bluuebunny/crossref_metadata_2025 huggingface dataset:
Efficacy and Safety of Isavuconazole in the Treatment of Invasive Fungal Disease in Patients with Hematological Disorders
Authors: Yang Nan , Pan Pan , Pang Aiming , Jiang Erlie
Published: December 2024

Introduction:

Invasive fungal infections (IFIs) are a significant contributor to morbidity and mortality in hematological malignancies (HM) patients and stem cell transplant (SCT) recipients. The FDA approved Isavuconazole (ISA) as a primary treatment for invasive aspergillosis and mucormycosis. This study aims to investigate the practical application of Isavuconazole in patients with hematological malignancies (HM) and assess their clinical outcomes and safety profiles.

Patients & methods:

We conducted a retrospective study of patients with probable or possible mold infections who were treated with isavuconazole for a period of at least 10 days between May 2022 and May 2024. A total of 67 patients were included in the study, 20 of whom were receiving chemotherapy, and 47 who had undergone hematopoietic stem cell transplantation (HSCT). Of these, 44 were male and 23 were female, with a median age of 48 years (ranging from 34.5 to 52). Clinical and radiological assessments were carried out at baseline and at 6 and 12 weeks of follow-up.

Results:

The overall success rate was 51% (CR of 30% and PR of 21%) at 6 weeks, 75% (CR of 46% and PR of 28%) at 12 weeks. The 100-day all-cause mortality rate was 19.1%[8.71-28.3%]. Multivariate logistic regression analysis showed that persistent neutropenia, male sex and Invasive Mucormycosis were independently associated with inferior responses to ISA. Isavuconazole was used as monotherapy in 22% of the cases and in combination in 78%. Adverse events potentially related to isavuconazole were reported in three patients (4%), two of whom exhibited elevated creatinine levels, and one patient who exhibited a rash associated with isavuconazole . ISV was employed as the 1st line therapy in 16 (24%) patients and as a subsequent therapy in 51 (76%) patients.

Conclusion:

This study provides encouraging data regarding the efficacy and safety of ISA in HM patients. Neutropenia, male sex, Invasive Mucormycosis are possibly associated with inferior responses to ISA therapy.
A Novel, Universal Targeting Receptor-Adaptor CAR Platform Demonstrates Versatile, Flexible, and Controlled Tumor Inhibition in an Ipsc-Derived T-Cell
Authors: Devaney Andrew , Carton Jill , Hoffman Hunter , Millar Hillary J. , Walker David , Wheeler John , Miller Mike , Dower Chris , Morse Barry , Levitsky Hy , Naso Michael
Published: December 2024

Despite positive outcomes in some hematological oncology patients, several barriers to current autologous CAR T-cell therapies exist that prevent broader clinical success. These include tumor antigen escape, on-target off-tumor toxicities, and limited patient access to treatment. To increase safety and efficacy of CAR T-cell therapies, we are developing a Universal CAR (UCAR) that is activated through a soluble tumor targeting protein that pairs specifically with the UCAR. The UCAR approach enables a single CAR T-cell to target many different antigens through alternative targeting proteins to address antigen escape and tumor heterogeneity. Additionally, greater control of the activity of the cells within a patient is possible since effector activity is regulated through the dosing of the soluble protein. UCAR platforms are particularly interesting for inclusion into allogeneic cell therapy products, as the combination of these technologies enables a fully universal and “off-the-shelf” cell therapy cell line.

Here, we describe the development and functional assessment of the Universal Targeting Receptor-Adaptor Platform (UTRAP), an adaptable, highly specific, and developable system comprised of a novel UCAR and a soluble Bispecific Anti-Idiotype Targeting (BAIT) protein. We demonstrate that T-cells engineered with UTRAP have tunable activity that elicits robust cytotoxicity and tumor control, equivalent to a conventional CAR T-cell in vitro and in vivo. Our results show the flexibility of the antibody-based platform including multi-specific targeting and antigen swapping. Finally, we demonstrate that UTRAP is fully functional in allogeneic iPSC-derived gamma delta T-cells. These results pave the way for UTRAP to be used in any cell type advancing towards a truly universal, off-the-shelf cell therapy product that can adapt to target any tumor type.
A Single-Center, Single-Arm, Phase I Clinical Trial of Anti-BCMA/GPRC5D Bispecific CAR T-Cells in Patients with Refractory and Relapsed Multiple Myeloma with Extramedullary Disease
Authors: Qiu Ling , Ren Shihui , He Ying , Chen Dan , Lai Sihan , Dou Baitao , Li Mengjiao , Shu Rou , Chang Alex H. , Su Yi , Yao Hao , Fan Fangyi
Published: December 2024

Background: Despite the promising outcomes of chimeric antigen receptor (CAR) T-cell therapy in treating relapsed or refractory multiple myeloma (RRMM), addressing extramedullary disease (EMD) in this context remains a therapeutic challenge. This study aimed to evaluate the safety, efficacy, and activity of anti-BCMA/GPRC5D bispecific CAR T-cells in RRMM patients with EMD.

Methods: A Phase I, single-arm, open-label clinical trial was conducted at the Western Theater Command Hospital of the Chinese People's Liberation Army. Eligible patients were RRMM adults aged 18-75 years with ECOG performance status 0-3 and EMD. Patients received escalating doses of anti-BCMA/GPRC5D bispecific CAR T-cells, starting at 1.0×10⁶ CAR T-cells per kg. The primary endpoints were safety and efficacy, while secondary endpoints encompassed assessment of activity. Safety and activity analyses encompassed all patients who received the CAR T-cell product per protocol. The study protocol was approved by the institutional ethics committee (2024EC1-ky033).

Results: Between July 2023 and May 2024, nine patients with RRMM and EMD were enrolled and underwent leukapheresis. Three patients were subsequently excluded due to rapid disease progression. Six patients received the anti-BCMA/GPRC5D bispecific CAR T-cell infusion. With a median follow-up of 6.5 months (IQR 2-12), the median age was 58 years (IQR 48-67), and the cohort included three males (50%) and three females (50%). Genetic risk factors were present in three patients, undetected in two, and absent in one. The maximum tolerated dose was established as 2.0×10⁶ CAR T-cells per kg. Hematological toxicity, excluding lymphopenia, was the most frequent grade 3 or higher adverse event (100%). Cytokine release syndrome (CRS) occurred in 62.5% of patients, all of which were grade 1 or 2. The overall response rate (ORR) among evaluable patients was 100%, with three patients (50%) achieving complete response (CR), including one with subsequent disease progression. One patient (17%) achieved very good partial response (VGPR), and two (33%) achieved partial response (PR), both of whom subsequently died due to disease progression.

Conclusion: The findings of this Phase I trial suggest that anti-BCMA/GPRC5D bispecific CAR T-cell therapy may represent a promising therapeutic option for patients with refractory and relapsed multiple myeloma complicated by extramedullary disease. Further investigation in larger, controlled trials is warranted to confirm these preliminary observations and refine treatment strategies.
Analysis of Anxiety and Depression Status and Influencing Factors in Multiple Myeloma Patients
Authors: Gao Mengran , Zhang Wei , Fu Rong
Published: December 2024

OBJECTIVE

To explore the incidence and influencing factors of anxiety and depression in multiple myeloma patients. METHODS: 509 patients with multiple myeloma in the Department of Hematology, General Hospital of Tianjin Medical University from September 2021 to March 2024 were selected for the study. The Generalized Anxiety Disorder Scale (GAD-7) and Depression Self-assessment Scale (PHQ-9) were used to assess the patients' anxiety and depression, and the Verbal Pain Rating Scale ( VPRS ) and Total Neuropathy Scale Reduced (TNSr) were used to assess the degree of pain and peripheral neuropathy. Clinical data such as patients' personal information, laboratory tests, DS/ISS/RISS staging, treatment line, and efficacy assessment were improved to analyze the incidence and influencing factors of anxiety and depression in patients with multiple myeloma. RESULTS: The incidence of anxiety and depression in patients with multiple myeloma was 22.2% and 31.4%, respectively . Comorbid pain and/or peripheral neuropathy were independent risk factors for the development of anxiety and depression and were significantly associated with the severity of both. CONCLUSION: The incidence of anxiety and depression in patients with multiple myeloma is high, and clinicians should pay attention to the rational intervention of pain and peripheral neurologic symptoms and the early assessment of emotional problems during the peri-therapeutic period of patients!
Rituximab, Lenalidomide, and Zanubrutinib (ZR2) with Sequential CAR-T Cell As First-Line Therapy for High-Risk Large B Cell Lymphoma
Authors: Zhang Mingming , Hu Yongxian , Wei Guoqing , Fu Shan , Feng Jingjing , Hong Ruimin , Chang Alex H. , Huang He
Published: December 2024

Introduction

CAR-T cell therapy has made a breakthrough in refractory or relapsed large b cell lymphoma (LBCL), but data as a first-line therapy is still scarce. ZUMA-12 study showed significant efficacy of CAR-T cell as first-line therapy for high-risk LBCL. However, some patients have difficulty tolerating chemotherapy prior to CAR-T cell therapy, especially elderly and frail patients. Here we report the results of rituximab, lenalidomide, and zanubrutinib (ZR2) with sequential CAR-T cell as first-line therapy for high-risk LBCL.

Methods

We conducted a single-group trial of first-line therapy in adults with newly diagnosed high-risk LBCL. High risk LBCL included high-grade B-cell lymphoma (HGBL) with MYC and BCL2 and/or BCL6 translocations, HGBL not otherwise specified, and DLBCL not otherwise specified with an IPI score of ≥3. Patients received two cycles of ZR2 (rituximab 375 mg/m2 intravenous once on day 1, lenalidomide 25 mg once per day on days 1-21, and zanubrutinib 160 mg twice daily continuously of each 28-day cycle), and those with efficacy assessment of partial response or stable disease received sequential CD19 CAR-T cell therapy. The primary endpoint was complete remission (CR) rate after CAR-T cell therapy. Here we report initial results from this study.

Results

At the cut-off date of 31 June 2024, 23 patients were enrolled. The median age was 68.5 (range 50-84) years old. All the 23 patients finished 2 cycles of ZR2 which was well tolerated. Among them, 18 patients have received sequential CD19 CAR-T cell therapy. During CAR-T cell therapies, only 5 grade 1 cytokine release syndrome was observed, and no grade 2 or higher cytokine release syndrome or Immune effector cell-associated neurotoxicity syndrome was observed. The most common adverse events were hematological, which were thought to be associated with lymphodepleting chemotherapy and recovered in a short time without blood transfusion. After CD19 CAR-T cell therapy, 17/18 (94.4%) patients achieved CR. One patient with stable disease after CAR-T cell therapy received several lines of salvage chemotherapy, and finally achieved CR. After a median follow-up of 24 months, 2 patients experienced disease relapse at 6 and 16 months after CAR-T cell therapy, respectively. Both 2 patients received chemotherapy of RCHOP, and achieved CR again. As of date cut-off, all the 18 patients after CAR-T cell therapy remained in CR. The overall survival rate was 100% and the progression free survival rate at 1 year was 85.7%.

Conclusions

ZR2 with sequential CAR-T cell therapy has enabled chemo-free treatment in newly diagnosed high-risk LBCL. This treatment is characterized by high CR rate, high long-term survival, and low toxicity.
IGF2BP1 Orchestrates Post-Transcriptional Control of Globin Switching through Direct Binding to Globins and Their Regulators
Authors: Coyne Steven , Wu Ting , Vinjamur Divya , Hossain Mir , Bauer Daniel E.
Published: December 2024

Despite over a century of work on hemoglobinopathies, a comprehensive understanding of hemoglobin switching and factors regulating hemoglobin expression remains incomplete even as the first CRISPR/Cas9 gene therapy has obtained approvals for the treatment of transfusion-dependent β-thalassemia and sickle cell disease.

We studied factors related to the heterochronically silenced m6A RNA-binding protein IGF2BP1 which has been identified as a positive regulator of fetal hemoglobin expression by directly suppressing BCL11A expression. In contrast to this observation, post-transcriptional control by IGF2BP1 generally acts through improving the translation and mRNA stability of IGF2BP1 target mRNAs in an m6A-dependent manner. To better characterize the IGF2BP1-mediated mechanism responsible for control of fetal hemoglobin expression, we assessed whether BCL11A was required for IGF2BP1-mediated regulation of fetal hemoglobin and discovered a direct BCL11A-independent mechanism. Additionally, we characterize a heretofore poorly understood BCL11A-dependent mechanism for IGF2BP1 in regulating fetal hemoglobin expression.

We developed a luminescent transgenic reporter to disentangle BCL11A-dependent and newly identified BCL11A-independent mechanisms and used this system to discover that IGF2BP1 directly supports the translation of HBG mRNA in a manner requiring intact stop codon-proximal m6A motifs which are absent from the HBB mRNA.

Hemoglobin switching relies on a set of heterochronically regulated transcription factors, miRNAs, and RNA-binding proteins. This work identifies a novel mode of direct post-transcriptional regulation of HBG mRNA by a heterochronically regulated RNA-binding protein, IGF2BP1, that was previously described to regulate HBG indirectly via BCL11A. This work shows that HBG mRNA is bound directly by IGF2BP1 which supports HBG translation. HBB, not normally directly regulated by IGF2BP1, becomes responsive to IGF2BP1 overexpression upon addition of the HBG-derived stop codon-proximal m6A sites to an HBB transgene, a strategy with potential to be exploited for the improved expression of globin transgenes for hemoglobinopathy gene therapy.
Optimizing Chimeric Antigen Receptor Endodomains for Effector Function and Cell Expansion in iPSC-Derived T-Cell Therapies
Authors: Dower Chris , Heron Sean , Devaney Andrew , Wheeler John , Naso Amanda , Mohamed Wisam , Naso Michael , Carton Jill
Published: December 2024

Autologous chimeric antigen receptor (CAR) T-cell therapies are demonstrating success in the treatment of some hematological malignancies. Building on these clinical successes, we are developing engineered, allogeneic, iPSC-derived, iNK- and iT-cells to complement autologous cell therapies. In both hematological and solid tumor settings, antigen-driven expansion and maintenance of cytotoxic potential are important functional phenotypes to achieve therapeutic responses. CAR-mediated signaling is a primary and direct mechanism impacting these functionalities. To optimize CAR-driven function in our iPSC-derived platform cells, we generated a diverse panel of 45 different CAR formats that will be fully described within the data set and represents combinations of key immune effector cell signaling modalities, including: costimulatory domain families, cytokine support, ITAM multiplicity, hinge, transmembrane domain variants, and CD3 family signaling. This CAR panel was transduced via lentivirus into iPSC-derived T cells and assessed in vitro for their impact on the cell differentiation process, cytotoxic persistence in serial killing assays, and antigen-driven expansion. Through this, we identified a family of signaling components that mediate target-driven expansion and improved, persistent, cytotoxic function. This allowed for subsequent rational design of lead CAR constructs that combine signaling domain hits to further improve CAR-mediated expansion. The lead designs were characterized in a mouse tumor inhibition model and selected endodomains are incorporated into therapeutic designs for preclinical programs.
Real-World Use and Outcome of Ropeginterferon ALFA-2B in Polycythemia VERA Patients: A Multicenteric Study
Authors: Pugliese Novella , Di Perna Maria , De martino Ugo , Amico Valeria , Annunziata Mario , Martorelli Maria Carmen , Abagnale Davide Pio , Russo Roberta , Ferrentino Maria , Scarpa Alessia , Muriano Francesco , Martinelli Vincenzo , Pane Fabrizio
Published: December 2024

Ropeginterferon alfa-2b (Ropeg) emerges as a novel, long-lasting mono-pegylated IFN approved in Italy for treating polycythemia vera (PV) in adult patients without symptomatic splenomegaly and reimbursed for PV patient intolerant to hydroxyurea (HU), women of childbearing potential, and individuals with a history of skin cancer (SC).

Ropeg proved effective and safe in phase 3 trials, but more data is needed to understand its real-world use.

The study collected clinical data to assess the safety and efficacy of Ropeg in PV patients across five hematological centers in Campania. Ropeg was administered as per the data sheet, complete hematological response (CHR) was evaluated according to ELN criteria. Additionally, HCT, PLT, WBC and molecular responses were analyzed separately.

A total of 47 PV patients were enrolled, with a median age at Ropeg start of 60 years (range 28-90), and 70% (33/47) were male.

Thirty patients switched from prior HU after a median time of 11.8 months (range 1-299), due to intolerance. Three patients also transitioned from ruxolitinib, and one from peg-IFN. Ropeg was used as first-line therapy for 3 women of childbearing age, 2 patients with prior SK, and 8 male patients who were intolerant to phlebotomies and declined HU. The median time from PV diagnosis and Ropeg initiation was 45 months (range 1-345). In the year prior to starting Ropeg, the median number of phlebotomies was 2 (range 1-5). The median time on Ropeg was 12 months (range 1-21), with the median starting dose being 125 mcg every two weeks (range 50-150). At 12 months CHR rate was 64.7% for the 17 evaluable patients, HCT response was 82.4%. Ten patients (21.2%) required additional phlebotomy sessions during the treatment. At the same time point PLT and WBC responses were 82.4% and 100%, respectively and all but one had JAK2V617F allele burden reduction. CHR rate, HCT, PLT and WBC response increased over treatment as well as the median Ropeg dose that at 12 months was 200 mcg (range 100-250).

Three patients had grade 1 liver function test increases, which resolved after dose reduction, and one patient developed autoimmune thyroiditis without discontinuing treatment. After three months, four patients discontinued Ropeg for the following reasons: suspected Ropeg-related depression, gastrointestinal bleeding, worsening splenomegaly, autoimmune disease; one patient died from an unrelated cause.

Our findings indicate that Ropeg is generally well-tolerated and shows improvements in hematologic parameters. Although some response rates may vary, the overall trend is positive, with several measures reaching complete or near-complete response over time.
Distribution of ABO Blood Groups and Rhesus Factor in Critical Care Patients and Its Association with Mortality and Morbidity
Authors: Abuhaimed Jawahir , Abbas Amina , Bahatheg Mohammed , Altwaim Abdullah , Aloraier Hind , Almutairi Sulaiman , Hussein Tasneem , Al Gahtani Farjah Hassan
Published: December 2024

Background: ABO blood group and RhD factor have been associated with various health conditions, but their impact on critically ill patients remains unclear. This study aims to explore the distribution of these blood groups among critical care patients and their association with morbidity and mortality outcomes. Methods: We conducted a retrospective analysis of ICU patients at King Saud University Medical City. Patient data, including blood group, RhD status, and clinical outcomes, were collected from medical records. Statistical analyses were performed using the Kruskal-Wallis test, chi-square test, and multivariable regression models to assess associations, controlling for age and gender. Results: The study included patients from various ABO blood groups and RhD statuses. Our findings indicate no statistically significant differences in 28-day and 90-day mortality rates among the different blood groups. Other clinical outcomes, including days alive and free of invasive breathing, days alive and free of circulatory assistance, length of ICU stay, and SOFA scores, also showed no significant variations across ABO and RhD groups. The multivariable regression analysis confirmed the lack of significant associations between these blood types and mortality or morbidity outcomes. Conclusion: The study suggests that ABO blood group and RhD factor are not significant predictors of mortality or morbidity in critically ill patients. These findings highlight that other factors, such as underlying health conditions and severity of illness, are more critical in determining patient outcomes in the ICU. Further research in larger, multicenter settings is recommended to confirm these results and explore potential underlying mechanisms.
Immunomodulatory Effects of MEK Inhibition in Multiple Myeloma
Authors: Acharya Chaitanya , Jayasinghe Reyka G. , Hamilton Mark , Gnjatic Sacha , Kim-Schulze Seunghee , Cho Hearn Jay , Kumar Shaji , Mulligan George
Published: December 2024

Introduction

As part of the Multiple Myeloma Research Foundation (MMRF) MyDRUG umbrella clinical trial, we compared immune cell transcriptomic and proteomic signatures of multiple myeloma bone marrow samples from patients assessed by single-cell RNA sequencing (scRNA-seq) and mass cytometry (CyTOF). The goal of our study is to evaluate the effects of cobimetinib on the immune repertoire over the treatment course at single-cell resolution.

Methods

Bone marrow samples were collected from patients prior to therapy with cobimetinib (baseline/BL), after 2 cycles of cobimetinib therapy (EOC2), after 2 cycles of combination cobimetinib with IPd (EOC4) and at the end of treatment or at disease progression (EOT). All samples were processed into CD138+ myeloma cell and CD138- “immune cell” fractions for single-cell analysis. In this study, the CD138- “immune cell” fractions were transcriptionally profiled by single-cell RNA-sequencing (scRNA) at up to four time points.

Results

Transcriptome profiles of approximately 50,000 cells were analyzed across 22 samples from 9 patients with tumor MAPK Pathway mutations including: BRAF (p.K483Q, p.V600E, n=2); KRAS (p.Q61H, p.G12V, p.G12R, n=3); NRAS (p.Q61H, p.Q61R, p.Q61K, n=4). A subset of paired samples were subject to CyTOF with two immune related panels to supplement the single-cell transcriptomic findings. Focusing on one immune panel, 4 samples were profiled at baseline and end of cycle 2 while two samples were profiled only at baseline capturing over 600,000 cells spanning both timepoints. Our immune focused CyTOF panel contained 41 markers including T and NK cell markers (CD4,CD8A,CD3, GZMB, CD56), Myeloid lineage markers (CD14), B/Plasma markers (CD19, CD20, SLAMF7, CD38) among others. The largest populations captured by CyTOF included 260,000 neutrophils, 139,000 monocytes, 130,000 CD4/CD8 T cells, 43,000 NK cells, 40,000 dendritic cells and 22,000 B/plasmablasts/myeloma cells. Exhaustion markers, including LAG3 and TIGIT, are increased in CD8 and CD4 T cells after MEK inhibition, while PD1 is downregulated. PD-L1 is upregulated in both T cell compartments (CD4/CD8), monocytes and in residual myeloma cells profiled by both CyTOF and scRNA-seq. Both the IFNG and STAT1 pathways are concurrently upregulated post MEK inhibition in transcriptionally distinct CD8 T cell subsets profiled by scRNA-seq, suggesting a mechanism influencing PD-L1 expression.

Conclusions

These data indicate IFNG pathway activation by CD8+ T cells post MEK inhibition promotes PD-L1 expression on residual myeloma cells, suggesting the PD-L1 and IFNG signaling axes is activated in relapsed refractory myeloma patients after MEKi treatment. Here we present immunophenotypic and transcriptional profiling of longitudinal samples pre/post-MEK inhibition with multiple myeloma patients which can inform precision guided treatment options.
Analysis of Spatial and Temporal Dynamics of the Myeloma Immune Microenvironment
Authors: Fooksman David , Akhmetzyanova Ilseyar
Published: December 2024

Immune cell microenvironment is a critical component regulating myeloma progression and efficacy of various immunotherapies. Myeloma grows in complex foci or clusters within the bone marrow, and the composition and organization of these structures is poorly understood. Interestingly, we have found non-malignant long lived plasma cells also form dynamic clusters in the bone marrow suggesting a common survival niches. In patients, analyses of myeloma tumor microenvironment (TME) typically rely on imprecise bone marrow aspirates, which can collect random samplings of the marrow parenchyma as well as peripheral blood, and may contain variable and unknown proportion of myeloma foci or the relevant TME. To precisely profile immune microenvironment precisely within myeloma foci, we inoculated photoactivatable-GFP (PA-GFP) expressing immunocompetent mice with Vk*MYC GFP cells, and using two-photon microscopy in the tibia, we photo-labeled immune cells within the foci and analyzed by flow. We discovered the composition within foci was distinct compared to bulk bone marrow within the same marrow, which mimicked naïve marrow composition, suggesting unique TME were limited to myeloma foci volume. We also conducted longitudinal bulk analyses of total BM with tumor growth and found that distinct changes in myeloid cell composition with tumor burden, suggesting multiple phases in TME evolution. Finally, using genetic ablation of myeloid cells using DTR expressing mice, we found that macrophage and monocytes promoted tumor spreading to other bones, while plasmacytoid dendritic cells suppressed spreading. These findings highlight the dynamics and precision of the myeloma TME and how it regulates tumor progression.
Inferior Vena Cava Filters for Pulmonary Emboli in the Absence of Residual Deep Venous Thrombosis: A Survey of Current Physician Practices in Nova Scotia, Canada
Authors: Rasmussen Shannon , Manuel Eric Guy , Sandila Navjot , Shivakumar Sudeep
Published: December 2024

Introduction: There is a lack of evidence for the role of inferior vena cava (IVC) filters for pulmonary embolism (PE) in the absence of deep vein thrombosis (DVT). We surveyed physicians in Nova Scotia, Canada, to assess how lower limb ultrasound findings affect the decision to place an IVC filter in patients with pulmonary emboli and a contraindication to anticoagulation.Methods: A web-based survey comprised of five scenarios with fictional patients who had confirmed PE and different contraindications to anticoagulation was sent to 153 physicians who may see patients with venous thromboembolism. Each scenario had four sub-scenarios with different ultrasound findings including DVT, two SVTs varying in length, and no DVT. For each sub-scenario, physicians had to choose if they would insert an IVC filter or not. Results:The survey response rate was 33%. A total of 48 surveys were included, with 2 surveys excluded due to a survey error. Respondents were mostly general internists (n=20), followed by hematologists (n=9). Across total responses for each sub-scenario (n=204 DVT, n=412 SVT, n=207 no DVT), physicians would insert an IVC filter in 87% (n=177) of patients with a DVT, 64% (n=263) of patients with SVT, and 46% (n=96) of patients with no DVT or SVT. Conclusion:The majority of physicians included in this survey would place an IVC filter in patients with a DVT or SVT, whereas the absence of a DVT or SVT is reassuring to slightly over half of physicians. Further studies are required to guide recommendations on placement of IVC filters in patients with an acute PE without DVT or SVT and a contraindication to anticoagulation.
Development of an Early and Complete Examination Protocol for Women and Girls with Inherited Bleeding Disorders (WGBD)
Authors: Halimeh Susan , Daoud Mohamad Ayman , Schmitz Derya , Habermann Bjoern
Published: December 2024

Women and girls with bleeding disorders (WGBD) share the same bleeding symptoms and limitations in quality of life (QoL) as men, but they also confront unique physical and psychological challenges.1

Physically, WGBD experience joint problems, heavy menstrual bleeding (HMB), postpartum haemorrhage, excessive postoperative bleeding, epistaxis, easy bruising, and oral bleeding, also in haemophilia carriers with factor levels ≥ 40 IU/ml.2

Psychologically, they grapple with feelings of shame regarding HMB and guilt about bearing children with haemophilia.3These challenges can severely diminish QoL, yet they remain underappreciated.4

The primary physiological challenge lies in joint problems.5 WGBD, including haemophilia carriers, are at risk for subclinical joint bleeding, necessitating early detection.6 Inflammation can cause joint dysfunction before structural damage occurs, leading to pain, altered movement, and muscle degeneration, which destabilizes joints further and exacerbates pain.7

Early identification of joint issues is therefore crucial. Although ultrasound can detect joint problems, it is limited to recognizing clinical synovitis, a sign of already advanced joint problems.8

This creates two main barriers to comprehensive diagnosis and early treatment of WGBD: a) incompleteness, as diagnoses have previously concentrated on physiological aspects alone, and b) lateness, due to the absence of tools to identify joint changes before synovitis becomes clinically evident. Consequently, many with WGBD remain undiagnosed and untreated.

To address these barriers, we are creating a comprehensive assessment protocol for WGBD, evaluating all bleeding symptoms and their QoL impact. This protocol integrates new tools with existing diagnostic tools such as the Pictorial Blood Loss Assessment Chart (PBAC), Haemophilia Joint Health Score, ISTH Bleeding Score, and Haemophilia Early Arthropathy Detection with Ultrasound (HEAD-US).

The first new tool is a psychometric QoL questionnaire for WGBD, developed collaboratively with patients and health professionals based on established scientific questionnaire methodologies.

The second tool is the Noraxon system, which employs validated sensing methods, including inertial measurement units (IMUs), electromyography (EMG), and paedobarography. This system is highly regarded in biomechanical research, with over 400 peer-reviewed publications supporting its efficacy. A 2018 literature review highlighted 11 studies on biomechanical analysis in paediatric and adult patients with hemophilia, showing significant correlations with functional tests, ultrasounds, and MRI scans.

Interim test results are already available and show promising results for a smaller sample size of 26 women. Once our new psychometric questionnaire is validated, we will correlate its results with the other assessment toosl, test the whole protocol on a larger sample size and publish the results for peer review.
Non-Factor Therapy Reduces Burden of Care for Caregivers of Children with Severe Hemophilia a: A Longitudinal Cohort Study Using the Hemophilia Family Impact Tool (H-FIT v1.1)
Authors: O'Rourke Roxy , Bouskill Vanessa , Wakefield Cindy , Carcao Manuel , Abad Audrey , Blanchette Victor , Malcolmson Caroline
Published: December 2024

Background: A child's hemophilia diagnosis has an impact on the caregiver. Caregivers often need to administer routine prophylactic therapy to prevent bleeds. Historically, in patients with severe hemophilia A (PWH-A), this consisted of giving intravenous (IV) clotting factor concentrate (CFC) multiple times per week. With the advent of novel non-factor therapies (e.g., emicizumab), there is an option to deliver prophylaxis subcutaneously (SC) less frequently. The Hemophilia Family Impact Tool (H-FITv1.1) is a caregiver-reported questionnaire that measures the burden of caring for a child with hemophilia. This validated tool can be used to evaluate how the burden of care changes when switching from IV to SC therapy.

Aim: To evaluate the impact of emicizumab, as compared to IV clotting factor, on caregiver burden of care for PWH-A.

Methods: Caregivers of PWH-A (0-18 years) on routine prophylaxis completed the H-FIT at four time points: baseline (day of switch to emicizumab), and at 3, 6, and 12-months post-switch to emicizumab. Institutional research ethics board approval and informed consent from all participants were obtained. The H-FIT is scored from 0 to 100, with 100 representing the lowest family burden.

Results: Caregivers (N=35) of PWH-A (Mage=12.43 years, SD=4.15, 100% male, 97% infusing prophylaxis >twice/week pre-switch) completed the H-FIT. There was a non-statistically significant trend of improved burden of care following a switch to emicizumab (t11=-.81, CI=-14.86-6.86). Age stratification revealed this trend was consistent in children over the age of seven; however, the trend of improvement was not seen in the <7 years of age cohort.

Conclusion: There was a trend of improved burden of caring for PWH-A in caregivers following the child's switch from IV to SC therapy. This trend was not seen in the under 7 cohort, however sample size was limited, and further studies are needed to elucidate age-related differences in care.
Neurofilament Light Chain: A Potential Biomarker for Chemotherapy Induced Peripheral Neuropathy in Pediatric Adolescent Young Adults
Authors: Belsky Jennifer , Orgel Etan , Carter Allie , Lustberg Maryam , Roth Michael E. , Rhodes Steven D. , Dage Jeff , Schneider Bryan P
Published: December 2024

Background: Chemotherapy-induced peripheral neuropathy (CIPN) is a common treatment side effect in both pediatric and adolescent young adult patients (AYAs). CIPN causes significant clinical challenges resulting in chemotherapy dose reductions in attempt to improve painful symptoms, and additionally can result in decreased health-related quality of life and potentially impacting disease outcomes. CIPN detection in children and AYAs is a challenge, and current tools for CIPN detection rely on moderate to severe CIPN phenotypes in patients >12 years of age, with no patient reported outcomes (PROs) validated for younger children. Earlier, more sensitive tools for CIPN detection are need for younger patients. Neurofilament light chain (NF-L) is an emerging biomarker that has shown correlation with axonal damage in rat models, with the potential to serve as an early and sensitive biomarker of CIPN. This pilot study aimed to describe changes in NF-L in children and AYAs with CIPN.

Methods: This single-center, prospective study examined patients aged >10 years with leukemia or lymphoma who received a backbone chemotherapy regimen which included a tubulin toxin. CIPN was assessed by PROs with the FACT-GOG-ntx. Outcomes were evaluated at baseline prior to chemotherapy, day 1 of each chemotherapy cycle, and 6-12 weeks after last dose of chemotherapy. Values were reported as the Normalized Protein eXpression (NPX) per OLINK standard reporting on a log2. Negative N-FL values were excluded from percent change calculations. Values were calculated at baseline, first follow up, and at 90 days or the closest visit to 90 days.

Results: A total of 11 participants (aged 12-24 years) who received tubulin toxin chemotherapy backbones including to treat B-lymphoblastic leukemia (n=3), Hodgkin lymphoma (n=7), and mature B-cell lymphoma (n=1) were analyzed. 10 participants had a median % change in baseline NF-L of 67.94%. Mean NF-L levels increased from baseline timepoint (mean 2.32, median 2.16) to time point 2 (mean= 3.28, median 3.47) and remained increased at the follow-up visit closest to 90 days (mean 3.85, median 4.38). All participants had CIPN as evidenced by FACT-GOG-ntx scores, with majority of scores changing overtime from timepoint 1 (mean=1.27, median=0) to timepoint 2 (mean=3.55, median=3.00).

Conclusions: We report the first investigation of N-FL in the pediatric and AYA cancer setting as a potential novel biomarker for early and sensitive CIPN detection. These results are hypothesis generating and serve as the foundation for larger, future studies to continue investigating N-Fl as an early and objective detection measure of CIPN in this unique patient population.
CRISPR-TOF Platform: A Single-Cell Approach for Studying Kinase-Dependent Immune-Signaling Dynamics in Human Primary T Cells
Authors: Holzem Alessandra M. E. , Rovira-Clave Xavier , Drainas Alexandros , Reinhardt Hans Christian , Sage Julien
Published: December 2024

Introduction

Chimeric antigen receptor (CAR) T-cell therapies have improved the treatment of hematologic malignancies, but challenges such as activation and persistence limit broader application. Recent insights into the T-cell receptor (TCR) pathway revealed promising strategies for modulating activation kinetics, crucial for enhancing the efficacy of CAR T-cells. Here, we developed a single-cell approach for studying kinase-dependent signaling dynamics in primary human T cells, which unravels critical mechanisms of the T-cell activation cascade to better understand the genetic programs that determine T-cell response to (tumor) antigens.

Methods

We set up a platform called “CRISPR-TOF”, in which we combined arrayed CRISPR-Cas9 gene editing, an enhanced mass tag live-cell barcoding approach, and single-cell mass spectrometry (CyTOF). We knocked out nearly 1,000 kinases in primary human T cells and tracked the phosphorylation of 23 different proteins over 30 minutes, at 20 time points, post-stimulation with anti-CD3/CD28.

Results

The expected signaling patterns were observed in the analyzed components of the TCR pathway. Initially, proximal kinases such as SLP-76, ZAP70, and VAV1 exhibited rapid phosphorylation, which decreased after one minute. In contrast, phosphorylation of MEK1/2 and ERK1/2 emerged at 2 minutes, and the S6 ribosomal protein, peaked at nine minutes post-activation. Upon knocking out specific kinases, our study identified a range of novel targets that alter downstream TCR signaling. We categorized genes that either prolong or attenuate activation and identified those that modify signaling relationships or influence individual pathway components through independent mechanisms.

Conclusions

We have generated a dynamic, high-resolution map of T-cell signaling, facilitating targeted investigations of specific pathway components. Our results represent one of the most comprehensive assessments of T-cell signaling dynamics to date. This platform allows focusing on key signaling molecules and is thus a powerful complement to platforms such as Pertub-seq in which RNA changes are measured. It opens avenues for massively multiplexed single-cell assays for drug target discovery, delving deeper into immune signaling by studying entire signaling pathways. In the future, functional analyses of specific gene knockouts in samples will be essential to fully elucidate their roles in TCR activation, signal transduction, and resultant cellular functions. More broadly, these insights could be harnessed to optimize and expand cellular therapies.
Evaluation of the New Minion Chemistry and Duplex Basecalling for Immunoglobulin Gene Analysis in Chronic Lymphocytic Leukemia
Authors: Martinez-Serra Jordi , Nastasi Julieta , Ferrer Lores Blanca , Serrano-Alcalá Alicia , Moquera Adrian , Sampol Mayol Antonia , Terol Castera Maria Jose , Gutiérrez Antonio
Published: December 2024

Introduction:

The analysis of somatic hypermutation (SHM) in the immunoglobulin heavy chain variable region (IGHV) is crucial for managing chronic lymphocytic leukemia (CLL). Traditional methods such as Sanger sequencing (SS) or second-generation sequencing (NGS) are reliable but labour-intensive. This study aims to evaluate the efficacy and accuracy of the new MinION Kit 14 chemistry, nanopore sequencing (NS), using POD5 files and duplex basecalling with the Dorado software, compared to standard methods (SS).

Methods:

CLL patient samples were analysed using a custom pipeline that includes conversion of POD5 to BAM files with Dorado, BAM to FASTQ conversion, optional quality filtering, clustering by similarity with VSEARCH, generating consensus sequences, and querying IMGT/V-Quest for mutation status assessment. Statistical analysis for comparison between the NS and SS methods was performed using Spearman correlation test.

Results:

We included 15 CLL patient samples comparing our new NS method with SS. Results showed high concordance between NS and SS in terms of the percentage of SHM of IGHV (ρ=0.99; p<0.001) and 100% agreement in mutated/unmutated status definition, with NS detecting additional productive subclones. Even using Flongle flow cells, the quality of sequencing was sufficient to identify correct rearrangements with a single sample, ensuring high accuracy. The Kit 14 chemistry, which includes the E8.2 motor protein and R10.4.1 flow cells, achieved Q12-Q16+ accuracy with simplex data, significantly improving with duplex basecalling, making it suitable for SHM IGHV testing even for rapid results from individual samples. Generating 50,000 sequences per sample ensured sufficient coverage for reliable SHM detection and subclonal analysis, unlike Additionally, we provide results in less than 24 hours including just 1 hour sequencing, enabling a low-cost procedure that compares favorably with SS or second generation NGS.

Conclusions:

The new MinION chemistry and duplex basecalling offer a precise, faster, and cost-effective alternative to traditional SHM analysis methods. This approach enhances the detection of SHM and subclonal populations in CLL in the short-term, allowing for affordable real-time clinical decisions.
A Novel Small Molecule Drug-Based Approach for Treating Beta-Globinopathies
Authors: Liu Jun , Shen Ziyang , Dong Yanhua , Yu Na , Chai Li , Tenen Daniel G. , Bauer Daniel E. , Herman Adelaide , Pate Braden , Vanuytsel Kim
Published: December 2024

Beta-globinopathies, such as β-thalassemia and sickle cell disease, present significant global health challenges. Existing treatments, including γ-globin inducers, blood transfusions, gene therapies, and bone marrow transplantation, face hurdles of cost, efficacy, and tolerability. In addition to the known function of BCL11A, recent research underscores the crucial role of ZBTB7A/LRF, another transcriptional factor, in suppressing γ-globin during the erythroid globin switch. While transcription factors have historically been deemed “undruggable”, recent advancements in therapeutic modalities, like molecular glue degraders, have spurred interest in targeting them. Leveraging nearly two decades of expertise in transcription factor biology, access to compound libraries, and collaborations, we identified and characterized a novel small molecule drug that is capable of downregulating both ZBTB7A and BCL11A, two known γ-globin repressors.

Through in silico and phenotypic screens, we discovered SH6, which was originally characterised as a novel small molecule ZBTB7A degrader but later found to also downregulate BCL11A levels. The globin induction effect of SH6 was evaluated and validated using various erythroid cell models, including HUDEP2 cells, normal CD34 cell-derived erythroid cells and sickle cell patient iPSC models. γ-globin induction was assessed by qPCR, flow cytometry, and clinical grade HPLC analyzer. Mechanistic studies explored SH6's dependence on the proteasome machinery and the Cereblon E3 ligase system to explore the drug's mechanism of action as a molecular glue degrader. Synergistic effects with other disease-modifying agents, such as hydroxyurea, decitabine, and pomalidomide, were investigated.

Treatment with SH6 (0.25uM) led to a 6-fold increase in γ-globin expression in HUDEP2 cells compared to DMSO, surpassing other γ-globin-inducing treatments and without causing significant toxicity or erythroid differentiation. In healthy donor CD34 stem cell-derived erythroid cells, SH6 induced significant functional hemoglobin F without affecting differentiation markers at 0.25uM (11% vs 34% HbF, compared to DMSO by HPLC). Preliminary evidence suggests SH6's potential efficacy in hydroxyurea-unresponsive sickle cell patients, with increased trend of γ-globin expression observed with flow cytometry. SH6 effectively downregulated/degraded ZBTB7A/BCL11A in a proteasome-dependent manner within 24 hours of treatment, and its effect is abrogated in a CRBN-E3 ligase knockout HUDEP2 cell line. Synergistic effects with Decitabine were notable, showing a 4-fold increase in mRNA expression compared to SH6 alone and a 9-fold increase compared to Decitabine alone. Additionally, SH6 demonstrated the unique ability to reactivate more “primitive” forms of globins, epsilon and zeta globins, suggesting potential relevance for treating both alpha and beta thalassemia.
An Evaluation of Characteristics Associated with Academic Hematology-Oncology Fellowship Placement
Authors: Beltran Santiago , Matos Noboa Constangela , Yan John , Dhanaraj Nitya , Wei John X , Castellucci Enrico , Sharma Amy
Published: December 2024

Background:

Hematology is a discipline growing rapidly with advances in research and therapeutics. Thus, engagement with research, the understanding and conduct of it, is extremely important in the education and development of future hematologists. Academic fellowship programs affiliated with medical schools traditionally promote the development of academic hematologists. And so, understanding the factors associated with placement into academic fellowship programs is essential for tailoring training and support. This study aims to explore pre-fellowship characteristics that are associated with placement into academic hematology-oncology fellowships in the United States (US).

Methods:

We conducted a sampling of graduated fellow profiles available on the public websites of US hematology-oncology programs. To minimize over-representation from programs, even sampling of all available graduated fellows by year was done with a maximum of 10 per program. Characteristics analyzed included residency type (academic vs community), sex, international medical graduate (IMG) status, and pre-fellowship research productivity (number of Google Scholar publications in the 3 years prior to fellowship). Academics was defined as training in an institution associated with a medical school. Logistic regression modeling was utilized to identify characteristics associated with higher odds of initial academic fellowship placement.

Results:

We analyzed 519 fellows who graduated between 2007 and 2023, data was available from 63 out of the 181 programs in the US, representing 35% of all programs. Fellows of academic fellowships were more likely to have graduated from academic residencies (OR 13.7, 95% CI 7.39-26.4, p < 0.001). IMGs were more likely to be academic fellows compared to non-IMGs (OR 2.34, 95% CI 1.24-4.54, p = 0.010). Pre-fellowship scholarly productivity did not reach statistical significance (Tertile 2: OR 1.84, 95% CI 0.93-3.69, p = 0.081; Tertile 3: OR 1.80, 95% CI 0.86-3.80, p = 0.12). Sex did not show a significant association with fellowship placement (OR 1.14, 95% CI 0.63-2.05, p = 0.7).

Conclusions:

Our findings indicate that academic fellows had higher odds of being IMGs and coming from academic residencies. Pre-fellowship research productivity did not show a significant association with academic fellowship placement. An explanation is that the quantity of research does not necessarily imply quality; further investigation is needed to establish its significance. Additionally, IMGs might seek academic placements more frequently than community placements. These insights highlight critical areas where fellowship programs can enhance support for aspiring academic oncologists.