individuals received a fourth dose. We evaluted the clinical outcome of breakthrough SARS-CoV2 infections in HM who previously received a fourth dose of anti-SARS-CoV-2 vaccine, in particular HM from a multi-national European registry.

Methods: Patients (pts) from the EPICOVIDEHA registry were eligible as long as they could fulfil these criteria: a) active HM within the last five years (ys) before COVID-19 diagnosis, b) pts ≥18 ys old, c) laboratory-based diagnosis of SARS-CoV-2 infection, and d) reception a fourth anti-SARS-CoV-2 dose before COVID-19 diagnosis.

Results. As of August 2023, 6867 HM pts with SARS-CoV-2 infection have been registered in the EPICOVIDEHA registry. Out of these, 51 (0.7%) were diagnosed with COVID-19 after having received a fourth vaccine dose. Most of those (30/51, 58.8%) were male; median age was 71 ys of age (IQR 65-73), with only three pts below the age of 50 ys of age. Twelve pts (12/51, 23.5%) did not present any baseline underlying conditions besides the HM. Lymphoproliferative malignancies were prevalent (47/51, 92%). Most pts had a controlled HM at the time of COVID-19 diagnosis (30, 58.8%). Pts had their fourth vaccine dose at a median of 32 days (IQR 13-54) before COVID-19 diagnosis, almost exclusively mRNA based (50/51, 98%). COVID-19 remained asymptomatic or mild in almost all pts (49/51, 96%), with only one patient with critical infection (2%) requiring intensive care. Hospital admission rate was 47.1% with a median hospital stay of 9 days (IQR 5-14). Only 26 pts (51%) received a specific treatment for SARS-CoV-2, with almost all of them (18/26, 69.2%) receiving monoclonal antibodies. Pts were followed up for a median of 65 days (IQR 26-86) and only two died (3.9%) due to COVID-19.

Conclusions. With the caution derived from the limited number of pts and the intrinsic nature of this study, our data show favourable clinical presentation and outcome of breakthrough SARS-CoV2 infections in HM who previously received a fourth dose of anti-SARS-CoV-2 vaccine. The data suggests that a second vaccine booster may be of particular importance to protect this vulnerable HM patient population from severe or potentially life-threatening COVID-19.
Euroflow-Based Ultrasensitive NGF for the Evaluation of Bone Marrow Minimal Residual Disease in Multiple Myeloma
Authors: Tabares Gaviria Paula , Schmiedgen Katharina , Dhanraj Jayabhuvaneshwari , Dagdeviren Hüsniye , Wilnit Amy , Ulbrich Maria , Paiva Bruno , Kortüm K. Martin , Rasche Leo , Einsele Hermann , Beilhack Andreas , Waldschmidt Johannes M
Published: December 2023

Background:

Euroflow next-generation flow cytometry sensitively assesses minimal residual disease (MRD) in multiple myeloma (MM). It is widely adopted in labs worldwide for tracking disease progression, therapy efficacy, and decision-making in MM trials. Euroflow MRD significantly enhances MM patient outcomes, alongside advancements in anti-cancer approaches like BiTEs, CAR-T cells, and ADCs, resulting in increased overall survival rates. However, blood-based methods do not always align with bone marrow PC analysis, requiring heightened sensitivity to detect residual malignant cells post-therapy. The recent BloodFlow method, achieves remarkable sensitivity up to 10 -8 by enriching CD138 + PCs from 50 mL peripheral blood. In this study, we compare conventional Euroflow BM-MRD with the ultrasensitive method using CD138 enrichment of BM cells before analysis.

Methods:

BM-MRD followed Euroflow group guidelines. For ultrasensitive assessment, CD138 + plasma cells were isolated using CD138 MicroBeads. Data analysis was performed with Infinicyt software. Automatic gate and identification tool was used for conventional MRD, while data was manually analyzed for ultrasensitive MRD assessment.

Results:

We compared conventional and ultrasensitive MRD in the bone marrow of 70 MM patients. All positive MRDs using conventional NGF remained positive with ultrasensitive MRD. Notably, among 40 patients with negative conventional NGF-MRD, 5 cases turned positive after ultrasensitive MRD, by detecting aPCs from 0,0003 to 0,002% in all nucleated cells in the bone marrow. Patients 1, 2, and 3, diagnosed between 2020 and 2022, were in CR during assessment. Patients 4 and 5, diagnosed in 2016 and 2014 respectively, underwent multiple therapies, two autologous hematopoietic cell transplants, and patient 5 also had an allogeneic hematopoietic cell transplant. Ultrasensitive MRD for patients 4 and 5 was done after anti-BCMA CAR T cell therapy. The CD138 isolation approach allowed evaluating BM-MRD at detection levels of 10 -8 cells. Further studies using ultrasensitive MRD will prospectively evaluate the predictive value of the assay in patients with NDMM, especially with high and ultra-high-risk disease.

Conclusions:

This study demonstrates the significant advantages of direct CD138 enrichment over bulk lysis for detecting residual malignant PCs in MM. Ultrasensitive MRD offers superior sensitivity compared to conventional MRD, a critical factor in the era of more effective therapies leading to deeper responses. Currently, we are prospectively assessing the clinical benefits in a larger patient cohort.
Phase Separation Mediates RUNX1- Mutation Leukemic Transformation
Authors: Su Xiuhua , Sun Tao , Xia Yuan , Li Mingying , Wang Dongmei , Lu Fei , Ye Jingjing , Ji Chunyan
Published: December 2023

B ackground :

The high degree of genetic heterogeneity in acute myeloid leukemia (AML) leads to refractory or relapse in some patients, so it is important to deepen the mechanistic studies and find specific drug targets.As a classical regulatory molecule associated with AML, the absence or down-regulation of RUNX1 can lead to the development of AML. Liquid-liquid phase separation is closely related to the occurrence and development of the disease, and it is not clear whether RUNX1 can participate in the pathogenesis of AML through phase separation. This study is the first to explore the involvement of RUNX1 in the pathogenesis of AML from the perspective of phase separation.

Methods:

The fluorescent expression of RUNX1 was detected by immunofluorescence. The extracellular phase separation of RUNX1 and the mutants were studied by prokaryotic expression of the target protein RUNX1. The intracellular phase separation of RUNX1 was studied by cellular transfection and live-cell imaging. The cellular differentiation ability after phase separation of AML cell lines was investigated by using Rachel Giemsa staining and flow cytometry. The specific mechanism of RUNX1 phase separation regulation of downstream target molecules was examined by Co-IP coupled with mass spectrometry and Western Blot. The effect of RUNX1 phase separation on the pathogenesis of AML was studied by using the AML mouse model.

Results:

In this study, we firstly confirmed that RUNX1 showed obvious spot aggregation during hematopoietic differentiation. We then confirmed that RUNX1 can perform liquid-liquid phase separation both in vitro and intracellularly. Furthermore, based on the structural domain of RUNX1 and the RUNX1 mutation sites in AML patients, it was confirmed that the IDR region of RUNX1 drove occurrence of phase separation, while the RHD region had an inhibitory effect on phase separation, and the RUNX1 point mutants associated with poor prognosis of AML had abnormal phase separation.The occurrence of phase separation of RUNX1 could regulate the differentiation function of AML cells, and the differentiation of AML cells in the phase separation-deficient type was inhibited. Transcriptome sequencing results showed that RUNX1 phase separation promoted the enrichment of signaling pathways such as cell differentiation, and it was also found that RUNX1 phase separation promoted downstream target molecules to participate in the regulation of AML cell function. In addition, we demonstrated that aberrant RUNX1 phase separation was involved in the pathogenesis of AML in the mouse model of AML.

Conclusion:

This study reveals for the first time that RUNX1 liquid-liquid phase separation is involved in the process of AML occurrence and development, and refines the functional regulation mechanism of RUNX1, providing new ideas for AML targeted therapy.
Bcor Truncation Mutations Collaborate with a U2AF1 S34F Mutant to Promote Clonal Hematopoiesis and MDS
Authors: Chung Yang Jo , Wokasch Anthony , Onozawa Masahiro , Aplan Peter D.
Published: December 2023

Mutations in spliceosome genes such as U2AF1 have been identified across major cancer types, including myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Mutations involving the U2AF1 gene, mostly commonly involving amino acids 34 or 157 have been reported in up to 20% of patients with MDS. To investigate how mutant U2AF1 contribute to hematologic malignancy, we generated transgenic mice that expressed a mutant U2AF1 S34F mutant under control of hematopoietic specific Vav1 regulatory elements. Sanger sequencing demonstrated that expression of the mutant U2AF1 was similar to that of wild-type (WT) U2AF1 in bone marrow and spleen. U2AF1 S34F mice showed decreased long-term hematopoietic stem cells (HSC) compared with age matched wild type WT mice. Hematopoietic stem cell transplantation (HSCT) showed that U2AF1 S34F HSCs repopulated recipients less efficiently than WT HSCs, consistent with previous reports. However, U2AF1 S34F mice did not develop evidence of hematologic malignancy, suggesting that additional mutations were required to develop MDS or AML. BCOR, a corepressor for BCL6, has been reported to be frequently co-mutated in MDS patients with U2AF1 S34F, suggesting that these two mutations might collaborate to produce MDS. To test the hypothesis, we introduced a BCOR truncation mutation into hematopoietic stem and progenitor cells (HSPC) from U2AF1 S34F or WT mice, using CRISPR-Cas9, followed by transplant to WT recipient mice. Primary recipients of U2AF1 S34F and Bcor double mutant bone marrow (U2B) showed decreased engraftment compared to WT-Bcor mutant (WB) recipients, consistent with a competitive disadvantage of U2AF1 S34F HSPC observed previously by several labs. We performed secondary HSCT to evaluate the long-term repopulation pattern of U2B HSPC. Secondary transplant recipients from U2B primary mice showed anemia and leukopenia, consistent with MDS. U2B secondary recipients typically showed low engraftment initially (reflecting ineffective hematopoiesis) followed by gradually increasing proportion of U2B cells in the peripheral blood. At necropsy, all secondary recipients of U2B bone marrow showed evidence of clonal hematopoiesis involving cells with 1 or 2 distinct, unique Bcor truncation mutations. We then sequenced bone marrow DNA from the U2B mouse used as donor for the secondary transplant and identified 14 independent Bcor mutations. All 14 Bcor mutations were out of frame, and encoded a truncated Bcor protein, indicating that cells with a U2AF1 S34F and a Bcor truncation had a competitive advantage in vivo. These results suggest that loss of normal Bcor function can collaborate with a U2AF1 S34F mutant, resulting in clonal expansion of abnormal HSC.
Efficacy and Prognostic Analysis of Percutaneous Vertebroplasty/Kyphoplasty in Patients with Newly Diagnosed Multiple Myeloma
Authors: Zhang Fujing , Liu Shuzhong , Zhou Xi , Liu Shuangjiao , Liu Yong , Zhuang Junling
Published: December 2023

Objective To evaluate the efficacy and prognosis of percutaneous vertebroplasty/ kyphoplasty (PVP/PKP) in patients with newly diagnosed multiple myeloma (NDMM).

Methods Clinical data of NDMM patients who underwent PVP/PKP during front-line regimen at Peking Union Medical College Hospital from January 1 2003 to June 30, 2023, were retrospectively analyzed. Patients with comparable severity of bone disease and no orthopedic surgery were selected as controls. Visual analogue scale (VAS) score, progression-free survival (PFS), and overall survival (OS) were compared. Statistical methods mainly adopt χ 2-test, t-test, Kaplan Meier, and COX regression analysis.

Results Baseline characteristics were matched between the surgical group (n=51 with 56 surgeries) and non-surgical group(n=102), including gender, age, paraprotein type, serum M protein level, International Staging System(ISS), numbers of lytic lesions, cytogenetic abnormalities, first-line treatment, and autologous stem-cell transplantation (ASCT)（ P＞0.05). Bone lesions for PVP/PKP were located at thoracic vertebrae (53.6%, 30/56) or lumbosacral vertebrae (46.4%, 26/56). The postoperative VAS score was significantly improved (2.25±0.81 vs 5.92±1.05 , P<0.001). The median follow-up time was 51[38,70] months. Kaplan-Meier survival analysis suggested that PFS (37[17,89] vs 23[12.61] months, HR 0.648, 95%CI 0.431-0.973, P=0.047) and OS (not reached vs 66[28, NR] months, HR 0.519, 95%CI 0.296-0.910, P=0.045) in the surgical group were significantly improved. COX multivariate analysis suggested that PVP/PKP was an independent prognostic factor for PFS ( P=0.021, HR 0.589，95%CI 0.376-0.922)and OS( P=0.038, HR 0.496，95%CI 0.255-0.963), while not receiving ASCT or ISS stage II/III were independent risk factors. Subgroup analysis confirmed that patients with ISS II/III or non-ASCT achieved better PFS and OS in the surgical group(PFS: P=0.033, P=0.040; OS: P=0.024, P=0.018), while patients with ISS I or receiving ASCT had similar survival outcome（PFS P＞0.05; OS P＞0.05) between two groups.

Conclusion For NDMM patients, PVP/PKP is associated better quality of life due to alleviating bone pain. Meanwhile, it improves the PFS and OS of patients with ISS stage II/III or non-transplant eligible, which suggests that shortening the gap from symptom onset to diagnosis by surgical pathology will improve response in MM.
Biological and Therapeutic Implications of NPM1-Mutated AML Receiving Allogeneic Hematopoietic Cell Transplantation
Authors: Yuan Xiaolin , Wu Yibo , Lai Xiaoyu , Yang Luxin , Shi Jimin , Liu Lizhen , Yu Jian , Zhao Yanmin , Zheng Weiyan , Wu Wenjun , Zhao Yi , Cai Zhen , Huang He , Luo Yi
Published: December 2023

Background: The nucleophosmin 1( NPM1) gene mutations are the most common gene alteration in acute myeloid leukemia (AML) patients, occurring in approximately 30%-35% of adult AML cases. However, molecular and response to therapy heterogeneity are existed in this distinct group. Response to therapy, differing patterns of co-mutation and the prognosis of NPM1-mutated AML patients receving allogeneic hematopoietic stem cell transplantation (allo-HSCT) treatment were unknown.

Aims: To evaluate prognostic factors in the NPM1-mutated AML who underwent allo-HSCT treatment.

Methods: A total of 115 consecutive NPM1-mutated AML patients who underwent allo-HSCT from 2016 to 2022 were enrolled in this study. The primary endpoint was the cumulative incidence of relapse (CIR).

Results: The median patient age was 46 (IQR: 36-50) years. The most frequent co-mutations were FLT3-ITD(58.3%) and DNMT3A (40.0%), followed by IDH2(18.3%), NRAS (13.9%) and IDH1 (11.3%). The overall 3-year cumulative incidence of relapse (CIR) and leukemia-free survival (LFS) was 17.3% and 77.2%. The 3-year CIR of patients with FLT3-ITD was significantly higher than that without FLT3-ITD (24.3% vs. 7.0%; P=0.023). The 3-year CIR of patients with DTA mutations ( DNMT3A, TET2, and ASXL1) was significantly higher than that without DTA mutations (24.7% vs. 11.2%; P=0.030). Patients who received matched sibling donor (MSD) allo-HSCT had the highest CIR compared with unrelated donor (URD) allo-HSCT and haploidentical (HRD) allo-HSCT (46.2%, 10.0% and 11.2%, P=0.002). The 3-year CIR in patient with pre-HSCT measurable residual disease negative (MRD-), MRD+ and non-remission was 10.9%, 30.0% and 72.2%(P<0.001). After multivariable adjustment, DTA mutations, pre-MRD+ or non-remission, MSD allo-HSCT remained statistically significantly associated with higher CIR as well as lower LFS and OS. In univariate and multivariate analyses, the cell of origin of AML did not impact the outcomes of NPM1-mutated AML patients accept allo-HSCT treatment .

Conclusion: The DTA mutations ( DNMT3A, TET2, and ASXL1) were identified a distinct group of patients with adverse prognosis in NPM1-mutated AML patients who received allo-HSCT. Our results indicate that NPM1-mutated patients accept HRD/URD donors can achieve lower CIR than MSD donors in transplantation. HRD allo-HSCT may suitable for high risk patients within this group.
Preliminary Results of a Phase Ⅱ Study of Lenalidomide Combined with Immunochemotherapy in Patients with Primary Central Nervous System Lymphoma
Authors: Yao Shuna , Yao Zhihua , Yan Zheng , Wang Haiying , Chu Junfeng , Zhao Shuang , Zhao Xinyang , Liu Yanyan
Published: December 2023

Conclusion

Introduction：PCNSL is a diffuse large B cell lymphoma (DLBCL) occurring exclusively in the brain, spinal cord, cranial nerves, leptomeninges and/or eyes. Pathophysiology is incompletely understood, although a central role seems to comprise immunoglobulins binding to self-proteins expressed in the central nervous system (CNS) and alterations of genes involved in B cell receptor, Toll-like receptor and NF-κB signalling. Standard of care includes methotrexate-based polychemotherapy followed by age-tailored thiotepa-based conditioned autologous stem cell transplantation, whole-brain radiotherapy or singledrug maintenance。Lenalidomide, an immunomodulatory agent that penetrates the CNS, has shown promise in relapsed/refractory (R/R) aggressive NHL, with well-demonstrated singleagent activity and tolerability. We hypothesized that lenalidomide combined with immunochemotherapy(MTX-based regimen)might be a favorable option for PCNSL pts. Here, we present preliminary data from the phase 2 trial of lenalidomide combined with immunochemotherapy in PCNSL pts (NCT04737889).

To investigate the efficacy of lenalidomide combined with immunochemotherapy in untreated PCNSL.The primary objective is 2-year progression free survival(PFS). The secondary objectives are objective response rate (ORR), incidence of grade 3/4 adverse event (AE), and overall survival (OS).

aged 18-75 years untreated PCNSL and adequate organ function are being enrolled.Lenalidomide(25mg qd po,totally 10 days/cycle) plus R-MT(Rituximab-Methotrexate+Temozolomide) was administered in TN PCNSL. A totally of 6 cycles of treatment was planned for pts. Completed responders(CR) undergo autologous stem cell transplantation (ASCT) 、reduced dose whole brain radiotherapy or lenalidomide maintenance for 24 months, based on the patient's fitness and preference.

RESULTS：From March 2019 to 18 April 2023, 24 pts have been enrolled . Their baseline characteristics are displayed in Table 1.

By 30 May 2025, all pts had been evaluated for response. There were 19 CRs (79.2%), 3 PRs (12.5%) and 2 PDs(8.3%) . The median follow-up time was10 (1.2 - 51.4) months . One pt withdrew informed consent due to grade 3 renal toxicity , One died from non-tumor-related disease. Two year progression-free survival (PFS) and overall survival was 62% and 67%，respectively,the median PFS and OS not achieved.

The most common adverse events were hematologic toxicities. Grade 3/4 AEs occurred in ≥20% pts were neutropenia, leukopenia .Grade 3/4 non-hematologic AEs occurred in ≥20% pts was hepatic injury.

Lenalidomide combined with immunochemotherapy showed encouraging activity and acceptable tolerance in pts with TN PCNSL.The study is still ongoing and it is worth looking forward to updating the long-term survival data.
Measuring Minimal Residual Disease in Plasma of Multiple Myeloma Patients By Intact Mass Spectrometry
Authors: Genovesi Luca , Schirm Michael , Dupuis Nick , Pottiez Gwenael
Published: December 2023

Multiple Myeloma (MM) is a hematological cancer that is currently treatable but incurable, and is characterized by the accumulation of malignant plasma cells in bone marrow. Clinical studies have shown that post-cancer treatment, the minimal residual disease (MRD) status is a prognostic factor indicative of the extent of disease remission. MRD assessment has been primarily assessed in bone marrow (BM) using flow cytometry or next generation sequencing. Not only these measurements may lead to false-negative results due to spatial heterogeneity in the BM, hemodilution and/or extramedullary disease as indicated by Derman et al. in Blood Cancer Journal ((2021), 11:19) but the procedure for BM collection is invasive and painful for the patient. These authors present a peripheral blood (PB)-based method that potentially can overcome the barriers in assessing for residual disease in MM patients.

We hereby present a non-invasive blood-test for measurement of MRD using mass spectrometry (intact mass approach) for the detection of Ig product in MM patients. We will describe the assay development, analytical characterization of assay performance, and data from the analysis of plasma/serum samples from MM patients enrolled in a clinical trial.

A very low volume of patient plasma/serum sample (10 μL) is required to perform this assay. The plasma/serum sample is diluted in purification buffer, applied to Melon gel resin and incubated for 5 min, followed by centrifugation and reduction with DTT. The supernatant is then analyzed by LC-MS on a Q-Exactive mass spectrometer. Deconvolution of the intact protein m/z spectra is performed to yield intact mass of the specific protein target species of interest.

Overall, the method can process up to 96 samples in parallel within 3 hrs, making this procedure highly scalable for clinical applications.

Data obtained from the assay characterization indicate that the assay is sensitive with an LOD <500 ng/mL and differentiates MM patient from a healthy donor as well allows for the monitoring of MM patient before and after treatment.

In conclusion, although still an exploratory analysis, the blood-based mass spectrometry method can be used to screen for MRD in patients who have undetectable disease by flow cytometry or next generation sequencing.
Prognostic Risk Factors for Patients with Candida Tropicalis Fungemia Infections and Hematological Malignancies
Authors: Zhao Haisu , Fu Shan , Zhu Yuanyuan , Huang He , Zheng Weiyan
Published: December 2023

Background: Candidemia is one of the most common nosocomial infections, contributing significantly to morbidity and mortality. The most commonly reported non-albicans Candida species is Candida tropicalis, which is currently spreading around the world. Clinical studies have suggested a particular susceptibility to C. tropicalis infection among patients with hematological malignancies, particularly leukemia.

Methods: We retrospectively analyzed 106 patients with both hematological malignancies and invasive candidiasis. Patient age, sex, medical history (basic disease and medication history), length of hospital stay, laboratory test results, and other clinical features were recorded. Drug sensitivity information was available for 90 of the 106 patients. Then we used logistic regression and some machine learning models to identify the prognostic factors related to patient death.

Findings: Of 106 patients, 35 (33.1%) died within one month after being diagnosed. Most patients (105/106) were diagnosed with C. tropicalis fungemia in the myelosuppressive phase, and were mainly concentrated in the first (35/106, 33%) and second (18/106, 17%) chemotherapy courses. Drug sensitivity information was available for 90 of the 106 patients. Resistance to voriconazole and fluconazole were observed in 52 and 51 patients, respectively. In addition, 73.3% of the patients were resistant to itraconazole. Multivariate logistic regression showed that the use of amphotericin B(OR=0.295 ,95% CI 0.100-0.877), sex(OR=0.263 ,95% CI 0.082-0.847), duration of fever before diagnosis(OR=4.053 ,95% CI 1.281-12.825), combined bacterial infection(OR=5.309 ,95% CI 1.701-16.573), decannulation(OR=0.0922 ,95% CI 0.013-0.636), and serum creatinine levels(OR=1.020 ,95% CI 1.001-1.038) were determined to be predictors of death(p < 0.05). The random forest model, with an AUC value of 0.793, which showed the best performance of five machine learning models, suggested that serum creatinine level was the most important prognostic factor for candidemia.

Conclusion: In this study, we finded that the main predictors of death were found to be the use of amphotericin B, sex, duration of fever before diagnosis, combined bacterial infection, decannulation, and serum creatinine levels.
Immune Checkpoint Expression Patterns on T Cell Subsets in Light-Chain Amyloidosis: VISTA, PD-1, and Tigit As Potential Therapeutic Targets
Authors: Wang Jinghua , Zhao Yujie , Liao Yujie , Li Yangqiu , Zhong Liye , Chen Shaohua
Published: December 2023

Abstract

Background: Systemic immunoglobulin light chain amyloidosis (AL amyloidosis) is a rare plasma cell dyscrasia with a poor prognosis. T-cell exhaustion can lead to impaired T cell-mediated antitumor immunity, but there is limited information available regarding the T-cell immunodeficiency status in AL amyloidosis. This study aims to investigate the T-cell immune checkpoint expression patterns in AL amyloidosis and its relationship with clinicobiological characteristics.

Methods: Using multi-color fluorescent flow cytometry, we examined the expression levels of V-domain immunoglobulin suppressor of T cell activation (VISTA), programmed death 1 (PD-1), T cell immunoglobulin and mucin-domain-containing-3 (Tim-3), T cell immunoreceptor with Ig and ITIM domains (TIGIT) on CD3+, CD4+, CD8+ T-cell, and regulatory T cell (Treg) in peripheral blood (PB) and bone marrow (BM) from 19 patients with newly diagnosed AL amyloidosis. We used 36 PB and BM samples from patients with newly diagnosed multiple myeloma (MM) and 19 PB and 10 BM samples from healthy individuals (HIs) as controls.

Results: Our results showed that PB from patients with AL amyloidosis had significantly higher percentages of VISTA+ and PD-1+ subsets within CD3+, CD4+, CD8+ T cells and Tregs when compared to HIs. No statistical differences for frequency of VISTA+, PD-1+, Tim-3+ and TIGIT+ T-cell in BM were observed between the two groups. The percentages of double-positive VISTA+ PD-1+, VISTA+ Tim-3+, VISTA+ TIGIT+, PD-1+ Tim-3+ and PD-1+ TIGIT+ T cells in PB from the patients with AL amyloidosis were also considerably higher than those in the HIs. Moreover, we discovered that the patients with renal involvement had greater percentages of PD-1+ or TIGIT+ subsets within CD3+, CD4+ T cells and Tregs than the individuals without renal involvement. Additionally, PD-1+CD3+%, PD-1+CD4+%, and PD-1+Treg% were found to be positively correlated with 24-hour proteinuria levels. Furthermore, we observed that the AL amyloidosis patients had significantly higher counts of PD-1+ Treg in PB than the MM patients, while the MM patients had significantly higher counts of TIGIT+ subsets within CD3+, CD4+ and CD8+ T cells than the AL amyloidosis patients.

Conclusions: Our findings suggest that there are elevated proportions of VISTA+ and PD-1+ T cells in PB of AL amyloidosis patients, which are indicators of an immunosuppressive milieu. The rise in PD-1+ or TIGIT+ T cells was associated with renal damage. Notably, MM and AL amyloidosis have very distinct immunosuppressive patterns based on immune checkpoint alteration. Overall, VISTA, PD-1, and TIGIT may be considered as potential targets for reversing T cell exhaustion and enhancing T cell activity in AL amyloidosis.