Venetoclax (VEN) combined with hypomethylating agents (HMA) is a standard treatment for unfit acute myeloid leukemia (AML) patients, but the conventional 28-day cycle often leads to prolonged cytopenias and infection risks. Recent studies suggest shorter VEN regimens may improve safety without compromising efficacy, but this remains controversial. To this end, we performed a systematic review and meta-analysis to compare the impact of VEN duration in patients receiving VEN-HMA. A comprehensive literature search was conducted in PubMed and Embase up to May 28, 2025. The Cochran Q test and I-squared statistics were used to identify the heterogeneity among the included studies. The network meta-analysis was performed by employing The mtc.model and mtc.run functions of the gemtc R package. We performed all statistical analyses using R 4.0.3. Six studies with 728 patients were included. Meta-analysis showed no significant difference in remission rates between shorter (≤ 14 days) and longer (> 14 days) treatments (RR = 1.07, 95

BackgroundMyelodysplastic syndromes (MDS) are common hematologic malignancies among older adults. With the rapid growth of short-video platforms, patients increasingly seek MDS-related health information online; however, the quality of MDS-related health information available on short-video platforms has not been systematically evaluated.ObjectiveThis study assessed the quality and reliability of MDS-related short videos across three major Chinese platforms and examined the relationship between content quality and user engagement.MethodsBetween June 25 and June 30, 2026, a total of 210 MDS-related videos were systematically collected from TikTok (Chinese version: Douyin), Bilibili, and Xiaohongshu (70 videos per platform). Video metadata were extracted, and content quality was independently evaluated using three validated instruments: the modified DISCERN (mDISCERN), the Journal of the American Medical Association (JAMA) benchmark criteria, and the Global Quality Scale (GQS). An overall quality score (SUM) was calculated by combining the three assessment tools. Spearman correlation analysis was performed to examine the associations between user engagement metrics and content quality.ResultsBilibili videos demonstrated the highest overall educational quality (mean SUM score: 9.79 ± 2.35), whereas TikTok generated the greatest user engagement (median number of likes: 81.00). Videos produced by professional creators achieved significantly higher scores across all quality assessment instruments than those produced by non-professional creators (mean SUM score: 8.81 vs. 5.03, p < 0.001). No significant associations were observed between any user engagement metric and content quality. Video duration was the only variable positively correlated with content quality (r = 0.332, p < 0.01).ConclusionsUser engagement was disconnected from information quality in MDS-related short videos. Professional creators provided greater educational value without consistently receiving commensurate engagement. Digital health communication should prioritize expert-led content, transparent source attribution, and quality-informed dissemination.

The efficacy of anti-CD19 Chimeric Antigen Receptor (CAR) T-cell therapy has been demonstrated in patients with relapsed or refractory B-cell non-Hodgkin lymphoma (B-NHL). However, therapy-induced senescence (TIS) has been identified as a novel resistance mechanism, potentially exacerbating immunosuppression and impairing CAR-T cell function. Raji and SU-DHL-2 cells were treated with 40 nM doxorubicin (Dox) for 72 h to induce TIS, and CCR7 was inhibited with 10 µg/mL Cap-100. Cellular senescence was assessed by SA-β-Gal staining and flow cytometry; proliferation and apoptosis by CCK-8 and Annexin V/PI, respectively. CAR-T cytotoxicity was evaluated via flow cytometry. CCR7 expression on T cells from B-NHL patients and healthy controls was measured. Lentiviral transduction regulated CCR7 expression; protein expression and interactions were analyzed by Western blot and Co-IP. NF-κB and ARHGAP/RhoA pathways were inhibited using GS143 and Y27632, respectively. CCR7 overexpression enhanced SASP, while knockdown attenuated it. In the Dox-induced TIS model, KMT2D decreased, whereas H3K9me3, CCR7, and LGALS9 increased; CCR7 inhibition reversed these changes. Cap-100 improved CD19 CAR T killing by alleviating exhaustion in co-culture. Mechanistically, Cap-100 stabilized IκBα to inhibit NFκB, and inhibition of NFκB or ROCK reversed pro-senescence. Cap-100 reduced T cell proliferation but did not affect apoptosis or exhaustion. Clinically, CCR7 was lower on T cells from B NHL patients than healthy controls, and higher CCR7 correlated with better T cell quality. Transcriptomics showed Dox activated TIM3/Galectin 9 and PD- 1/PD-L1 pathways, while Dox + Cap-100 suppressed them. Co-IP confirmed interactions among CCR7, KMT2D, and LGALS9. Collectively, Dox induced TIS to upregulate CCR7, increasing SASP and LGALS9, which interacted with TIM-3, causing immune cell exhaustion and reducing killing efficiency; CCR7 blockade reversed this state and enhanced cytotoxicity. Our findings demonstrate that blockade of KMT2D-CCR7-mediated senescence enhances CD19 CAR-T cell anti-tumor activity in B-NHL.

BackgroundCurrent genetic risk stratification systems for acute myeloid leukemia (AML), including the 2017 European Leukemia Network (ELN), 2022 ELN, and 2023 China (CN) stratifications, inconsistently categorize key genetic variants such as CEBPA and FLT3-ITD. These systems also demonstrate limited applicability to Chinese patients, complicating clinical decision-making. This study optimized genetic stratification for Chinese AML patients and developed a multi-omics prognostic model by integrating immunophenotypic and clinical characteristics to enhance outcome prediction.MethodsWe retrospectively enrolled 378 Chinese AML patients (298 for training, 80 for external validation) and compared the prognostic performance of the 2017/2022 ELN and 2023 CN systems (both integrating molecular genomic and cytogenetic data). Controversial genetic variants were re-evaluated to establish a novel 2023 CN (n-2023 CN) classification. A multi-omics model was constructed by combining n-2023 CN classification with CD15 positivity (leukemic immunophenotype), del(7q) (cytogenetic abnormality), and age ≥ 60 years (clinical factor). The model underwent external validation, was assessed for use in guiding hematopoietic stem cell transplantation (HSCT), and was compared with a published model.ResultsThe 2023 CN demonstrated superior efficacy in predicting AML relapse and 3-year relapse-free survival compared to 2017/2022 ELN systems. The n-2023 CN reclassified CEBPAbZIP as favorable-risk, t(8;21)/inv(16) with KITD816/FLT3-ITDlow as intermediate-risk, and FLT3-ITDhigh as adverse-risk among patients not receiving FLT3 inhibitors (regardless of NPM1 co-mutation). CD15 positivity, del(7q), and age ≥ 60 years were independent adverse prognostic factors for overall survival (OS). The model stratified patients into three risk groups with significantly distinct complete remission (CR) rates (93.2%, 71.2%, 35.7%) and OS (P < 0.001), with a concordance index of 0.729 (internal) and 3-year OS AUC of 0.752 (external). High-risk patients receiving HSCT exhibited longer OS. The model outperformed a published comparator in predicting CR and 1- and 3-year OS (all P ≤ 0.001).ConclusionsThe n-2023 CN resolves uncertainty in genetic risk stratification for Chinese AML patients. The novel multi-omics model incorporates immunophenomics and clinical and cytogenetic factors, captures comprehensive AML characteristics, enhances prognostic precision, and provides reliable guidance for personalized treatment, including HSCT selection.

BackgroundAlthough CD19 CAR-T cell therapy has improved outcomes in relapsed or refractory diffuse large B-cell lymphoma (DLBCL), responses are variable and predictive biomarkers are lacking. The triglyceride glucose-body mass index (TyG-BMI)—a surrogate of insulin resistance—has shown prognostic value in solid tumors, but its relevance to DLBCL and CAR-T efficacy remains unknown.MethodsWe retrospectively analyzed 204 newly diagnosed DLBCL patients treated with rituximab-containing regimens, stratified by median TyG-BMI. Prognostic associations were assessed by Kaplan-Meier and multivariate Cox regression, and a nomogram was constructed. In an exploratory sub-analysis, 20 patients who received CD19 CAR-T therapy were assessed. CAR-T cell persistence was measured by flow cytometry one week post-infusion and correlated with pre-infusion TyG-BMI by Pearson analysis. CRS and ICANS were graded per ASTCT criteria. Post-CAR-T survival was compared by Kaplan-Meier analysis stratified by median pre-infusion TyG-BMI.ResultsHigh TyG-BMI was independently associated with poorer PFS (HR, 2.146; 95% CI, 1.092–4.214; P = 0.027) and OS (HR, 2.531; 95% CI, 1.084–5.913; P = 0.032), and further stratified risk within IPI risk groups and across disease stages. A nomogram incorporating TyG-BMI, IPI, and bone marrow involvement predicted 1-, 3-, and 5-year OS (AUC: 0.886, 0.799, 0.772). In the CAR-T subcohort (n = 20), elevated pre-infusion TyG-BMI was significantly inversely correlated with CAR-T cell persistence at one week post-infusion (r = -0.570, P = 0.009). Patients with high pre-infusion TyG-BMI experienced inferior responses to CAR-T therapy, with shorter median PFS (5.0 vs. 19.5 months) and OS (39.0 vs. 81.0 months) compared with the low TyG-BMI group.ConclusionTyG-BMI is an independent prognostic factor in DLBCL that refines IPI-based stratification, and elevated pre-infusion TyG-BMI is associated with attenuated CAR-T cell persistence and inferior outcomes. This readily available metabolic index may aid risk assessment and treatment planning in DLBCL.

OBJECTIVES:B-cell lymphoma exhibits significant clinical heterogeneity, necessitating improved biomarkers for risk stratification. C-C chemokine receptor 7 (CCR7) and trimethylation of histone H3 lysine 9 (H3K9me3) are implicated in cellular senescence and tumor invasion. While the clinical significance of their co-expression in lymphomagenesis remains unclear. This study aims to define the expression profiles of CCR7 and H3K9me3 in B-cell lymphoma, explore their correlation with aggressive clinical indicators, and evaluate their combined prognostic value. METHODS:The expression of CCR7 and H3K9me3 in tumor tissues from B-cell lymphoma patients was analyzed by immunohistochemical (IHC) double-staining. The mechanistic association between the two was verified by co-immunoprecipitation assays and Western blot (WB) experiments detecting changes in cellular H3K9me3 levels following CCR7 ligand stimulation. The association between co-expression and patient clinical parameters, tumor burden, and progression-free survival (PFS) was evaluated through correlation analysis, Kaplan-Meier survival curves, and Cox regression analysis. RESULTS:H3K9me3 expression was predominantly nuclear, whereas CCR7 was expressed on the cell membrane. Both markers were significantly upregulated in aggressive lymphomas and positively correlated with LDH, β2-microglobulin, and neutrophil percentage. An interaction between CCR7 and H3K9me3 could be demonstrated in that CCR7 ligand stimulation resulted in an upregulation of H3K9me3 expression. Enhanced H3K9me3 expression was associated with bone marrow infiltration. High expression of CCR7 was associated with poorer progression-free survival (PFS), whereas high expression of both CCR7 and H3K9me3 identified patients with the worst prognosis. Univariate and multivariate Cox regression analysis indicated that the combined expression was a potential prognostic biomarker for B-cell lymphoma. CONCLUSION:Co-elevated CCR7 and H3K9me3 expression defines a high-risk B-cell lymphoma subgroup with high tumor burden, bone marrow infiltration, and poor prognosis, highlighting their potential as biomarkers for risk stratification and candidate therapeutic targeting.

Background: Cytarabine-based regimens remain the backbone of AML induction and consolidation therapy, yet drug resistance and relapse continue to hinder long-term outcomes. Vincristine is rarely used in AML, and its potential synergy with cytarabine has not been fully elucidated. Methods: IC50 values were determined via CCK-8 assay, followed by assessment of apoptosis, cell cycle, and senescence. Drug synergy was evaluated using the combination index. Immunofluorescence was used to examine microtubules and ARHGAP18 expression. In vivo efficacy was assessed using a xenograft AML model with bioluminescence imaging and survival analysis. Flow cytometry quantified leukemic blasts in bone marrow, peripheral blood, and spleen. RNA-seq and Western blotting were conducted to investigate underlying mechanisms. Results: Therapy-induced senescence contributed to cytarabine resistance in AML cells both in vitro and in vivo. Co-treatment with vincristine enhanced cytarabine sensitivity by attenuating cellular senescence. This combination markedly reduced leukemic burden in the bone marrow, peripheral blood, and spleen (Fig. 1A–B), and significantly prolonged the survival of tumor-bearing mice (Fig. 1C). Mechanistically, vincristine decreased the microtubule-associated expression of ARHGAP18, thereby inhibiting the initiation and progression of senescence. Further analysis indicated that ARHGAP18-induced senescence and resistance were mediated via activation of the PI3K/AKT signaling pathway. Conclusion: Vincristine synergizes with cytarabine to overcome therapy-induced senescence in AML by targeting the ARHGAP18/PI3K/AKT axis, offering a promising strategy to enhance therapeutic efficacy. This work was supported by grants from General Projects of Natural Science Foundation of Anhui Province (No.2208085MH217) and Anhui Clinical Medical Research and Translational Project (No.202427b10020036).

Background Resistance to existing therapies is a major cause of treatment failure in patients with refractory and relapsed B-cell non-Hodgkin’s lymphoma (r/r B-NHL). Therapy-induced senescence (TIS) is one of the most important mechanisms of drug resistance.Methods This study used single-cell RNA sequencing to analyze doxorubicin-induced senescent B-NHL cells. C-C chemokine receptor 7 (CCR7) expression in patients with aggressive B-NHL was assessed using immunohistochemistry and flow cytometry. Lentiviral transfection was used to target CCR7 expression in Raji and SU-DHL-2 cells. Protein localization was visualized through immunofluorescence, while western blotting and co-immunoprecipitation were used to analyze protein expression and interactions. Cell proliferation was measured with the Cell Counting Kit-8 assay, and senescent cells were detected using senescence-associated β-galactosidase staining. The stemness of cells was evaluated through colony and sphere formation assays. Transwell assays assessed cell migration and invasion. Finally, inhibitors GS143 and Y27632 were used to examine the effect of IKBα and ARHGAP/RhoA inhibition on B-NHL-TIS.Results Here we identified a distinct group of TIS, composed of memory B-cell population characterized by strong positive expression of CCR7, which was significantly elevated in TIS population compared with normal proliferating and autonomously senescent lymphoma cell populations. Additionally, CCR7 expression was significantly upregulated in patients with r/r B-NHL, and was an independent prognostic factor in B-NHL, with high CCR7 expression being strongly associated with poor prognosis. In vitro results indicated that CCL21 induced migration and invasion of B-NHL cells via CCR7, while blocking CCR7 reduced doxorubicin-induced migration and invasion of these cells. Furthermore, B-NHL-TIS regulated by CCR7 and exhibited enhanced phenotypic and functional stemness features, including the upregulation of stemness markers, increased colony-forming, invasive and migratory capabilities. Mechanistically, blocking CCR7 reversed the stemness characteristics of senescent B-NHL cells by inhibiting the activation of ARHGAP18/IKBα signaling.Conclusions Together, TIS promotes the stemness of B-NHL cells via CCR7/ARHGAP18/IKBα signaling activation and targeting CCR7/ARHGAP18 might overcome the chemoresistance of senescent B-NHL cells by inhibiting stemness acquisition and maintenance.

Cellular senescence plays a critical role in the relapse of acute myeloid leukemia (AML), yet the underlying mechanisms remain incompletely understood. Here, we investigated the function of ribosomal protein L5 (RPL5) in mediating cellular senescence and its impact on AML relapse. In relapsed AML patients, the proportion of senescent cells and the expression of RPL5 were elevated, compared to patients at newly diagnosised or in complete remission, suggesting a potential link between RPL5 and AML relapse. Following chemotherapy induction, a cellular senescence model was constructed using KG-1 A cells, with RPL5 expression significantly elevated. Knockdown of RPL5 suppressed cellular senescence and enhanced apoptosis, possibly due to different regulatory mechanisms for cell proliferation and senescence. Moreover, downregulation of RPL5 mitigated chemotherapy-induced senescence and improved the response of AML cells to chemotherapy drug. Mechanistically, RPL5 regulated cellular senescence via the p53-p21-pRb pathway and its downregulation led to a reduction in senescence-related protein expression levels. These findings suggest that RPL5 plays a critical role in mediating cellular senescence and chemotherapy response in AML, providing insights into novel therapeutic strategies for overcoming chemotherapy resistance and preventing disease relapse. Future studies may explore RPL5-targeted therapies and assess their clinical applicability in AML.

OBJECTIVE:To investigate the effect of non-chemotherapy strategy of retinoic acid (ATRA) combined with arsenic trioxide (ATO) on the survival of patients with acute promyelocytic leukemia (APL). METHODS:The data of APL patients with complete information diagnosed in the hematology department of our hospital from June 2009 to November 2024 were retrospective analyzed. All patients in the non-CHT group received ATRA-ATO induction, consolidation and maintenance therapy. Patients in the CHT group received ATRA-ATO+chemotherapy induction therapy, followed by 3 cycles of ATRA-ATO+CHT consolidation therapy and 6-10 cycles of ATRA-ATO maintenance therapy. The primary endpoint was event-free survival (EFS). Secondary endpoints included overall survival (OS), remission rate, differentiation syndrome (DS) and safety. RESULTS:There were 182 patients with APL and 15 patients with early death (ED), accounting for 8.24%, which was related to age and risk stratification. There was no significant difference in remission rate between the non-CHT group and the CHT group (P =0.486). As of February 2025, the median follow-up time of patients was 39.5 months. The EFS of the non-CHT group was significantly better than that of the CHT group (P =0.038). There was no significant difference in OS between the two groups (P =0.442). Subgroup analysis showed that EFS in the non-CHT was longer in standard-risk patients (P =0.012). There was no significant difference in EFS (P =0.585) and OS (P =0.473) between the CHT and non-CHT groups in high-risk patients. The incidence of mild DS was 23.6% in the non-CHT group and 23.1% in the CHT group, respectively, with no statistically significant difference(P =0.937). Compared with CHT group, the incidence of serious adverse events was lower in the non-CHT group. CONCLUSION:The non-chemotherapy regimen of ATRA combined with ATO is a feasible method to cure APL patients.

Acute myeloid leukemia (AML) lacks effective prognostic markers. While lymphocyte subsets are recognized as valuable predictive indicators in hematologic malignancies, their role in AML remains largely unexplored, particularly during different stages of AML. Our study analyzed the levels and changes of lymphocyte subsets in AML patients at newly diagnosed (ND) and first complete remission (CR) status, and explored the correlation between lymphocyte subsets and prognosis in different disease stages. Flow cytometry detected peripheral blood lymphocyte subsets in 145 ND AML patients, 125 CR AML patients, and 47 healthy controls (HCs). Dynamic testing was conducted on 28 AML patients at both ND and CR status. Our study found significant differences in lymphocyte subsets between ND, CR, and HCs, with notable changes in CD3+T, CD4+T, CD8+T, effector T (Teff), B, and natural killer (NK) cells between ND and CR status. Low frequencies of CD8+T below HCs thresholds and high regulatory T cell (Treg) frequency above HCs thresholds in the ND group, were independent risk factors for non-response to treatment. ROC curves evaluated the prognostic value of lymphocyte subsets and established cutoff values. Lymphocyte subsets in the ND group were not significantly associated with relapse or survival. Low absolute counts of CD3+T, B, and NK cells in the CR group were linked to AML relapse, and a low NK cell count was an independent predictor of overall survival (OS). Lymphocyte subsets can act as prognostic biomarkers, and their dynamic monitoring predicts treatment response, relapse, and survival in AML.

Purpose: CD19 chimeric antigen receptor T-cell (CAR-T) therapy demonstrates remarkable efficacy in patients with B-Cell Acute Lymphoblastic Leukemia (B-ALL), with six US Food and Drug Administration–approved products available. Despite high initial response rates, most patients eventually relapse. Relapse after CD19 CAR T therapy remains a significant challenge to survival outcomes. As outcomes for patients with disease recurrence after CD19 CAR-T therapy are not comprehensively studied, this analysis aims to define optimal treatment strategies. Methods: We conducted a retrospective multi-institutional study involving 74 patients with B-ALL who relapsed following CD19 CAR-T cell therapy, 67 of whom received salvage treatment. This study analyzed clinical characteristics, salvage regimens, and survival outcomes in relapsed/refractory (R/R) B-ALL patients who experienced relapse following CD19 CAR-T cell treatment. Complete remission (CR), relapse-free survival (RFS), overall survival (OS), and potential influencing factors were analyzed using IBM SPSS Statistics version 27 and R software version 4.4.2. Results: A total of 67 patients with R/R B-ALL (31 males, 36 females; age range 6 to 67 years) were included in the salvage therapy analysis. At initial diagnosis, 16 patients (23.9%) exhibited high-risk cytogenetic abnormalities, while 51(76.1%) were standard-risk. The BCR::ABL1 fusion gene was detected in 21 patients (31.3%). Twenty-two patients (32.8%) had undergone prior allogeneic hematopoietic stem cell transplantation (allo-HSCT) or umbilical cord blood transplantation before salvage therapy, including 9 who received consolidative transplantation after achieving CR from initial CAR-T therapy.At relapse, leukemic blasts were CD19+ in 46 cases (68.7%), CD19-/dim in 15 (22.4%), and had undetermined CD19 status in 6 (8.9%). Relapse sites included bone marrow (n=48, 71.6%) and extramedullary disease (EMD; n=14, 20.9%): 8 with central nervous system (CNS) involvement and 6 with non-CNS EMD. Half of EMD cases (7/14) had pre-existing extramedullary involvement. Five patients (7.5%) lacked documented relapse site assessments. With a median follow-up of 8.07 months post-progression, 32 patients (47.8%) achieved complete remission (CR/CRi) following salvage therapy. CR rates differed significantly by treatment modality. The response rates were 61% for targeted treatment (CAR-T reinfusion 61%, monoclonal antibodies 71%, chemotherapy combined with TKi 50%), 19% for chemotherapy (P=0.0035). CR/CRi rates showed no significant association with common high-risk factors (age, bone marrow tumor burden, molecular genetics, EMD), BCR::ABL1 status, CD19-negative relapse, or pre-relapse transplantation. However, CNS relapse demonstrated significantly higher CR rates than non-CNS EMD (100% vs. 17%; P=0.003). Median overall survival (OS) was 8.6 months (95% CI: 4.2–13.1), and median relapse-free survival (RFS) was 7.6 months (95%CI: 4.8–10.4). Patients receiving targeted salvage therapies showed superior OS compared to chemotherapy (P=0.004). CNS relapse was associated with superior OS than non-CNS EMD (P=0.043). Multivariate analysis confirmed targeted therapy as an independent predictor of improved OS (HR: 2.63; 95%CI: 1.17–5.92; P=0.019). Seven patients underwent consolidative allo-HSCT post-salvage (first transplant: n=6; second transplant: n=1). Of these, one patient died from transplant-related complications, two experienced relapse, and four remained in CR at final follow-up. Conclusion:Patients with relapsed B-ALL following CD19 CAR-T therapy exhibit universally poor prognoses. Salvage therapies involving molecular targeting—including CAR-T reinfusion, monoclonal antibodies, and TKIs—demonstrate superior efficacy over salvage chemotherapy. These approaches provide critical opportunities for bridging to transplantation and improving overall survival. Notably, extramedullary relapse involving the central nervous system (CNS) confers a more favorable prognosis compared to non-CNS extramedullary disease. Trial registration: The source data of this study is from the following registered clinical trials: NCT04532268,NCT02735291,NCT04605666. Disclosures: No relevant conflicts of interest to declare.

Abstract Background Acute myeloid leukemia (AML) is an aggressive blood cancer with limited treatment options and poor outcomes. Progress in AML immunotherapy is hampered by the scarcity of tumor-specific antigens. Mannose receptor C-type 2 (MRC2), largely restricted to the myeloid lineage, contributes to immune regulation, but its utility as a tumor-restricted surface antigen and immunotherapeutic target is yet to be determined. Methods Membrane proteomics was performed on leukemia blasts from AML patients and normal hematopoietic cells from healthy donors. Potential membrane targets were then characterized by flow cytometry, immunofluorescence, and immunohistochemistry across AML cell lines, primary AML blasts, and normal counterparts. Their clinical relevance was further corroborated by integrated analyses of patient-derived datasets. Then, functional experiments experiments were conducted in vitro and in vivo. Results MRC2 was identified as a significantly overexpressed membrane protein in AML, a discovery corroborated in both primary patient-derived AML blasts and established AML cell lines. MRC2 expression remained negligible across diverse normal blood cells, hematopoietic stem cells, and was entirely undetectable in healthy human tissues. Elevated cell membrane surface MRC2 levels were associated with complex chromosomal karyotypes, adverse molecular risk stratification, diminished complete remission rates, and shortened relapse-free survival in AML cohorts. Functional assays confirmed MRC2's critical role in sustaining AML cells proliferation, clonogenic potential, and migratory capacity. Conclusion MRC2 is a functionally protein exhibiting restricted expression on AML cell membranes, suggesting its utility as an immunotherapy target. Future strategies to improve the prognosis of AML patients may include CAR-T therapy or antibody therapy targeting the cell surface protein MRC2. This work was supported by grants from General Projects of Natural Science Foundation of Anhui Province (No.2208085MH217) and Anhui Clinical Medical Research and Translational Project (No.202427b10020036).

Background: CAR-T cells therapy has achieved breakthrough success in acute lymphoblastic leukemia (ALL), but its efficacy remains poor in acute myeloid leukemia (AML). Current research efforts are focused on enhancing CAR-T cells persistence, overcoming the immunosuppressive tumor microenvironment (TME), and improving metabolic fitness to boost therapeutic efficacy in AML. Among the components of the TME, serum-derived factors have emerged as critical modulators of CAR-T cells function. Our study investigates serum-derived molecular differences between ALL and AML patients that may account for differential CAR-T cells therapy responses. Methods: This study first investigated the impact of serum-free co-culture conditions and co-culture conditions supplemented with serum from healthy donors (HD), ALL patients, or AML patients on the anti-tumor activity of CD19 CAR-T cells and CD33 CAR-T cells in vitro. Subsequently, secretome analysis was used to identify and compare differences in serum secretory proteins between ALL and AML patients. The effect of targeted intervention of the identified target protein on the anti-tumor function of CD19 CAR-T cells and CD33 CAR-T cells was then assessed. Results:(1) CD19 CAR-T cells exhibited poor killing efficacy against co-cultured ALL cells in serum-free medium or medium supplemented with AML patient serum, but demonstrated potent killing in medium supplemented with HD serum or ALL patient serum. (2) CD33 CAR-T cells exhibited poor killing efficacy against co-cultured AML cells in serum-free medium or medium supplemented with AML patient serum, but demonstrated potent killing in medium supplemented with HD serum or ALL patient serum. (3) Secretome analysis revealed significantly lower expression of mitochondrial ribosomal proteins (MRPs), particularly mitochondrial ribosomal large subunit protein 57 (MRPL57), in AML patient serum compared to ALL patient serum. (4) Inhibiting MRPL57 in serum from ALL patients significantly downregulated the anti-tumor killing ability of CD19 CAR-T cells against ALL cells. (5) Supplementing MRPL57 into serum from AML patients significantly upregulated the anti-tumor killing ability of CD33 CAR-T cells against AML cells. (6) MRPL57 supplementation enhanced the proportion and proliferative potential of stem cell-like central memory T cells in CAR-T populations via mitochondrial translation, and promoted the release of granzyme B and perforin. Conclusion: AML patients exhibit a significant deficiency in endogenous MRPL57 production that leads to suboptimal therapeutic outcomes with CAR-T cells therapy, which highlights a novel metabolic and microenvironmental axis to improve CAR-T cells efficacy in AML.

Background: Relapsed/refractory acute B lymphoblastic leukemia (R/R B-ALL) has a poor prognosis and low overall survival rate. Some patients experience rapid clinical relapse after achieving complete remission (CR) with chimeric antigen receptor T-cell (CAR-T) treatment. Therefore, predicting clinical relapse and implementing preemptive therapy after CAR-T treatment is of great significance for improving patient prognosis. Methods: The clinical data of 55 patients with R/R B-ALL who received CD19 CAR-T treatment were retrospectively analyzed. Survival analysis was conducted on peripheral blood B-cell count, bone marrow minimal residual disease (MRD), bone marrow CAR gene copy number, and the choice of preemptive therapy in patients who achieved CR. Results: The CR rate among 55 R/R B-ALL patients who received CD19 CAR-T treatment was 81.8%. For patients achieving CR, the median relapse-free survival was 290 days (95% CI 163-416), and the median overall survival was 870 days (95% CI 656-1084). All patients who achieved CR exhibited undetectable peripheral blood B-cell counts (0/μL) following CAR-T treatment. Survival analysis demonstrated that clinical relapse was highly correlated with peripheral blood B-cell early regeneration (within 45 days post-CAR-T infusion, but not after 45 days post-CAR-T infusion) (p<0.01), and was not associated with bone marrow MRD positivity or loss of CAR-T copy number at the same time. Furthermore, when peripheral blood B-cell early regeneration occurred, the choice of preemptive anti-leukemia therapy (including chemotherapy, targeted agents, or secondary CAR-T infusion) resulted in a significantly better relapse-free survival compared to immunomodulatory therapy (including interferon, interleukin-2) (p<0.05). Conclusion: B-cell early regeneration within 45 days after CD19 CAR-T treatment highly predicts clinical relapse in R/R B-ALL patients. Initiating anti-leukemic therapy promptly upon B-cell early regeneration can significantly extend relapse-free survival. This work was supported by grants from General Projects of Natural Science Foundation of Anhui Province (No.2208085MH217) and Anhui Clinical Medical Research and Translational Project (No.202427b10020036).

Neutrophil extracellular traps are essential in the development and advancement of multiple myeloma (MM). However, research investigating the prognostic value with NET-related genes (NRGs) in MM has been limited. Patient transcriptomic and clinical information was sourced from the gene expression omnibus database. Cox regression analysis with a univariate approach was employed to explore the link between NRGs and overall survival (OS). Kaplan-Meier methods were applied to assess variations in survival rates. A nomogram integrating clinical data and predictive risk metrics was crafted using multivariate logistic and Cox proportional risk model regression analyses. Additionally, we investigated the disparities in biological pathways, drug sensitivity, and immune cell involvement, and validated differential levels of two key genes through qPCR. We identified 148 differentially expressed NRGs through published articles, of which 14 were associated with prognosis in MM. Least absolute shrinkage and selection operator Cox regression model established a nine-gene NRG signature-comprising ANXA1, ANXA2, ENO1, HIF1A, HSPE1, LYZ, MCOLN3, THBD, and FN1-that demonstrated strong predictive power for patient survival. The Cox regression model with multiple variables demonstrated that the risk score independently predicted OS, showing that those with a high score had worse survival rates. Furthermore, a nomogram incorporating patient age, LDH levels, the International Staging System, and NRGs was developed, demonstrating strong prognostic prediction capabilities. Drug sensitivity correlation analysis also offered valuable guidance for future immuno-oncological therapies and drug selection in MM patients. The NRGs signature was a reliable biomarker for MM, effectively identifying high-risk patients and forecasting clinical outcomes.

Chemotherapy incorporating the proteasome inhibitor bortezomib (BTZ) has improved outcomes for patients with multiple myeloma (MM); however, resistance to chemotherapy and disease relapse remain significant challenges, closely associated with cellular senescence. This study investigated the key drivers of myeloma cell senescence and its role in MM progression. Flow cytometry assessed senescence-associated β-galactosidase (SA-β-gal) activity in myeloma cells from bone marrow samples of MM patients. BTZ was used to establish cell senescence model. RNA-seq identified key genes regulating myeloma cell senescence, which were verified by qPCR. After CBX7 knockdown and overexpression, SA-β-gal staining, CCK-8 assay, cell cycle assay, and colony formation assay were performed to investigate its effects on senescence. RNA-seq further identified downstream target genes and pathways, and small interfering RNA and ERK inhibitor were used to explore their effects. A xenograft mouse model was used to validate CBX7’s effect on myeloma cell senescence. Our results demonstrated that BTZ-based chemotherapy induces senescence in myeloma cells, with SA-β-gal activity linked to malignant proliferation. CBX7 was identified as a critical regulator of cellular senescence in myeloma cells. Elevated CBX7 levels were observed in newly diagnosed MM, decreased during remission, and increased again at relapse. CBX7 levels positively correlated with blast counts, creatinine, and β2-microglobulin levels, and negatively correlated with SA-β-gal activity. Functionally, CBX7 knockdown promoted BTZ-induced myeloma cell senescence and senescence-like growth arrest, whereas CBX7 overexpression had the opposite effect. Mechanistically, CBX7 regulates senescence-like growth arrest in myeloma cells via the ERK/STAT3/MIX1 axis. Silencing PIM1 or the ERK inhibitor U0126 mitigated CBX7-mediated myeloma cell senescence and enhanced the inhibitory effects of BTZ on cell viability and clone formation. In vivo, CBX7 knockdown enhanced BTZ-inhibited xenograft tumor growth. CBX7 is a pivotal target for regulating cellular senescence in myeloma cells, operating through a novel CBX7/ERK/PIM1 regulatory axis. Targeting CBX7 and its downstream pathways may augment the efficacy of standard chemotherapy.

INTRODUCTION:Hematopoietic stem cell transplantation (HSCT) and chimeric antigen receptor T (CAR-T) cell are effective treatments for acute lymphoblastic leukemia (ALL). Various forms of intra- and extramedullary relapses have been reported after HSCT and CAR-T-cell therapy for ALL; however, no reports have investigated isolated central nervous system (CNS) relapse after HSCT and CAR-T-cell therapy. Hence, no clinical treatment has been established for such rare patients. CASE PRESENTATION:An 18-year-old male patient with B-cell ALL suffered from isolated CNS relapse after HSCT and CAR-T-cell therapy. Conventional systemic intravenous and intrathecal chemotherapies were ineffective and intolerable. A unique immunosuppressive microenvironment of decreasing NK cell percentage and increasing IL-8 concentration and CAR-T-cell exhaustion had been illustrated in the cerebrospinal fluid. Finally, the patient received immunomodulatory therapy with lenalidomide and obtained complete remission. CONCLUSION:Lenalidomide might be a therapeutic strategy for isolated CNS relapse after HSCT and CAR-T-cell therapy.