In this systematic review, we aimed to assess the diagnostic test accuracy (DTA) of abdominal fat pad sampling and bone marrow biopsy for immunoglobulin light-chain (AL) amyloidosis using organ biopsy as the reference standard. We searched the literature using PubMed, Embase, and CENTRAL databases from inception through March 2024. We used QUADAS-2 to assess the risk of bias and the GRADE approach to assess the certainty of evidence (COE). After deduplication, 29,237 records were screened, and 31 studies were included. Twenty-five studies (n=4,649) reported on fat pad sampling, with a pooled sensitivity of 0.77 (95%CI 0.72-0.81), while twenty studies (n=2,771) reported on bone marrow biopsy, with a pooled sensitivity of 0.55 (0.46-0.63). All studies enrolled patients with confirmed disease, specificity was not reported in the included studies. Fat pad aspiration (15 studies, n=1,590) and surgical or punch biopsy (4 studies, n=222) showed similar sensitivities of 0.80 (0.76-0.83) and 0.77 (0.61-0.88), respectively. In a single study (n=612), performing both tests together yielded a sensitivity of 0.89. COE was judged to be Moderate for fat pad sampling and Low for bone marrow biopsy.
Unfolded protein response (UPR) promotes protein homeostasis under endoplasmic reticulum stress. UPR signaling has numerous functions in metabolism, cancer, immunology, and neurodegenerative diseases. Recent studies also showed that UPR signaling has important roles in hematopoietic stem and progenitor cell biology. However, whether UPR signaling regulates hematopoietic lineage fate decision remains elusive. Here, we found that FcgR- MPP3 generates erythroid lineage and Jak2V617F mutation leads to overproduction of erythroid cells by expanding FcgR- MPP3. We showed that UPR signaling increases myeloid cell production through promoting FcgR- MPP3 transition to granulocyte/macrophage progenitor producing FcgR+ MPP3 at the expense of erythroid lineage via the XBP1 pathway. Under a disease condition, UPR signaling cooperates with Jak2V617F mutation and exacerbates disease phenotype in a mouse model of polycythemia vera (PV) through the ATF4 pathway. Activation of UPR signaling also increased myeloid output in healthy donor bone marrow MPP cells while skewing the output towards erythroid lineage in PV patient bone marrow MPP cells. Together, our results identify a novel function of UPR signaling in hematopoietic lineage specification and provide critical insights into targeting UPR signaling in hematological malignancies.
Sickle cell disease (SCD) is a hemoglobinopathy affecting more than 8 million people worldwide. Pain, acute and chronic, is the most common and debilitating symptom of SCD. The use of standardized and well-established tools and protocols by researchers and clinicians for data collection can facilitate analyzing data from across different studies and, potentially, uncover previously unknown aspects of SCD pain. The PhenX (consensus measures for Phenotypes and eXposures) Toolkit (https://www.phenxtoolkit.org) is a web-based catalog of recommended measurement protocols and associated bioinformatics tools that facilitate study design and promote cross-study data integration and analyses. In 2019, the National Heart, Lung, and Blood Institute provided co-funding to the PhenX Toolkit to expand its collection of SCD-related protocols, strengthening the framework for data sharing across research projects. In 2021, a Working Group of 10 researchers and clinicians with expertise in SCD pain was assembled to recommend protocols for inclusion in the Toolkit. Using a consensus-driven approach that incorporated input from the scientific community, the SCD Pain Working Group selected protocols based on availability, researcher/participant burden, and validation status and prioritized well-established and broadly validated measures. Released in May 2022, the final selection included 22 protocols covering key dimensions of SCD pain, such as intensity, sensory characteristics, location, interference, physical mobility, impact on daily activities, and coping strategies. Consistent use of these protocols will improve data quality and comparability and will support meta-analyses. Adoption of these protocols could facilitate clinical guidelines and enhance comparative effectiveness research and implementation science for SCD pain management.
Venous thrombo-embolism is a serious complication of sepsis despite standard thromboprophylaxis. Mast cells (MCs) promote deep vein thrombosis (DVT) in sterile murine models, but their contribution during sepsis remains unknown. We explored the role of MCs in DVT in lipopolysaccharide (LPS)-induced sepsis in mice. We show that LPS treatment does not alter DVT prevalence or size in control C57BL/6 mice but increases intra-thrombus inflammatory mediators including interleukin-1b (IL-1β) and citrullinated histone 3 (CitH3), a hallmark of neutrophil extracellular traps (NETs). MC ablation (KITW-sh mice) protected animals from sterile DVT for up to one week, while in endotoxemia settings, KITW-sh mice produced thrombi similar to controls. Thrombi from KITW-sh mice were more susceptible to lysis by a combination of tissue plasminogen activator and DNase I and contained less CitH3 than controls. Septic DVT exacerbated pulmonary thrombo-inflammation characterized by large fibrin deposits and high levels of CitH3 and IL-1b in control but not KITW-sh mice. MC deficiency during septic DVT was associated with decreased plasma angiopoietin-2 levels, suggesting improved endothelial stability and reduced vascular inflammation. Thus, in septic conditions, MCs are dispensable for thrombus initiation but promote NET-rich thrombus stability, resistance to lysis and pulmonary thrombo-inflammation, the major life-threatening complication of DVT.
Vaccine-induced immune thrombocytopenia and thrombosis (VITT) and "VITT-like" cases are an emerging cause of severe thrombocytopenia and thrombosis associated with IgG antibodies targeting platelet factor 4 (anti-PF4). These syndromes are characterized by more frequent and severe thrombosis than heparin-induced thrombocytopenia (HIT), which is typically caused by antibodies targeting heparin-modified PF4 (anti-PF4/H). In this study, we compared the intrinsic prothrombotic properties of anti-PF4 and anti-PF4/H IgG antibodies to identify mechanisms that may contribute to the severity of VITT/VITT-like syndromes. Using the monoclonal IgG1 antibodies 1E12 (anti-PF4) and 5B9 (anti-PF4/H), we showed that anti-PF4 IgG exhibited an increased propensity to induce the formation of fibrin-rich platelet/leukocytes aggregates in a microfluidic model of whole blood (WB) thrombosis. This phenomenon was related to a strong potentiation of thrombin generation in plasma by 1E12, but not by 5B9, and only in the presence of platelets and coagulation factors V and XI. We also demonstrated a substantially faster and stronger ability of 1E12 compared to 5B9 to activate platelets using the serotonin release assay, confirmed with human VITT and HIT samples. Furthermore, 1E12 was more likely than 5B9 to induce procoagulant platelets (CD62P+/phosphatidylserine+) in platelet-rich plasma (PRP) and to produce procoagulant microvesicles in WB and PRP. In conclusion, platelets are activated faster and exhibit more procoagulant activity in response to anti-PF4 antibodies than to anti-PF4/H antibodies, which may contribute to the severity of VITT and VITT-like syndromes.
Congenital Dyserythropoietic Anemia type I (CDA-I) is an autosomal recessive disease characterized by anemia due to ineffective erythropoiesis and results primarily from mutations in CDAN1, which encodes CODANIN1. Research efforts to understand the CDA-I pathogenesis have been impeded by the embryonic lethality of germline Cdan1 deleted mice as well as mice deleted for Cdan1 in the erythroid compartment, using the constitutively active EpoR-Cre allele. To study the function of CODANIN1 in adult erythropoiesis, we generated mice with inducible erythroid-specific biallelic Cdan1 deletion using the Gata1-CreERT2 allele. Following tamoxifen administration to adult mice, Cdan1 is excised, resulting in features of CDA-I, including anemia, impaired erythroid differentiation, disturbances in erythroblast cell cycle progression, and the finding of 'spongy' heterochromatin in bone marrow erythroblasts. These findings confirm a critical role for CODANIN1 in effective adult erythropoiesis and demonstrate the successful generation of an inducible CDA-I mouse model, which serves as a valuable platform for testing novel therapies for this orphan disease.
Teclistamab is the first approved B-cell maturation antigen×CD3 bispecific antibody with weight-based dosing for triple-class-exposed relapsed/refractory multiple myeloma (RRMM). We evaluated the safety and efficacy of teclistamab combined with the anti-CD38 monoclonal antibody daratumumab in the phase 1b TRIMM-2 study. Eligible patients had RRMM (≥3 prior lines of therapy [LOT] or were double-refractory to a proteasome inhibitor and immunomodulatory drug); prior anti-CD38 exposure was permitted. Patients received subcutaneous daratumumab per approved schedule plus weight-based or fixed-dose subcutaneous teclistamab. The primary endpoint was safety; secondary endpoints included overall response rate (ORR) and duration of response (DOR). Progression-free survival (PFS) was an exploratory endpoint. Sixty-one patients received the weight-based recommended phase 2 doses (RP2D; teclistamab 1.5 mg/kg QW or 3.0 mg/kg Q2W); median number of prior LOTs was 5 (range, 1-14). Median follow-up was 12.0-months. The most common treatment-emergent adverse events (TEAEs) were infections, cytokine release syndrome, neutropenia, and anemia; grade 3/4 TEAEs occurred in 93.4% and 7 died from TEAEs. No dose-limiting toxicities occurred. ORR was 68.9% (complete response or better, 44.3%); median DOR was not reached. Median PFS was 26.3 months. A cohort exploring fixed-dose teclistamab (100-300 mg) ended prematurely after a safety signal for fatal infections was identified; out of an abundance of caution, all patients were switched to weight-based dosing. In conclusion, the fully immune-based combination of weight-based RP2D teclistamab plus daratumumab demonstrated deep and durable responses, with a well-characterized safety profile. Results highlight the importance of infection management, including early immunoglobulin replacement. Registered at ClinicalTrials.gov: NCT04108195.
Neutrophils are the most abundant circulating leukocytes in humans and act as key effectors of innate immunity. Severe congenital neutropenia (SCN) encompasses a heterogeneous group of disorders characterized by early onset of recurrent infections. Mutations in Jagunal homolog 1 (Jagn1) cause SCN, yet the mechanisms linking Jagn1 deficiency to defective neutrophil function and neutropenia remain unclear. Here, we identify Jagn1 as a critical regulator that connects intracellular protein trafficking and surface glycosylation during neutrophil development. Using neutrophils derived from Hoxb8-ER-immortalized progenitors, we show that Jagn1 expression is restricted to early progenitor stages and transcriptionally controlled by the transcription factor C/EBP-α. Loss of Jagn1 delays neutrophil differentiation, resulting in progenitor accumulation. Functionally, Jagn1-deficient murine neutrophils exhibit defective E-selectin-dependent rolling and chemokine-induced adhesion in ex vivo and in vitro flow chambers, mirrored by impaired rolling, adhesion and transmigration in vivo during TNF-induced inflammation. Mechanistically, lectin-binding and static adhesion assays revealed reduced sialylation of adhesion-relevant receptors on Jagn1-deficient neutrophils leading to impaired E-selectin and CXCL8 binding. In addition, NLRP3-dependent S100A8/A9 release from Jagn1-deficient neutrophils was reduced upon E-selectin stimulation. These findings define Jagn1 as a key determinant of neutrophil protein sialylation through regulating ER-Golgi posttranslational glycosylation, ensuring proper sialylation of adhesion relevant molecules including E-selectin ligands and CXCR2. This mechanism provides a unifying explanation for the clinical and cellular phenotype in Jagn1-associated SCN, which might offer new therapeutic approaches in the treatment of patients with loss of function mutation of Jagn1.
B-cell targeting CAR-T cells (CAR-T) therapies achieve high remission rates, yet durable responses occur in fewer than 40% of patients. Deletion of negative T-cell regulators such as PTPN2, a key inhibitor of TCR and cytokine signaling, represents a promising strategy to enhance CAR-T efficacy. While transfer of PTPN2-knockout (KO) T cells has demonstrated antitumor benefits in murine models, their impact on human-derived CAR-Ts and, importantly, the associated in vivo efficacy and toxicity remain unclear. Here, we demonstrate that PTPN2-KO human CD19 CAR-Ts exhibit enhanced cytokine production, proliferation, cytotoxicity, TCR and CAR functional avidity and signaling, which together lead to superior in vitro elimination of leukemic cells with low CD19 expression. In an in vivo dose-escalation study in a non-human primate (NHP) model of CD20/B-cell-targeting CAR-T therapy, we demonstrated that PTPN2-KO CAR-Ts exhibited superior in vivo expansion and B-cell depletion in a dose-dependent manner than PTPN2-WT CAR-Ts. PTPN2-KO CAR-T expansion was associated with increased toxicities, particularly neurotoxicity/ICANS, compared to WT CAR-Ts, driven by enhanced CNS-infiltration. Transcriptional profiling revealed a dominant effector and proliferative signature, with cytotoxic CNS-infiltrating CD8+ PTPN2-KO CAR-Ts implicated in ICANS pathogenesis. This study details the comprehensive evaluation of PTPN2-KO CAR-Ts in an immunocompetent primate model, demonstrating their enhanced on-target functionality, while highlighting increased toxicity risks, underscoring the value of rigorous preclinical assessment of potent genetic modifications in CAR-T therapy.
Acute kidney injury (AKI) develops in 20-40% of critically injured patients and contributes to poor outcomes. In sepsis and critical illness, dysregulated coagulation and fibrinolysis are linked to microangiopathic kidney dysfunction, but similar relationships in trauma are not well characterized. We performed a secondary analysis of a single-center randomized controlled trial of severely injured adult trauma patients. Plasma thrombin-antithrombin (TAT), plasmin-2-antiplasmin (PAP), and fibrinogen were measured within 2 hours of injury and 6 hours later. The primary outcome was AKI, defined by the RIFLE (Risk, Injury, Failure, Loss, End-stage kidney disease) criteria. Adjusted logistic regression models evaluated biomarker associations with AKI. Receiver operating characteristic analyses identified discriminatory thresholds. Among 149 patients, 30 (20.1%) developed AKI. Patients with AKI had higher TAT and PAP concentrations and lower fibrinogen levels at both time points. The 6-hour TAT-to-fibrinogen ratio was independently associated with AKI (Adjusted OR 7.17; 95% CI 2.34-21.9, p = 0.001, AUC = 0.88). Supplemental analyses demonstrated consistent associations for TAT and fibrinogen individually, whereas PAP was not independently predictive. A 6-hour TAT level < 99 pg/mL identified patients unlikely to develop AKI (negative predictive value [NPV] 93.9%). Fibrinogen levels > 173 mg/dL showed a low likelihood of AKI (NPV 88.2%). Elevated TAT and reduced fibrinogen measured 6 hours after injury were independently associated with AKI. This suggests that early coagulation imbalance may reflect evolving microvascular kidney dysfunction in injured patients and potentially warrants the evaluation of the TAT-to-fibrinogen ratio as an early marker of trauma-related AKI. (NCT02535949).
Infection is a major cause of morbidity and mortality in myelodysplastic syndromes (MDS), yet infection risk remains incompletely defined in the contemporary treatment era. We conducted a retrospective study of 708 patients with MDS to characterize the incidence, microbiology, temporal dynamics, and predictors of infection-related hospitalization over two decades. Overall, 78.8% (n=558) of patients required hospitalization, of which 69.9% were infection related. Infection-related hospitalization was independently associated with inferior overall survival. In multivariable Cox proportional hazards model, comorbidity burden, red blood cell transfusion dependence, higher IPSS-R risk, exposure to chemotherapy or stem cell transplantation, and severe neutropenia independently predicted infection. Among azacitidine-treated patients, 71.4% experienced infection-related hospitalization, with nearly three-quarters occurring within the first six cycles, identifying a critical early vulnerability window. Neutropenia remained the dominant driver of infection risk; however, immune dysfunction independently increased susceptibility. Low monocyte counts, cytokine dysregulation, and TP53 mutations identified high-risk patients despite preserved neutrophil counts, reflecting impaired myeloid reserve. Consistent with this dynamic vulnerability, recent infection or severe neutropenia (<0.5×10⁹/L) increased subsequent infection risk by 2.47-fold. We observed a concerning shift in antimicrobial resistance, with 14% of Gram-negative infections producing extended-spectrum beta-lactamases, 21% of Pseudomonas aeruginosa isolates resistant to piperacillin-tazobactam, and vancomycin-resistant enterococci prevalence increasing from 14% to 46%. Together, these findings identify infection in MDS as a dynamic, prognostically important complication driven by cytopenia, treatment, and immune dysfunction. This supports time-adapted risk stratification, targeted prevention, and antimicrobial stewardship, particularly during early treatment and high-risk disease phase.
Thalidomide has emerged as a fetal hemoglobin-inducer with potential to reduce transfusion burden in transfusion-dependent thalassemia (TDT). However, optimal dosing remains undefined A prospective, randomized, open-label, multicentric clinical trial was conducted at four centres in India to compare efficacy and safety of thalidomide at 1 mg/kg/day (Group 1) versus 2 mg/kg/day (Group 2) in patients with TDT aged ≥12 years. The primary endpoint was reduction in transfusion requirement at week 24, categorized as good (>50%), moderate (25-50%), or no (<25%) response. Responders underwent stepwise dose tapering during weeks 25-72 to evaluate response sustainability. Safety assessments were performed every 4 weeks. Of 188 enrolled patients (94 per group), 82.4% completed the week-24 evaluation. The overall response rate (ORR) was 58.5%, significantly higher in Group 1 than Group 2 (67.1% vs 50.0%, p=0.012). Good, moderate, and no response were observed in 20.5%, 37.8%, and 41.6% of patients, respectively, with transfusion independence in 9 patients (5.6%). Clinical benefit was observed within 12 weeks in 71.8% of good responders and 100% moderate responders. Sustained response at week 72 was seen in 49% and 57.5% of initial responders in Groups 1 and 2, respectively. Adverse effects were mostly grade 1; drug discontinuation due to toxicity was required in 10.6 % participants. These findings suggest that Thalidomide at 1 mg/kg/day was non-inferior to 2 mg/kg/day in reducing transfusion burden in patients with TDT, with an acceptable safety profile. Low-dose thalidomide appears to be a feasible treatment option in resource-limited settings. ICMR trial registry (Trial no. CTRI/2022/05/042781).
The decision whether to continue patients diagnosed with venous thromboembolism (VTE) on anticoagulation after the initial treatment period is often a nuanced decision. Many patients will have one or more transient risk factors associated with a time-limited increased risk for VTE. Transient risk factors are further divided into major, such as joint replacement, or minor, such as prolonged travel, which defines the magnitude of risk for an initial VTE. In addition, major transient risk factors generally carry a lower risk for recurrent VTE compared to minor transient risk factors. However, many patients will also have one or more persistent VTE risk factors, which further increase the risk for recurrence. Some persistent risk factors, such as inflammatory bowel disease, are associated with a high enough risk for recurrent VTE that guidelines recommend indefinite anticoagulation in this setting. There are multiple other persistent risk factors that carry a lower risk for recurrent VTE, however, such as chronic renal insufficiency and obesity. The High-Risk Patients with Provoked Venous Thromboembolism (HI-PRO) trial sought to assess the efficacy and safety of extended thromboprophylaxis for the prevention of recurrent VTE in patients with an initial provoked VTE and at least one persistent risk factor. The study found a lower risk of recurrent VTE in the thromboprophylaxis cohort, challenging the traditional time-limited treatment for provoked VTE, but heterogeneity of the study population and small sample size preclude identification of which subsets of patients could safely discontinue anticoagulation. We consider the conundrum of "provoked VTE" in this review.
This paper highlights a critical challenge in the FACT peer-accreditation system: a growing imbalance between the rising number of accredited and applicant organizations and the limited pool of volunteer inspectors. This imbalance places a disproportionate burden on current inspectors, concentrates accreditation expertise within a small subset of programs, and threatens the efficiency of the peer-review model. The paper presents compelling evidence that organizations contributing to the inspectorate gain measurable benefits. These include stronger audit performance in renewal cycles, access to education in quality standards and best practices, and broader professional networks. The paper concludes with a call to action: accredited organizations should share responsibility and contribute equitably to the FACT inspectorate to maintain the quality and sustainability of the accreditation process.
Blinatumomab is increasingly incorporated into frontline therapy for B-cell acute lymphoblastic leukaemia (B-ALL), yet risk stratification remains based on chemotherapy-era factors that may not apply in the immunotherapy setting. We analysed a national cohort to define determinants of relapse following frontline blinatumomab. Children and young people (1-24 years) in the UK and Ireland diagnosed with B-ALL between 2018 and 2025 who were chemotherapy-intolerant or resistant received blinatumomab in place of selected components of the frontline chemotherapy backbone. Outcomes were analysed according to conventional prognostic variables, genetic features, and response to blinatumomab. Among 225 patients, 195 received chemotherapy following blinatumomab (Blin-CT) and 30 underwent first-remission HSCT. In the Blin-CT cohort, traditional risk factors, including age, white cell count, high-risk genetics, and pre-blinatumomab end-of-induction measurable residual disease (MRD) did not predict relapse. On univariable analysis, relapse risk was increased with detectable MRD after blinatumomab cycle 1 (C1-END; hazard ratio HZR 6.61, p<0.001), IKZF1plus (HZR 3.64, p=0.04), JAK-STAT abnormalities (HR 3.56, p=0.05), and DUX4 rearrangements (HZR 4.24, p=0.03). In multivariable modelling, C1-END MRD and IKZF1plus remained independently associated with relapse, whereas DUX4-rearranged cases were strongly associated with persistent MRD at C1-END. An integrated model incorporating C1-END MRD, IKZF1plus, and DUX4-r defined a high-risk subgroup (33%) with an 18% 2-year relapse rate versus 0% in remaining patients (bootstrapped HZR 12.82, p=0.001, C-index=0.81). Relapse following frontline blinatumomab is determined by early treatment response and genetic subtype rather than conventional chemotherapy-derived risk factors. These findings support immunotherapy-specific risk stratification to guide treatment intensity.
Given the uncertainty regarding the optimal management of molecular relapse in patients with CBF or NPM1-mutated AML, we retrospectively analyzed the outcome of 121 adults from 12 centers with CBF (n=28) or NPM1-mutated (n=93) AML and first molecular relapse according to the salvage strategy used (upfront allogeneic HCT [n=19], intensive chemotherapy [IC; n=21], venetoclax and azacitidine [VEN-AZA]; n=70), and other strategies (n=11; including AZA, gemtuzumab ozogamicin, selective inhibitors). At three years, OS was not statistically different between the four groups (84% for upfront allo vs. 81% for IC vs. 79% for VEN-AZA vs. 64% for other, P=0.31). Allogeneic HCT was received by 98 patients (81%) with a cumulative incidence of allogeneic HCT at 12 weeks of 100% for upfront allo, 71% for IC, 73% for VEN-AZA, and 82% for other (P<0.001) with better outcomes in transplanted patients. In patients who received allogeneic HCT, type of salvage therapy was not statistically associated with post-HCT relapse, relapse-free survival, or OS. Our data suggests that upfront allogeneic is a valuable option, if feasible, while other salvage strategies are associated with favorable outcomes and relatively low non-relapse mortality after allogeneic HCT.
The impact of research and the development of therapeutics for blood cancer must be measured to direct future studies. Here, we used a life-years saved (LYS) methodology to quantify that impact. We estimate 25.8 million cumulative LYS across major blood cancer types in the US since 1949. The majority of LYS (~20 million) were for Hodgkin lymphoma (HL), acute lymphoblastic leukemia (ALL), and non-Hodgkin lymphoma (NHL). NHL stands out due to a high incidence, while HL and ALL have accumulated LYS over the longest period. LYS were lowest for acute myeloid leukemia and intermediate for multiple myeloma, chronic lymphocytic leukemia, and chronic myeloid leukemia. Progress has also been uneven across ages and racial groups. Black patients had worse survival outcomes than White patients for all blood cancers except multiple myeloma. The LYS methodology may be used to assess unmet needs and track progress, particularly where progress lags.
Adult B-cell precursor acute lymphoblastic leukemia (B-ALL) is genetically heterogeneous, comprising numerous subtypes. Standard methods leave many patients unclassified, and routine implementation of whole-transcriptome sequencing (WTS) or whole-genome sequencing (WGS) is limited. We evaluated a combined workflow of Optical Genome Mapping (OGM) and targeted next-generation sequencing (t-NGS) for genetic characterization of 100 adult BCR::ABL1-negative B-ALL patients enrolled in PETHEMA-LAL-19. This approach was compared with protocol genetic testing. The integration of OGM and t-NGS enabled the classification of 84% of BCR::ABL1-negative patients into 15 distinct subtypes, reducing the proportion of B-other/unclassified case from 50% to 16%. OGM was useful in identifying BCR::ABL1-like (26% of cases), associated with post-induction minimal residual disease (MRD) positivity (72% vs. 34%, p=0.0011), while t-NGS contributed to the detection of mutation-defined subtypes (PAX5 P80R and PAX5alt), correlated with MRD negativity (91% vs. 51%, p=0.012). OGM and t-NGS identified genetic patterns of certain subtypes, including a higher prevalence of IKZF1plus in BCR::ABL1-like (62% vs. 12%, p<0.001), JAK-STAT mutations only in patients with CRLF2-rearrangements and a high prevalence of PAX5-biallelic events (92% vs. 2%, p<0.001) in PAX5-subtypes. Therefore, the integration of OGM and t-NGS represents a feasible approach that enables comprehensive genetic characterization of adult BCR::ABL1-negative B-ALL, supporting its implementation in routine diagnostic workflows.
