HemaSphere最新文献

Flow cytometry (FCM) is a co-criterion in myelodysplastic neoplasms (MDS) diagnostics, currently not used for prognostication. This study aimed to develop an FCM-score predicting overall survival (OS) in MDS to improve early clinical patient prognostication. FCM of bone marrow samples was performed for diagnostic purposes in 509 therapy-naïve MDS patients and 77 healthy donors. The following methodology was used: (1) uni- and multivariate Cox proportional hazards regression and Kaplan-Meier curves for OS to assess FCM-parameters' prognostic value; (2) receiver operating characteristic (ROC) curves to test the prognostic superiority of FCM-parameters versus established FCM-scores and clinical risk-scores; and (3) development of a FCM-prognostic score (FCM-PS) based on six FCM-parameters with independent prognostic impact. The final FCM-PS included aberrancies of progenitor cells (increased CD45 mean fluorescence intensity [MFI]-ratio of lymphocytes and myeloid progenitor cells, decreased % of lymphatic progenitor cells), granulopoiesis (increased CD33 MFI, decreased sideward scatter [SSC]-ratio of granulopoiesis and lymphocytes), lymphocytes (increased % of B-lymphocytes), and plasmacytoid dendritic cells (increased %). FCM-PS outperformed established scores for OS (hazard ratio [HR] 4.08 [95% CI 2.54-6.55] vs. Ogata-score: 2.44 [1.61-3.70], International Prognostic Scoring System-Revised [IPSS-R]: 2.37 [1.61-3.49], International Prognostic Scoring System-Molecular [IPSS-M]: 0.816 [0.303-2.196]). Patients in the FCM-PS low score category showed significantly better OS (P < 0.0001). Further, FCM-PS allowed discrimination within IPSS-R area under the curve [AUC]: 0.70 vs. 0.62) and IPSS-M (AUC: 0.75 vs. 0.48) subgroups. Validation of the prognostic FCM-PS in an independent patient cohort confirmed good discrimination performance (AUC: 0.70). We introduce a unique, easy-to-use prognostic FCM-PS score (panel: CD45/CD34/CD117/CD33/CD19/CD123/HLA-DR) for OS in MDS, allowing refined risk stratification for IPSS-R subgroups.

Flow cytometry-based measurable residual disease (MRD) testing is routinely used in acute myeloid leukemia (AML), but methodologies need refinement to optimize assay characteristics. Here, we examined 1215 adults with AML or myelodysplastic syndrome/AML allografted in morphologic remission to study how the type(s) of leukemic blasts and degree/number of immunophenotypic abnormalities could improve MRD testing. Among 233 patients with pre-hematopoietic cell transplantation (pre-HCT) MRD, 80 (34%) had non-stem cell-like (NSC-like) leukemic blasts, 109 (47%) had stem cell-like (SC-like) leukemic blasts, and 44 (19%) had NSC- and SC-like leukemic blast cell populations. Across all MRD^pos patients, a higher degree/number of immunophenotypic abnormalities was associated with increased relapse risk and worse relapse-free survival (RFS) and overall survival (OS). Maximally selected rank statistics estimated a total MRD immunophenotype score cut point of ≤4.5 (n = 63 [27% of MRD^pos patients]) vs. >4.5 (n = 170 [73% of MRD^pos patients]) as optimal for RFS discrimination. After multivariable adjustment, a high score was associated with a significantly increased relapse risk (hazard ratio [HR] = 4.99 [95% confidence interval: 3.92–6.36]; P < 0.001), shorter RFS (HR = 3.88 [3.15–4.78]; P < 0.001), shorter OS (HR = 2.99 [2.42–3.70]; P < 0.001), and higher risk of NRM (HR = 1.78 [1.07–2.81]; P = 0.014) relative to MRD^neg patients. In contrast, there was no significant relapse risk or RFS difference between patients with low total MRD immunophenotype score and those without MRD. While requiring validation, our data suggest considering the type and degree/number of immunophenotypic abnormalities may refine MRD testing and identify a significant subset of MRD^pos patients with outcomes like MRD^neg patients.

Chronic myelomonocytic leukemia (CMML) shows marked prognostic heterogeneity. Although leukocytosis is a recognized adverse prognostic factor, the contribution of its individual components remains insufficiently defined. In a cohort of 240 patients classified according to International Consensus Classification (ICC) and World Health Organization (WHO) 2022 criteria-including 23% with oligomonocytic CMML-we evaluated the prognostic impact of neutrophil and monocyte percentage, along with surrogate markers of their relative increase, including relative lymphopenia (<20%) and elevated monocyte-to-lymphocyte ratio (MLR > 1). Both relative lymphopenia and MLR > 1 emerged as independent adverse prognostic factors, correlating with adverse mutations (TP53, RAS pathway) and high-risk clinical features. Notably, MLR > 1 identified a subset of patients with dysplastic CMML with molecular and clinical profiles resembling proliferative CMML (MP-CMML). These variables retained their prognostic impact after adjustment for established prognostic models (CMML-specific prognostic scoring system [CPSS], CPSS with the addition of the variable platelet < 100 × 10⁹/L [CPSS-P], and Mayo prognostic model), and their addition improved predictive performance. Based on these and other objective variables readily available from a routine complete blood count (CBC), we developed the objective prognostic index for CMML (OPIC), which integrates hemoglobin < 11 g/dL, platelets < 100 × 10⁹/L, MP-CMML, and MLR > 1. OPIC stratified patients into four risk categories with distinct survival outcomes (median overall survival [OS]: 104, 66.6, 34.3, and 18.3 months), demonstrating strong discriminatory power. Variable selection was performed using stepwise and elastic net regression, and random survival forests. Model performance metrics, including the C-index, time-dependent area under the receiver operating characteristic curve, and the Brier Score, were internally validated using bootstrapping-based resampling methods and externally validated in a cohort of 250 patients. OPIC provides a robust, accessible tool for CMML risk stratification, supporting its integration into routine clinical workflows and early therapeutic decision-making.

Studying how microenvironmental cues influence metabolic reprogramming can uncover mechanisms driving tumor progression. Using an in vitro model with proliferative stimuli of the in vivo lymph node niche (LN)-including interleukin-21 (IL-21)-we examined metabolic rewiring in chronic lymphocytic leukemia (CLL) cells. We found that the metabolic intermediates of upper glycolysis and its branching pathways are key in fulfilling metabolic demands of proliferating CLL cells. Among branching pathways, the pentose phosphate pathway (PPP) was the most transcriptionally upregulated in proliferating CLL cells. Increased expression of PPP genes was detected ex vivo at the bulk and single-cell level in the LN-resident and -emigrating CLL cells, with more consistency across enzymes of the nonoxidative PPP branch. Expression of the latter correlated with shorter failure-free survival in CLL patients. At the cellular level, metabolomics and 13C-glucose tracing confirmed high activity of the non-oxidative PPP in proliferating CLL cells. IL-21 regulated the expression of PPP enzymes, with STAT3 serving as the primary downstream effector. CRISPR/Cas9-mediated silencing of PPP enzymes revealed that, in vitro, proliferating CLL cells from most patients were not dependent on these enzymes. In contrast, silencing transketolase (TKT)-the rate-limiting enzyme of the non-oxidative PPP-abolished tumor engraftment in vivo, demonstrating that CLL cells rely on this pathway within the tumor microenvironment. These findings uncover a CLL-specific metabolic reprogramming wherein IL-21-STAT3 drives PPP activity and identify the nonoxidative PPP as a critical in vivo vulnerability of leukemic cells in the murine CLL model.

Hereditary anemias encompass a genetically heterogeneous spectrum of disorders, often involving multi-locus inheritance, which can complicate clinical management and worsen disease severity. This study investigates the impact of the co-inheritance of SEC23B loss-of-function pathogenic variants, which lead to congenital dyserythropoietic anemia type II (CDA II), and PIEZO1 gain-of-function pathogenic variants, associated with dehydrated hereditary stomatocytosis type I (DHS1), on hematological parameters and iron metabolism. Among 583 patients with suspected hereditary anemia, 13 were found to carry both SEC23B and PIEZO1 variants, leading to a dual diagnosis of CDA II and DHS1. Compared to those with isolated CDA II, these patients exhibited a significantly higher absolute reticulocyte count and bone marrow responsiveness index, alongside an increased prevalence of elevated ferritin levels. Functional studies in Hep3B human hepatoma cells confirmed that SEC23B knockdown combined with PIEZO1 gain-of-function led to marked ferritin accumulation and reduced hepcidin expression, driven by altered BMP/SMAD signaling and ERK1/2 MAPK pathway. These findings demonstrate how multi-locus inheritance can modify disease severity, particularly by exacerbating iron overload. Our results underscore the clinical relevance of comprehensive genetic testing for enhanced risk stratification and personalized management of hereditary anemias.

Pediatric regimens improve outcomes in adolescent and young adult (AYA) acute lymphoblastic leukemia (ALL) patients. End-consolidation (time point 2 [TP2]) minimal residual disease negativity (MRD^neg) is associated with improved survival. In this study, standard consolidation chemotherapy was replaced with blinatumomab to improve TP2 MRD^neg—a key survival surrogate in B-lineage ALL. From 2019 to 2022, 55 patients constituted the intention-to-treat (ITT) cohort, median age 25 (range, 16–39) years. Using a Simon's 2-stage design, blinatumomab replaced standard consolidation chemotherapy cycles with TP2 MRD^neg as the primary endpoint. Blinatumomab was associated with an improved TP2 MRD^neg rate of 70.8% (95% CI, 55.9%–83.0%) versus the null hypothesis of 60% (P = 0.037). When compared to our previous ALL06 study, median time from protocol I commencement to next treatment phase was 84 versus 97 days (P = 0.0001), with 82.7% versus 45.1% (P < 0.0001), commencing protocol M or high-risk block therapy by day 94. Induction mortality was 1.8%. Blinatumomab was well tolerated. Median follow-up was 42.9 (range, 1.9–54.7) months, with 3-year disease-free survival (DFS) 88.6% (95% CI, 76.3%–94.7%) and 3-year overall survival (OS) 90.5% (95% CI, 78.6%–95.9%) in the ITT cohort. Higher than medium-risk patients had poorer DFS but not OS. Standard genomic risk patients had 100% 3-year DFS and OS. Adverse genomic risk stratified by TP2 MRD^pos predicted poorer DFS but not OS. Blinatumomab consolidation for de novo B-lineage AYA ALL was associated with high MRD^neg rates and excellent survival, particularly in standard-risk disease. Genomics may assist in predicting response to blinatumomab in de novo ALL (ACTRN12618001734257).

Erythropoiesis is a finely regulated process ensuring continuous red blood cell production to maintain oxygen delivery. Disruptions in this process give rise to defective erythropoiesis, characterized by impaired lineage commitment and progenitor development, and ineffective erythropoiesis (IE), marked by expansion of erythroid progenitors with intramedullary apoptosis of late precursors. These abnormalities underlie anemia in diverse hematological disorders, including β-thalassemia, sickle cell disease (SCD), myelodysplastic syndromes (MDSs), and emerging entities such as vacuoles, E1 enzyme, X-linked, autoinflammatory, somatic (VEXAS). This review synthesizes recent insights into the molecular regulators of erythropoiesis, emphasizing the roles of GATA1, its chaperone HSP70, caspase activity, and extrinsic signals such as growth differentiation factor 11 (GDF11) and inflammatory cytokines. We highlight how globin-chain imbalance, inflammasome activation, oxidative stress, and clonal mutations converge on common pathways that disrupt erythroid maturation. Advances in therapy now directly target these mechanisms: Luspatercept, a TGF-β ligand trap, has transformed care in β-thalassemia and MDS by restoring late-stage differentiation, while anti-inflammatory strategies, metabolic modulators, and gene editing approaches are being actively explored. Together, these discoveries reframe IE as a modifiable process rather than an inevitable consequence of disease. By distinguishing defective from IE and mapping their mechanistic underpinnings, this review outlines how novel therapeutic strategies can improve anemia, reduce systemic complications, and ultimately enhance outcomes in patients with erythroid disorders.

Hematopoietic stem cell transplantation (HSCT) is a curative treatment for high-risk childhood leukemia but may lead to chronic graft-versus-host disease (cGvHD), a severe long-term complication. This study analyzed data from the LEA cohort, including 446 childhood leukemia survivors treated with allogeneic HSCT. The standardized cGvHD evaluation, using the NIH consensus criteria, was conducted 8.7 ± 0.3 years post-HSCT. Long-term cGvHD was reported in 21% of patients (9% mild, 7% moderate, 5% severe), primarily affecting eyes, skin, lungs, and mouth, with some cases involving multiple organs. Most patients with long-term cGvHD were untreated (84%), while 11% received systemic and 5% local treatments. cGvHD was associated with other long-term complications. Administration of anti-thymocyte globulin was a significant determinant (OR 0.6, 95% CI 0.4–0.99, P = 0.045). Long-term cGvHD showed a marked detrimental effect on the quality of life (QoL), even after adjusting for the other long-term complications. The SF-36 physical and psychological adjusted composite scores in patients with versus without cGvHD were 50 ± 2 versus 55 ± 1 (P = 0.01) and 38 ± 2 versus 43 ± 1 (P = 0.01), respectively. Even mild and moderate forms significantly affected the QoL, especially on psychological dimensions. These findings support standardized cGvHD evaluation and management to improve long-term outcomes of transplanted childhood leukemia survivors.