bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical Feature and Predictive Model for Transplanted Patients with Functional High-Risk Multiple Myeloma
Authors: Yu Tongyong , Li Juan
Published: December 2024

Objective: Despite these therapeutic advancements, a subset of patients continues to exhibit poor outcomes, highlighting the complexity and heterogeneity of the disease. These subsets of patients with MM who relapsed early, even without any high-risk features at baseline, were defined as functional high-risk (FHR) MM. And the prognosis of FHR MM was worse than high-risk (HR) MM. Therefore, it is important to distinguish patients with FHR MM from patients with standard risk (SR). In this study, we explored risk factors for clinical features to identify through comparing FHR with SR. Additionally, we sought to develop a prognostic model to predict FHR MM patients with transplantation according to risk factors.

Methods: We conducted a retrospective analysis of clinical data from MM patients diagnosed between January 1, 2007, and March 22, 2024, who underwent autologous hematopoietic stem cell transplantation (ASCT) with a minimum follow-up of 24 months after transplantation. The cases were randomly divided into a training set (151 cases, 70%) and a validation set (65 cases, 30%). Univariate and multivariate analyses were performed to identify independent risk factors for FHR MM. A predictive model was constructed using logistic regression analysis and internally validated.

Results: A total of 357 MM patients were included in our study, with 49 (13.73%) classified as FHR, 167 (46.78%) as SR, and 141 (39.50%) as GHR. The FHR groups had the shortest survival compared with the HR and the SR groups, including PFS and OS. The median PFS was 22.5 months, 49.7 months, and 111.2 months in the FHR, HR, and SR groups, respectively. The median OS was 39.7 months, 71 months, and 132.1 months in the FHR, HR, and SR groups, respectively. Univariate and multivariate analyses revealed that LDH ≥190U/L (OR=2.70, P=0.015), PET-CT SUVmax≥7.5 at onset of MM (OR=3.13, P=0.035), the proportions of PET-CT SUVmax decline after induction therapy ≥80% (OR=4.79, P=0.044), PLT <80*10^9/L (OR=10.55, P=0.012), and ferritin ≥650μg/L (OR=4.76, P=0.031) were independent risk factors for FHR MM. Additionally, Pattern B of M-protein decline (OR=0.39, P=0.012) and achieving CR after ASCT (OR=0.37, P=0.009) were identified as independent protective factors. A predictive model incorporating these factors demonstrated areas under the ROC curves of 0.753 for the training set and 0.857 for the validation set. Calibration plots, Decision Curve Analysis (DCA) curves, and Hosmer-Lemeshow test results indicated that the model has good predictive accuracy.

Conclusion: This study developed a predictive nomogram model for FHR MM patients with transplantation, which shows promising predictive capabilities.
Different Large Granular Lymphocyte Count Drew Distinguish Characteristics in Large Granular Lymphocytic Leukemia
Authors: Yu Ying , Li Yuxi , Cui Rui , Sun Jingwen , Lyu Rui , Wang Tingyu , Wenjie Xiong , Yan Yuting , Qiu Lu-Gui , Yi Shuhua
Published: December 2024

Introduction

Large granular lymphocytic leukemia (LGLL) is a rare disorder characterized by the clonal expansion of large granular lymphocytes (LGLs). While, an LGL count greater than 2×10^9/L is common, some researchers consider > 0.5×10^9/L necessary. However, the WHO and other studies do not specify a clear LGL count, leading to ambiguity. Diagnosing patients with low LGL counts, especially < 0.5×10^9/L, remains challenging. Establishing a precise clinical diagnostic parameter for LGLL is urgently needed.

Methods

From July 2005 to July 2022, 175 patients were diagnosed with LGLL at Blood Hospital of CAMS & PUMC.，excluding patients with aplastic anemia (AA) or other hematologic malignancies. Then patients categorized into low (<0.5× 10^9/L), intermediate (0.5-2× 10^9/L), and high (>2× 10^9/L) LGL count groups (27.4%, 31.4%, and 41.1% of patients, respectively).

Results

Among the 175 enrolled LGLL patients, 20 (11.4%) had NK-LGLL and 155 (88.6%) had T-LGLL. The median proportion of LGL cells was 55.8% (range: 4.1%-98.2%), with a median LGL count of 1.1 × 10^9/L (range: 0.10-71.21 × 10^9/L). The low LGL count group had 48 patients (27.4%), the intermediate group had 55 patients (31.4%), and the high group had 72 patients (41.1%). Anemia (75.0% vs. 83.6% vs. 48.6%, P<0.001), thrombocytopenia (27.1% vs. 20.6% vs. 5.6%, P<0.001), and pure red cell aplastic anemia (29.2% vs. 43.6% vs. 13.9%, P=0.001) were significantly more prevalent in the low and intermediate LGL count groups than in the high LGL count group (Table 1). Notably, Treg cell proportion was higher in the low and intermediate LGL count group compared to the high LGL count group (P=0.015). Overall, the low and intermediate LGL count groups exhibited lower overall response rates (ORR) and worse overall survival (OS) compared with the high LGL count group (ORR: 86.8% vs. 64.3% vs. 91.4%, P=0.005; OS: 3-year OS rate 86.5% vs. 90.9% vs. 100%, P=0.029). However, no evidence of a difference in ORR and OS among the three LGL count groups was observed under treatment subgroups.

Conclusions

Generally, while LGL counts reveal unique clinical and biological features, they should not be obligatory for diagnosing LGLL.
Busulfan-Based Intensified Myeloablative Conditioning Regimen Allogeneic Hematopoietic Cell Transplantation for Juvenile Myelomonocytic Leukemia-Single Center Study
Authors: Wang Yun , Sun Ruijuan , Zhao Yanli , Wei Zhijie , Xiong Min , Liu Deyan , Lu Peihua , Lu Yue
Published: December 2024

Background: Juvenile myelomonocytic leukemia (JMML) is a life-threatening myeloproliferative neoplasm, allogeneic hematopoietic stem cell transplantation(allo-HSCT) is the only curable method for it, but disease recurrence and graft rejection are major causes of treatment failure, thus,new conditioning regimen are needed to improve the survival outcomes in patients with JMML. Method:We retrospectively analyzed 14 patients who diagnosis with JMML(according to 2022 WHO) and received Busulfan(BU)-based intensified Myeloablative conditioning regimen(MAC) allo-HSCT,evaluated its prognosis.Results: 9 males and 5 females; the median age were 4 years (10 months to 10 years); 8 cases with PTPN1 mutation positive, 3 cases NF1 mutation positive, 1 NRAS , 1 KRAS, 1 CBL; 5 of them with another gene mutation abnormality, 3 of them with three or more gene mutation abnormalities. 3 patients had chromosome -7. 5 of them had hepatosplenomegaly.Donor type: 9 patients received related haploidentical donors, 4 received matched sibling donors, and 1 received unrelated matched donors. The median proportion of blast cells in bone marrow before transplantation was 11% (range:3.4-21.2). 9 patients received demethylating drug treatment before transplantation.All patients received BU-based intensified MAC, Seven patients received Decitabine/Cytarabine (Ara-c) /BU/ Cyclophosphamide (Cy) or Fludarabine (FLU) /Melphalan (MEL) / ATG (n=7), 3 cases received FLAG + BU/Cy or FLU /ATG (n=3); 4 received Ara-c/BU/CY/MEL /ATG (n=4). All patients achieved stem cell engraftment, the median neutrophils and platelet engraftment times were 15 (range:10-18) days and 12 (range:9-87) days respectively. Six patients developed II-IV aGVHD, three patiens developed mild cGVHD, and two developed severe cGVHD. Eleven patients survived, the median survival time were 36 (12-108) months; the 5-year overall survival(OS) and relapse-free survival(RFS) were 78.6±11.1% and 78.6±11.1%, respectively, and the 5-year cumulative incidence of relapse(CIR) was 15.4±10.0%; the 5-year non relapse mortality(TRM) was 8.1±6.9%. Three patients died until last follow-up time, two of whom died of disease recurrence (six months and eight months after transplantation, respectively), and one died of aGVHD after transplantation three months. Conclusion: Allo-HSCT with busulfan-based intensified myeloablative conditioning regimen for JMML achieved all stem cell engraftment, lower CIR and encouraging survival.
Impact of Mono- and Bi-Allelic TP53 Aberrations on Transformation of Human HSPCs
Authors: Lind Karin , Dutta Sayantanee , Cosenza Marco , Eder Thomas , Foßelteder Johannes , Pabst Gabriel , Schöll Wolfgang , Prokesch Andreas , Reinisch Andreas , Woelfler Albert , Zebisch Armin , Grebien Florian , Korbel Jan O. , Sill Heinz
Published: December 2024

Introduction

Clonal hematopoiesis with mono-allelic TP53 mutations confers a high-risk for progression to myeloid neoplasms, particularly in the context of exposure to cytotoxic stress. During leukemic transformation, bi-allelic TP53 aberrations frequently evolve - however, the exact molecular mechanisms driving transformation are not well understood. Here, we use genetically engineered human HSPCs to investigate the impact of mono- and bi-allelic TP53 aberrations on HSPC function and genomic integrity.

Methods

Cord blood derived HSPCs were engineered using a CRISPR-Cas9 and adeno-associated viral vector-mediated knock-in strategy to generate TP53R175H/WT, TP53R175H/KO, TP53R273H/WT, TP53R273H/KO, TP53KO/WT and TP53KO/KO aberrations, respectively. To control for gene editing, the AAVS1 safe harbor locus was targeted. To understand the functional impact of different TP53 allelic states, cell cycle analysis, colony formation and serial replating as well as bulk mRNA sequencing were performed. The emergence of structural chromosomal variants was investigated by single-cell template strand sequencing upon doxorubicin exposure.

Results

Successful editing of HSPCs at the TP53 locus was confirmed at the genomic DNA, RNA and protein levels, respectively. Bi-allelic TP53 aberrant HSPCs showed an increase of cells in S-phase and formed significantly more colonies with a high proportion of BFU-E as compared to control cells. Mono-allelic TP53 aberrant cells lost their replating capacity after three passages whereas bi-allelic ones were not exhausted after five replatings. Transcriptome analysis revealed the induction of an inflammatory signature in cells with both, mono-and bi-allelic TP53 aberrations, with a skewed expression rate of erythroid and myeloid genes. Single-cell template strand sequencing indicated that mono-allelic TP53 aberrations, especially in conjunction with doxorubicin treatment, already induced a substantial increase of structural genomic variants including chromothripsis.

Conclusions

Our data indicate that already mono-allelic TP53 aberrations induce an inflammatory signature and structural chromosomal alterations in human HSPCs. Alterations of the second TP53 allele increase the proliferative and self-renewing capacity of HSPCs further promoting their leukemic transformation and progression. These data contributes towards a better understanding of human, TP53 aberrant leukemogenesis.
Single-Cell Analysis Reveals Altered Tumor Microenvironments of Relapse/ Refractory and Remission-Associated Acute Myeloid Leukemia Treated By ABC-14 Regimen
Authors: Huang Xin , Li Minming , Wu Ping , Deng Chengxin , Lu Zesheng , Wang Yulian , Geng Suxia , Luo Chengwei Luo , Lai Peilong , Du Xin , Weng Jianyu
Published: December 2024

The preliminary study (2023 ASH Poster #1522) demonstrated that ABC-14 regimen (Azacitidine, Venetoclax, and Chidamide) was comparable to “3+7” regimen in the induction therapy of newly diagnosed AML, while overcoming the limitations of drug resistance of VA regimen (Azacitidine and Venetoclax), even with better for the subtype of AML-M4/M5, and reducing the toxicity of the traditional “3+7” regimen. However, it is important to identify deeper characterization of the disease that can be used to better guide ABC-14 regimen therapy decisions and improve clinical outcomes. Here, we utilize single-cell RNA sequencing (scRNA-seq) to analyze AML bone marrow (BM) samples from diagnosis (6 cases), remission (3 cases), short time relapse (2 cases), and refractory (4 cases) timepoints, which provide insights into AML tumor and microenvironment.

Bone marrow samples aspirates from AML patients and healthy individuals were obtained from the Guangdong Provincial People's Hospital under ethical guidelines with informed consent. scRNA-seq data analysis including cell clustering and cell-type annotation, CNV estimation in single cells, differentially expressed genes (DEGs) testing, GO/KEGG enrichment analysis.

We focused on the distinct clusters of primary refractories, short time relapse, and continuous complete remission (CCR) associated AML blasts, monocytes, T cells, and macrophages with differential expression of genes. Relapse-associated group have more CD16+Mono, exhausted TIGIT+CD8+T cells and C1QB+macrophages, while CCR-associated group have more CD14+IFIT1+Mono, CD14+S100A12+Mono, CD14+HLA-DPB1+Mono, CD14+AQP9+Mono. Post-therapy residual blasts and primary refractory group exhibits downregulation of signal transduction by p53 class mediator genes. Also, a post-therapy T-cell cluster associated with relapse group exhibits downregulation of antigen processing and presentation of peptide antigen via MHC Class II and T-cell activation genes.

Altogether, this study characterizes ABC-14 regiment treated AML relapsed /refractory and CCR associated groups to provide insights into the BM microenvironment landscape. Our results will facilitate a better understanding and the development of novel therapeutic strategies for relapsed/refractory AML patients.
Impact of CD3 Expression on Outcome of Pediatric Anaplastic Large Cell Lymphoma
Authors: Obeid Ahmed , Hafez Hanafy , Semary Samah Fathy , Nageb Eman , Soliman Sonya Fahmy , Zaky Iman , Kamal Mohammed , Lehmann Leslie E. , Elhaddad Alaa
Published: December 2024

Background: Anaplastic large cell lymphoma (ALCL) constitutes 10-15% of childhood non-Hodgkin lymphoma. Despite different treatment strategies, event-free survival has remained stable. CD3 expression, a T-cell marker, can be lost or diminished in some ALCL cases, which may affect the prognosis, especially in relapse settings according to some studies. The purpose of this study is to determine the effect of CD3 expression on survival and its relation to the other prognostic variables in pediatric ALCL.

Methods: A retrospective analytical study was conducted on 89 newly diagnosed pediatric ALCL patients (under 18 years old) treated at Children's Cancer Hospital Egypt (CCHE-57357) from July 2007 to December 2019. Immunohistochemistry was utilized to confirm the diagnosis and determine CD3 expression in tumor cells. Survival results - event-free survival (EFS), relapse-free survival (RFS), and overall survival (OS) - were analyzed concerning CD3 expression and other risk factors.

Results: The median age was 10.7 years with a male-to-female ratio of 1.8-1. The majority of patients (85.4%) were ALK positive. CD3 was positive in 31 (34.8%) patients. The median follow-up period was 60 months. The five-year OS, EFS and RFS rates for the entire group were 84.3%, 73.1%, and 81.5%, respectively. CD3 positivity was linked to more central nervous system (CNS) involvement (p=0.03) but did not significantly correlated with other patient features or survival outcomes.

Conclusion: This study indicates that CD3 expression may not affect the survival in newly diagnosed pediatric ALCL. However, stage, B symptoms, and particular organ involvement were linked to lower relapse-free survival. Additional studies are needed to validate these findings in larger cohorts.
Construction and Verification of Prediction Model of Frailty Risk in Elderly Patients with Hematological Neoplasms
Authors: Zhao Jinying , Qianqian Zhang , Liu Yating , Li Bing , Xie Wenjun , Jiang Erlie , Wang Xin
Published: December 2024

Objective: Develop and validate a nomogram for the prediction of frailty among elderly patients with HN.

Methods: A total of 505 elderly patients with HN from 17 hospitalsin China from April 2023 to November 2023 were included in the study. Characteristic features and disease-related information were collected, and geriatric assessments were conducted.Patients were divided by the Geriatric 8 questionnaire into frail and non-frail.The Chi-square test and Mann Whitney U test were used to compare the characteristics of the two groups. Univariate analysis and binary logistic regression were used to explore factors affecting frailty, and a risk prediction model was established in the form of a nomogram.Internal validation was carried out by the bootstrap using receiver operating characteristics, calibration curve,and decision analysis curve.

Results: The incidence rate of frailty in elderly patients with HN was 56.6%(286/505).The nomogram model (including eyelesof chemotherapy, self care ability, nutritional status, and hemoglobin level ) showed good predicting performance in both training and validation sets. In the validation cohort, the AUC was 0.867(95%C10.833~0.897), Youden index was 0.557, and the sensitivity and specificity were 85.71% and 69.95%, respectively. The average absolute error of the coincidence degree betwcen the predicted value and the real value was 0.01.

Conclusion: The nomogram showed good predicting value on elderly patients with HN, which can provide a reference for clinical doctors and nurses to identifythe frailty risk.
MagnetisMM-32: Evaluation of Elranatamab Vs EPd, PVd, or Kd in Patients with Relapsed or Refractory Multiple Myeloma and Prior Anti-CD38-Directed Therapy
Authors: Schuster Steven , Ito Satoshi , Hoyle Margaret , Yver Anne , Hong Fangxin , Finn Gregory
Published: December 2024

BACKGROUND

Elranatamab (ELRA), a humanized B-cell maturation antigen (BCMA)-CD3 bispecific antibody, has shown efficacy and manageable safety as a monotherapy in patients with relapsed or refractory multiple myeloma (RRMM). This study will evaluate ELRA monotherapy vs elotuzumab-pomalidomide-dexamethasone (EPd), pomalidomide-bortezomib-dexamethasone (PVd), or carfilzomib-dexamethasone (Kd) in patients with RRMM to determine whether ELRA can provide superior clinical benefit.

STUDY DESIGN AND METHODS

MagnetisMM-32, a phase 3, open-label, multicenter, randomized study, will enroll ≈492 patients. Patients will receive ELRA (Arm A) or investigator's choice of EPd, PVd or Kd (Arm B), until disease progression, unacceptable toxicity, withdrawal of consent, loss to follow-up, or study termination. Patients treated with ELRA will receive 2 step-up priming doses followed by weekly doses in a 28-day cycle. Patients who achieve a PR or better for ≥2 months will be eligible for reduced dosing frequency. Patients will be randomized 1:1 (stratified by prior line of therapy [1 vs 2 vs 3/4] and International Staging System disease stage [1/2 vs 3]).

Key inclusion criteria include age of ≥18 years, prior multiple myeloma diagnosis with measurable disease (per IMWG criteria), evidence of progressive disease or failure to achieve a response to last line of multiple myeloma therapy, 1 to 4 prior lines of therapy including an anti-CD38 antibody-containing regimen (for ≥2 consecutive cycles) and a lenalidomide-containing regimen (for ≥2 consecutive cycles), adequate bone marrow function, and an ECOG performance status of ≤2. Key exclusion criteria include stem cell transplant ≤12 weeks prior to enrollment; active, uncontrolled infection; prior BCMA-directed or CD3-redirecting therapy; and unable to receive EPd, PVd, or Kd.

The primary and key secondary endpoints are progression-free survival (PFS) by blinded independent central review (BICR) per IMWG criteria and overall survival (OS), respectively. Other secondary endpoints include PFS by investigator, objective response rate, duration of response, time to response by BICR, PFS on next line of therapy per IMWG criteria, MRD negativity rate, safety, pharmacokinetics of ELRA, immunogenicity, and health-related quality of life outcomes. The primary endpoint and OS will be compared statistically between treatment arms by stratified log-rank tests. Clinical trial information: NCT06152575. Study funding: Pfizer.
Aberrant Mitochondrial Iκbα/NF-Κb-Signaling in Lung Cancer Cells Mediates Endothelial Cell Activation and Platelets Aggregation, Facilitating Lung Cancer Metastasis and Cancer-Associated Thrombosis
Authors: Carra' Giovanna , Vacca Rita , Basile Roberta , Menga Alessio , Morotti Alessandro
Published: December 2024

Cancer presents a significant risk of blood clot formation in cancer patients, impacting patient outcomes. The intricate relationship between cancer-associated thrombosis and metastasis involves multifaceted mechanisms, including a hypercoagulable state, endothelial dysfunction, platelet activation, and interaction with the coagulation system, alongside inflammatory and immune responses. Here, we aim to unveil the non-cell autonomous pathways driving endothelial activation, platelet aggregation, and pro-thrombotic terrain for metastatic fomentation in a murine model of lung cancer.

The inhibitor IκBα, a regulator of the p65-NF-κB signaling pathway, plays a pivotal role in cancer metastasis. Our previous research identified IκBα overexpression in lung cancer, highlighting its potential role in the tumor's pathogenesis. To delve deeper into the role of mitochondrial IκBα, we engineered a construct specifically targeting it to the mitochondria (IκBα-MTS) of cancer cells. Intriguingly, lung cancer cells expressing IκBα-MTS displayed increased aggressiveness, as measured by their capability to increase proliferation, reducing apoptotic response and inducing migration.

Additionally, mitochondrial IκBα led lung cancer cells to activate endothelial cells (ECs), promoting an inflammatory and adhesive state, ultimately priming ECs at the metastatic site to facilitate cancer cells intravasation. This mechanism may be attributed to the metabolic shift observed in lung cancer cells with IκBα-MTS compared to those with wild-type IκBα, related to the induction of mitochondrial dysfunction.

In response to this environment, ECs were conducive to cancer-associated thrombosis, characterized by a hypercoagulable state resulting from the release of the procoagulant factors vWF, and favoring platelet activation and aggregation. Simultaneously, this facilitated cancer cell intravasation and metastasis.

Therefore, our findings offer crucial insights into “thrombo-oncotic” mechanisms and underscore the significance of ECs activation, microthrombi and metastasis formation as potential therapeutic targets in cancer treatment.
A Simple Pathway to Identify Duffy-Null Associated Neutrophil Count (DANC) Early on Hematology Consult Process
Authors: Ribeiro Maria-Jose
Published: December 2024

Introduction: The normal range for neutrophil counts in individuals with Duffy-null RBCs is often lower than commonly used laboratory reference ranges. This is a genetically determined baseline neutrophil count. The Duffy-null phenotype is caused by a variant in the ACKR1 promoter that results in loss of expression of Duffy antigens (Fy) on RBCs.

Method: Primary care submits electronic consults for patient who have white blood cell count below the laboratory normal range. The hematology service consultant recommends peripheral blood smear review, differential count, B12, folate, copper, HIV screen and HCV screen the last two according to CDC guidelines. The hospital blood bank committee created a process that include the routine type and screen with a comment that Duffy a and Duffy b report required.

Results: Forty-five patients have been tested on this pathway from November 2023 to July 2024 because the hematologist suspected DANC, from those 43 had the confirmation of Fy(a) and Fy(b) negative. Two were Fy(a) negative but Fy(b) positive. All had normal B12, folate, copper, negative HIV, and hepatitis c screening. On the referral for those patients, several have been told that they were being referred for possible myelodysplastic syndrome and for bone marrow biopsy. Once the test results were available, the patients who wanted to have a meeting to discuss the results were either scheduled on a clinic with medical students or residents with purpose of educating trainees and expand the knowledge of this normal variant. The individuals who wanted included their self-identified race. The incidence on different races were discussed only when the patients inquired.

Conclusion: This test is simple, produces rapid results, is relatively inexpensive and is easily incorporated on day-to-day practice. An education material was submitted to primary care providers, hematology fellows, residents, and students. Future direction is to develop education material for patients. Test consideration will be discussed at our center for patients who initiate chemotherapy with counts lower than the reference range or who have prolonged neutropenia during treatment to avoid unnecessarily holding therapy. Plan to develop studies and guidelines specific for patients with DANC based in developing literature.
Clinical Efficacy of Avatrombopag in the Treatment of Patients with Post-Transplantation Thrombocytopenia
Authors: Jian Xing , Xu Ziwei , Jiang Daozi , Wang Huafang
Published: December 2024

Post-transplantation thrombocytopenia (PT) is a common, serious complication following allogeneic hematopoietic stem cell transplantation (allo-HSCT) and can lead to increased transplantation-related mortality. The current treatment regimen for PT based on immune thrombocytopenia (ITP) has a limited effectiveness. Avatrombopag, a second-generation oral, small-molecule thrombopoietin receptor agonist (TPO-RA), has shown favourable therapeutic effect in ITP, chemotherapy-induced thrombocytopenia and thrombocytopenia associated with chronic liver disease, and may be an alternative choice for patients sufferring from PT. The clinical data of 34 cases with PT treated with avatrombopag at the Union Hospital of Huazhong University of Science and Technology from June 2021 to April 2023 were retrospectively analysed to investigate the efficacy and safety of the treatment.The study endpoints were overall response rate (ORR) of avatrombopag response and adverse drug reactions at week 8 of the treatment phase. platelet count level above 50×109/L for 7 consecutive days without the need for platelet transfusions was considered as avatrombopag response. In this study, 34 cases with PT were included, of which 85.3% (N = 29) were secondary failure of platelet recovery (SFPR), 14.7% (N = 4) were delayed platelet engraftment (DPE). Avatrombopag was administred at a median time of 75.5 days post-transplantation. The overall response rate was 79.4% (N = 27), which was slightly higher in the SFPR group than that in the EPD group (82.8% vs 60.0%, P = 0.268). We observed 3 patients who were still effective after a second administration of avatrombopag. The mild reversible hepatic dysfunction (11.8%, N = 4) and thrombosis (2.9%, N = 1) were documented adverse events in the study period. Additionally, some cases have developed pulmonary infections (11.8%, N = 4), hemorrhagic cystitis (2.9%, N = 1) cytomegalovirus viremia (2.9%, N = 1), limited chronic graft versus-host disease (11.8%, N = 4) during avatrombopag treatment. Consequently, avatrombopag might offer a treatment option for patients with thrombocytopenia following allo-HSCT, which was effective and safe.
Senescence of Mesenchymal Stromal Cells in Myelofibrosis Tumor Microenvironemnt Is Mediated By Glutamate-Induced Intracellular Fumarate Accumulation
Authors: Giallongo Sebastiano , Giallongo Cesarina , Longhitano Lucia , Ferrigno Jessica , Amorini Angela Maria , Calvo Deborah , Li Volti Giovanni , Romano Alessandra , Di Raimondo Francesco , Palumbo Giuseppe Alberto , Tibullo Daniele
Published: December 2024

Primary myelofibrosis (PMF) is a type of myeloproliferative neoplasm characterized by ineffective blood cell production. The growth of malignant cells is influenced by their interaction with the tumor microenvironment (TME), which is marked by significant fibrosis and inflammation. In this setting, mesenchymal stromal cells (MSCs) are reprogrammed to become senescent, releasing proinflammatory cytokines as part of the senescence-associated secretory phenotype (SASP), thereby worsening the inflammatory state of the PMF-TME. Additionally, the PMF-TME is rich in various metabolites that reshape the microenvironment to support the growth of malignant cells.

To investigate the metabolic changes in PMF, we analyzed several metabolites in peripheral blood using HPLC and found an accumulation of Glutamate (Glu) and its byproduct fumarate. Both Glu and fumarate are known to induce cell senescence. To explore their impact on the mesenchymal compartment, we used an in vitro model of healthy MSCs. Treatment with Glu and fumarate led to increased accumulation of reactive oxygen species (ROS), causing elevated DNA damage and fumarate-associated epigenetic modification H3K36me2. Moreover, fumarate inhibited cell proliferation, as shown by clonogenic assays and triphosphate accumulation. Importantly, supplementation with Glu and fumarate induced cell senescence, as evidenced by increased β-Galactosidase activity. This senescent phenotype was further supported by the accumulation of phospho-p53 and increased expression of SASP-associated genes IL6, TNF, and TGFβ. Senescent cells also showed increased 5mC levels, similar to those observed in primary PMF-MSCs. Additionally, fumarate supplementation led to enhanced expression of Col1a1, highlighting its role in promoting the characteristic fibrotic environment of the PMF-TME.

In conclusion, our findings reveal that Glu metabolism plays a critical role in the onset of PMF. The conversion of Glu to fumarate induces a senescent profile in MSCs, contributing to the fibrotic TME typical of PMF. Future studies aiming to inhibit Glu uptake could lead to novel therapeutic strategies against PMF.
Elucidating the Interplay between ddx41 and cux1 mutations in Hematopoiesis in Danio Rerio
Authors: Awad Valerie , Bowman Teresa V.
Published: December 2024

Myelodysplastic syndromes (MDS) are a family of myeloid cancers involving defective hematopoietic stem and progenitor cells (HSPCs). In MDS, there is clonal expansion of defective HSPCs over healthy cells, which impacts cell maturation and increases the risk of transformation to acute myeloid leukemia (AML). Somatic and germline mutations contribute to MDS/AML formation by altering HSPC proliferation, differentiation, and inflammatory signaling. Heterozygous germline mutations of DEAD-box helicase 41 (DDX41) predispose individuals to hematologic malignancies, especially MDS/AML. However, patients typically do not develop MDS until later in life. This clinical observation suggests secondary factors play key roles in driving disease. Researchers identified a strong enrichment for co-occurrence of mutations in DDX41 and CUT-like homeobox 1 (CUX1), which encodes for a non-clustered homeodomain transcription factor involved in regulating gene expression and DNA repair. To decipher if co-mutations in DDX41 and CUX1 could promote hematopoietic dysfunction, we generated a zebrafish system that mimics germline ddx41 mutations and somatic cux1 mutations. In humans, the CUX1 locus has two divergent splice isoforms that encode for two distinct proteins, CUX1 and CASP. In zebrafish, these factors are encoded by two paralogous genes, cux1a and cux1b. As zebrafish ddx41 homozygous loss-of-function mutant embryos expand their HSPCs and develop macrocytic anemia, we assessed these two cell types in ddx41 and cux1-paralog co-mutated embryos. Transient mutagenesis of cux1a or cux1b elevated cmyb+ HSPC levels in ddx41 homozygous mutants beyond non-mutagenized mutants. Similarly, mutagenesis of cux1a or cux1b worsened anemia in ddx41 homozygous mutant embryos and induced anemia in ddx41 heterozygous embryos. Of note, cux1a mutagenesis had no impact on erythrocyte levels in ddx41 wildtype embryos, suggesting genotype-selective effects of mutating this CUX1 homolog. These findings indicate that combined insufficiencies in DDX41 and CUX1 act synergistically or additively to promote hematopoietic dysfunction and provide a platform to discover the mechanism underlying how this might contribute to the development of MDS/AML.
Enhanced Potency of BCMA/CD19 Dual-Targeting Allogeneic CAR-T Cells for Relapsed/ Refractory Multiple Myeloma with 4-1BB/TACI Intracellular Domains
Authors: Wang Steven , Pang Iris , Chang Claudia , Burrascano Michelle , DeMarco Nick , Mendoza Danny , Reyes Sonia , Barcenas Arturo , Nieto Pastor , Diaz Jose , Arauz Garrett , Lynn Chris , Nguyen Tony , Reed Connor , Connerney Jeannette , LE Quy , Coronella Julia , Shedlock Devon J
Published: December 2024

B-cell maturation antigen (BCMA)-directed chimeric antigen receptor (CAR) T cell therapy has shown high initial response rates in relapsed/refractory multiple myeloma (RRMM), but a majority of patients eventually relapse with progressive disease. Previous studies have identified a less differentiated, CD19-positive progenitor subpopulation of MM cells that contributes to resistance and poor survival. Furthermore, it has been reported that anti-CD19 CAR-T cells given after high-dose melphalan and stem cell transplant improve patient survival and can eliminate MM progenitor cells that are resistant to BCMA-directed CAR-T cells in ex vivo assays.

P-BCMACD19-ALLO1 is a fully allogeneic CAR-T therapy expressing two full-length CARs targeting BCMA and CD19, each with an optimized intracellular domain (ICD) for enhanced potency. P-BCMACD19-ALLO1 is a T stem cell (TSCM)-rich product derived from healthy donor pan-T cells using a nonviral transposon-based system for transgene delivery, with knockout of TRBC1/2 and B2M. Clinical data from Poseida and others have demonstrated safety and efficacy advantages associated with TSCM CAR-T, including for P-BCMA-ALLO1, our BCMA-targeting allogeneic CAR-T for RRMM (NCT04960579).

We show that replacing the standard 4-1BB co-stimulatory domain with an ICD derived from the tumor necrosis factor (TNF) receptor family member transmembrane activator and CAML interactor (TACI) enhances in vivo potency against a CD19 +/BCMA+ xenograft model when TACI is placed in the CD19 CAR. In contrast, the configuration of CD19 CAR with 41BB + BCMA CAR with TACI underperformed compared to the dual 4-1BB configuration.

The BCMA CAR was constructed using two unique fully human single-domain heavy-chain variable domains (VHs) against BCMA joined by a GS4 linker in tandem, independently binding BCMA. The binding of two unique BCMA epitopes may prevent the emergence of BCMA escape mutants. The anti-CD19 CAR binder comprises a fully human single-domain heavy-chain VH against CD19. Both CARs are expressed from a single transgene containing BCMA and CD19 CARs, an inducible Caspase-9 safety switch, and a dihydrofolate reductase mutein, separated by T2A sequences for independent expression. P-BCMACD19-ALLO1 CAR-T cells demonstrated potent in vitro cytotoxicity against tumor cells expressing BCMA, CD19, or both antigens, while single BCMA and CD19 CAR-T cells only recognized tumor cells that are BCMA and CD19 positive, respectively.

In this study, we report the enhanced in vivo activity of P-BCMACD19-ALLO1 CAR-T cells using 4-1BB/TACI ICD combination. These findings underscore the therapeutic potential of P-BCMACD19-ALLO1 CAR-T cells to be evaluated in clinical trials.
Comparing BAT and Beighton Scores in Patients with ITP: A Retrospective Observational Cohort Study
Authors: Gaston Farrah N. , Barclay Dominique , Kreitzer Sarah , Tarantino Michael D
Published: December 2024

Introduction

Immune thrombocytopenic purpura (ITP) is an acquired thrombocytopenia that leads to clinically significant bleeding in many. The Bleeding Assessment Tool (BAT) is a validated tool used to assess the degree of bleeding with scoring directly proportional to the severity of bleeding. Predicting the occurrence of bleeding is challenging in patients with ITP. Hypermobility syndromes have been associated with higher rates of clinically significant bleeding. Joint hypermobility, assessed by the Beighton Scoring System, has been shown to be associated with more severe bleeding phenotypes in persons with hemophilia, von Willebrand disease, and in some platelet function disorders, but has not been studied in patients with ITP.

Methods

We conducted a retrospective observational cohort study to investigate the relationship between joint hypermobility and the degree of bleeding by comparing BAT scores and age/sex-specific Beighton scores in patients with ITP at least 6 years age at the Bleeding and Clotting Disorders Institute. Correlations were determined using nonparametric analyses to test the difference between two groups: (a) abnormal BAT versus Beighton scores and (b) abnormal BAT versus degree of thrombocytopenia or sex. The Independent-Samples Kruskal-Wallis Test was used to determine the impact of age and platelet count on BAT and Beighton scores, if one existed.

Results

Data from 174 patients were obtained, though 122 were excluded due to a lack of BAT or Beighton score, or both. Of the 52 patients remaining, 34 (65.4%) were female and 18 (34.6%) were male, with ages ranging between 12-81 years. BAT scores ranged between 0-18 with a median of 5 and mean of 5.8+4.4, while Beighton scores ranged between 0-9 with a median of 3.5 and mean of 3.4+3.4. A moderate correlation (r values between 0.5-0.7) was found between overall BAT and Beighton scores (r=0.66), while an inverse correlation was found between Beighton scores and platelet counts (r= -0.89). All 12 patients with positive Beighton scores had abnormal (high) BAT scores (r=1.00) and were all female. Overall, there was no significant association (p=0.681) between gender, BAT, and Beighton scores. Further, there was no significant difference in BAT and Beighton scores across age and platelet count.

Conclusions

Hypermobility was associated with more bleeding based on higher BAT scores. The degree of bleeding was not predicted by the platelet count or age. Our data also suggest that females with ITP and hypermobility are more likely to have more clinically significant bleeding, however results comparing hypermobility and platelet count across the categories of age and sex were unrevealing. Broader investigation is needed to determine the full impact of the association of joint hypermobility and bleeding severity in patients with ITP.
Haploidentical Bone Marrow Transplantation for Wiskott-Aldrich Syndrome
Authors: Wang Yingjie , Zhang Mengmeng , Zhang Chengtao , Kang Zhijie , Liu Junli , Hu Jiong , Yan Jinsong
Published: December 2024

Introduction: Wiskott-Aldrich syndrome (WAS) is an X-linked disorder caused by mutations in the WAS gene, which encodes the WAS protein involved in actin polymerization. WAS is a rare syndrome affecting approximately 1 in 100,000 live births. Hematopoietic stem cell transplantation is the mainly curative option for WAS. In this study, the outcome of Haplo-BMT in four patients with WAS was investigated.

Methods: Four patients with WAS, who were confirmed with frameshift mutation in the WAS gene, were treated with haploidentical bone marrow transplant (haplo BMT) in second hospital of Dalian Medical University (China) from May 2018 to July 2024. The follow-up period was up to July 31, 2024. Flu/Bu/Cy/ATG was used as conditioning regimen, containing fludarabine (Flu, 30 mg/m2) once a day for 4 consecutive days, Busulfan (Bu, 0.8 mg/kg) four times a day for 4 days, cyclophosphamide (Cy, 30 mg/kg) once a day for 2 days, and rabbit ATG (SangStat, France) of 2.5 mg/kg once a day for 4 days. G-CSF-primed HSCTs plus marrow grafts were infused into recipients. Father served as blood stem cell donor.

Results: Four patients with were retrospectively collected, who had median age of 1.8 (range, 0.4-14) years. They received a median mononuclear cell number of 18.7 (range, 14.2-24.0) × 108/kg of body weight, or median CD34+ cell number of 12.9 (range, 10.3-14.9) × 106/kg of body weight. They were all successfully engrafted with a median time for neutrophil or platelets of 12 (range, 11-16) days and 12 (range, 11-14) days, respectively. One developed limited chronic graft versus host disease, and he well recovered by treating with methylprednisolone (1 mg/kg/day) and tacrolimus. One patient died of transplantation associated thrombotic microangiopathy at 8 months post-transplantation. Four patients with overall survival is 75%, with a median follow-up time of 26 (range, 2.5-48) months.

Conclusion: This retrospective investigation indicated haplo-BMT is feasible for a long-term overall survival in pediatric patients with WAS.
Toward a circular economy in Italian agri-food: upstream partners in insect biorefineries
Authors: Cattaneo Arianna , Padula Cecilia , Meneguz Marco , Mileto Carola , Barbero Silvia , Dabbou Sihem
Published: December 2024

The insect biorefinery (IB) is central to recycling nutrients from food losses and waste (FLW) in an emerging circular bioeconomy approach. The sustainable leadership of the European insect industry and its integration into the bioeconomy depend on implementing symbiotic production models through site selection and partnerships. Only 8 studies, including 3 in the EU, have examined stakeholder acceptance. This pilot study is the first to focus on potential upstream symbiosis partners (USPs) in the EU, examining the factors that influence the willingness of potential USPs in suitable Italian locations to adopt IB. Preliminary analyses identified a final sample of 31 respondents who completed an online-survey divided into four sections. After data elaboration, a principal component analysis was conducted, considering 27 questions and their correlations with the first two principal components. The analysis revealed that the presence of internal research and development within the company, along with the company’s role in the supply chain and the seasonality of production, had a minor influence on explaining the variance of the data. Interestingly, company size was negatively correlated with the willingness to become part of the insect-rearing supply chain or receive information about this technology. On the contrary, interest in innovative production and self-management of by-products were strongly related. Finally, funding opportunities could positively influence both the Italian and EU contexts, encouraging innovative practices among small and medium-sized enterprises. In contrast, barriers to implementing insect farming include misinformation, anticipated negative media impacts on branding, and resistance to changing by-product management practices. This pilot study serves as a foundational exploration, providing insights into the acceptability of insect farming among potential USPs and offers valuable insights for future studies, thereby shaping the discourse on the acceptability and integration of insect farming within the circular bioeconomy.
The host genotype actively shapes its microbiome across generations in laboratory mice
Authors: Benga Laurentiu , Rehm Anna , Gougoula Christina , Westhoff Philipp , Wachtmeister Thorsten , Benten W. Peter M. , Engelhardt Eva , Weber Andreas P. M. , Köhrer Karl , Sager Martin , Janssen Stefan
Published: December 2024

Background

The microbiome greatly affects health and wellbeing. Evolutionarily, it is doubtful that a host would rely on chance alone to pass on microbial colonization to its offspring. However, the literature currently offers only limited evidence regarding two alternative hypotheses: active microbial shaping by host genetic factors or transmission of a microbial maternal legacy.

Results

To further dissect the influence of host genetics and maternal inheritance, we collected two-cell stage embryos from two representative wild types, C57BL6/J and BALB/c, and transferred a mixture of both genotype embryos into hybrid recipient mice to be inoculated by an identical microbiome at birth.

Conclusions

Observing the offspring for six generations unequivocally emphasizes the impact of host genetic factors over maternal legacy in constant environments, akin to murine laboratory experiments. Interestingly, maternal legacy solely controlled the microbiome in the first offspring generation. However, current evidence supporting maternal legacy has not extended beyond this initial generation, resolving the aforementioned debate.

Graphical Abstract
Jail-based interventions to reduce risk for opioid-related overdose deaths: Examples of implementation within Ohio counties participating in the HEALing Communities Study
Authors: Sprunger Joel , Brown Jennifer , Rubi Sofia , Papp Joan , Lyons Michael , Winhusen T. John
Published: December 2024

Background

Opioid-related overdose is a leading cause of death for criminal legal-involved individuals and, although naloxone distribution and medications for opioid use disorder (MOUD) are effective means for reducing post-release overdose death risk, jail-based availability is limited. This case report describes the challenges faced by three Ohio communities as they implemented evidence-based practices (EBPs) in jails to combat post-release opioid overdose deaths.

Method

We present case examples of how barriers were overcome to implement jail-based EBPs in three Ohio communities (two urban and one rural) as part of the HEALing Communities Study (UM1DA049417; ClinicalTrials.gov Identifier: NCT04111939). Of the 18 participating Ohio HEALing Communities Study counties, we highlight 3 communities for the novelty of their EBPs implemented, the challenges that they faced, and their rural/urban status. We present descriptive data regarding the EBPs that they implemented and discuss the challenges identified by HEALing Communities Study staff with first-hand experience facilitating their implementation.

Results

Newly implemented interventions included overdose education and direct provision of naloxone to incarcerated individuals upon release (2 of 3 communities), initiating MOUD prior to release (3 of 3), linkage to ongoing MOUD treatment in the community (2 of 3), peer support-facilitated treatment retention efforts (2 of 3) and emergency housing (1 of 3) in the immediate post-incarceration period. Common challenges that emerged included skepticism about the need and feasibility of implementing EBPs to reduce overdose and death, lack of knowledge about the options available and whether external agencies may assist, and difficulty engaging stakeholders to overcome inertia.

Conclusions

Creative flexibility, calm persistence, technical facilitation, and collaboration with community service providers were assets that helped these Ohio jails implement evidence-based strategies that combat the opioid epidemic and reduce the likelihood of post-incarceration overdose and death in a high risk, formerly incarcerated population.