bluuebunny/crossref_metadata_2025 huggingface dataset:
A Retrospective Analysis of Orelabrutinib-Containing Regimen for Marginal Zone Lymphoma
Authors: Wang Hua , Liang Yang , Liang Dong , Feng Demei
Published: December 2024

Background: Marginal zone lymphoma (MZL) is the second most prevalent B-cell lymphoma in China, with a proportion of 12.4%. To date, there is no established standard systemic treatment for MZL. Traditional immunochemotherapy regimens such as BR and R-CHOP are commonly used, but they are associated with a high incidence (60-70%) of grade 3 or higher adverse events, which severely limits patient benefit. There is an urgent need for safer and effective treatment strategy. Orelabrutinib, a novel Bruton's tyrosine kinase inhibitor, offers higher kinase selectivity and a favorable safety profile. A phase II study of orelabrutinib monotherapy for relapsed/refractory MZL showed an objective response rate (ORR) of 58.9%, with a median duration of response of 34.3 months, and orelabrutinb has been approved in China for the treatment of previously treated MZL. This study retrospectively analyzed the efficacy and safety of treatment orelabrutinib-containing regimens for MZL.

Methods: This retrospective study included 15 MZL patients treated with orelabrutinib-containing regimens at our center between April 2023 and June 2024. Each treatment cycle was 28 days. The regimens included orelabrutinib monotherapy (O), orelabrutinib combined with rituximab (OR), orelabrutinib combined with rituximab and lenalidomide (OR2), and orelabrutinib combined with rituximab and pomalidomide (POR), every 28 day for a cycle. The specific dosages were: orelabrutinib 150 mg, once daily; rituximab 375 mg/m2, d1; lenalidomide 20 mg, d1-d21; pomalidomide 4 mg, d1-d21. The primary endpoint was objective response rate (ORR), and secondary endpoints included complete response rate (CR), progression-free survival (PFS), overall survival (OS), and safety.

Results: Fifteen patients (6 males, median age 60, range 33-69) were included in the study, comprising 14(93.3%) cases of extranodal MZL and 1(6.7%) case of nodal MZL. Nine(60.0%) patients were in stages III-IV, 3(20%) patients had an MZL-IPI score ≥3, 3(20%) had Hepatitis B virus, 1(6.7%) had Epstein Barr virus, and 1(6.7%) had Helicobacter pylori(Hp). One(6.7%) patient had received local treatment (anti-Hp therapy), and 6(40%) had received systemic treatment. For the current regimens, 2(13.3%) patients received orelabrutinib monotherapy, 4(26.7%) received OR treatment, 7(46.7%) received OR2 treatment, and 2(13.3%) received POR treatment. Among the 10 evaluable patients, the ORR was 80.0% (5 CR, 3 PR), with 2 (20.0%) cases of disease progression. Three (20.0%) patients experienced adverse events: 1 had neutropenia (Grade 3) and lymphopenia (Grade 2), 1 had a rash (Grade 2), and 1 had a hemoptysis (Grade 2). No BTKi-related adverse events were observed.

Conclusion:This retrospective study preliminarily confirmed that orelabrutinib-containing regimens are effective and safe for treating MZL. Further long-term follow-up and additional studies are anticipated to validate these findings.
Exploring Red Blood Cells As an Antigen Delivery System to Modulate the Immune Response Towards FVIII in Hemophilia a
Authors: Miranda Mariarosaria , Bradsma Eelke , Robben Lotte , Van Dender Helena , Van Alphen Floris , Fijnvandraat Karin , van den Biggelaar Maartje , Lacroix-Desmazes Sebastien , van Bruggen Robin , Voorberg J. J.
Published: December 2024

The main complication in hemophilia A treatment is the development of inhibitory antibodies against factor VIII (FVIII). Immune tolerance induction, the gold standard for eradicating anti-FVIII antibodies, is efficient in only 60-80% of cases. This underscores the need for more efficient induction of tolerance in hemophilia A patients with FVIII inhibitors. In this study we explored whether red blood cells (RBCs) can be utilized as antigen delivery system to modulate the immune response against FVIII. Two promiscuously HLA-DR presented peptides derived from the A2 and C1 domains of FVIII were fused to the TAT-cell penetrating peptide and incubated with RBCs. Biotinylated TAT-A2 and TAT-C1 peptides were found to interact with RBCs as shown by flow cytometry and imaging flow cytometry. Moreover, macrophages efficiently phagocyted TAT-FVIII peptide-treated RBCs. Using mass spectrometry based immunopeptidomics we established that TAT-FVIII peptides were presented on MHC class II of macrophages that phagocyted TAT-peptide pulsed RBCs. Specifically, the TAT-A2 peptide exhibited efficient processing and presentation on HLA-DR molecules. Importantly, incubation of TAT-C1 peptide treated RBCs loaded macrophages with a FVIII-specific T cell hybridoma led to a significant increase in IL-2 production, suggesting functional presentation of TAT-C1 derived peptides by macrophages. Our findings indicate that RBCs can serve as effective vehicle for the delivery of FVIII derived peptides to antigen presenting cells. The successful display of T cell epitopes on APC using ex vivo loaded RBC may be potentially utilized to modulate pathogenic immune responses such as observed in a subset of patients with hemophilia A.
Treatment with a Novel Small Molecule Dual Factor IIa/Xa Inhibitor Protects Against Coagulopathy and Organ Dysfunction in a Baboon Model of Staphylococcus Aureus Sepsis
Authors: Silasi Robert , Keshari Ravi Shankar , Abe Tomohiro , Regmi Girija , Lupu Cristina , Simmons Joe , Pfaff Nils , Heitmeier Stefan , Lupu Florea
Published: December 2024

Background: Sepsis-associated coagulopathy and disseminated intravascular coagulation (DIC) significantly contribute to organ failure and mortality in sepsis patients. Factors IIa and Xa are pivotal in the coagulation cascade, and their over-activation in sepsis leads to systemic hypercoagulable states. We hypothesized that in intensive care settings, comprehensive and controllable anticoagulation and organ protection can be achieved by targeting these two key proteases in the common coagulation pathway with the short-lived dual inhibitor BAY 3389934.

Objective: To assess the efficacy and safety of BAY 3389934, a novel short-lived, small molecule dual FIIa/Xa inhibitor, in reducing coagulopathy and protecting organ function in a baboon model of Staphylococcus aureus (S. aureus) sepsis.

Methods: The study employed a baboon model infused with heat-inactivated S. aureus (3.3 x 1010 bacteria/kg bw). Baboons were treated with BAY 3389934 (1-2 mg/kg bw, continuous IV infusion up to 8 hours post-bacterial challenge) starting at T0 (before) or 2 hours post-bacterial challenge. Various parameters, including markers of coagulation, fibrinolysis, organ function, and bleeding time, were assessed. Animals were kept in anesthesia for 9 hours and subsequently followed up to 48 hours after the start of bacterial challenge.

Results: S. aureus infusion induced robust activation of coagulation, as reflected by plasma levels of protease-serpin complexes (FXIIa-AT, Kal-AT, FXIa-AT, FVIIa-AT, and T-AT) and mirrored by fibrinogen and platelet consumption. S. aureus infusion led to organ dysfunction, as indicated by elevated vital signs, increased plasma ALT, total bilirubin, pancreatic amylase, plasma creatinine, and blood urea nitrogen, as well as histological evidence of kidney damage.

Treatment with BAY 3389934 significantly decreased the coagulation markers, blocking the activation of coagulation when given at T0 and strongly reducing it when given 2 hours after the challenge. Importantly, the treatment mitigated fibrinogen and platelet consumption without causing bleeding throughout the experiment. ROTEM analysis confirmed these findings.

Additionally, BAY 3389934 protected against organ dysfunction, as evidenced by reduced markers of liver (ALT), pancreas (amylase), and kidney (BUN, creatinine) damage, and decreased lactate levels. Histological analysis showed that the treatment prevented fibrin deposition in the kidney and lungs and necrosis of the kidney's tubular epithelium.

Conclusion: In a model of sepsis-induced coagulopathy in baboons, the dual FIIa/Xa inhibitor BAY 3389934 reduced the hypercoagulable state strongly and protected from organ damage. Further studies are warranted to explore its full therapeutic potential in the treatment of sepsis-associated DIC.
CRISPR Cas-9 Editing for Treating Sickle Cell Disease: A Systematic Review and Meta-Analysis
Authors: Desai Karna , Croce Phillip , Dadhania Nain , Mirza Wasique , Hanif Ahmad , Thalji Mousa , Sharma Rohit
Published: December 2024

Introduction

Exagamglogene autotemcel (exa-cel) represents a novel nonviral cell therapy aimed at reactivating fetal hemoglobin synthesis through the ex vivo application of clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 gene editing. This process targets autologous CD34+ hematopoietic stem and progenitor cells (HSPCs) at the erythroid-specific enhancer region of BCL11A. We performed a systematic review and meta-analysis of all clinical studies related to this therapy to evaluate its efficacy and safety in treating sickle cell disease (SCD).

Methods

A comprehensive search strategy was employed, incorporating keywords such as “CRISPR,” “exagamglogene autotemcel,” and “sickle cell disease.” The databases PubMed, Embase, and Medline were utilized to identify relevant studies. Inclusion criteria were based on clinical trials involving CRISPR/Cas9 gene editing for SCD treatment.

Results

A total of 34 patients were included in the analysis, comprising 26 males and 8 females with an average age of 25.51 years. The predominant genotype was βS/βS, observed in 30 patients. Key findings included an annualized rate of severe vaso-occlusive crises of 4.14, an average total hemoglobin level of 8.3 g/dL, and an average fetal hemoglobin level of 5.5 g/dL. The median number of mobilization cycles was 2. At 12 months, 32 out of 34 patients were free from vaso-occlusive crises, and 33 out of 34 were free from hospitalizations. The p-value for these outcomes was less than 0.0001, assuming a null hypothesis of a 50% response rate.

Discussion

Sickle cell disease (SCD) is a prevalent monogenic blood disorder characterized by severe pain, organ damage, and reduced life expectancy. Treatment options are notably limited, with hematopoietic stem cell transplantation from a matched related donor being the only curative approach. The ex vivo modification of autologous hematopoietic stem and progenitor cells, followed by transplantation of these genetically modified cells, offers a potential permanent cure applicable to all patients, regardless of donor availability or the risk of graft-vs-host disease. This review emphasizes the application of CRISPR/Cas9 gene editing in treating SCD, focusing on the correction of the β-globin (HBB) mutation and the induction of fetal hemoglobin to counteract cell sickling. We highlight significant accomplishments and challenges within this field, providing insights into the future potential of gene-editing therapies for curing SCD.
Investigating the Biomarker Role of Long Non-Coding RNAs in Pediatric Precursor B-Cell Acute Lymphoblastic Leukemia
Authors: Almási Laura , Horváth Ágoston , Oláhné Szabó Rita , Csanádi Krisztina , Pálóczi Krisztina , Kutszegi Nóra , Péterffy Borbála , Hegyesi Hargita , Erdelyi Daniel J
Published: December 2024

Introduction:

The diagnosis of childhood acute lymphoblastic leukemia (ALL) typically relies on cell detection from bone marrow (BM) and cerebrospinal fluid (CSF). Existing BM-based methods, while sensitive, are invasive, time-consuming, and laborious.

Central nervous system (CNS) involvement in ALL negatively impacts prognosis. However, routine CNS diagnostic methods yield a high percentage of false negative results. While blasts primarily adhere to the meninges in CNS involvement, molecules secreted by blasts might escape and diffuse more easily into the CSF. Long non-coding RNAs (lncRNAs) are non-coding polynucleotides with specific expression patterns for different cell types, maturation stages, and diseases.

Methods:

De novo precursor-B cell (pre-B) ALL patients under 18 years of age, diagnosed between 2016 and 2023 and treated as per the ALL IC-BFM 2009 protocol were enrolled. Samples from bone marrow aspirate (BM), peripheral blood (PB), and cerebrospinal fluid (CSF) were collected, centrifuged, and separate cell-containing and cell-free aliquots were stored.

An 89-target screening QPCR plate was built after a comprehensive literature search to find lncRNAs significant in ALL and B-cell development. Multiple RNA isolation methods were tested to suit the different body fluids. The lncRNA expression was measured in two experimental setups: 1) The QPCR screening was executed on 11 patients' day 0 and day 33 cell-free BM samples. 2) Five lncRNAs significantly overexpressed on day 0 were selected for droplet digital PCR (ddPCR) measurements on cell-free BM, PB, and CSF samples.

Results:

In the screening, five lncRNA targets had significantly higher expression in day 0 cell-free BM samples compared to day 33 samples, namely LINC01013 (p=0.007), AC002464.1 (p=0.013), AC002454.1 (p=0.035), BX293535.1 (p=0.006), LINC00958 (p=0.036). The 5-lncRNAs were well detected by ddPCR in cell-free BM and PB samples, but no significant signals were gained from the 300-700 μl volume CSF samples.

Conclusions:

The transcriptomic exploration in cell-free BM identified five lncRNAs as potential pre-B ALL biomarkers for further research. Low CSF lncRNA concentrations question their suitability as biomarkers in this compartment.
Dysregulation of Transcription Machinery Drives the Acquisition of BTK Inhibitor and Venetoclax Treatment Resistance in Mantle Cell Lymphoma
Authors: Gao Jing , Qiu Kevin , Danappa Jayappa Kallesh , Yin Qing , Wang Michelle , Hu Allen , Llaneza Danielle C , Blackburn Loryn L , Craig Jeffrey W , Shah Bijal D , Portell Craig A. , Williams Michael E , Zang Chongzhi , Tao Jianguo
Published: December 2024

Mantle cell lymphoma (MCL) is an incurable B-cell malignancy with current front-line treatment regimens. Dysregulated B-cell receptor signaling and resistance to apoptosis are major hallmarks in MCL pathobiology. Bruton's tyrosine kinase inhibitors (BTKi), such as ibrutinib, and BCL-2 inhibitors, such as venetoclax, have been used to treat refractory and relapsed (R/R) MCL. While these agents show clinical efficacy, patients frequently develop drug resistance with consequent poor clinical outcomes. We previously characterized the molecular mechanisms of drug response and resistance to BTKi and venetoclax single-agent therapies in MCL and identified the combination of BTKi and venetoclax as a novel approach for overcoming drug resistance and providing a more durable and synergistic response in MCL. Translation of our preclinical model was achieved through the establishment of a randomized multicenter phase 3 clinical trial with BTKi-venetoclax vs BTKi-placebo. Although this trial showed improved responses to BTKi-venetoclax therapy, the majority of patients ultimately developed resistance to this regimen. Thus, there is an urgent need to identify new therapeutic targets and to develop newer therapeutic agents and strategies to overcome and prevent the emergence of therapy resistance. With the goal of eradicating residual resistant tumor subclones and sustaining complete remissions, we here characterize the molecular genetic and epigenetic resistance mechanisms to BTKi-venetoclax combination therapy using multi-omics approaches including scRNA-seq and scATAC-seq in MCL. Our integrative multi-omics single-cell analysis reveals that epigenetic chromatin accessibility alterations underpin BTKi-venetoclax combination treatment resistance-associated transcriptome reprogramming and resistance evolution. We also validate the functional role of epigenetic regulation and transcription activation determinants (CDK7, CDK9) in BTKi-venetoclax resistance evolution. Furthermore, we establish that BTKi-venetoclax treatment reshapes the tumor microenvironment (TME) in MCL in favor of immune-responsiveness, which is evidenced by an enhanced susceptibility to immunotherapy. Taken together, this work demonstrates that the adaptive transcriptional machinery activation and immune-responsive TME state transition induced by BTKi-venetoclax are therapeutic vulnerabilities that may be effectively exploited by both targeted- (e.g., CDK9 inhibitors) and immune-therapies (e.g., CD19 CAR-T) to overcome MCL therapy resistance. This highly translational work has the potential to have an immediate impact on MCL treatment approaches through the rational design of new combination regimens.
Primary Pulmonary Follicular Lymphoma - a Rare Presentation of Non-Hodgkin Lymphoma
Authors: Panchal Megha , Arunachalam Janani , Memon Ahsan , Pilathodan Hashwin , Sripathi Keerthi , Rawal Shalin , Bagga Arindam , Sandhu Jujhar , Varma Samyukta , Atodaria Kuldeepsinh , Yarlagadda Bhavya Raja Gopal , Biswas Navanita , Parvataneni Tarun
Published: December 2024

INTRODUCTION

Primary pulmonary lymphoma (PPL) is an uncommon form of non-Hodgkin lymphoma that begins in the lungs. PPL originates directly within the lung tissue, unlike more prevalent lymphomas that frequently metastasize to the lungs from other sites. Lung primary non-Hodgkin lymphoma (NHL) is an exceptionally uncommon condition, representing approximately 0.4% of all malignant lymphomas. Only 3-4% of non-Hodgkin lymphoma (NHL) that does not start in lymph nodes is primary pulmonary lymphoma (PPL). This is less than 1% of all NHL cases and 0.5-1% of all primary lung cancers. Here we present a rare case of primary pulmonary follicular lymphoma characterized by bilateral pleural effusion.

AIM

The main objective in managing primary pulmonary lymphoma (PPL) is to promptly identify, precisely diagnose, and efficiently treat the condition to enhance patient outcomes. This entails increasing knowledge, employing sophisticated diagnostic methods, precisely determining the stage of the disease, and creating personalized treatment strategies.

CASE REPORT

A 74-year-old male patient presented with shortness of breath that was progressively worsening for 3 to 4 weeks. The patient denied having fever, chills, sweating, unexpected weight loss, or lymphadenopathy in the armpit, neck, or groin. The patient had no prior history of lung disease, and he did not use home oxygen or inhalers. The patient smoked up to 1 pack of cigarettes daily for 10 years but quit in 1995. He was initially treated with antibiotics, but the dyspnea persisted. A chest X-ray and CT angiogram were done at the time, which displayed bilateral pleural effusions, moderate on the right and small on the left, and no evidence of pulmonary thromboembolic disease. Therapeutic and diagnostic aspirations had been made. The cytology of the fluid was suggestive of exudative and found negative for malignant cells. Despite initial negative cytology for malignant cells, the patient returned 2 weeks later with dyspnea and increased bilateral pleural effusions from prior studies. A second thoracocentesis and analysis of the fluid, using flow cytometry and FISH, showed a single group of B cells that were positive for lambda CD10. This is a sign of follicular lymphoma, which is supported by changes in BCL6, BCL2, and t(14:18)/IGH-BCL2.

CONCLUSION

In summary, this case adds to the existing knowledge about the various ways NHL can present and supports the need for a comprehensive approach to caring for patients with pleural effusion. It also highlights the importance of early detection and prompt initiation of treatment for unexpected cancer cases, improving results through focused therapeutic approaches.
Molecular Genetics of Fletcher and Fitzgerald Traits
Authors: Adenaeuer Anke , Pinheiro Alessandro S. , Shim Young Jun , McCrae Keith R. , Häuser Friederike , Lämmle Bernhard , Schmaier Alvin H. , Rossmann Heidi
Published: December 2024

Deficiencies of the plasma protease zymogen prekallikrein (PK) and its substrate, high molecular weight kininogen (HK), were discovered in 1965 and 1975, respectively. Initially, they were referred to as Fletcher and Fitzgerald traits after the first patients identified. Shortly thereafter, other deficient subjects were discovered, such as Williams and Reid. Williams trait is a kininogen null plasma, whereas Fitzgerald trait contains low molecular weight kininogen but no HK coagulant activity. In 2003, Fitzgerald trait was found to have a small HK, missing domain 6. Its DNA was sequenced, and a deep intronic InDel of unknown significance in KNG1 was hypothesized to be causative. Due to technical progress and new scientific insights, we were now able to sequence and reevaluate cell-free DNA from these historic plasma samples using pyro- and next generation sequencing techniques. As a control, Williams plasma was confirmed to carry the homozygous KNG1 variant c.586C>T, p.Arg196*. Fletcher trait was found to be KLKB1 c.451dupT (rs560588447), the most prevalent African variant, leading to PK deficiency. Fitzgerald and Reid traits were found to have the same variant, c.1609_1610delCA p.Gln537Aspfs*25 (rs758429575), in KNG1s exon 10. This variant likewise only occurs in people of African descent, which is consistent with Reid's and Fitzgerald's reported ancestry. On immunoblots, Fitzgerald and Reid plasmas show a small HK with antibodies to HK's domain 5, but none with antibodies to domain 6, rendering the present frameshift mutation solely responsible for the residual HK in Fitzgerald trait. Thus, the first cases of PK and HK deficiency discovered have now been genetically clarified. They confirm that no missense mutations are currently known to cause HK deficiency. Further, they emphasize the comparatively increased prevalence of PK and HK deficiency in sub-Saharan Africans. This finding suggests that widespread diseases may have influenced progression of these mutations in this population.
Brentuximab Vedotin-Based Treatment in Patients with CD30-Positive Lymphoma: A Retrospective Analysis
Authors: Li Yixue , Zeng Ruolan , Xia Chen , Li Yajun , He Yizi , Wang Caiqin , Zhou Hui
Published: December 2024

Background:

Brentuximab vedotin (BV) is an antibody-drug conjugate targeting CD30 which has been approved for the treatment of adult patients with CD30-positive lymphomas, specifically relapsed or refractory systemic anaplastic large cell lymphoma (sALCL) and classical Hodgkin lymphoma (cHL). This study presents a retrospective analysis of BV's efficacy and safety in CD30-positive lymphoma patients at our institution.

Methods:

This study included CD30-positive lymphoma patients treated with BV from July 2020 to April 2024. The primary endpoint was the overall response rate (ORR). The secondary endpoints were complete response (CR) rate, progression-free survival (PFS), overall survival (OS) and adverse events (AEs). Efficacy was assessed based on the 2014 Lugano Conference, and treatment-related AEs were evaluated according to CTCAE5.0.

Results:

A total of 66 patients were analyzed. The median age at diagnosis was 40 years (range: 15-79 years), with 43 (65.1%) being male. The patient cohort included 39 (59.1%) with cHL, 26 (39.4%) with peripheral T-cell lymphoma (PTCL), and 1 (1.5%) with primary mediastinal large B-cell lymphoma (PMBL). Two (3.0%) had previously undergone stem cell transplantation. The median number of prior chemotherapy lines was 0.5 (range: 0-7). B symptoms were present in 24 (36.4%). 5 (7.6%) had an ECOG score ≥ 2 and 43 (65.2%) exhibited extranodal disease. 22 patients (33.3%) had spleen involvement and 38 (57.6%) had elevated erythrocyte sedimentation rate at diagnosis. Disease progression was noted in 26 (39.4%) prior to BV treatment. BV was combined with chemotherapy in 65 patients (98.5%), while 1 patient (1.5%) received BV monotherapy. The median follow-up time was 13.75 months. There were 6 deaths due to tumor progression, 1 due to low platelet count, and 1 patient due to pulmonary embolism. The median PFS and OS were not reached. The ORR was 89.4% (59/66) and the CR rate was 66.7% (44/66). Common AEs included leukopenia (30.3%) and anemia (10.6%), with other AEs being abnormal liver function, nausea, fatigue, gastrointestinal reactions, numbness of the limbs, rash, decreased appetite, etc. Grade 3-4 AEs comprised of leukopenia (15.1%).

Conclusions:

BV shows significant efficacy in treating CD30-positive lymphoma with acceptable safety, supporting its role as a valuable therapeutic option for these patients.
Pathway-Specific Dysregulation in Follicular Lymphoma Correlates to Histological Transformation
Authors: Enemark Marie Hairing , Wolter Katharina , Engelbrecht Hybel Trine , Andersen Maja Dam , Sørensen Emma Frasez , Hindkær Linnea Meier , Lauridsen Kristina Lystlund , Madsen Charlotte , Plesner Trine , Hamilton-Dutoit Stephen , Honoré Bent , Ludvigsen Maja
Published: December 2024

Follicular lymphoma (FL) is the most common low-grade non-Hodgkin lymphoma. Despite its indolent nature, FL carries an inherent risk of histological transformation (HT) to a more aggressive lymphoma. Existing clinicopathological features and biomarkers are insufficient to predict HT, indicating the need for more robust biological predictors. Previously, we used proteomics to identify differentially expressed proteins in FLs with and without subsequent HT. This study sought to further evaluate these by investigating identified proteins acting in important cellular pathways that could be divided into five major classes: (i) apoptotic regulation (BID), (ii) cell cycle progression (CDC26, CDK6, SRSF1, SRSF2), (iii) GTPase signaling (IQGAP2, MEK1), (iv) cytoskeletal rearrangement and cellular migration (ACTB, CD11a, MMP9, SEPT6), and (v) immune processes (CD81, IgG, MPO, PIK3AP1). We analyzed protein expression in pre-therapeutic lymphoma samples from 48 FL patients stratified as either non-transforming FL (nt-FL, n=30) or subsequently-transforming FL (st-FL, n=18) with histologically-confirmed transformation to diffuse large B-cell lymphoma. Paired transformed lymphomas (tFL, n=18) were also analyzed. We used immunohistochemistry and digital image analysis to quantify protein levels. In all five pathway classes, several proteins were differentially expressed between samples (p<0.05). Interestingly, we found correlation between expression levels of several proteins, indicating a complex interplay between the pathways. Differential expression of most proteins was also associated with shorter transformation-free survival (p<0.05). These findings emphasize underlying differences in FL biology, highlighting deregulation of interconnected cellular pathways, and suggest the potential mechanistic importance or predictive role of impaired cellular homeostasis, and altered microenvironment in FL transformation.
Bone Marrow Endothelial Progenitor Cells Are Involved in Resistance to Tyrosine Kinase Inhibitors in Chronic Myeloid Leukemia
Authors: Yu Shunjie , Yuan Mengyao , Bao Mei , Zhang Xiaoshuai , Jiang Qian
Published: December 2024

Background Tyrosine kinase inhibitor (TKI) resistance is a significant challenge in chronic myeloid leukemia (CML). Data of endothelial progenitor cells (EPCs) from bone marrow microenvironment in TKI resistance are limited.

Aims To explore whether EPCs are associated with TKI resistance in patients with CML.

Methods EPCs were isolated from TKI-sensitive or resistant patients with CML using Ficoll-Hypaque density-gradient centrifugation for co-culture with K562 cells ± TKIs. The number of EPCs and apoptosis level of K562 were detected by flow cytometry. Tube formation and migration assays were performed to evaluate the functions of EPCs. RNA sequencing was employed to analyze gene expression profiles of EPCs and K562 cells.

Results Compared with those from TKI-sensitive patients, the number of EPCs from TKI-resistant patients was reduced (0.07% ± 0.01% vs. 0.11% ± 0.02%, p = 0.139), tube formation (17005 ± 3561 vs. 41084 ± 3693, p = 0.003) and migration ability of the EPCs were impaired (103.2 ± 17.41 vs. 202.0 ± 14.59, p < 0.001). Moreover, EPCs from TKI-resistant patients exhibited higher level of reactive oxygen species (2683 ± 158 vs. 2170 ± 320, p = 0.211). Co-culture experiments demonstrated that contact-dependent endothelial-mediated resistance to imatinib was observed in the EPCs from TKI-resistant patients, characterized by reduced K562 apoptosis (22.51% ± 1.52% vs. 36.58% ± 1.88%, p < 0.001) and G0/G1 phase (31.30% ± 3.05% vs. 43.10% ± 3.59%) compared with that from TKI-sensitive patients, and similar phenomenon also occurred when nilotinib or ponatinib replaced imatinib. KEGG pathway enrichment analyses showed that cell adhesion, immune and inflammatory related pathways were enriched in EPCs from TKI-resistant patients and K562 cells co-cultured with TKI-resistant EPCs.

Conclusions Our findings suggested that EPCs from TKI resistant CML patients were dysfunctional which might contribute to TKI resistance.
Ptcy Overcomes HLA Mismatch in Unrelated Donor Transplant for Acute Lymphoblastic Leukemia: A Study from the ALWP of the EBMT
Authors: Sanz Jaime , Ferhat Allain-Thibeault , Kulagin Aleksander , Kröger Nicolaus , Rovira Montserrat , Lazzari Lorenzo , Arat Mutlu , Montoro Juan , Versluis Jurjen , Reményi Péter Pál , Sica Simona , Blaise Didier , Saif Muhammad , Vydra Jan , Fagioli Franca , Nur Erfan , Giebel Sebastian , Piemontese Simona , Mohty Mohamad , Ciceri Fabio
Published: December 2024

Background: Post-transplant cyclophosphamide (PTCy) seems to mitigate the disparity in outcomes between use of matched (MUD) or mismatched Unrelated Donors (MMUD) hematopoietic stem cell transplantation (HSCT), but there are no specific studies in patients with acute lymphoblastic leukemia (ALL).

Methods: We retrospectively analyzed the characteristics of 8/8 MUD and 7/8 MMUD that may affect transplant outcomes in patients with ALL in complete remission (CR) who received a first peripheral blood allogeneic HSCT with PTCy between January 2010 and December 2022, registered in the EBMT database. The primary endpoint was leukemia-free survival (LFS).

Results: Overall, 350 patients were included with a median age of 37 years (range, 18-76), and 231 (66%) were in first CR. 280 (80%) patients had B-cell ALL of which 101 (29%) were Philadelphia positive. Regarding donors, median age was 28.8 years (range, 18-57), 237 (68%) were MUDs and 113 (32%) were MMUDs. Patient, donor and transplant characteristics were equally distributed across donor types except for older donor age, more frequent recipient male sex (58%) and use of TBI containing myeloablative conditioning in MMUD transplants (55%). The 180-day cumulative incidence of acute graft-versus-host disease (GVHD) grade II-IV and III-IV was 28% (95% CI 23-33) and 9% (95% CI 6-12), respectively, while the 2-year cumulative incidence of chronic and chronic extensive GVHD was 28% (95% CI 23-34) and 12% (95% CI 9-17), respectively. After a median follow-up of 24 months, the cumulative incidence of relapse and non-relapse mortality and the probabilities of LFS, overall survival (OS) and GVHD-free, relapse-free survival (GRFS) were 24% (95% CI 19-29), 15% (95% CI 11-19), 62% (95% CI 56-67), 73% (95% CI 67-78) and 48% (95% CI 42-54), respectively. The use of MUD or MMUD did not have a significant impact on LFS (hazard ratio [HR] 0.84; 95% CI 0.56-1.24) or any other transplant outcomes. Considering other donor related variables, the use of female donor to male recipient increased the risk of acute GvHD grade II-IV (HR 1.78; 95% CI 1.05-3, p=0.03) and the use of older donor age increased grade III-IV (HR 1.7; 95% CI 1.1-2.64, p=0.02). Additionally, CMV-neg donor for CMV-neg recipient was associated with improved LFS (HR 0.55; 95% CI 0.32-0.96, p=0.034) and overall survival (HR 0.52; 95% CI 0.28-1,p=0.49).

Conclusions: For patients undergoing HSCT from an unrelated donor with PTCy for ALL in CR the use of MMUDs does not negatively affect outcomes. Other donor related variables such as age, gender and CMV serostatus might be prioritized over HLA match considerations.
Comparative Efficacy of Carfilzomib, Lenalidomide and Dexamethasone (KRD) Vs. Daratumumab, Bortezomib, Lenalidomide and Dexamethasone (DVRD) in Newly Diagnosed Multiple Myeloma: A Systematic Review and Meta-Analysis
Authors: Wang Yuqi , Huan Chen , Hanzhen Zhang , Zhang Shanshan , Yuan Youhai , Feng Ru , Wei Yongqiang , Wei Xiaolei
Published: December 2024

Background: Daratumumab in combined with bortezomib, lenalidomide and dexamethasone (DVRD) or carfilzomib, lenalidomide and dexamethasone (KRD) has been recommended as first-line induction regimen in patients with newly diagnosed multiple myeloma (NDMM). However, the superiority of these two regimens remains unknown. Given the lack of head-to-head clinical trials, this study systematically reviewed the clinical studies regarding DVRD and RVD regimen in the treatment of NDMM to explore the better choose of the first-line therapy.

Methods: We searched in the Cochrane Library, PubMed, Embase, and Web of Science databases to collect studies containing DVRD or KRD for NDMM, then we performed pooled and meta-analysis to compare overall response rate (ORR), complete remission(CR) rate, stringent complete remission (sCR) rate, minimal residual disease (MRD)-negative status, progression-free survival (PFS), overall survival (OS) and treatment-related discontinuation rate, grade 3-4 adverse events rate between DVRD and KRD regimen by using R 4.3.3 software.

Results: Of 3570 studies screened, 6 randomized controlled trial and 8 single-arm clinical trial were eligible including 483 patients treated with DVRD and 1509 treated with KRD. We found a statistically non-significant difference in therapeutic response including ORR, CR, sCR and MRD-negative status between DVRD and KRD regimen. Compared to KRD regimen, patients treated with DVRD were associated with improved PFS (not reached vs. 58.24 months, pool HR 0.41; 95%CI 0.32-0.53) and OS (both were not reached, pool HR 0.64, 95%CI 0.45-0.90). The grade 3-4 adverse events rate between DVRD and KRD regimen was non-significant statistical different while the therapy discontinuation rate led by adverse events was significantly lower in patients treated with DVRD than those treated with KRD (2% vs. 6%, P=0.01).

Conclusion: This meta-analysis indicated that DVRD regimen as first-line induction treatments could bring patients with NDMM with better outcomes and lower treatment-related discontinuation rate than KRD regimen.
Red Cell Alloimmunization Rate in Patients with Sickle Cell Disease
Authors: Asante Akua , Henrichs Kelly , Blumberg Neil , Love Tanzy
Published: December 2024

Background: Sickle cell disease(SCD) is a single gene disorder resulting in dysfunctional hemoglobin tetramers that polymerize causing vaso-occlusion (VOC), hemolysis, endothelial damage and inflammation leading to pain and end-organ damage. Red blood cell (RBC) transfusions are vital despite recent advances in treatment. RBC alloimunization complicates treatment with rates of up to 47% limiting availability of compatible RBC units. The etiology of alloimmunization is multifactorial; including donor recipient racial differences, transfusion volume, non-leukoreduced RBCs, infection or VOC during transfusion, age, and pregnancy. Two studies have shown decreased alloimmunization rates in leukemia patients post implementation of universal leukoreduction and ABO identical (ABO-ID) blood product transfusions. The aim of this study is to analyze the impact of ABO-ID policy on alloimmunization rates in the SCD population.

Methods: A retrospective chart review was performed during June 1, 2004 to Dec 31, 2023. 437 patients with SCD were identified. Data was gathered on transfusion history, alloimmunization status, and antibodies identified.

Results: 257 patients were transfused and of that 211 were transfused prior to ABO-ID policy implementation in April 2005 (pre ABO-ID) and 49 after (post ABO-ID). In regard to SCD genotype, 67% were SS, 22% SC, and 11% SBthal. The alloimmunization rate for pre ABO-ID is 30% and post ABO-ID is 4% (p=0.0006). Among the pre ABO-ID group, 182 received ABO-ID transfusions with an alloimmunization rate of 27% compared to a rate of 48% for those transfused with ABO non-identical (ABO non-ID). The post ABO-ID group received significantly less transfusions and were younger compared to those transfused with ABO-ID RBC from the pre ABO-ID group (p<0.05 and p<0.05 respectively).

Conclusion: The alloimmunization rate in patients transfused prior to the ABO-ID policy was higher compared to those transfused after. These results suggest that avoiding the infusion of ABO non-ID antibody, even in small amounts, as with group O RBC transfused to non-O patients, may reduce the rate of alloimmunization in patients with SCD. The increased alloimmunization rate in patients transfused with ABO-ID RBCs prior to ABO-ID policy can be explained by record review limitations due to patients seeking care at outside facilities where transfusion history access was not possible, older age, and higher number of RBC units transfused. This study highlights the need for a better understanding of the risks and pathophysiology of alloimmunization in patients with SCD. Prospective multicenter studies are needed to further evaluate the long term effect of ABO-ID transfusions in this patient population.
Evaluating Clinical Endpoints for C5 Inhibitors
Authors: Ma Mark
Published: December 2024

Using free C5 levels as surrogate endpoints in complement therapies, especially for marketed Eculizumab and Ravulizumab, is grounded in the drug's specific mechanism of action and the correlation between free C5 levels and clinical outcomes. In studies targeting PNH, trials have shown that Ravulizumab maintains free C5 levels below the established threshold (<0.5 μg/mL).

RLYB116 is an affibody with a high affinity for blocking C5. To determine the free C5 levels from post-dose time points, a free C5 assay was developed on the Gyrolab platform. In the RLYB116 first-in-human (FIH) study, the measured free C5 levels seemingly deviated from the predicted post-dose free C5 levels. This discrepancy was further characterized by spiking varying concentrations of RLYB116 in human serum with endogenous C5. The results indicated that free C5 levels were overestimated when compared to free C5 concentration quantified using the same technology for Ravulizumab studies. Compared to Ravulizumab, the faster Koff for RLYB116-C5 complex likely resulted in the assay capture reagent outcompeting for C5 from the RLYB116, resulting in an apparently higher concentration of unbound C5 (i.e., free C5).

Additional experiments were conducted to determine the correlation between hemolysis levels and free C5 levels in samples treated with RLYB116. Experiments included spiking serial concentrations of RLYB116 or Ravulizumab into 100% C5 human serum and then measuring the free C5 and hemolysis levels and, adding serial concentrations of C5 and RLYB116 or Ravulizumab into C5-depleted human serum and measuring the hemolysis levels. Despite a higher measured free C5 value in RLYB116-treated samples, a similar level of hemolysis reduction (>90%) from baseline was observed, indicating complete complement inhibition is achieved in samples treated with RLYB116 or Ravulizumab.

The experimental evidence suggests that the free C5 levels obtained from RLYB116 study were significantly overestimated and it did not reflect the true complement inhibition by RLYB116. Furthermore, based on the totality of data from ex-vivo and clinical results, we conclude that RLYB116 demonstrated comparable or better potency compared to Ravulizumab, with a once weekly/self-administered dose. Additional pharmacodynamic markers, instead of free C5, should be used to support RLYB116 clinical development.
Development of Novel Peripheral Blood Count-Based Prognostic Index for Multiple Myeloma
Authors: Ferri Grace M. , Yildirim Cenk , Park Joseph , Do Nhan V. , Brophy Mary T. , Munshi Nikhil C. , Fillmore Nathanael R. , Edwards Camille V.
Published: December 2024

In multiple myeloma (MM), clonal plasma cells evade immune surveillance and proliferate within an inflammatory bone marrow microenvironment. During pathogenesis, monocyte-derived tumor-associated macrophages help myeloma cells to avoid apoptosis; effector T cell lymphocytes undergo senescence to enable preferential upregulation of their immunosuppressive regulatory T cell counterparts; and myeloma cells interact with hematopoietic stem cells to inhibit downstream megakaryocyte differentiation into platelets.1 Our prior research has suggested a prognostic role for the absolute monocyte count (AMC), absolute lymphocyte count (ALC), and platelet count as biomarkers. We have shown that abnormal AMC and decreased ALC and platelet count are associated with decreased overall survival (OS) among patients with MM at diagnosis and follow-up. Based on the easy accessibility of these peripheral blood markers and need for dynamic prognostication among patients with MM undergoing novel therapies, we created a categorical peripheral blood-based prognostic index for MM incorporating AMC, ALC and platelet count. We retrospectively evaluated this index in 10790 patients diagnosed with MM between 2000 and 2019 at Veterans Administration hospitals using cell counts obtained closest to diagnosis and up to 2.5 years thereafter. Patients were stratified into 3 levels based on a logarithmic risk score, where those with the lowest counts across all cell lines (AMC, ALC, and platelet count) were in level III and those with counts approaching normalcy were in level I. At diagnosis, 8% of patients were in level III and had the highest risk of death (hazard ratio [HR] 2.04; 95% confidence interval [CI] 1.85-2.24; P < .01) compared to those in levels I (HR 1.15; 95% CI, 1.08-1.23; P < .01) and II (HR 1.37; 95% CI, 1.28-1.46; P < .01). Moreover, the risk of inferior OS associated with level III status persisted during treatment and follow-up and existed independently of other known risk stratification tools (including Charlson comorbidity index, lactate dehydrogenase, International Staging System classification). With further validation, our findings support the use of our novel prognostic index, which potentially reflects the underlying tumor microenvironment, as an available tool for risk stratification during the MM disease course.
Efficacy and Safety of BeEAM Regimen for Autologous Hematopoietic Stem Cell Transplantation in Lymphoma Patients: A Retrospective, Multicenter Clinical Study
Authors: Hu Renzhi , Liu Jun , Li Qiying , Liu Yao
Published: December 2024

Objective: To evaluate the efficacy and safety of the BeEAM (Bendamustine + Etoposide + Cytarabine + Melphalan) conditioning regimen for autologous hematopoietic stem cell transplantation (ASCH) in lymphoma patients.

Methods: This study is a retrospective, multicenter clinical study, analyzing the clinical data of 167 lymphoma patients (NHL=152, HL=15) who underwent ASCH after BeEAM conditioning regimen from January 2021 to March 2024 in five tertiary teaching hospitals including the Cancer Hospital affiliated to Chongqing University. The clinical characteristics, transplant-related adverse reactions, hematopoietic reconstitution, efficacy, and survival status of the patients were assessed.

Results: The median age of the patients was 49 years (11-71 years), mainly male (58.0%). Most patients had aggressive B-cell lymphoma, including diffuse large B-cell lymphoma (53.2%), Burkitt's lymphoma (3.0%), as well as mantle cell lymphoma (4.2%), Hodgkin's lymphoma (8.9%), follicular lymphoma (1.2%), and peripheral T-cell lymphoma (16.7%). Among them, 132 patients (78.90%) were in stages III/IV, 111 patients (66.40%) had intermediate-high/ high risk IPI scores, and 60 patients (36.00%) had large tumor masses. Before the intensive therapy, the median number of chemotherapy courses was 2 times (1-9 times), and 78 patients (46.6%) received transplantation after second-line chemotherapy. The median number of CD34+ cells infused after conditioning was 5.14 (2.04-15.68). After transplantation, 8 patients had poor platelet engraftment, and the rest achieved hematopoietic reconstruction. The median time for good engraftment of neutrophils and platelets in peripheral blood post-transplant was 10.42 days (6-30 days) and 11.88 days (5-34 days), respectively.

The median follow-up time of the patients was 15.0 months (7.0, 48 months). During the follow-up period, 71 patients had PD (progressive disease), 31 patients died, the 2-year PFS (progression-free survival) was 56.1%, and the 2-year OS (overall survival) was 84.1%. Among the 167 patients during the conditioning process, 31 patients (18.5%) had infections, 13 patients (7.8%) had severe gastrointestinal reactions, 6 patients (3.6%) had acute kidney injury, and 3 patients (1.8%) had cardiac toxicity.

Conclusion: The BeEAM regimen is effective and well-tolerated for conditioning in autologous hematopoietic stem cell transplantation for lymphoma patients.
Inhibition of p300/CBP Overcomes BTK Inhibitor Resistance in MCL By Counteracting IL-6/JAK/STAT3 Signaling
Authors: Tashiro Yusuke , Shirakawa Kotaro , Arima Hiroshi , Nomura Ryosuke , Horisawa Yoshihito , Emani Stanford , Takeuchi Suguru , Caroline Jelagat , Kazuma Yasuhiro , Matsui Hiroyuki , Matsumoto Tadahiko , Yusa Kosuke , Takaori-Kondo Akifumi
Published: December 2024

Introduction: Although Bruton's tyrosine kinase inhibitors (BTKi) are effective against relapsed refractory mantle cell lymphoma (MCL), their limited efficacy and resistance remain clinical issues. Previously, we conducted CRISPR/Cas9 screening and found that p300/CBP, lysine acetyltransferases, are the genes responsible for one mechanism of BTKi resistance. A p300/CBP inhibitor A-485 with ibrutinib synergistically induced apoptosis and cell cycle arrest in ibrutinib-resistant MCL cells.

Methods: To elucidate the synergistic mechanism, we conducted RNA sequencing using an ibrutinib-resistant MCL cell line, Granta519, and compared the transcriptomic profiles of Granta519, treated with ibrutinib, A-485, or both.

Results: Genes related to the G2M checkpoint were upregulated when treated with the combination therapy. Pathway enrichment analysis revealed that the combination therapy downregulated pathways including Myc targets, TNF-α signaling via NF-κB, E2F targets, mTORC1 signaling, and IL-2/STAT5 signaling activation. Moreover, we observed that addition of A-485 restored the efficiency of ibrutinib to suppress downstream BCR signaling in Granta519 cells. We also detected IL6/JAK/STAT3 signaling activation by ibrutinib monotherapy, and the addition of A-485 significantly counteracted IL6/JAK/STAT3 signaling activation, suggesting that IL6/JAK/STAT3 signaling contributes to ibrutinib resistance.

Pathway analysis showed that Myc was the most suppressed target by the combination therapy, reducing Myc mRNA levels by 90%. Using the MTS assay, we confirmed that Myc overexpression impaired the therapeutic effect of A-485 and significantly reduced the synergistic therapeutic effect of the combination therapy. This impairment suggests that the combination therapy achieves therapeutic effects by attenuating Myc function. We next focused on IL6/JAK/STAT3 signaling and tested ruxolitinib to inhibit JAK1/2. Interestingly, the IC50 value of ruxolitinib was lower in ibrutinib-resistant cells than ibrutinib-sensitive cells. To confirm that A-485 suppresses IL6/JAK/STAT3 signaling, we also confirmed that A-485 inhibited the nuclear translocation of STAT3 and the acetylation of STAT3 by Western blotting.

Conclusion: p300/CBP inhibition overcomes BTKi resistance in MCL through IL-6/JAK/STAT3 signaling inhibition and attenuation of Myc transcription. Thus, p300/CBP inhibition could be a novel strategy for treating BTKi-resistant MCL.