BACKGROUND:With life expectancies of patients with chronic myeloid leukemia (CML) now approaching those of the general population under tyrosine kinase inhibitor (TKI) therapy, understanding long-term treatment-related toxicities becomes paramount. Although imatinib's nephrotoxic potential is established, the renal safety profiles of second-generation TKIs-dasatinib, nilotinib, and flumatinib-remain incompletely characterized. METHODS:This single-center retrospective study analyzed renal function trajectories in 348 patients with CML receiving first-line imatinib, dasatinib, nilotinib, or flumatinib. Renal function was assessed using serum creatinine and estimated glomerular filtration rate (eGFR). Mixed-effects models incorporating treatment group, time, and their interaction as fixed effects were used to formally compare eGFR trajectories across treatment groups. Chronic renal adverse events and acute kidney injury were also evaluated. RESULTS:Formal comparison of eGFR trajectories across treatment groups revealed a significant time × treatment interaction, indicating differential renal function changes among the 4 TKIs. Significant serum creatinine elevation and eGFR decline were observed in imatinib and flumatinib groups, contrasting with stable creatinine and divergent eGFR patterns in dasatinib (slight increase) and nilotinib (decline) groups. The incidence of chronic renal adverse events varied substantially across cohorts-12.3% (imatinib), 2.6% (nilotinib), 4.3% (dasatinib), and 9.6% (flumatinib). Notably, among 3 documented acute kidney injury cases, all occurred during flumatinib treatment. Furthermore, treatment-free remission achievement did not confer significant renal function recovery, suggesting potential irreversibility of established renal damage. CONCLUSION:TKIs exhibit differential renal safety profiles, underscoring the critical importance of individualized TKI selection based on baseline renal status and implementation of rigorous renal monitoring protocols throughout treatment duration.

Background:T-lineage acute leukemias are aggressive malignancies for which treatment options are limited. Objectives:This study evaluated the efficacy and safety of a non-cytotoxic regimen combining venetoclax, azacitidine, and dexamethasone (VAD) as first-line induction therapy for T-cell acute lymphoblastic leukemia (T-ALL) and T/myeloid mixed-phenotype acute leukemia (T-Myeloid MPAL). Design:We retrospectively analyzed the clinical parameters and survival data of 11 patients with newly-diagnosed T-ALL or T-Myeloid MPAL who received the VAD regimen. Methods:The complete remission (CR) rate and adverse events were assessed. Kaplan-Meier curves were constructed to evaluate survival outcomes. Results:Among 11 patients (7 with early T-cell precursor [ETP] ALL, 2 with T-Myeloid MPAL, and 2 with non-ETP T-ALL), the overall CR rate after one cycle of VAD therapy was 90.9%, with 45.5% achieving measurable residual disease (MRD) negativity by flow cytometry. The historically poor-prognosis ETP-ALL subgroup showed an 85.7% CR rate (6/7), while both T-Myeloid MPAL cases attained MRD negativity. No tumor lysis syndrome or treatment-related mortality occurred. Over a median follow-up of 358 days, all 5 patients who received transplantations remained relapse-free. Conclusion:The VAD regimen demonstrated a high response rate and favorable safety, enabling effective transplantation bridging with avoidance of conventional cytotoxic therapy.

Objective:To evaluate the long-term efficacy and safety of high-dose idarubicin plus busulfan (I-Bu) conditioning followed by autologous stem cell transplantation (ASCT) compared to intermediate- to high-dose cytarabine (Ara-C) consolidation in young acute myeloid leukemia (AML) patients with favorable- or intermediate-risk in first complete remission (CR1). Methods:We retrospectively analyzed clinical data from 59 young AML patients (aged ≤ 65 years) with favorable- or intermediate-risk disease who received the I-Bu conditioning regimen followed by ASCT between December 2004 and December 2021 (ASCT group). Clinical outcomes were compared with 57 favorable- and intermediate-risk AML patients treated with intermediate- to high-dose Ara-C consolidation chemotherapy alone (chemotherapy group). Overall survival (OS) and disease-free survival (DFS) were evaluated, and univariate and multivariate analyses were performed to identify prognostic factors associated with OS. Results:A total of 116 patients were included with a median follow-up of 79.5 months. Median OS was not reached in either group. The ASCT group achieved a significantly higher 2-year OS rate (84.5% vs 59.7%, P=0.0018) and sustained a significant OS benefit in the 10-year OS rate (P=0.0017). DFS rate also showed superiority in the ASCT group at 2-year (77.7% vs 53.5%, P=0.0037) and maintained this benefit in the 10-year DFS rate (P=0.0038). Multivariate Cox regression identified treatment modality as an independent prognostic factor for OS (HR = 3.12, 95% CI: 1.48-6.59, P=0.0028). In subgroup analysis, ASCT significantly improved OS (P<0.001) and DFS (P=0.0014) in favorable-risk patients, whereas no differences were observed in intermediate-risk patients (OS, P=0.13; DFS, P=0.21). Conclusion:The I-Bu conditioning regimen followed by ASCT provides durable survival benefits and a favorable safety profile for young, favorable-risk AML patients in CR1, representing a potential post-remission therapeutic option. Its role in intermediate-risk AML requires further validation.

This study aims to investigate the genetic characteristics of Acute Myeloid Leukemia (AML) patients and identify which patients derive the greatest benefit from a low-intensity regimen of decitabine combined with modified Cytarabine + Aclarubicin + Granulocyte Colony-Stimulating Factor (D-CAG) or intensive chemotherapy (IA regimen). We retrospectively analyzed cytogenetic and molecular data of 331 newly diagnosed AML patients and examined the associations between genetic characteristics, risk status, treatment approaches, and clinical outcomes. The median follow-up duration was 45 months (range: 2-120 months). Patients receiving IA therapy achieved higher complete remission (CR) rates (79.3

Immunogenic cell death (ICD) represents a specific form of regulatory cell death that initiates an adaptive immune response. We aimed to investigate the significance of immunogenic cell death-related genes (ICDGs) in AML, utilizing a combination of bioinformatics analysis, consensus clustering, functional enrichment analysis, and experimental validation in cell lines and animal models. Here, we identified 34 ICDGs, and single-cell analysis indicated that CD8+ T cells and NK cells exhibited gene expression patterns closely associated with ICD. Consensus clustering revealed two distinct subtypes of AML (ICD-high and ICD-low), with the former correlating with more favorable clinical outcomes and heightened infiltration of immune cells. A predictive model was established through LASSO regression, yielding a risk signature comprising six key ICDGs (FGFBP2, GZMB, ALPK2, NELL2, OPTN, FCGR2B), which successfully categorizes patients into high-risk and low-risk cohorts based on overall survival outcomes. Notably, HSPA6 emerged as a critical ICDG, with its knockdown in OCI-AML3 cells significantly inhibiting proliferation and inducing apoptosis, suggesting its potential as a therapeutic target. In summary, our research emphasizes the importance of ICD-related genes in predicting the prognosis of AML and initiates the development of a prognostic risk signature that may pave the way for personalized treatment strategies while highlighting the need for further validation and exploration of HSPA6 in AML.

Acute myeloid leukemia (AML) commonly affects the elderly with a poor prognosis. Body water composition analysis provides a new perspective for biomedical research. This study aims to develop and validate a simple nomogram for predicting overall survival (OS) in AML survivors. A total of 291patients were enrolled and randomly divided into a training cohort and an internal validation cohort. The median duration of follow-up was 32.2 months.The LASSO regression was used to screen predictors of survival in the training cohort, and the multivariate Cox model was used to establish a nomogram. The discrimination and calibration of the nomogram were evaluated using the C-index, area under the time-dependent receiver operating characteristic curve (time-dependent AUC), and calibration plots. The net benefits of the nomogram at different threshold probabilities were quantified. Five predictors of AML survival were identified: Age, Extracellular water/Intracellular water (ECW/ICW) ratio, European Leukemia Net Risk, Sarcopenia, and WBC. The nomogram showed good performance in both the training cohort (C-index 0.755, 95

Purpose:This study evaluated the efficacy and safety of a 14-day blinatumomab-venetoclax (BV) regimen as induction therapy for newly diagnosed Ph-negative B-cell acute lymphoblastic leukemia (B-ALL), focusing on rapid remission and tolerability in unfit patients. Patients and Methods:Thirteen patients received venetoclax (100 mg on day 1, 200 mg on day 2, 300 mg on day 3, and 400 mg from days 4 to 14) with blinatumomab (9 to 28 ug/day) for 14 days. Bone marrow assessments were performed at days 14-21. Primary endpoints were complete remission (CR) rate, minimal residual disease (MRD) negativity by flow cytometry, and adverse events. Results:The CR rate after one cycle of BV regimen was 92.3% (12/13), and all patients achieved MRD-negativity; 91.7% (11/12) achieved MRD clearance by day 21. Grade 1-2 cytokine release syndrome occurred in 46.2% (6/13; 1 grade 3). Hematologic toxicity included grade 3-4 neutropenia (92.3%) and thrombocytopenia (46.2%), with only 30.8% febrile neutropenia. All AEs resolved rapidly with supportive care, allowing therapy to continue without interruption. At median follow-up of 283 days, 1-year relapse-free survival rate and overall survival rate were 60.6% and 83.3%. Conclusion:The 14-day BV regimen induced rapid deep remission (91.7% MRD-negative by day 21) with manageable toxicity in Ph-negative B-ALL. Synergistic T-cell activation by venetoclax may explain enhanced efficacy.

Philadelphia chromosome-negative B-cell acute lymphoblastic leukemia (Ph-negative BCP-ALL) accounts for a significant portion of adult cases. Blinatumomab, a bispecific T-cell engager, has shown efficacy in relapsed or refractory BCP-ALL, but its role in induction therapy with reduced-dose chemotherapy is being explored. In this retrospective study, 35 newly diagnosed Ph-negative BCP-ALL patients received reduced-dose chemotherapy followed by two weeks of blinatumomab (RDC-Blinatumomab-2W) as part of our previous clinical trial. These patients were compared with a propensity score-matched historical control group of 35 patients treated with the hyper-CVAD regimen. The primary endpoint was composite complete remission (CRc); secondary endpoints included minimal residual disease (MRD) negativity, adverse events, and survival outcomes. After matching, both groups had 17 patients (49

BACKGROUND:Splicing factor 3b subunit 1 (SF3B1) mutations have been implicated in hematologic malignancies, but the clinical significance of distinct SF3B1 mutation variants remain unclear. METHODS:The objective of this study was to evaluate clinicopathologic features, mutational profiles, and outcomes of 1691 patients with hematologic malignancies, including myelodysplastic syndromes (MDS; n = 402), acute myeloid leukemia (AML; n = 758), and chronic lymphocytic leukemia (CLL; n = 531). RESULTS:The frequency of SF3B1-mutated (SF3B1MUT) MDS, AML, and CLL was 70 of 402 patients (17.4%), 23 of 758 patients (3.0%), and 45 of 531 patients (8.5%), respectively. p.K700E was the most prevalent SF3B1MUT variant and was identified in 43 of 70 of patients with MDS (61.4%), in seven of 23 patients with AML (30.4%), and in 19 of 45 patients with CLL (42.2%). In MDS and AML, TET2 was the most frequent co-mutated gene in patients with SF3B1MUT disease (20 of 70 patients with MDS [28.6%]; 11 of 23 patients with AML [47.8%]). For patients with SF3B1MUT CLL, the most common co-mutated genes were ATM (11 of 45; 24.4%) and TP53 (11 of 45; 24.4%). Kaplan-Meier analysis indicated that the SF3B1 p.K700E variant was significantly associated with improved overall survival (OS) and progression-free survival (PFS) in patients who had MDS (p < .001 and p = .016, respectively) but with worse OS and PFS in those who had AML (p = .006 and p = .006, respectively) compared with those who had wild-type SF3B1. In patients who had CLL, p.I704F was associated with reduced OS (p < .001) and p.K700E was associated with a shorter time-to-first treatment (p = .028) compared with those who had wild-type SF3B1. Multivariable analysis identified p.K700E as an independent protective factor for OS in patients with MDS (p = .048) but as an independent risk factor for both OS and PFS in patients with AML (p = .036 and p = .035, respectively) and for the time to first treatment in patients with CLL (p = .033). CONCLUSIONS:Specific SF3B1 variants should be incorporated into prognostic stratification for hematologic malignancies.

BACKGROUND:T/myeloid mixed-phenotype acute leukemia (T/My MPAL) is a malignant disease characterized by co-expression of lymphoid and myeloid features. The lack of molecular classification of T/My MPAL results in highly heterogeneity in treatment responses and clinical outcomes. Identifying molecular subtypes and developing subtype-specific treatment strategies are crucial for improving prognosis and enabling personalized therapies. METHODS:We constructed a single-cell transcriptomic landscape of T/My MPAL, acute myeloid leukemia (AML), T-cell acute lymphoid leukemia (T-ALL), and normal donors by analyzing nearly 275,000 cells. Malignant cells were identified using lineage-specific markers and healthy reference datasets. By comparing the whole transcriptomic profiles of T/My MPAL malignant cells with those of AML and T-ALL, we defined three distinct subpopulations and uncovered both intra- and inter-tumoral heterogeneity. Subpopulation-specific molecular markers were identified and validated using immunohistochemistry and independent datasets. These markers were further linked to clinical outcomes. Finally, potential subpopulation-specific therapeutic drugs were identified by correlating gene signatures with IC50 values. RESULTS:Malignant cells in T/My MPAL display distinct lineage characteristics and experience differentiation arrest at a more primitive stage compared to other leukemias. Biphenotypic and bilineal MPAL subtypes defined by flow cytometry exhibit similar transcriptomic profiles, indicating the traditional classification based on a limited set of lineage markers is insufficient. Instead, we define three subpopulations of malignant cells in T/My MPAL, including AML-like, T-ALL-like, and a unique subpopulation that shows distinct transcriptional characteristics neither similar to AML nor to ALL. Markers for each subpopulation are identified and further validated by independent datasets and immunohistochemistry. The unique subpopulation exhibits higher stemness and quiescence, with elevated HOPX expression. Notably, patients with higher levels of the unique subpopulation have significantly poorer prognoses. We further computationally screen potential drugs targeting each subpopulation and indicate that Venetoclax could effectively inhibit the unique MPAL subpopulation and help patient achieve complete remission. CONCLUSIONS:Our study provides new insights into the molecular heterogeneity and offers personalized diagnostic and therapeutic targets for T/My MPAL patients. These findings offer valuable insights for enhancing patient outcomes and developing personalized treatment strategies.

Flumatinib is a novel second-generation tyrosine kinase inhibitor (2G-TKI), which was approved in November 2019 in China. A previous phase III study evaluated the efficacy and safety of flumatinib as a first-line therapy for patients with chronic phase chronic myeloid leukemia (CML-CP). However, randomized trials comparing flumatinib with other 2G-TKIs remain lacking. To assess the efficacy and safety of flumatinib versus nilotinib as a first-line treatment for CML-CP. A multicenter retrospective study. We retrospectively analyzed 101 and 64 patients treated with flumatinib and nilotinib during the same period, respectively. Patients in the flumatinib group were significantly older than patients in the nilotinib group (median age, 44 vs 37 years; p = 0.004). The optimal response and treatment failure rates at 24 months were comparable between the two groups. At 12 months, 85.1% and 88.2% of patients in the flumatinib and nilotinib groups, respectively, achieved a major molecular response (MMR; p = 0.648). By 24 months, 9.9% and 12.5% of patients suffered treatment failure in the flumatinib and nilotinib groups, respectively (p = 0.602). At 9, 12, and 24 months, the rate of MR4 (a BCR::ABL1 transcript level ⩽0.01%) achievement was significantly higher in patients treated with nilotinib than in those treated with flumatinib (26.0% vs 53.7%, p = 0.007; 40.4% vs 60.8%, p = 0.044; and 41.7% vs 80.8%, p = 0.042, respectively). In addition, elevated alanine aminotransferase or aspartate aminotransferase (ALT/AST), glucose, and serum lipid; hyperbilirubinemia; rash; and alopecia were more frequent among patients receiving nilotinib, whereas diarrhea was more frequent in those receiving flumatinib. Flumatinib is a suitable alternative as a first-line treatment for patients with CML-CP to achieve a fast MMR with better tolerability.

Acute myeloid leukemia (AML) cells and activated T cells rely on aerobic glycolysis for energy metabolism. The TP53-induced glycolysis and apoptosis regulator (TIGAR) inhibits glycolysis and protects AML cells from apoptosis. Preliminary studies suggest that combining TIGAR inhibition with the glycolysis inhibitor 2-deoxy-D-glucose (2-DG) may offer a therapeutic strategy for AML. However, it remains unclear whether silencing TIGAR can enhance T cell function and thereby improve AML prognosis. This study aims to investigate whether TIGAR silencing in host can eliminate AML cells and rejuvenate dysfunctional T cells with mouse models. TIGAR knockout mice on the C57BL/6J background were generated and AML mouse models were established through intravenous injection of C1498 cells. We found that TIGAR depletion enhanced CD8+ T cell counts and raised CD4/CD8 ratio, downregulating CD44 and immune checkpoints CTLA-4, LAG-3, PD-1 on cell surface of CD8+ T cells. TIGAR depletion boosted cytokine secretion (IFN-γ, perforin, granzyme B, TNF-α) by CD8+ T cells and IL-2, TNF-α by CD4+ T cells, improving cytotoxicity against AML cells, proliferation, and reducing apoptosis. TIGAR suppression in host with 2-DG prolonged AML mouse survival, decreased tumor burden, and leukemic infiltration. TIGAR suppression restored thymic T cell development and peripheral immune balance. Single-cell RNA sequencing analysis also revealed that high TIGAR expression influences the glycolysis pathway, and correlates with markers of T cell exhaustion. This study indicates that blocking TIGAR prevents CD8+ T cell dysfunction and induces anti-AML immunity.

Introduction: Mixed-phenotype acute leukemia (MPAL) is a rare and heterogeneous group of malignant diseases, accounting for 2%-5% of acute leukemia. MPAL can be further classified as B/myeloid (B/My), T/myeloid (T/My), rare types, acute leukemia of ambiguous lineage, not otherwise specified or acute undifferentiated leukemia based on the immunophenotype, according to the WHO criteria. The treatment of acute leukemia has seen significant advances over the last three decades, partly due to individualized treatment. However, clinical prognosis for MPAL patients is worse than both acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML), with a 5-year survival rate of 47-75% in children and 20-40% in adults. Given the rarity of MPAL, historically poorly understanding in the pathogenesis and biology and a lack of prospective trial data to guide therapy, there are currently no established standard treatment guidelines for MPAL to date. The diversity in treatment sensitivity arises from the heterogeneity and unique molecular characteristics of malignant cells. Therefore, there is an urgent need to clarify the molecular subtypes of MPAL and identify potential therapeutic methods to provide personalized treatment strategies and improve the prognosis of the patients with MPAL. Methods: We conducted an in-depth analysis of single-cell transcriptome data from nearly 260,000 derived from the bone marrow of patients with AML, T-cell acute lymphoblastic leukemia (T-ALL), or T/myeloid MPAL, comparing these data with healthy donors. We employed various analytical methods, including cell subpopulation analysis, pseudotime analysis, cell type mapping, stemness evaluation, and functional enrichment analysis, to identify novel molecular subtypes associated with T/My MPAL. The findings were subsequently validated through external bulk transcriptome data and immunohistochemistry (IHC) staining. Furthermore, we assessed survival prognosis and drug sensitivity differences among patients with these subtypes. Results: Our study revealed that T/My MPAL malignant cells exhibited unique biphenotypic characteristics, significantly different from traditional AML or T-ALL. Although clinical differences between biphenotypic and bilineal T/My MPAL may exist, they were not significant at the whole transcriptome level. Using a topological manifold learning algorithm, we identified three subpopulations of MPAL malignant cells at the single-cell level, including AML-like, ALL-like, and a unique subpopulation that highly expressed the HOPX gene and exhibited higher stemness and quiescence. Notably, patients with a high prevalence of the unique subpopulation had significantly poorer prognosis. Additionally, we identified specific molecular markers for each subpopulation and validated these findings in multiple independent public datasets. Furthermore, we screened potential therapeutic drugs for each subtype, with venetoclax showing significant efficacy potential for the unique T/My MPAL subtype. Conclusions: Our research provided new insights into the molecular heterogeneity of MPAL and offered personalized diagnostic and therapeutic targets for patients. By identifying and validating different molecular subtypes and their specific markers, we screened potential therapeutic drugs for each subtype, with venetoclax showing promising therapeutic potential, particularly for patients with the unique T/My MPAL subtype. These findings are expected to improve the prognosis of MPAL patients and advance the development of personalized treatment strategies.

Given the extremely high inter-patient heterogeneity of acute myeloid leukemia (AML), the identification of biomarkers for prognostic assessment and therapeutic guidance is critical. Cell surface markers (CSMs) have been shown to play an important role in AML leukemogenesis and progression. In the current study, we evaluated the prognostic potential of all human CSMs in 130 AML patients from The Cancer Genome Atlas (TCGA) based on differential gene expression analysis and univariable Cox proportional hazards regression analysis. By using multi-model analysis, including Adaptive LASSO regression, LASSO regression, and Elastic Net, we constructed a 9-CSMs prognostic model for risk stratification of the AML patients. The predictive value of the 9-CSMs risk score was further validated at the transcriptome and proteome levels. Multivariable Cox regression analysis showed that the risk score was an independent prognostic factor for the AML patients. The AML patients with high 9-CSMs risk scores had a shorter overall and event-free survival time than those with low scores. Notably, single-cell RNA-sequencing analysis indicated that patients with high 9-CSMs risk scores exhibited chemotherapy resistance. Furthermore, PI3K inhibitors were identified as potential treatments for these high-risk patients. In conclusion, we constructed a 9-CSMs prognostic model that served as an independent prognostic factor for the survival of AML patients and held the potential for guiding drug therapy.

Introduction: Although intensive pediatric chemotherapeutic regimens have shown good efficacy in the treatment of Philadelphia Chromosome-negative (Ph-negative) B-cell precursor acute lymphoblastic leukemia (BCP-ALL) in adults, the high incidence of chemotherapy-related toxicities limits their use. Blinatumomab, a CD3/CD19 bispecific T-cell antibody, has shown high efficacy in relapsed/refractory or minimal residual disease (MRD) positive BCP-ALL; however, the efficacy of single-agent blinatumomab does not meet the treatment needs of adult patients. Therefore, combining low-intensity chemotherapy with blinatumomab could be a better induction regimen for newly diagnosed adult patients with BCP-ALL. Methods: This multicenter, single-arm, phase 2 trial (NCT05557110) enrolled adult patients (aged 15-59 years) with newly diagnosed Ph-negative BCP-ALL. The induction regimen comprised reduced intensity chemotherapy (idarubicin 8 mg/m 2 on day 1, vindesine 4 mg on day 1, and dexamethasone 9 mg/m 2/day from day 1-7) followed by 2 weeks of blinatumomab (9 ug/day from day 8-14, 28 ug/day from day 15-21). The primary endpoint was to assess the overall response rate (ORR=complete remission [CR] + CR with partial hematologic recovery [CRh] + CR with incomplete hematologic recovery [CRi]) of this induction regimen. Secondary endpoints were MRD negativity (<1x10 -4 by flow cytometry), event-free survival (EFS), overall survival (OS) and safety. Bone marrow evaluation and one intrathecal injection chemotherapy were performed on day 22±2. Patients not achieving ORR continued blinatumomab for another 2 weeks (28 ug/day from day 1-14), followed by bone marrow re-evaluation. Consolidation therapy was given after achieving ORR with recommended multidrug combination chemotherapy (including high-dose methotrexate or cytarabine combined with asparaginase) or alternating with blinatumomab (28 ug/day for 28 days). If allogeneic hematopoietic stem cell transplantation (allo-HSCT) is not performed, consolidation therapy needs at least 4 courses before 2 years of maintenance therapy. Results: Interim results are presented here.By July 6, 2023 (data cut-off), 25 patients were enrolled with a median age of 42 years (range: 15-53); 68% were females. The Eastern Cooperative Oncology Group Performance Status score ranged from 0-1. Eight (32.0%) patients had poor cytogenetic risk factors, including 3 with P2RY8::CRLF2, 2 with hypodiploidy, 2 with KMT2A rearranged and 1 with alterations of IKZF1 (Table 1). The bone marrow evaluation done in 19 patients on day 8 prior to blinatumomab treatment showed that none of the patients had got ORR. All 21 (100%) patients evaluable for the primary endpoint attained CR/Cri - 19 at the end of 2 weeks of blinatumomab and the remaining 2 after another 2 weeks of blinatumomab (Table 2). The MRD negativity rate was 90.5% (19/21) after 2 weeks of blinatumomab which increased to 95.5% (20/21) after another 2 weeks of treatment with blinatumomab. In 21 patients evaluated for safety, blinatumomab-related cytokine release syndrome was reported in 10 (47.6%) patients (grade 1: n=6, grade 2: n=3 and grade 3: n=1). No incidence of neurotoxicity was reported. Febrile neutropenia was reported in 4 (19%) patients (all ≥grade 3). The non-hematological AEs reported were fever in 7 (33.3%) patients (any grade), increased ALT in 10 (47.6%) patients (any grade) and infections in 4 (19%) patients, including pneumonia in 2 and sepsis in 2 (all ≥grade 3). The median duration of neutrophil deficiency (<0.5×10 9/L) and platelet deficiency (<20×10 9/L) during induction therapy was 9 days (range: 0-20) and 1 day (range: 0-18), respectively. The median follow-up time was 4 months and no relapse or leukemia-related deaths were observed. The median EFS and OS were not yet reached. All enrolled patients are alive except one who died of infection after multidrug combination consolidation chemotherapy. The median cycles for consolidation were 2 (range: 1-5) with 3 patients undergoing allo-HSCT. Conclusion: Thesepreliminary results indicate that the reduced intensity chemotherapy followed by blinatumomab is an effective regimen in adults with newly diagnosed Ph-negative BCP-ALL, resulting in favorable response rates, deep remission, and low-grade treatment-related AEs. These promising findings may provide a rationale for integrating blinatumomab into future induction therapy recommendations.

Chronic myeloid leukemia (CML) is a type of hematologic malignancies caused by BCR-ABL chimeric oncogene. Resistance to tyrosine kinase inhibitors (TKIs) leads to the progression of CML into advanced stages. Selinexor is a small molecule inhibitor that targets a nuclear transporter called Exportin 1. Combined with imatinib, selinexor has been shown to disrupt nuclear-cytoplasmic transport signal of leukemia stem cells, resulting in cell death. The objective of this study was to investigate the mechanism of drug resistance to selinexor in CML. We established K562 cell line resistant to selinexor and conducted single cell dynamic transcriptome sequencing to analyze the heterogeneity within the parental and selinexor resistant cell populations. We identified specific gene expression changes associated with resistance to selinexor. Our results revealed differential expression patterns in genes such as MT2A, TFPI, MTND3, and HMGCS1 in the total RNA, as well as MT-TW, DNAJB1, and HSPB1 in the newly synthesized RNA, between the parental and drug-resistant groups. By applying pseudo-time analysis, we discovered that a specific cluster of cells exhibited characteristics of tumor stem cells. Furthermore, we observed a gradual decrease in the expression of ferroptosis-related molecules as drug resistance developed. In vitro experiments confirmed that the combination of a ferroptosis inducer called RSL3 effectively overcame drug resistance. In conclusion, this study revealed the resistance mechanism of selinexor in CML. In conclusion, we identified a subgroup of CML cells with tumor stem cell properties and demonstrated that ferroptosis inducer improved the efficacy of selinexor in overcoming drug resistance.

OBJECTIVE:To investigate the clinical characteristics and influence of co-mutated gene on acute myeloid leukemia patients (AML) with FMS-like tyrosine kinase-3 (FLT3) mutations. METHODS:A total of 273 FLT3+ AML patients were enrolled, and the co-mutation gene data of the patients were collected to further analyze the prognosis of the patients. FLT3 and other common mutations were quantified by PCR amplification products direct sequencing and second-generation sequencing (NGS). RESULTS:When patients were divided into FLT3- ITD +, FLT3- TKD +, FLT3- ITD ++TKD + and FLT3- ITD -+TKD - group according to the type of FLT3 mutations, it was found that the frequencies of TET2, GATA2, NRAS and ASXL1 mutation were significantly different among the 4 groups (all P < 0.05). When patients were divided into allelic ratio (AR) ≥0.5 and <0.5 group, it was found that the frequencies of FLT3- ITD +, FLT3 -ITD - +TKD -, NPM1, NRAS and C-kit were significantly different between the two groups (all P < 0.05). When patients were divided into normal and abnormal karyotype group, it was found that the frequencies of FLT3- ITD +, FLT3- TKD +, NPM1, GATA2 and C-kit were significantly different between the two groups (all P < 0.05). The median overall survival (OS) of AML patients with FLT3 -TKD + (including FLT3- ITD ++TKD +) was longer than that of patients with FLT3- ITD + alone (P < 0.05). The OS and relapse-free survival (RFS) of AML patients with FLT3++TET2+ were both shorter than those of patients with FLT3++TET2- (both P < 0.05). CONCLUSION:The mutation frequencies of co-mutated genes are correlated with subtypes of FLT3, karyotype and AR. AML patients with FLT3 -TKD + have longer OS than patients with FLT3- ITD + alone, and patients with co-mutation of TET2 have shorter median OS and RFS.

Background: Acute myeloid leukemia (AML) patients with various nucleophosmin 1 (NPM1) mutations are controversial in the prognosis. This study aimed to investigate the prognosis of patients according to types of NPM1 mutations (NPM1mut). Methods: Bone marrow samples of 528 patients newly diagnosed with AML, were collected for morphology, immunology, cytogenetics, and molecular biology examinations. Gene mutations were detected by next-generation sequencing (NGS) technology. Results: About 25.2

The objective of this study was to analyze the correlation between skeletal muscle mass and the distribution of peripheral blood lymphocytes and natural killer (NK) cells, as well as their impact on prognosis in patients with acute myeloid leukemia (AML). A retrospective analysis was conducted on 211 newly diagnosed AML patients, evaluating skeletal muscle index (SMI), NK cell proportion, and absolute value, along with relevant clinical data. Linear regression and Spearman's correlation coefficient were used to assess the relationship between various indicators and SMI, followed by multiple linear regression for further modeling. Univariate and multivariate Cox proportional hazards regression models were used to identify independent predictors for overall survival (OS). Among the 211 AML patients, 38 cases (18.0%) were diagnosed with sarcopenia. Multiple linear regression analysis included weight, fat mass, ECOG score, body mass index, and peripheral blood NK cell proportion, constructing a correlation model for SMI (R2 = 0.745). Univariate analysis identified higher NK cell count (> 9.53 × 106/L) as a poor predictor for OS. Multivariate Cox proportional hazards regression model indicated that age ≥ 60 years, PLT < 100 × 109/L, ELN high risk, sarcopenia, and B cell count > 94.6 × 106/L were independent adverse prognostic factors for AML patients. Low skeletal muscle mass may negatively impact the count and function of NK cells, thereby affecting the prognosis of AML. However, further basic and clinical research is needed to explore the specific mechanisms underlying the relationship between NK cells and SMI in AML.

Introduction: The optimal treatment of acute myeloid leukemia (AML) in elderly patients remains a significant challenge, underscoring the urgent need for more feasible and safer treatment strategies. Microtransplantation (MST) has emerged as a novel therapeutic approach, involving the infusion of HLA-mismatched donor hematopoietic stem cells (HSC) to establish microchimerism, which has been shown to improve clinical outcomes. Umbilical cord blood (UCB) serves as a readily available source of allogeneic HSC, possessing intrinsic anti-tumor effects, immune modulation capabilities, and providing hematopoietic support. Furthermore, previous studies have indicated that interleukin-2 (IL-2) can bolster the proliferation and cytotoxic activity of both T cells and natural killer (NK) cells, potentially enhancing the immune response against AML. In light of these findings, we conducted an investigator-initiated, prospective study (ChiCTR-OPC-1900024089) to evaluate the efficacy and toxicity of a regimen combining decitabine, granulocyte colony-stimulating factor (G-CSF) priming, low-dose aclarubicin, and cytarabine (DCAG) chemotherapy followed by unrelated HLA-mismatched UCB microtransplantation (UCB-MST) and interleukin-2(IL-2) in elderly AML patients. Methods: A total of 100 patients with a median age of 68-years-old (60~85 years) who received DCAG combined with UCB-MST and IL-2 (MST, n=50) or DCAG regimen alone (non-MST, n=50) as induction and consolidation therapy were enrolled in the study. The patients in MST group received a DCAG regimen followed by an intravenous infusion of HLA-mismatched UCB after 24 to 72 hours without graft-versus-host-diseases (GVHD) prophylaxis. Additionally, subcutaneous injections of IL-2 at a dose of 1 million IU were administered every other day for 6 to 12 months after UCB infusion. Importantly, none of the patients received allogeneic or autolougous hematopoietic stem cell transplantation. Following UCB infusion, we performed real-time quantitative polymerase chain reaction (RQ-PCR) to assess microchimerism in the peripheral blood of 22 patients, with samples collected between 21 to 35 days post-transfusion. The immune cell populations and phenotypic characteristics of peripheral blood were analyzed in 13 patients (7 from the MST arm and 6 from non-MST arm) using single-cell mass cytometry, both before and after the first induction treatment. Results: Our data revealed that the patients in the MST group had significantly better complete remission (CR) rate (84% vs.66%, P=0.0377) and median overall survival (OS) (26 vs. 13.5months, P=0.0116) compared to those in non-MST group after one cycle of induction, with a median follow-up of 42 months. Notably, the CR rate after one induction cycle was significantly higher in the intermediate- and adverse-risk patients of the MST group compared to the non-MST group (77.8% vs. 52.9%, P=0.0287). Among the MST group, 2 patients experienced severe infusion-related adverse reactions, which were effectively managed. Both groups exhibited grade III to IV hematological toxicities, including neutropenia and thrombocytopenia. Importantly, no early deaths (defined as deaths occurring within 8 weeks post-treatment) were reported in the MST group, whereas 2 patients in the non-MST group succumbed to early mortality due to severe infections resulting from disease progression. Among the 22 patients who underwent microchimerism assessment, 17 displayed microchimerism, with values ranging from 0.002% to 0.034%. No definitive cases of acute or chronic GVHD were observed in all the patients. Additionally, single-cell mass cytometry analysis indicated that patients achieving CR in the MST group demonstrated significantly increased proliferation of naive T cells (TN), central memory T cells (Tcm), effector memory T cells (Tem), and natural killer (NK) cells in peripheral blood 21 to 28 days post-treatment, compared to CR patients in the non-MST group. Conclusions: Our clinical study demonstrates that DCAG combined with UCB MST and IL-2 treatment might enhance the patients' immune function and serve as a promising therapeutic option for elderly patients with newly diagnosed AML.