British Journal of Haematology

Background: Growing evidence suggests persistent thrombotic endothelial damage (together with elevated (fibrinaloid) microclot complexes (FMCs)) and immune dysfunction in the pathophysiology of Long COVID. Recently we proposed that there are different FMC phenotypes. Here we seek to determine the nature of these FMCs and aggregates in platelet-poor plasma (PPP) by using different markers, as well as thromboelastography (TEG) to assess for hypercoagulability of samples. Material and Methods: Whole-blood and PPP from control (n=19) and Long COVID (n=20) participants were assessed by thromboelastography. FMCs were quantified by imaging flow cytometry of Thioflavin-T (ThT)-stained PPP, 10X diluted PPP, and resuspended PPP pellets. The resuspended pellets were separately stained with a CD62P-PE antibody or Hoechst 33342 to label aggregates and FMCs containing amyloid, platelet, and nuclear material. ThT and CellMask Red were co-stained for confocal microscopy. ThT and myeloperoxidase (MPO), and ThT, Congo Red, and Hoechst were co-stained for fluorescence and polarized microscopy. Whole-blood smears were imaged by scanning electron microscopy (SEM). Results: Long COVID samples showed pronounced hypercoagulability in both whole blood and PPP, with shortened R, K and TMRTG and elevated alpha-angle and MRTG, but unchanged MA and TTG, indicating altered clotting kinetics. Persistence of this phenotype in PPP implicates soluble plasma constituents. ThT-positive FMCs were significantly increased in Long COVID across undiluted, diluted, and resuspended pellet samples; counts were processing-sensitive and a substantial ThT-positive population remained in the supernatant after centrifugation, indicating heterogeneity in density. Across probes, leukocyte material was the most abundant, then platelet material, and ThT-positive FMCs were the least abundant, with the three populations exhibiting unique morphology and occupying distinct size domains. Platelet-derived material was significantly elevated in Long COVID, whereas nuclear material was not. Co-stained samples subject to confocal, fluorescence, and polarized microscopy imaging showed that FMCs are heterogeneous, including events positive for ThT, CellMask, Hoechst, MPO, and Congo Red, and also a distinct subset of membrane-free, ThT-only events. Conclusion: In this Long COVID cohort, plasma is characterised by hypercoagulability and an increased burden of ThT-positive FMCs that are numerically minor relative to, and morphologically distinct from, aggregates and amyloidogenic FMCs marked with platelet- and leukocyte-derived material. The increased burden of platelet debris in PPP is likely indicative of persistent platelet activity. The existence of membrane-free, ThT-only FMCs, in addition to FMCs associated with cellular material, confirms an amyloid-dominated FMC population. Furthermore, positive Congo Red signal further confirms the amyloid nature of FMCs in PPP.

Background: Invasive fungal infection (IFI) is well recognised in children with acute leukaemia and allogeneic haematopoietic stem-cell transplantation but is poorly characterised in children with brain tumours. Children receiving intensive therapy for embryonal brain tumours (EBTs) have multiple potential risk exposures including corticosteroids, central venous access, neurosurgical devices, mucosal injury and myelosuppressive chemotherapy with, in selected protocols, autologous stem-cell rescue. Methods: We performed a single-centre retrospective cohort study of children aged 0-18 years treated for EBTs between 2015-2025. IFIs were classified as proven, probable, possible, or modified possible using EORTC/MSGERC and TERIFIC criteria. Clinical characteristics, treatment exposures, timing, microbiology and outcomes were described. IFI prevalence was calculated using exact binomial confidence intervals. Exploratory Cox proportional hazards analyses assessed associations with clinical and treatment factors. Results: Seventy-seven patients were included. Fourteen patients experienced 15 IFI episodes, giving a patient-level IFI prevalence of 18.2% (95% CI, 10.3-28.6%). Proven or probable IFI occurred in seven patients (9.1%; 95% CI, 3.7-17.8%). Nine episodes had microbiological evidence. Non-mould pathogens predominated, accounting for six of nine identified pathogens. Treatment on ACNS0334/ACNS0333 was associated with a lower hazard of proven/probable IFI compared with SJMB12 (HR 0.062; 95% CI, 0.002-0.78; p=0.031). Two patients had chemotherapy delays exceeding one month, one had persistent infection at 12 months; no deaths were directly attributed to IFI. Three patients received antifungal prophylaxis. Conclusion: Rates of IFI following intensive embryonal brain tumour therapy were comparable to those in other high-risk oncology populations. Local consideration of antifungal prophylaxis is warranted.

Background: In 2026, the US Food and Drug Administration (FDA) identified uveitis as a potential signal of serious risk for the B-cell maturation antigen (BCMA) x CD3 bispecific antibodies teclistamab and elranatamab. Uveitis in this setting may reflect opportunistic infection or immune-mediated inflammation, which require opposite management. Methods: We analyzed FDA Adverse Event Reporting System (FAERS) reports from July 2020 to June 2026 (8,690,601 reports after deduplication) using a case-non-case design, calculating reporting odds ratios (RORs) and the information component for primary-suspect reports. Comparators were belantamab mafodotin, talquetamab and CD3 bispecific antibodies used outside myeloma. Every primary-suspect uveitis report (n = 76) was classified as infectious, probable immune-mediated (hereafter immune-mediated) or indeterminate by a physician blinded to a rule-based algorithm. Results: Uveitis was reported in 18 of 2,995 teclistamab and 29 of 1,691 elranatamab reports (ROR 4.10 [95% CI 2.58-6.52] and 11.85 [8.21-17.12]). Of these 47 cases, 20 were infectious (18 cytomegalovirus chorioretinitis; median onset 92 days), and infection-specified uveitis was disproportionately reported for both agents and for CD3 bispecific antibodies used outside myeloma. Thirteen were immune-mediated, arose early (median onset 4 days) and were co-reported with cytokine release syndrome in six; immune-mediated uveitis showed a signal only for elranatamab (8 cases; ROR 3.59, 1.79-7.18), not for teclistamab or any comparator. Six of the eight elranatamab cases originated from Japan. Agreement between physician and algorithm was 98.7%. Conclusions: The FDA uveitis signal appears to comprise a late infectious uveitis shared by most T-cell-redirecting bispecific antibodies examined and an early immune-mediated uveitis that showed a signal only with elranatamab. These hypothesis-generating findings support prompt ophthalmological assessment of new visual symptoms and exclusion of cytomegalovirus retinitis before corticosteroids are started.

Purpose: Adults with high- or very high-risk acute leukemia (AL) or myelodysplastic syndrome (MDS) face substantial relapse risk after allogeneic hematopoietic stem-cell transplantation. We evaluated single-unit cord blood (CB) transplantation after ex vivo expansion with UM171 in this population. Patients and Methods: Two prospective, single-arm phase II trials at four centers enrolled 64 adults with high- or very high-risk AL or MDS; 60 received a UM171-expanded CB transplant and comprised the analysis population. CB units were preferentially selected at a 5/8 HLA match to maximize the graft versus leukemia effect. Patients received intermediate- or high-intensity conditioning with tacrolimus/mycophenolate mofetil graft-versus-host-disease (GVHD) prophylaxis. Endpoints included safety, feasibility, non-relapse mortality (NRM), relapse-free survival (RFS), overall survival (OS), GVHD, GVHD-free relapse-free survival (GRFS), chronic GVHD-free relapse free survival (CRFS). Results: Thirty-two percent of patients had undergone previous transplantation, 17% of patients with AL were not in remission and 24% of those with AML/MDS had TP53 mutations. Of 62 patients who remained eligible for transplantation, 60 had a graft successfully manufactured and infused. Median times to neutrophil and platelet engraftment were 17 and 38 days, respectively. NRM was 5.1% at day 100 and 15.2% at 1 year. Two-year cumulative incidence of relapse was 22.3%. Two-year OS and RFS were 63.9% and 60.4%, respectively. Grade III-IV acute GVHD incidence was 20.3% at 1 year and moderate-to-severe chronic GVHD incidence was 6.8% at 2 years. Conclusion: UM171-expanded CB transplantation was feasible and provided prompt engraftment, durable disease control, and infrequent clinically significant chronic GVHD in adults with high- and very high-risk AL/MDS. Comparative studies are warranted to define its role relative to contemporary donor platforms.

Mantle cell lymphoma (MCL) is a B-cell non-Hodgkin lymphoma characterized by heterogeneous clinical courses despite a common pathobiological initiating event. In this work we explore the genomic variants that characterize MCL and integrate transcriptomic data to comprehensively describe MCL biology. We performed whole exome sequencing (WES) on 28 tumor-normal pairs (lymph node and skin, respectively), as well as whole genome sequencing (WGS) and RNA sequencing on subsets of samples. We used established DNA and RNA analysis pipelines to detect single-nucleotide variants (SNV) and indels, structural variants, copy-number alterations, and RNA fusions. The canonical t(11;14)(q13;q32) CCND1::IGH translocation was detected in 8 of 10 WGS samples. Structural variant analysis additionally identified recurrent rearrangements involving KMT2A and PAFAH1B2. SNV and indel analyses revealed frequent mutations in ATM, TP53, CCND1, IGH, and NOTCH1. ATM exhibited diverse variant classes, including missense mutations, frameshift mutations, deletions, and duplications, while all detected NOTCH1 mutations were predicted loss-of-function frameshift variants. Copy-number analysis identified recurrent losses affecting DNA damage response genes, including TP53 and ATM, and recurrent gains involving transcriptional regulators and oncogenic signaling genes. Integrated pathway analysis demonstrated enrichment of transcriptional misregulation, DNA repair, PI3K/AKT signaling, and interleukin signaling pathways. We also identified recurrent alterations in candidate genes, including ASXL1, suggesting additional mechanisms of epigenetic dysregulation in MCL. Together, these findings provide a comprehensive description of somatic alterations in MCL and demonstrate that diverse genomic lesions converge on common pathways involved in genomic instability, transcriptional regulation, and tumor survival.

Background/Objectives: Many myeloid malignancies are defined by recurrent structural variants (SVs) and copy-number alterations (CNAs) under the current World Health Organization (WHO) and International Consensus Classification (ICC) frameworks. Detecting these abnormalities typically requires several standard-of-care (SOC) assays, each with its own blind spots, and then laboratories and clinicians are left to piece the results together. Genome mapping has emerged as a mainstream technology with the potential to serve as a high-resolution alternative to several standard-of-care cytogenetic assays such as karyotyping (KT), fluorescence in situ hybridization (FISH) or chromosomal microarray (CMA). Our laboratory at Augusta University has published numerous publications on the analytical and clinical utility of optical genome mapping (OGM) as a first-line assay for hematological malignancies. Electronic genome mapping (EGM) pursues the same goal with an electronic rather than optical readout. The objective of this study was to demonstrate the performance and utility of EGM as a valuable tool to detect clinically relevant SVs and CNAs found by SOC testing. Cytogenomic characterization of hematologic malignancies commonly requires multiple complementary assays, including karyotyping, FISH, and chromosomal microarray, each with distinct analytical limitations. Electronic genome mapping (EGM) provides genome-wide interrogation of structural variants and copy-number alterations using electronic single-molecule detection. We evaluated the feasibility and analytical performance of EGM in previously characterized myeloid malignancies. Methods: EGM was performed on the Nabsys OhmX platform using five previously characterized bone marrow aspirates from cases with acute myeloid leukemia (AML) or chronic myeloid leukemia (CML). EGM calls were compared with findings from karyotyping (KT), fluorescence in situ hybridization (FISH), and OGM. Results: All five samples passed quality control, with contig N50 values ranging from 1.8 to 3.8 Mb and more than 90% of reference labels represented in the assembled contigs. EGM detected every SOC-reported abnormality including two t(9;22)(q34.12;q11.23)/BCR::ABL1 rearrangements, trisomy 4, trisomy 21, and monosomy 7. In the most complex case, EGM resolved a del(20q) reported by KT and FISH into an intrachromosomal fusion, fus(20;20)(q11.21;q13.12), showing not only what was lost but where the chromosome rejoined. Like OGM, EGM identified an approximately 1.8 Mb loss at 7q22.1 involving 45 of genes of which only CUX1 is associated with cancer. This 7q loss was not detected by KT or FISH. Interestingly, EGM also detected the KMT2A-PTD variant previously detected by OGM. Collectively, EGM demonstrated 100% concordance (6/6 SOC-reported abnormalities; 9/9 OGM calls). Conclusions: In this pilot study on five hematological malignancy specimens, EGM matched SOC and OGM across translocations, aneuploidies, insertions and deletions, including BCR::ABL1 rearrangements in both AML and CML, and added high-resolution breakpoint detail beyond karyotype resolution. These results support larger studies across more subtypes to establish EGM as a first-line genome-wide cytogenomic tool for hematological malignancies. Keywords: Genome mapping, Electronic genome mapping, hematological malignancies, structural variants, copy number variants

Background: The clinical course of acute leukemias and related hematologic neoplasms is defined by aggressive clonal proliferation, severe hematopoietic disruption, and recurrent molecular alterations. Although serial bone marrow biopsies, multiparameter flow cytometry, and next-generation sequencing remain standard surveillance modalities, procedural invasiveness, high costs, and operational turnaround times limit their frequency. Minimally invasive, scalable biomarkers integrating routine hematologic indices, targeted gene expression, circulating oncoprotein levels, and patient compliance metrics may provide an accessible framework for dynamic monitoring. Methods: In this longitudinal cohort study of 66 patients across 272 serial encounters (discrete follow-up intervals: days 0, 14, 41, 71, and 78), we assessed the prognostic value of peripheral complete blood counts (white blood cell count [WBC], hemoglobin [Hb]), molecular indicators (FLT3, NPM1, and DNMT3A transcript levels; targeted soluble FLT3 quantification), and medication adherence. The primary clinical endpoint was longitudinal progression to critical deterioration or disease-related mortality. Generalized linear mixed-effects models (GLMM) with patient-specific random intercepts were constructed to account for intra-subject repeated measures. Predictors were standardized per 1-SD increment. Model performance was assessed via the Akaike Information Criterion (AIC) and the Concordance index (C-index). Results: Leukocyte expansion correlated inversely with hemoglobin concentration (r=-0.877, p<0.001), reflecting bone marrow infiltration and secondary erythroid suppression. Multivariable mixed-effects modeling confirmed that elevated WBC count (adjusted odds ratio [aOR] = 1.52 per 1-SD increase; 95% CI: 1.24-1.86; p<0.001) and reduced medication adherence (aOR = 0.58; 95% CI: 0.44-0.76; p<0.001) independently predicted adverse clinical transition. Integrating targeted molecular markers (FLT3 expression and soluble protein levels) with routine hematologic indices and adherence demonstrated superior calibration and discrimination over single-domain models ({Delta}AIC[&ge;]116; Harrells C-index = 0.81). Conclusions: Readily accessible peripheral hematologic indices, targeted molecular quantification, and medication adherence provide complementary prognostic information regarding disease activity. Integrating these low-burden parameters into clinical workflows offers an efficient, scalable paradigm for early risk stratification in hematologic malignancies.

Large genomically annotated acute myeloid leukaemia (AML) datasets exist, but population-based contemporary cohorts remain scarce. Here we report clinicopathological, genomic, and outcome data from Danish AML patients. 2,512 AML patients were identified between 2015-2022, of whom 33.8% had available NGS data (NGS+). In patients [&le;]70 years, baseline characteristics and outcomes were comparable between NGS+ and NGS- groups. In patients >70 years, more NGS+ patients received intensive treatment, but survival was similar among intensively treated patients. The distribution of mutations varied significantly by age and sex, with older age and male sex exhibiting higher frequencies of adverse-risk gene mutations. In intensively treated NGS+ patients, ELN2017 stratified 5-year OS: 58.4% (favorable), 43.4% (intermediate), and 28.2% (adverse), with hazard ratios (HRs) of 0.63 (favorable) and 1.45 (adverse) relative to intermediate. ELN2022 yielded corresponding OS rates of 56.9%, 51.8%, and 29.7%, with HRs of 0.78 and 1.86. The two models had comparable predictive performance for OS in a time-dependent model. In conclusion, outcomes of intensively treated AML patients were comparable irrespective of NGS status, underscoring the representativeness of the REFORM-AML database for the Danish AML population. Age and male sex correlated with adverse-risk mutations, and both ELN2017 and ELN2022 robustly predicted survival.

The genetic risk factors of immune cytopenias are unclear. Immune cytopenias have been reported in various genetic contexts: 1) inherited error of immunity genes, mainly due to rare germline variants, 2) systemic lupus erythematosus, associated with common germline variants, 3) hematological malignancies, and 4) clonal hematopoiesis, the latter two due to somatic variants. However, the respective contribution and interaction of these variants remain to be investigated. Here, we used two large biobanks with whole genome sequencing data to systematically investigate the genetic contribution to immune cytopenia. We found that the four types of genetic variants independently contribute to immune cytopenia risk. We notably found that carriers of variants in some autosomal recessive genes of inherited error of immunity had an increased risk of immune cytopenia. Additionally, common variant-mediated risk of systemic lupus erythematosus also increased the risk of immune cytopenia. Overall, a third to a half of patients with immune cytopenia carried at least one of the four genetic risk variants investigated. Combining the four variants allowed stratifying the risk of immune cytopenia in both general and high-risk population. In general population, the 10-year incidence of immune cytopenia in the lowest and highest risk groups was 0.08% and 1.5%, respectively. In sum, this work identified that different genetic risk factors can lead to immune cytopenia. A large proportion of individuals with immune cytopenia carried an underlying genetic risk factor. Finally, combining these genetic risk factors enabled risk stratification.

BACKGROUND: The transport and storage conditions of thawed plasma are not strictly regulated by the FDA and applying red blood cell transport standard of 1-10 degrees Celcius to recently thawed plasma often results in high discard rates. This study evaluated the extended 120-hour (5-day) coagulation factor stability and sterility of thawed plasma frozen within 24 hours (PF24) following a 6-hour transport cooler simulation. STUDY DESIGN AND METHODS: Fourteen PF24 units (8 group O, 6 group B) were thawed at 30-37 degrees Celcius and assigned as control (n=7, direct 1-6 degrees Celcius refrigeration) or experiment (n=7) units. Experiment units were held at room temperature for 30 minutes, stored in validated transport coolers for 6 hours, and then transferred to 1-6 degrees Celcius refrigeration. Measurements of temperature, prothrombin time (PT), Factor V (FV) activity, and Factor VIII (FVIII) activity were conducted at 0-, 6-, 24-, and 120-hour post-thaw. Sterility testing was performed at 0-hour and 120-hour using automated aerobic and anaerobic blood cultures. RESULTS: No statistically significant differences were observed between control and experiment units at 120-hour for mean PT (15.09 vs. 15.16 seconds, p = .44), FV activity (81.14 vs. 74.57%, p = .23), or FVIII activity (61.86 vs. 53.00%, p = .22). Delta analysis (120h-0h) confirmed equivalent factor decay rates between groups. All bacterial cultures showed no growth at 120-hour. CONCLUSION: A 6-hour cooler time of thawed PF24 does not accelerate coagulation factor degradation or compromise sterility over an extended 5-day shelf life. These findings validate flexible inventory return policies, allowing blood banks to reduce product waste.

BackgroundThe Philadelphia negative myeloproliferative neoplasms (MPN), including essential thrombocythemia (ET), polycythemia vera (PV) and primary myelofibrosis (PMF), are characterized by myeloid cell proliferation, thrombosis and inflammation. Suspicion of MPN arises from increased blood count in one or more lineages; however, knowledge on the MPN plasma proteome including biomarkers measurable in blood, are lacking. Comparing the plasma proteome of MPN patients to subjects with elevated blood counts but without MPN diagnosis, may provide an increased understanding of the MPN disease biology as well as diagnostic biomarkers measurable in blood. Patients and methodsWe performed plasma proteome profiling in 87 patients referred to the Department of Hematology due to elevated blood counts. Of these, 55 were diagnosed with MPN and 32 did not fulfill MPN diagnostic criteria and thus constituted the non-MPN control group. ResultsThe frequency of thrombosis was equal between the groups. We found 189 differentially expressed proteins between MPN and non-MPN, enriched for Hemostasis and Platelet activation proteins. Using Lasso multiple regression, we identified SORT1, GP1BA, PSPN, MMP1 and BAG6 separating MPN from non-MPN individuals, and TFRC and SEMA7A specific for MPN subtype PV. Interestingly, TFRC alone had a diagnostic accuracy for identifying PV of 89.3%, and when combined with serum erythropoietin it increased to 99.3%. Only two proteins, IL-6 and GH1, were increased in JAK2 mutant MPN compared to JAK2 wildtype MPN. The same trend was seen for JAK2 mutant ET compared to JAK2 wildtype ET, indicating that IL-6 is induced by JAK STAT activation. However, IL-6 levels did not differ between MPN and non-MPN patients. Discussion/conclusionIn conclusion, hemostasis and platelet activation are inherent to MPN disease whereas little difference was found in proinflammatory cytokines between MPN and non-MPN groups. We demonstrate novel potential blood biomarkers for MPN and MPN subtypes, in particular TFRC for identifying PV patients.

Clonal hematopoiesis (CH) can predict treatment outcomes in solid cancers, but the prevalence and clinical impact of CH in previously untreated, aggressive large B-cell lymphomas (LBCL) remains poorly characterized. Moreover, CH may be detected in cell-free DNA (cfDNA)-based biomarker assays, but the longitudinal dynamics of CH mutations during and after immunochemotherapy in LBCL remain unknown. Hence, we characterized the spectrum of CH mutations in 247 patients with high-risk primary LBCL from three phase II clinical trials. We identified CH mutations in 23% of patients. Compared with healthy controls, TET2 mutations were enriched in peripheral blood (PB) samples of LBCL patients (P=0.01). CH clone size showed high concordance between PB and cfDNA (R2=0.59, P<0.0001) but was not associated with circulating tumor DNA (ctDNA) concentration (P-values 0.25-0.86). In contrast with prior reports in the context of recurrent lymphoma, CH mutation status was not associated with overall survival (OS) (P=0.78) or progression-free survival (PFS) (P=0.89) in our primary LBCL cohort. CH clones showed variable longitudinal dynamics during immunochemotherapy, with 18% of evaluable patients developing mutant CH clones, mostly with TP53/PPM1D mutations. Together, CH mutations can be readily detected in cfDNA of lymphoma patients, but they do not predict survival in primary high-risk LBCL.

BackgroundFebrile neutropenia requires rapid empirical antibiotic therapy, with regimen selection shaped by local prescribing practice and pathogen patterns. We described starting treatment, later drug changes, microbiological findings, and inpatient outcomes. MethodsThis ambispective cohort comprised 96 episodes from 90 patients admitted to a tertiary-level teaching hospital. Treatment and outcome analyses used each episode as the unit of observation. Recorded variables included the starting antibiotics, later regimen modification, microbiology, hospitalization length, invasive fungal infection (IFI) as captured in study records, and disposition. Comparisons based on escalation status were exploratory. ResultCefoperazone-sulbactam appeared in 62/96 initial regimens (64.6%), amikacin in 42 (43.8%), teicoplanin in 28 (29.2%), and meropenem in 14 (14.6%). Eighteen episodes (18.8%) required a change that broadened or substituted antibacterial therapy. Among 31 microbiologically documented infections, 22 (71.0%) yielded Gram-negative organisms and 9 (29.0%) were polymicrobial. Median length of admission was 18.5 days in the escalation group and 9 days in the remainder (p<0.001). Mortality was 27.8% (5/18) with escalation and 5.1% (4/78) without it (p=0.011), with nine inhospital deaths overall (9.4%). ConclusionAntibacterial management differed within this cohort, with treatment escalation required in 18 of 96 episodes. Gram-negative organisms comprised most microbiologically confirmed infections. Escalation marked a clinically complex course with prolonged hospitalization and greater observed mortality; the observational design does not establish comparative efficacy or causation.

Homozygosity for the HBB codon 8/9 (+G) frameshift (c.27dup (p.Ser10ValfsTer14)) causes transfusion-dependent {beta}-thalassaemia and is common in South Asia. Prime editing can reverse this insertion without double-strand breaks or donor DNA, but its efficiency depends on pegRNA design. We derived Sendai-reprogrammed iPSCs from a homozygous patient, optimised PEmax editing (spacer, pegRNA extension, secondary nick, MLH1dn) and compared patient, PEmax-treated and control iPSC-derived erythroid cells by flow cytometry, colony assays, RT-qPCR, western blotting and cation-exchange HPLC. Patient iPSCs had a normal 46,XY karyotype, expressed pluripotency markers, formed all three germ layers and were Sendai-free by passage 15. A PAM-disrupting spacer with a +68 nicking sgRNA gave the highest intended-edit frequency 8.4%; MLH1dn added little. PEmax-treated cultures matured and formed colonies more like control than patient cultures, restored HBB transcript (about 4-fold control iPSC-derived cells) and detectable {beta}-globin protein, and contained an HPLC fraction consistent with HbA ( undetectable in patient cells). Fetal haemoglobin (about 80%) and embryonic globin remained predominant. Prime editing corrects HBB c.27dupG in patient iPSCs and restores {beta}-globin expression, with partial restoration of adult haemoglobin in a fetal/embryonic-type erythroid background. Validation in additional donors, haematopoietic stem cells and in vivo is required.

Quantitative measurement of fetal hemoglobin (HbF) using high-performance liquid chromatography (HPLC) is standard but expensive and widely unavailable in low-income settings. The newer Gazelle method, which employs single-use alkaline electrophoresis cartridges, provides a portable and relatively inexpensive potential alternative. In the current study, we tested 3055 venous blood samples collected from 1780 children with sickle cell anemia who were enrolled into the H-PRIME trial by both Gazelle and the Bio-Rad D-10 HPLC instrument, before and during hydroxyurea treatment. Data reinterpretation was required to quantify accurately high HbF values derived using the D-10. Mean (standard deviation) baseline HbF values by HPLC and Gazelle (N=2042) were 11.4% (SD 5.9%) versus 16.3% (SD 7.5%), respectively, while mean on-treatment HbF values (N=1013) were 18.9% (SD 9.0%) versus 25.4% (SD 11.5%), respectively. While overall agreement was good (Lins concordance 0.76, 95% CI 0.75-0.78), HbF values derived using Gazelle were modestly higher than HPLC values (Bland-Altman agreement mean +5.4%, 95%CI -2.4% to +13.2%), with the difference between measures increasing at higher HbF percentages. With strong concordance and more easily interpretable HbF values, Gazelle represents an attractive and inexpensive alternative to HPLC for HbF quantification in the low-income settings where most patients with sickle cell anemia live.

Background: Translocation t(11;14) defines a biologically distinct subset of multiple myeloma (MM) enriched for BCL2 dependency. Treatment with venetoclax, selective BCL2 inhibitor, has shown varied responses in clinical trials and retrospective cohorts. Efficacy of venetoclax-based combination therapies and optimal timing of treatment in t(11;14) MM patients remain incompletely characterized. Methods: We have compared the overall survival of t(11;14) MM patients who received Venetoclax (N = 97) at any moment in time, to those who never did (N =284), in the largest retrospective cohort (N = 381) reported to date. We used longitudinal fluorescence in situ hybridization (FISH) to assess cytogenetic evolution and genomic complexity. We performed transcriptomic profiling of CD138 enriched tumors using RNA sequencing in a subset of samples and gene expression signatures (UAMS/HALLMARKS) were used to define molecular subtypes. Ex vivo drug sensitivity assays integrated with paired RNA sequencing were used to identify subtype specific therapeutic vulnerabilities and rational venetoclax based combination strategies. Results: Venetoclax exposure was associated with an improvement of median overall survival by nearly four years compared to non VEN exposed patients (p = 0.0003). Longitudinal cytogenetic analysis demonstrated stability of the primary t(11;14) translocation over time, while secondary abnormalities tend to accumulate, including those harboring high-risk secondary cytogenetic abnormalities such as del13q, amp/gain1q21, del17p, and del1p, resulting in increasing genomic complexity with disease progression. Transcriptomic analyses identified selective enrichment of CD1/CD2 signature (associated with t(11;14) NDMM) by single sample gene set enrichment analysis as a marker of prolonged progression free survival. However, patients with more than five prior lines of therapy were enriched for transcriptionally complex biology characterized by CD1/CD2 with either proliferative/hypermetabolic or inflammatory transcriptional programming which were associated with inferior outcomes with VEN based treatment. Ex vivo drug sensitivity profiling revealed subtype specific vulnerabilities, identifying daratumumab, lenalidomide, ixazomib, and panobinostat as rational partners for venetoclax depending on transcriptional context. Conclusion: Venetoclax-based therapy significantly improved overall survival in t(11;14) MM. Clinical benefit, defined by improved progression free survival, was greatest when venetoclax was administered earlier, preceding the emergence of transcriptomic reprogramming that reduces BCL2 dependency. These findings support transcriptomic biomarker guided, subtype specific venetoclax based treatment strategies to optimize outcomes in patients with t(11;14) MM.

Objective: To investigate the distribution frequencies of Rh blood group antigens (C, c, D, E, e) and phenotypes in the population of Hengyang, Hunan Province, and to analyze the production of Rh alloantibodies in repeatedly transfused patients, thereby providing a basis for developing precise transfusion strategies. Methods: Rh phenotyping, antibody screening, and antibody identification were performed on 3,635 hospitalized patients and 5,326 blood donors using Rh blood group typing cards. A blood transfusion management system was used to identify and track patients' historical specific antibodies, with automatic alerts for inconsistent results. Results: The antigen frequency distribution in patients was D (99.56%) > e (94.69%) > C (91.64%) > c (48.06%) > E (38.79%). The phenotypic distribution frequencies among Rh(D)-positive patients were as follows: CCDee (51.31%) > CcDEe (30.01%) > CcDee (9.37%) > ccDEE (5.00%) > ccDEe (2.79%) > CCDEe (0.80%) > ccDee (0.39%) > CcDEE (0.28%) > CCDEE (0.05%). From March to October 2023, after implementing Rh phenotyping and antigen-matched compatible transfusions for five antigens, the antibody screening positivity rate decreased to 0.97%, compared to 1.14% during the same period in 2022 (p < 0.05). Antibody identification in 276 antibody-positive samples revealed that alloantibodies against the Rh system accounted for the highest proportion (46.01%, 127/276), which was lower than the 55.21% observed in 2022 (p < 0.05). Unexpected antibodies in the Rh system were the primary cause of crossmatch incompatibility in clinical transfusions, accounting for 46.01%. Conclusion: Rh phenotyping and sustained antigen-matched compatible transfusions in repeatedly transfused patients can effectively prevent and reduce alloantibody production. Continuous tracking of specific antibodies and transfusion efficacy evaluation can be achieved through an efficient blood transfusion management system.

Sickle cell disease (SCD) results from homozygosity or compound heterozygosity for a single point mutation in the beta hemoglobin gene. Despite carrying the same underlying causal mutation, patients exhibit highly heterogeneous clinical outcomes. Acute chest syndrome (ACS) is one of the leading causes of mortality in SCD. In this study, we aimed to identify genetic variants associated with ACS occurrence and recurrence (>5 episodes) in two SCD cohorts (OMG-SCD and Walk-PHaSST) of predominantly African ancestry and characterize their downstream molecular consequences. Although none of the SNPs reached the genome-wide threshold in the occurrence meta-analysis, we identified 40 independent genetic loci with p <10-5. However, the recurrence GWAS in OMG-SCD, which compared individuals with no prior history of ACS to individuals with a history of >5 episodes, identified one genome-wide significant locus along with 38 other independent loci with p <10-5. Survival analysis suggested lower survival probability among young ACS recurrence cases compared to controls or occurrence cases. Furthermore, molecular QTL analysis of ACS recurrence identified 42 SNP-protein and eight SNP-metabolite associations with p <10-3. Pathway and enrichment analysis identified secretory, platelet/hemostatic, and complement-mediated pathways, confirming the complex and heterogeneous pathophysiology of ACS. This study highlights the importance of a well-defined phenotype for identification of risk loci and elucidates the complex interplay of hemolytic, inflammatory, and cellular organization pathways that warrants further evaluation through large-scale multi-omic studies of SCD patients. Ultimately, the more extreme phenotype of ACS recurrence may be a more informative phenotype for analysis, risk prediction, and therapeutic targeting.

The non-receptor protein-tyrosine phosphatases Shp1 (PTPN6) and Shp2 (PTPN11) play critical roles in hematopoietic signaling networks, yet their specific functions in human megakaryopoiesis and thrombopoiesis remain incompletely understood. While Shp2 is recognized in murine models as a positive regulator of thrombopoietin (Tpo)-mediated signaling through the Ras/MAPK and PI3K/AKT pathways, Shp1 has been implicated in RhoA-dependent cytoskeletal remodeling. However, the extent to which these roles translate to human megakaryocyte (MK) development and platelet production is not known. In this study, we systematically investigated the contributions of Shp1 and Shp2 to human MK development and function using CRISPR/Cas9-mediated gene deletion of PTPN6 and PTPN11 in CD34+ hematopoietic stem and progenitor cells (HSPCs), combined with pharmacological inhibition of Shp2 using the structurally-distinct allosteric inhibitors SHP099 and RMC-4550. Efficient gene editing of PTPN6 and PTPN11 resulted in efficient ablation of Shp1 and Shp2 in CD34+ HSPC-derived MKs. Genetic deletion or pharmacological inhibition of Shp2 markedly impaired MK proliferation, polyploidization, maturation, and proplatelet formation, whereas loss of Shp1 expression did not. Further, Shp2 inhibition significantly reduced platelet production in a 3-dimensional human bone marrow tissue model. Deletion and inhibition of Shp2 abrogated Tpo-induced ERK1/2 and AKT phosphorylation, confirming its essential role in Mpl receptor signaling. These findings demonstrate the distinct functional roles of Shp1 and Shp2 in MKs and establish Shp2 as a critical positive regulator of Mpl- mediated megakaryopoiesis and thrombopoiesis. Key PointsO_LIEfficient deletion of Shp1 and Shp2 in human CD34 progenitor cell-derived MKs using CRISPR/Cas9. C_LIO_LILoss of Shp2 expression impairs thrombopoietin-induced human MK maturation, proplatelet formation and Mpl signaling. C_LI

Large Language Models (LLMs) are increasingly used by clinicians and patients for medical queries, yet their accuracy and safety at the specialist level in hematology remain insufficiently characterised. We benchmarked ten frontier proprietary and open-weight LLMs across two generations on 1,477 board-style hematology multiple-choice questions (MCQs) derived from five educational datasets spanning nine disease areas and six clinical skill domains, including text-only and multimodal case vignettes. Claude Opus 5 had the highest mean accuracy (92.7% text, 76.9% multimodal), followed closely by Gemini-3.1 Pro (91.4% and 78.7%), Gemini-3.6 Flash (91.0% and 74.8%) and GPT-5.6 Sol (89.9% and 76.7%). Accuracy significantly correlated with model size both for text-only and multimodal MCQs. Between model generations, the largest improvements in accuracy were seen for open-weight models whereas proprietary models showed only marginal gains. In error analysis, top-performing models exhibited highly concordant failure patterns, suggesting shared limitations on challenging cases. Frontier LLMs exhibit substantial specialist hematology knowledge across diverse subspecialist domains and clinical skill sets. Yet, despite high accuracy on board-style questions in hematology, continuous expert-on-the-loop output monitoring is paramount.