The efficacy of FLT3 inhibitors in acute myeloid leukemia (AML) is severely limited by resistance mutations, particularly the recalcitrant gatekeeper F691L and activation loop D835V/Y variants. Herein, through the structure-guided optimization of our previously reported lead SILA-123, we identified FYJ-195, a highly potent type II FLT3 inhibitor capable of overcoming these recalcitrant mutants. FYJ-195 exhibited single-digit nanomolar potency against Ba/F3-FLT3-ITD-F691L (IC50 = 9.09 nM) and subnanomolar activity against Ba/F3-FLT3-ITD-D835Y (IC50 = 4.64 nM) and Ba/F3-FLT3-ITD-D835V (IC50 = 0.76 nM). In vivo, FYJ-195 induced profound tumor regression (TGI = 125%) in the MV4-11 xenograft model (10 mg/kg) and achieved robust tumor growth suppression (TGI = 68.6%) in the Ba/F3-FLT3-ITD-F691L model (50 mg/kg), where quizartinib was ineffective. Mechanistic studies confirmed that FYJ-195 effectively blocked FLT3 signaling and induced apoptosis without observable toxicity. Collectively, FYJ-195 represents a promising lead candidate for drug-resistant AML.

ObjectiveTo investigate the efficacy and safety of reduced-dose emicizumab (EMI) prophylaxis in infants under one year of age with severe hemophilia A (HA).MethodsMedical records of 11 infants aged ≤12 months with severe HA who received reduced-dose EMI in the Department of Hematology and Oncology, Shenzhen Children's Hospital, from July 2022 to June 2025 were retrospectively reviewed. Efficacy and safety were analyzed statistically.ResultsThe median age at EMI initiation was 7.9 months (range 4.2-11.2 months). The median loading dose (first 4 weeks) and median maintenance dose were 6.2 mg/kg per 4 weeks (range 2.8-12.6 mg/kg per 4 weeks) and 3.1 mg/kg per 4 weeks (range 2.0-4.8 mg/kg per 4 weeks), respectively. The median duration of EMI prophylaxis was 15.8 months (range 1.7-36.0 months). After treatment, the median (range) annualized bleeding rate, spontaneous annualized bleeding rate, and annualized treated bleeding rate were 0.5 (0-6.2), 0 (0-0.5), and 0 (0-3.0), respectively. All decreased compared with pre-treatment, and the reduction in spontaneous annualized bleeding rate was statistically significant (P<0.05). Overall, 64% (7/11) of patients experienced no bleeding events requiring treatment. No EMI-related adverse events were observed.ConclusionsReduced-dose EMI prophylaxis effectively controls bleeding in infants with hemophilia A and shows good safety.

Background:Asciminib, a novel BCR::ABL1 inhibitor that functions by specifically targeting the myristoyl pocket, has shown superior efficacy and favorable safety and tolerability compared with adenosine triphosphate-competitive tyrosine kinase inhibitors (TKIs) in patients with newly diagnosed chronic myeloid leukemia in chronic phase (CML-CP). Flumatinib, a second-generation TKI available exclusively in China, does not have a head‑to‑head comparison with asciminib till date with asciminib. Therefore, this study aimed to conduct an anchored matching‑adjusted indirect comparison using data from the ASC4FIRST and FESTnd trials to fill this evidence gap. Methods:Imatinib was the common comparator across the ASC4FIRST (NCT04971226) and FESTnd (NCT02204644) trials. To match the two populations, effect modifiers and baseline variables were identified. Adjusted estimates were derived from individual patient-level data (ASC4FIRST) for asciminib and imatinib and from aggregate data for flumatinib (FESTnd). Outcomes were compared based on early molecular response (EMR), major molecular response (MMR), and treatment discontinuation due to adverse events (AEs). Results:Asciminib demonstrated significantly higher EMR rates at 12 weeks (adjusted: 90.0%) than did flumatinib (82.1%), yielding a significantly higher odds ratio [odds ratio (OR): 1.95; 95% confidence interval (CI): 1.05-3.73; P=0.03]. MMR rates, both at 48 and 96 weeks, were significantly higher for asciminib (adjusted: 66.5% and 74.0%, respectively) than for flumatinib (an estimated 52.6% and 61.3%, respectively), with an OR of 1.79 (95% CI: 1.17-2.75; P=0.006). Safety analysis showed fewer discontinuations due to AEs at 48 weeks with asciminib (5.5%) than with flumatinib (10.2%), corresponding to a significantly lower risk of discontinuation due AEs (risk ratio: 0.29; 95% CI: 0.10-0.84; P=0.02). Conclusions:A robust statistical model indicated that asciminib provides consistently superior efficacy and safety over flumatinib, supporting its value as a first-line treatment option for patients with CML-CP.

Fms-like tyrosine kinase 3 (FLT3) mutations occur in approximately 30

Accurate inference of absolute copy numbers beyond simple gains and losses from single-cell chromatin accessibility (scATAC-seq) data remains challenging, thereby obscuring the distinction between genetic and epigenetically driven oncogenic dependencies. Here, we present TeaCNV, a computational framework that reconstructs clonal absolute copy number profiles and tumor clonal architectures from scATAC-seq data without matched DNA baselines. Through validation both in silico and against bulk whole-genome sequencing in renal cell carcinomas, TeaCNV resolved subclonal absolute copy number profiles with less than 10% error and detected copy number variations (CNVs) with 98.6% accuracy, outperforming existing methods. Applied to six cancer types including renal, breast, pancreatic, head and neck, colorectal, and ovarian cancers, TeaCNV delineated polyclonal architectures and revealed distinct chromatin accessibility patterns driven by CNVs in key driver genes, including AKT2, ZNF217, and SOX2. By enabling absolute copy number profiling and clonal deconvolution from epigenomic assays, TeaCNV bridges critical gaps in studying oncogenic dependencies and genotype-phenotype relationships at single-cell resolution.

Higher eukaryotes include a widely distributed serine/threonine phosphatase called PPP4C (protein phosphatase 4 catalytic subunit). Its function is to preferentially bind to regulatory subunits, forming a complex to carry out its molecular roles. Although its involvement in pancreatic, colorectal carcinoma and breast cancer has been established, the specific role of PPP4C in pan-cancer remains ambiguous. Through pan-cancer analysis, we identified PPP4C as broadly overexpressed and linked to poor outcomes. Our investigation unveiled diverse expression patterns, demonstrating notable correlations between PPP4C expression and tumor prognosis. We further explored the association between PPP4C expression and various immune-related factors, including microsatellite instability (MSI), tumor mutation burden (TMB), the immune microenvironment, immune cell infiltration, and immune checkpoint molecules. Moreover, with the use of the Genomics of Drug Sensitivity in Cancer (GDSC) database, we discovered pharmaceutical compounds that have a strong association with PPP4C. The CancerSEA database analysis revealed that PPP4C exhibited a negative correlation with metastasis in renal cell carcinoma (RCC). The prognostic prediction of RCC was effectively carried out by the risk signature of PPP4C-related metastasis genes (PrMGs), displaying AUC values of 0.728, 0.700, and 0.706 for the one, two, and three years, respectively. Furthermore, there were notable differences in the immunological state between the two risk groups. In conclusion, our research indicates that PPP4C shows promise as a prognostic biomarker, and novel gene signatures related to metastasis can be employed for prognostic prediction in ccRCC. The risk signature based on PrMGs effectively stratifies ccRCC patient prognosis and reveals distinct immune microenvironment characteristics, providing a foundation for future research into targeted strategies.

OBJECTIVE:To evaluate the prognostic value of PCR-based measurable residual disease (PCR-MRD) in pediatric KMT2A-rearranged (KMT2A-r) acute myeloid leukemia (AML). METHODS:Forty-six children with KMT2A-r AML were monitored by PCR-MRD and multiparametric flow cytometry-based measurable residual disease (MFC-MRD) at the end of induction courses 1 and 2 (EOI1, EOI2) and post first consolidation (EOC1). MRD ≥ 0.01% was defined as positive. Outcomes included overall survival (OS), leukemia-free survival (LFS), and cumulative incidence of relapse (CIR). RESULTS:The median PCR-MRD levels decreased from EOI1 to EOC1. Among the total 96 paired samples of PCR-MRD and MFC-MRD assessment from EOI1 to EOC1, 48.9% showed concordance within the same log unit, 13.5% of total samples exhibited significant divergence (>1 log difference). Univariate and multivariate analyses demonstrated that PCR-MRD positivity at EOI1 and EOI2 was independently associated with inferior LFS and CIR, whereas allo-HSCT in first complete remission reduced CIR. Notably, EOC1 PCR-MRD positivity was significantly associated with inferior OS (p = 0.006), LFS (p = 0.014), and CIR (p = 0.003). CONCLUSION:PCR-MRD assessment during induction and consolidation provides critical prognostic information and should be integrated into risk stratification to optimize treatment strategies in pediatric KMT2A-r AML. REGISTRY: https://www.chictr.org.cn, CHICTR1900027146, REGISTRATION DATE: 2 November, 2019; ChiCTR2400080307, Registration date: 25 January, 2024.

Acute myeloid leukemia (AML) is a hematological malignancy characterized by clonal expansion of myeloid progenitors in the bone marrow, resulting in impaired hematopoiesis. AML remains challenging due to frequent disease recurrence and limited treatment alternatives, highlighting the necessity for further research. Corilagin, an ellagitannin isolated from ethnopharmacological plants, displays potential anti-cancer activity, but its anti-leukemic effects and mechanism in AML are unclear. This study aimed to assess whether corilagin induces cell death in AML cells, a cell-derived xenograft (CDX) model, and primary AML cells, and to elucidate its therapeutic mechanism in AML. Cell viability was measured using the Cell Counting Kit-8 and proliferation was assessed with an EdU imaging kit. Flow cytometry analyzed PI-positive cells. Pyroptosis was evaluated by morphology and LDH release. mRNA and protein levels were measured by qPCR and western blot. Stable cell lines with gene knockouts were created using CRISPR-Cas9. The therapeutic efficacy of corilagin was also tested in CDX models. Corilagin treatment induced pyroptotic bubbles in AML cells, increasing LDH release and PI-positive cells. Corilagin activated caspase-3, which cleaved gasdermin E (GSDME) to form active GSDME-NT, promoting pyroptosis. Corilagin also activated thioredoxin-interacting protein (TXNIP), and TXNIP knockdown by siRNA rescued corilagin-induced pyroptosis. Corilagin reduced viability and promoted pyroptosis in primary AML cells. In an AML CDX mouse model, corilagin inhibited leukemia progression and prolonged survival. Our results suggested that corilagin induced pyroptosis in AML by activating the TXNIP/caspase-3/GSDME pathway, offering a potential therapeutic strategy for AML.

Background This study aimed to evaluate the efficacy and safety of the regimens combining venetoclax with either intensive chemotherapy (3 + 7, anthracycline administered for 3 days combined with cytarabine infusion for 7 days) or modified intensive chemotherapy (2 + 5, anthracycline administered for 2 days combined with cytarabine infusion for 5 days) as induction therapy for newly diagnosed acute myeloid leukemia (AML). Methods We retrospectively collected data from 151 newly diagnosed AML patients (excluding acute promyelocytic leukemia) treated at the Institute of Hematology & Blood Diseases Hospital between 2016 and 2025. Propensity score matching (PSM) at up to a 1:4 ratio with a caliper of 0.2 was used to compare the 3 + 7+V group (n = 22) with the standard 3 + 7 group (n = 110). Additionally, patients who received the 2 + 5+V regimen (n = 19) were directly compared with those in the 3 + 7+V group (n = 22) during the same period. The primary endpoints included composite complete remission (CR/CRi) rate, measurable residual disease (MRD) negativity rate, overall survival (OS), and event-free survival (EFS). Results The CR/CRi rate was significantly higher in the 3 + 7+V group than in the standard 3 + 7 group (OR = 8.033, 95% CI: 2.020–54.752, P = 0.00373), with significantly improved EFS (HR = 0.186; 95% CI: 0.0575–0.604; P = 0.00509), albeit with more severe hematologic toxicities. In the direct comparison between the 2 + 5+V and 3 + 7+V groups, there was no significant difference in CR/CRi rates (84.21% vs. 90.91%; P = 0.993). Notably, the MRD negativity rate among responders was higher in the 2 + 5+V group (93.75% vs. 80%), and non-hematologic adverse events (e.g., diarrhea) were less frequent. Furthermore, all 3 relapsed AML patients who received 2 + 5+V as re-induction therapy achieved CR/CRi, with 2 attaining MRD-negative remission. Conclusion Adding venetoclax to standard 3 + 7 induction chemotherapy significantly improves remission rates and prolongs EFS in newly diagnosed AML, particularly in ELN low-risk patients. The 2 + 5+V regimen demonstrated promising activity with a potentially favorable toxicity profile, suggesting it may serve as an effective induction option for older or less fit patients; however, prospective validation in larger cohorts is required.

The BCL-2 inhibitor venetoclax has shown promise in acute lymphoblastic leukemia (ALL), but its role in first-line therapy for newly diagnosed (ND) Philadelphia chromosome-negative (Ph⁻) ALL is undefined. In this prospective phase 2 study, 167 adolescents and adults (aged 14-60 years) with ND Ph⁻ ALL received venetoclax combined with pediatric-inspired chemotherapy. The primary endpoint was the rate of measurable residual disease (MRD) negativity by multiparameter flow cytometry (MFC) after induction. The complete remission rate was 91.0%, and 73.0% of responders achieved MFC-MRD negativity, meeting the primary endpoint. After a median follow-up of 19.3 months, median overall and disease-free survival were not reached; estimated 2-year survival rates were 78.5% and 76.7%, respectively. Propensity score-matched analysis confirmed superior survival compared with historical chemotherapy-only controls. Grade ≥ 3 adverse events were primarily hematologic toxicities and infections, with an incidence comparable to that of the historical cohort. These results demonstrate that adding venetoclax to pediatric-inspired chemotherapy significantly improves MRD response and survival outcomes in ND Ph⁻ ALL, with a manageable safety profile. This trial was registered with ClinicalTrials.gov under the identifier NCT05660473.

e15537 Background: Immunotherapy offers limited therapeutic benefit in microsatellite stable (MSS) metastatic colorectal cancer (mCRC). Priorstudies indicate that anti- epidermal growth factor receptor (EGFR) antibody combined with chemotherapy might enhance malignant tumor immunogenicity. This study aims to evaluate the safety and efficacy of Sintilimab, a PD-1 inhibitor, combined with Cetuximab and chemotherapy, as a first-line treatment for pMMR/MSS and RAS/BRAF wild-type mCRC. Methods: A phase Ib/II dose escalation and expansion trial was conducted. Patients received 6 cycles of Sintilimab 200mg ivgtt q3w, cetuximab and chemotherapy q2w for at least 8 cycles, followed by maintenance therapy with capecitabine ±cetuximab for those whose best response is complete response (CR), partial response (PR) or stabel disease (SD) until disease progression or intolerable toxicity. Primary endpoints included safety and objective response rate (ORR). In the dose escalation phase (3+3 design), patients received cetuximab (500 mg/m², Q2W) and investigator-chosen chemotherapy regimens (mFOLFOX6, CAPEOX), with Sintilimab tested at 100 mg, 150 mg, and 200 mg (Q3W). The recommended phase II dose was 200 mg Q3W. Results: Between December 26, 2023, and December 31, 2025, 25 eligible patients were enrolled (72.0% male, 16.0% ECOG PS 0, median age 54 years， 60 % liver metastasis, 48% peritoneal metastasis, 24% PD-L1 CPS≥5 ). In the dose escalation phase, 88.9% of patients experienced grade 3/4 treatment-related adverse events, with 55.6% requiring dose adjustments for chemotherapy-related issues. No immune-related grade 3/4 adverse events were observed. The overall ORR was 76.0%, with 68.4% for the 200 mg Sintilimab cohort. The median progression-free survival (PFS) was not reached. Surgical resection was feasible in 10 patients (40.0%, 1 planned ,9 R0 resection, including 1 ypT0N0). Treatment was well tolerated, with 72.0% of patients experiencing ≥ grade 3 adverse events, most commonly neutropenia (36.0%), rash (8.0%). Conclusions: The combination of Sintilimab with cetuximab and chemotherapy shows manageable safety and efficacy in advanced pMMR/MSS and RAS/BRAF wild-type mCRC. The overall ORR was 76.0%% and 40.0% of patients achieved surgical resectability. Further clinical data are needed to confirm these findings. Clinical trial information: NCT06776757 . Treatment outcomes and surgical conversion outcomes. Therapeutic outcome N=25(%) ORR (Objective response rate) 19(76) DCR (Disease Control Rate) 25(100) SCR (Surgical conversion rate) 8(32) Surgical outcome N=9(%) Complete resection rate R0 9(100) R1 0(0) R2 0(0) Tumor regression grade TRG1 3(34) TRG2 2(22) TRG3 3(33) TRG4 1(11)

The bone marrow (BM) niche serves as a critical protective environment for leukemia cells, particularly chemo-resistant leukemia cells, and plays a central role in driving therapeutic resistance and disease relapse in acute myeloid leukemia (AML). This specialized microenvironment not only promotes leukemia cell survival, but also inhibits T cell infiltration, which serves as a major obstacle to the effectiveness of CAR-T therapy in myeloid malignancies. To overcome this limitation, we targeted Rac1 GTPase, a central regulator of cytoskeletal dynamics that controls membrane protrusion and migration, by engineering primary human T cells and CD33 CAR-T cells to express constitutively active Rac1 (Rac1V12). Our results demonstrated that active Rac1 enhanced the migration of T cells and CD33 CAR-T cells and promoted their residence in the BM in vivo. Furthermore, CD33 CAR-T cells expressing Rac1V12 displayed enhanced cytotoxicity against leukemia cells in vitro, as demonstrated by transwell migration-dependent killing assays. Crucially, these engineered CAR-T cells achieved superior robust suppression of leukemia in vivo and significantly prolonged survival in xenograft models. Mechanistically, Rac1V12 CD33 CAR-T cells in the BM demonstrated enhanced immunological memory phenotype and lower tonic signaling, a combination that promotes T cell persistence and enhances anti-tumor efficacy in vivo. Our data suggest that active Rac1-engineered CD33 CAR-T cells represent a novel strategy for targeting BM leukemia cells, with the potential to eradicate AML cells.

ABSTRACT Background Real‐world data on low‐dose emicizumab in young children with hemophilia A are limited. Methods This multicenter retrospective study included 26 children (median age: 2.2 years) receiving individualized low‐dose emicizumab (2019–2025). Results Over 48 months, the annualized bleeding rate decreased from 5.31 to 0.60 (p < 0.001); the median maintenance dose was 0.81 mg/kg/week. FVIII equivalent levels correlated with dose (Quality of life [CHO‐KLAT]) improved from 57.2 to 69.8 (p < 0.001). Subclinical joint changes progressed despite bleeding control. No thrombotic events occurred. Conclusions Low‐dose emicizumab effectively reduces bleeding and improves quality of life, supporting its feasibility in resource‐limited settings. Trial Registration The authors have confirmed clinical trial registration is not needed for this submission.

To develop and validate a prognostic model based on palmitoylation‑related genes for risk stratification in acute myeloid leukemia (AML), and to explore its associations with the immune microenvironment, drug sensitivity, and single‑cell expression patterns. A five‑gene prognostic signature was constructed using univariate Cox regression and LASSO‑Cox analyses in the TCGA‑LAML training cohort (n = 131) and validated in an independent GEO cohort (GSE71014, n = 104). Risk scores were calculated, and patients were divided into high‑ and low‑risk groups. Overall survival (OS) was compared by Kaplan‑Meier curves, and model performance was evaluated by time‑dependent ROC. Immune cell infiltration, immune checkpoint expression, and drug sensitivity (predicted IC50) were analyzed. Single‑cell RNA‑seq data (GSE116256) were used to examine cell‑type specific expression of the model genes. The five‑gene model (LYPLA2, ZDHHC11, ZDHHC7, ABHD6, ABHD17C) significantly distinguished high‑risk patients with poorer OS in both training (P < 0.001) and validation (P < 0.001) sets, with AUCs of 0.72, 0.73, 0.75 (training) and 0.67, 0.70, 0.60 (validation) for 1‑, 3‑, and 5‑year survival. Multivariate Cox regression confirmed independent prognostic value (HR = 2.78, P < 0.001). High‑risk patients showed increased regulatory T cells, activated dendritic cells, and follicular helper T cells, along with positive correlations with PD‑1, TIM‑3, LAG3, and VISTA expression. Ninety‑eight compounds with differential predicted IC50 values were identified. Single‑cell analysis revealed LYPLA2 and ZDHHC7 enrichment in monocytes/macrophages and dendritic cells. The five‑gene palmitoylation‑associated prognostic model effectively stratifies AML patients by risk, reflects immune microenvironment dysregulation, and offers potential guidance for personalized therapy.

Chimeric antigen receptor T-cell (CAR-T) therapy has revolutionized the treatment landscape for hematological malignancies such as relapsed/refractory B-cell acute lymphoblastic leukemia (R/R B-ALL), with several CAR-T products now approved globally for R/R B-ALL. Despite high initial response rates, major challenges remain, including disease relapse due to antigen escape and the limited persistence of CAR-T cells; treatment-related adverse events such as cytokine release syndrome, immune effector cell-associated neurotoxicity syndrome, hematologic toxicity, and infections; and limited access to CAR-T therapy. In this review, we discuss the limitations of CAR-T and strategies to overcome them, specifically in the context of B-ALL, including the use of allogeneic CAR-T, dual-targeted CAR-T, and combination strategies with novel technologies and agents. Furthermore, we explored the role of consolidative allogeneic hematopoietic stem cell transplantation after CAR-T therapy and the potential of integrating CAR-T into the first-line treatment for B-ALL. Future research should aim to increase the efficacy of CAR-T, reduce their toxicity, improve their accessibility, and expand their use to earlier lines of therapy for B-ALL.