bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical Characteristics, Molecular Features and Transplantation Outcomes in MDS with Low Blasts and without Defining Genetic Abnormalities: An Analysis of MDS-h and MDS-LB Patients
Authors: Guo Wenjing , Jia Li , Mi Yingchang , Feng Sizhou 
Published: December 2024

Objective: According to the World Health Organization (WHO) 2022 classification, myelodysplastic neoplasms/syndromes (MDS) patients without increased blasts and defining genetic abnormalities are categorized as either hypoplastic MDS (MDS-h) or MDS with low blasts (MDS-LB). This study aims to evaluate the clinical characteristics, genetic and cytogenetic profiles, and outcomes of MDS-h and MDS-LB patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT).
Methods: We retrospectively analyzed data from 25 MDS-h and 64 MDS-LB patients who underwent allo-HSCT between January 2010 and January 2023. A one-to-one propensity score-matched analysis was performed to minimize confounding factors. Additionally, genetic mutations and chromosomal abnormalities were compared among 25 MDS-h, 64 MDS-LB and 205 severe aplastic anemia (SAA) patients.
Results: Most patients were classified into higher-risk group by Molecular International Prognostic Scoring System (IPSS-M) (MDS-h: 80.00% vs. MDS-LB: 71.88%, P=0.605). Pre-transplantation transfusion dependency was observed in 80.0% of MDS-h and 79.7% of MDS-LB patients (P=0.541). MDS-h patients had lower platelet counts (35×109/L vs. 59×109/L, P=0.034) and a higher incidence of severe cytopenia (56.00% vs. 34.38%, P=0.104). Both MDS-h and MDS-LB patients exhibited significantly more mutations and chromosomal abnormalities compared to SAA patients (P&lt;0.030). BCOR mutations were prevalent in MDS-h (23.81%), while U2AF1 (39.62%), ASXL1 (30.19%) and RUNX1 (20.75%) mutations were common in MDS-LB. The estimated 3-year overall survival (OS) was 76.0% (61.0%-94.7%) for MDS-h and 83.9% (75.2%-93.6%) for MDS-LB (P=0.328). Failure-free survival, transplant-related mortality and cumulative incidences of engraftment, graft-versus-host disease and infection were also similar between groups. Risk factors for worse OS included bloodstream infection within 100 days post-HSCT and FAT1 mutations in MDS-h, and RUNX1 mutations in MDS-LB. Donor type and the interval from diagnosis to HSCT had no significant impact on post-HSCT outcomes.
Conclusions: MDS-h and MDS-LB patients undergoing allo-HSCT exhibited slightly distinct clinical and genetic features, but both groups achieved favorable outcomes after transplantation. This study highlights the potential efficacy of allo-HSCT in mitigating adverse genetic mutations in MDS-h and MDS-LB patients, with no significant impact from donor type or diagnosis-to-HSCT interval on outcomes.
Disruption of the Retinoblastoma Lxcxe Region Induces Cell Death in AML By Activation of the Unfolded Protein Response Leading to NOXA Induction
Authors: Bhuniya Avishek , Lu Lily , Ploi Adi Narayana Reddy , Tcyganov Evgenii , Mendoza Maria De Grecia Cauti , Sharma Paridhima , Ortiz Adiana Ochoa , Zhigarev Dmitry , Salvino Joseph , Montaner Luis 
Published: December 2024

The Retinoblastoma (Rb1) protein is expressed in high amounts in leukemia and myeloid cells. While RB1 controls cell cycle progression, it also regulates cell death through protein binding regions such as the LxCxE binding cleft. To test whether blocking the Rb1 LxCxE domain would induce specific cell death in acute myeloid leukemia (AML) cells, we developed a small molecule (AP-3-84) that can selectively bind the LxCxE domain of Rb1 blocking protein-protein interactions. Statistical analysis was performed with GraphPad Prsim.
The activity of AP-3-84 was evaluated in vitro using multiple AML cell lines (THP-1, AML-193, Kasumi-1, MOLM-14) and in vivo using luciferase-transduced THP-1 cells implanted in NGS mice. AP-3-84 induced cell death in a concentration-dependent manner in all AML cell lines tested. In vivo treatment with AP-3-84 also reduced tumor growth as measured by luciferase signal using an IVIS platform. Importantly, AP-3-84 treatment did not induce cell death in PMA-differentiated THP1 cells or non-hematopoietic cancer cell lines. PROTAC-mediated Rb1 degradation reduced AP-3-84-induced cell death in THP-1 cells, confirming Rb1 as the target for AP-3-84.
Bulk RNA sequencing showed that 30 min treatment of THP-1 cells with AP-3-84 resulted in upregulation of the NRF2-mediated oxidative stress response and unfolded protein response (UPR) and induction of ATF4 and DDIT3 (CHOP). Further evaluation of the UPR pathway via western blotting indicated increased expression of pPERK, peIF2a, ATF4, ATF3, and NOXA in a sequential time-dependent manner. Notably, the levels of BAX and BAK remained unchanged at early time points (up to 6 hours), suggesting that the UPR pathway is the predominant pathway triggered by AP-3-84. In the same experiments we observed induction of lipid peroxidation. As lipid peroxidation induce ferroptosis, treatment with ferrostatin 1 reduced the effect of AP-3-84, supporting that induction of ferroptosis is the predominant cell death pathway induced by AP-3-84. We also observed an increase in cardiolipin levels consistent with lipid peroxidation changes after AP-3-84 treatment, supporting the contribution of mitochondria to activation of cell death pathways.
We conclude that AP-3-84 triggers cell death in AML by activating the UPR via PERK-eIF2a-ATF4-NOXA pathway involving mitochondrial regulation. Our results indicate that Rb1 LxCxE domain may represent a novel therapy target in AML.
Higher NOTCH1 Mutation Load Reveals Favorable Prognosis in Pediatric T-Cell Acute Lymphoblastic Leukemia
Authors: Zhang Senlin , Ji Qi , Zhang Zhiqi , Chen Ping , Chu Xinran , Meng Na , Zhang Xiao , Gao Wei , Liu Minyuan , Liu Yu , Hu Shaoyan 
Published: December 2024

Background: Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy, approximately 10-15% of which are derived from T cell precursors and/or immature T-cells. Previously, gene mutations were identified to be associated with the risk stratification, treatment response, and prognosis of pediatric T-ALL patients. NOTCH1 was the most common gene mutation in T-ALL patients. Previous studies only fucosed on the influence of NOTCH1 mutation on the prognosis of pediatric T-ALL, few studies explored the role of variant allele frequency (VAF) in NOTCH1 mutation.
Methods: We retrospectively reviewed children diagonsed as T-ALL in our center from April 2012 to November 2022. All included patients performed RNA sequencing (RNA-seq) using samples collected at diagnosis. Overall survival (OS) was calculated and compared by Kaplan-Meier analysis and log-rank tests. The primary objective of this study was to compare the influence of VAF in NOTCH1 mutation on the prognosis of pediatric T-ALL.
Results: A total of 123 children were enrolled. Among these patients, 98 were male and 25 were female, with a ratio of 3.9:1. The median age of all patients at diagnosis were 8.0 years (range 1.2-16.8 years). The median white blood cell (WBC) at diagnosis were 106.33 (range 1.05-750.20). The median hemoglobin and platelet count at diagnosis were 98.50 (range 37.00-159.00) and 63.00 (range 5.00-412.00), respectively. Regarding to the immunophenotype, there were 7 early thymic precursor (ETP) and 116 non-ETP. Sixty-one of 123 (49.5%) patients have NOTCH1 mutation, which did not influence the 5-year OS (77.1% ± 5.67% vs 74.2% ± 6.01%, P=0.81). Based on the maximally selected log-rank statistic, mean VAF was found to be the cutoff point for differentiating patient prognosis. Patients with a higher mutation load (VAF≥20%, n=37) had superior 5-year OS when compared to patients carrying a lower mutation load (VAF＜20%, n=24)(88.9% ± 5.23% vs 61.7% ± 10.1%, P=0.027).Conclusion: Our study indicated that NOTCH1 mutation did not influence the prognosis of children with T-ALL. While higher NOTCH1 mutation load may have a favorable prognosis, which may contribute to the risk stratification of pediatric T-ALL. Larger-scale and prospective randomized studies are warranted to further evaluate this conclusion.
Artificial Intelligence Versus Expert Physician Assessments of Real-World Hematology Cases: Implications for Clinical Practice
Authors: Main Olivia , Struck Matthew , Vaughn John L , Hsu Jingmei , Abu Zaid Mohammad , Saint Fleur-Lominy Shella , Braunstein Marc J 
Published: December 2024

Introduction: Artificial intelligence (AI) is increasingly being incorporated into the healthcare space as a tool to support clinical decision making and enhance patient care. Professional societies including ASH, as well as private companies such as Primum, Inc, offer free consultations with appropriate experts who provide personalized responses to clinician-submitted hematology cases. However, the potential of AI tools to supplement or supplant expert consultations remains uncertain. We report results of a study comparing AI versus expert physician responses to 107 real-world hematology cases.
Methods: Among 107 cases, inquiries included lymphomas (30), myeloma (24), leukemias (11), myeloid disorders (10), as well as classical hematology cases (32), assessed among 20 unique experts. Responses to these de-identified cases submitted by practicing clinicians to the Primum platform (www.primum.co/) between June 2022-July 2023 were compared to GPT-4 responses (openai.com/chatgpt/). The instructional prompt to GPT-4 was, “You are an expert oncologist conversing with another oncologist as a peer. You prefer to rely on guidelines and data published in reputable medical journals when responding.” Five expert faculty at our institution adjudicated the blinded comparative responses, including their preference, quality and practical value scores, and prediction of which response was AI generated. Comparison of scores was by t-test to generate P-values between expert and AI groups, and Pearson correlation was used for comparisons between adjudication scores.
Results: Expert responses were preferred by &gt;50% of adjudicators in 75% of cases (deviation ±25%). Randomized AI responses were correctly identified 90% of the time. Mean expert vs AI scores (Likert scale 0-4) for quality (2.0 vs 2.1, P=0.9) and practical value (2.1 vs 2.1, P=0.9) were equivalent. Interestingly, AI responses were preferred in 46% (n=15) of classical hematology and 31% (n=9) of lymphoma cases, largely due to being more concise. However there was no concordance between high practical value scores and disease subtype for either group.
Conclusions: Expert physician responses were preferred for most of the cases, suggesting an implicit value of personalized responses compared to AI. Results showed no significant differences in quality or practical utility between AI generated responses and those from experts, reflected a similarity in the information extracted from standardized guidelines. Our findings may be limited by the broad coverage of hematologic conditions for which experts and guidelines vary. Overall, these data suggest that while AI can supplement knowledge of management paradigms by providing basic management strategies, at present it cannot replace expert clinical consultation in clinical practice.
Sumoylation As a Novel Target for T-Cell Neoplasms
Authors: Milshteyn Larry , Lownik Joseph , Villamejor Anton , Merchant Akil 
Published: December 2024

Despite breakthroughs in our understanding of peripheral T-cell lymphoma (PTCL) through studies such as genetic profiling, this disease has proven to be difficult to manage compared to it B-cell counterparts. There have been few advances in treatment regimens since the initial trials with CHOP-based chemotherapy (Cyclophosphamide, Doxorubicin, Vincristine, and Prednisone) for many decades. In recent years, there have been more targeted therapies such as Brentuximab-Vedotin, however this is not applicable to most PTCL. Given the persistent poor outcomes in this patient population, novel therapeutic options are needed.
Similar to B-cell lymphomas, a subset of PTCL lymphomas exhibit alterations in cMYC expression. Recent studies have demonstrated a unique susceptibility of cMYC-driven B-cell lymphomas to the inhibition of SUMOylation. SUMOylation is a process of post translational modification (PTM), similar as ubiquitination. However, SUMOylation results in alteration of protein function, rather than explicit changes such as targeting for destruction. Using a recently descript PTCL model, LM-23, which overexpresses cMYC, we found that inhibition of SAE-1/2 with SB-4826, which are necessary for SUMOylation, led to decreased tumor growth in a subcutaneous tumor model, as well as cell death in in vitro models.
Here, we demonstrate a susceptibility of T-cell neoplasms (PTCL and T-lymphoblastic leukemia) to the inhibition of SUMOylation by targeting SAE-1/2 with SB-4826. Using multiple neoplastic T-cell lines, we demonstrate a low micromolar ICI50 for SB-4826 for neoplastic T-cell lines including LM-23 and Jurkat, similar to B-cell lymphoma cells lines. This ICI50 value is significantly lower than non-neoplastic T-cells, suggesting decreased effects on non-neoplastic counterparts of the functional immune system. Additionally, LM-23 tumor bearing mice treated with SB-4826 showed significant reductions in tumor volume compared to CHOP treated mice.
Additional studies on murine and human PTCL and T-ALL cell lines will be presented. While further preclinical studies are necessary, these studies identify SUMOylation as a novel therapeutic target in PTCL and T-ALL, especially cMYC- driven T-cell neoplasms.
Role of Alpha Granule Secretion in Platelet Glucose Metabolism
Authors: Prakhya Shravani , Whiteheart Sidney W. , Joshi Smita , Vekaria Hemendra , Sullivan Patrick 
Published: December 2024

Platelets play a critical role in hemostasis and thrombosis through their activation, shape change, secretion, and aggregation. These highly metabolically active cells shift from a low-energy resting state to a high-energy activated state. Platelet secretion is a key step in hemostasis, involving the release of cargo from alpha, dense, and lysosomal granules, which merge with the plasma membrane to modify the local environment to help form a clot achieving hemostasis.
Alpha granules, the most numerous and diverse, are crucial for platelet functions such as hemostasis, inflammation, immunity, wound healing, and angiogenesis. They contain numerous bioactive proteins, including integrins, immunoglobulin family receptors, and GLUT3, a glucose transporter.
In resting conditions, GLUT3 is found in alpha granules but translocates to the plasma membrane upon platelet activation, facilitating glucose uptake necessary for platelet function. GLUT3 deletion impairs platelet activation and function, highlighting the importance of glucose metabolism.
This study, using the V2Δ3Δ8-/- mouse model with deficient SNARE proteins, reveals that GLUT3 translocation is critical for platelet secretion and clot formation. We show clot contraction defect in V2Δ3Δ8-/- platelets that can be rescued by the addition of pyruvate (downstream intermediate metabolite whose destiny is determined by the availability of oxygen within the environment) indicating the metabolic aspect of platelet secretion. Further research is needed to understand GLUT3 levels in these mice. Our findings underscore the link between platelet secretion and metabolism, showing that GLUT3 translocation from granules to the surface is crucial for platelet function and metabolism.
Inflammasome Cytokine Secretion in Heparin-Induced Thrombocytopenia Is Supported By Platelet-Derived Mediators and Complement
Authors: Michael James V , Khandelwal Sanjay , Tawk Antonios , Lazar Sophia , Arepally Gow , McKenzie Steven E 
Published: December 2024

Background: Heparin-induced thrombocytopenia (HIT) is an adverse drug reaction that is caused by IgG immune complexes that activate platelets and monocytes through Fc Receptor for IgG IIA (FcγRIIA). These immune complexes also fix complement. Stimulated monocytes produce proinflammatory cytokines including inflammasome dependent IL-1β release, which has not been investigated in HIT.
Aims: Platelets have been shown to support monocyte inflammasome cytokine release during inflammation, primarily in microbial infection. This study sought to investigate the role of inflammasome cytokine production in HIT pathology.
Methods: We investigated the role of the NLRP3 inflammasome following immune complex (IC) challenge in healthy donor whole blood or immortalized monocyte-like cells. The contribution of inflammasome assembly and cytokine release on HIT pathology was assessed utilizing NLRP3 inhibitor MCC950 in the triple allele ‘HIT’ mouse model. Plasma IL-1β cytokine production was measured using ELISA, and complete blood counts were taken to assess thrombocytopenia. Furthermore, the contribution of complement was assessed using IC challenge of isolated primary blood mononuclear cells suspended in plasma or heat-inactivated plasma.
Results: Following human whole blood challenged or isolated mononuclear cell/platelet suspensions with two distinct immune complexes (heat aggregated IgG or heparin/PF4 complexes) we observed a significant increase in inflammasome cytokine production at 22 hours. Mononuclear/platelet cell fractions challenged with heparin/PF4 complexes had a significant increase in inflammasome cytokine secretion that was reduced with pretreatment with an FcγRIIA blocking antibody, or removal of complement with heat-inactivated plasma. We observed that administration of heparin and monoclonal αPF4/heparin antibody KKO to HIT mice resulted in thrombocytopenia and significantly increased circulating IL-1β, whereas mice treated with NLRP3 inflammasome inhibitor MCC950 protected mice from a platelet drop and cytokine production in our humanized mouse model of HIT in vivo.
Conclusions: These data demonstrate for the first time that HIT IC mediates the production and secretion of inflammasome cytokines, which is influenced by the presence of platelets in a FcγRIIA dependent manner as well as by complement. Furthermore, the NLRP3 inflammasome is shown in HIT model mice to be a critical component of HIT pathophysiology. These results provide a scientific rationale for exploring mechanisms that drive platelet/monocyte/complement communication and targeting the inflammasome as a possible therapeutic strategy in HIT.
Long Term Follow up of a Child with a Rare Hemoglobinopathy: Hemoglobin Little Venice
Authors: Abdulhai Safa , Narang Shalu , Herrick Jennifer L. 
Published: December 2024

We report the second known case in the world of a rare beta chain variant hemoglobinopathy Hemoglobin Little Venice [β42 Phe→Cys, HBB:c.128T&gt;G] originally found in 2009 in a 7 month-old Asian Indian female with severe chronic hemolytic anemia during infancy who is now 15 years of age. Her newborn hemoglobinopathy screen showed FA. She presented with jaundice at 1 month of age with a bilirubin of 15 mg/dL initially attributed to breast milk and was managed with supportive care. Hemoglobin electrophoresis with confirmatory DNA sequencing was performed by Mayo Clinic laboratories at 7 months of age to establish the diagnosis. At that time hemoglobin was 5.6 g/dL and bilirubin was 6.9 mg/dL. Hemoglobin electrophoresis showed Hb A 71.6%, Hb A2 2.5%, Hb F 24.5% and Hb variant 1.4%. Stability studies were abnormal. The testing revealed a previously undescribed beta 42 mutation of TTT&gt;TGT (Phe&gt;Cys). Our patient has been receiving blood transfusions every four weeks leading to iron overload at 5 years of age treated with an iron chelator. Her growth and development have been normal, similar to peers. At age 9, she had cholecystectomy for gallstones. At the age of 11, she had SARS COVID-19 infection that was treated with supportive care and she recovered well. She was noted to have mild thrombocytopenia ranging from 70-110's and had splenectomy at 13 due to increased frequency of blood transfusion and chronic splenomegaly. At 14, she was treated for asymptomatic babesiosis with atovaquone and azithromycin.
At present, she continues to require regular transfusions every 4 weeks due to severe hemolytic anemia and has significant thrombocytosis along with leukocytosis and nucleated red blood cells. She has significant transfusion-associated hemosiderosis and receives double chelation with deferasirox and deferiprone with marked history of noncompliance. Last MRI of Liver T2* in 2023 showed an approximate hepatic iron concentration of greater than 20 mg/g that is significantly elevated and MRI of heart T2*decay time of greater than 20 ms, which indicates low risk of clinically significant myocardial iron deposition. Both parents and two sisters have no evidence of hemolytic anemia and there are no other family members with chronic anemia or history of splenectomies.
The importance of phenylalanine in position 42 was described by Bartram et al in 2012 for the position's role in maintaining the solubility and stability of the β-hemoglobin chain. When replaced with cysteine, it disrupts oxygen delivery to the cells. This rare hemoglobinopathy requires multiple transfusions to treat vaso-occlusive episodes. At present, little is known regarding treatment choices for this rare hemoglobinopathy therefore, we will continue chronic blood transfusions and encourage compliance with iron chelation. A similar variant affecting the same codon, Hb Hammersmith [β42 Phe→Ser, HBB:c.128T&gt;C] has comparatively more data in the literature which may offer some general guidance.