This study aimed to evaluate the feasibility of CD72 as a complementary CD19-independent B-lineage gating marker for longitudinal measurable residual disease (MRD) surveillance in relapsed/refractory B-cell acute lymphoblastic leukemia (R/R B-ALL) following CD19 CAR-T therapy. Correlation analyses were performed in 66 B-ALL samples to compare MRD detection using CD72, CD19, and cytoplasmic CD79a. CD72 expression specificity was further evaluated in 129 leukemia patients. In addition, 129 patients with R/R B-ALL treated with autologous CD19 CAR-T therapy in registered clinical trials (ChiCTR-IIh-16008711; NCT03173417) between January 2021 and December 2022 were retrospectively analyzed, with follow-up continued until January 2025. CD72 gating showed excellent concordance with both CD19- and cCD79a-based strategies for MRD assessment. CD72 expression demonstrated high specificity in B-ALL, with a positivity rate of 95.77%, compared with 29.27% in AML and 23.53% in T-ALL. All 129 heavily pretreated patients achieved MRD-negative CR at day 28 after CAR-T infusion and subsequently underwent allo-HSCT, with a median interval of 54 days (range, 40-338). A total of 16 patients experienced MRD relapse during follow-up, including four clinically confirmed CD19-negative relapses that retained CD72 expression. Patients with pre-CAR-T MRD ≤1% showed earlier B-cell recovery than those with MRD >1% (median 30 [18-45] vs. 32 [26-79] days, p = 0.028). The MRD ≤1% cohort demonstrated significantly improved 3-year overall survival compared with the MRD >1% cohort (88.1% vs. 69.2%, p = 0.014). The 3-year cumulative incidence of MRD relapse was significantly lower in the MRD ≤1% cohort than in the MRD >1% cohort (3.96% vs. 17.95%, p = 0.019), while non-relapse mortality was also numerically lower in the MRD ≤1% cohort (5.92% vs. 17.95%, p = 0.053). Multivariate analysis identified KMT2A rearrangement, IKZF1 mutation, TP53 mutation, and elevated pre-CAR-T MRD as independent predictors of inferior outcomes. CD72 represents a feasible complementary B-lineage marker for longitudinal MRD surveillance following CD19 CAR-T therapy. Retention of CD72 expression in clinically confirmed CD19-negative relapses supports its potential utility when CD19 expression is lost after targeted therapy.

BACKGROUND:High-dose methotrexate (HD-MTX) is essential for pediatric acute lymphoblastic leukemia (ALL) but frequently causes hepatotoxicity. To address the critical lack of early prediction tools, this study investigated untargeted metabolomic profiles to identify predictive biomarkers and establish robust machine learning (ML) models for early clinical risk assessment and therapeutic optimization. METHODS:This study included 106 pediatric patients with ALL undergoing HD-MTX therapy. Of these, 56 developed moderate-to-severe liver injury. Untargeted plasma metabolomics was performed before and after treatment. Metabolic alterations were identified using multivariate analyses, with pathway annotation performed via KEGG enrichment. Key discriminatory metabolites were further screened via LASSO regression to identify potential biomarkers. Three ML algorithms (Random Forest, Support Vector Machine, and Bayesian logistic regression) were developed for risk prediction, integrating SHAP analysis for model interpretability and feature contribution evaluation. RESULTS:Patients with liver injury showed distinct metabolic signatures compared with controls. Prior to HD-MTX treatment, 13 metabolites-such as prominently elevated glycocholic acid-were identified as potential predictors of liver injury, with overarching pathway alterations primarily involving arginine biosynthesis and glutathione metabolism, among others. Post-treatment, 11 distinct metabolites-including arachidic acid-further distinguished the injury group, reflecting complex regulatory mechanisms and multi-pathway shifts. Crucially, all three ML models developed using pre- or post-treatment metabolic biomarkers exhibited high predictive and discriminative accuracy for hepatotoxicity (AUC > 0.900). CONCLUSIONS:These findings characterize the metabolic alterations underlying HD-MTX-induced liver injury, provide a metabolomics-based framework for early risk assessment, and facilitate the development of precision therapeutic strategies in pediatric ALL.

Background Relapsed or refractory (R/R) hematologic malignancies, including T-cell acute lymphoblastic leukemia/lymphoblastic lymphoma (T-ALL/LBL), remain a therapeutic challenge due to limited treatment options and poor clinical outcomes. Chimeric antigen receptor (CAR) T-cell therapy has emerged as a promising immunotherapeutic approach for hematologic malignancies. Previous studies have demonstrated that autologous anti-CD7 CAR-T cells generated using the “Natural Selection” (NS) platform (NS7CAR) exhibit manageable toxicity profiles and good efficacy in patients with R/R T-ALL/LBL and CD7-positive acute myeloid leukemia (AML). This pooled analysis evaluates the efficacy and safety of NS7CAR-T therapy based on data from 5 clinical trials (https://clinicaltrials.gov, NCT04572308, NCT05626400, NCT04916860, NCT04928105, and NCT04938115) involving patients with CD7-positive hematologic malignancies. Methods This study represents a pooled, cohort analysis of subjects enrolled in clinical trials evaluating NS7CAR therapy. Eligible participants included patients with R/R CD7-positive hematologic malignancies who received at least one infusion of autologous NS7CAR-T cells and were observed between November 9, 2020, and May 21, 2025. Baseline demographic and clinical characteristics, CAR-T cell dosing, and post-infusion outcomes were systematically collected. Efficacy and safety assessments included overall response rates (ORR), overall survival (OS), progression-free survival (PFS), and the incidence of adverse events. Descriptive statistics were used to summarize key variables. Results The analysis included a total of 334 patients with R/R CD7-positive hematologic malignancies. The cohort comprised 159 cases (47.6%) of T-ALL and 121 cases (36.2%) of T-LBL, mixed phenotype acute leukemia accounted for 24 cases (7.2%), AML for 21 cases (6.3%), and peripheral T-cell lymphoma (PTCL) for 9 cases (2.7%). The median age was 24 years (range: 2–67 years), with 255 males (76.3%) and 79 females (23.7%). At enrollment, 88 patients (26.3%) were relapsed from prior transplants, including 4 patients who relapsed after 2nd transplants. The median time to relapse post-transplantation was 265 days (range: 31-2432 days). Regarding baseline disease involvement before NS7CAR-T infusion, 128 patients (38.3%) showed isolated bone marrow (BM) involvement, 56 patients (16.8%) presented with isolated extramedullary disease (EMD), and the remaining 137 patients (41.0%) demonstrated concurrent involvements of both BM and EMD. Among those with BM involvement, 29.8% (79/265) had ≥20% blasts. Patients received three dose levels of NS7CAR-T cells, 15.3% (51/334) of 5×10⁵ cells/kg, 83.2% (278/334) of 1×10⁶ cell/kg, and 1.5% (5/334) of 1.5–2×10⁶ cells/kg. The median dose of NS7CAR-T cell infusion administered to patients was 1×10⁶ cells/kg. After NS7CAR-T therapy, among 329 evaluable patients as of August 1, 2025, the ORR reached 86.3% (284/329), with a complete remission/complete remission with incomplete hematologic recovery (CR/CRi) rate of 84.5% (278/329). Post-treatment evaluation revealed that 245 patients (73.4%) underwent consolidation/salvage transplantation following CAR-T therapy, with a median interval of 57 days (range: 28-227 days) between CAR-T infusion and transplantation. Survival analysis demonstrated a median PFS of 547.5 days (range: 45-1694 days) and median OS of 567.5 days (range: 45-1694 days). Safety assessment of the full cohort (n=334) showed cytokine release syndrome (CRS) occurring in 95.8% of patients (320/334), with the majority (85.6%, 286/334) experiencing grade 1-2 events and 10.2% (34/334) developing grade ≥3 CRS. Immune effector cell-associated neurotoxicity syndrome (ICANS) was observed in 3.3% of cases (11/334), including 0.9% (3/334) with grade 1-2 and 2.4% (8/334) with grade ≥3. Conclusions This large pooled cohort analysis demonstrates that NS7CAR-T therapy exhibits very promising clinically efficacy, achieving an 84.5% CR rate, along with a manageable safety profile in patients with R/R CD7-positive hematologic malignancies, including T-ALL/LBL, AML, MPAL and PTCL. While a small subset of patients experienced grade ≥3 CRS and ICANS, the robust response rates and survival outcomes position NS7CAR-T as a potentially transformative treatment option for R/R CD7-positive hematologic malignancies, including post-transplant relapsed patients.

Background: Patients with t(8;14)(q24;q11)/ TCRA/D::MYC-translocated T acute lymphoblastic leukemia/lymphoma (T-ALL/LBL) are a rare subgroup of T-ALL/LBL associated with induction failure and aggressive disease. But recently, we found that this type is not uncommon in refractory and relapsing T ALL. Aims: We hope to find the commonality of this kind of disease and explore effective treatment methods. Methods: All T-ALL/LBL patients were confirmed by comprehensive diagnosis of cytomorphology, immunophenotype, cytogenetics and molecular biology. The CD7 Chimeric Antigen Receptor T-Cell (CAR-T) therapy clinical trial for T-LBL/ALL patients was initiated October 2020 (NCT04916860). Results: We conducted a retrospective analysis of 14 patients with the t(8;14) translocation, who were all detected in refractory or relapsed patients. Five had the t(8;14) (q24.1;q11.2) in their primary tumor, and 9 at the time of relapse. Extramedullary invasion occurred in 9/12 patients. Four patients had a STIL::TAL1 translocation, 6 had a PTEN mutation, and 6 had CDKN2A/B gene loss. Seven patients who only received chemotherapy or allogeneic hematopoietic stem transplantation (HSCT) all died. The median overall survival (OS) was only 3 months (range: 2-7 months). Since the CD7 CAR-T therapy was used for refractory or relapsed T-ALL/LBL from Oct, 2020, 6 patients were treated with the CD7 CAR-T therapy and 5 achieved CR on Day 30. Three patients achieved minimal residual disease (MRD) negative status and two subsequently became MRD-positive. Four patients who bridged to HSCT after CAR-T have remained disease-free to date. Summary/Conclusion: The prognosis of the patients with t(8;14)/ TCRA/D::MYC-translocated T-ALL/LBL was very poor, we should pay enough attention to it. while CD7 CAR-T allows patients to achieve short-term CR. Allo-HSCT after CAR-T therapy should be considered for long survival. Table 1. Characteristic of the 14 T-ALL/LBL patients harboring the t(8;14)(q24;q11) translocation - Characteristic Value Gender (M/F) 10/4 Age Median (range) — yr 15 (2-35) 2-14yr 6 15-61yr 8 With EMD — No. CNSL 6/12 Diffused EMD 3/12 With other fusion gene 5 STIL::TAL1 4 MLL-PTD 1 Complex cytogenetic 10/12 Additional gene mutations PTEN/PTPN 6 NOTCH1 2 CCND3 3 TP53 1 CDKN2A/B deletion 6/14 Disease status — No. Refractory (3 median chemotherapy courses) 5 Relapsed (10 median chemotherapy courses) 9 Transplantation status — No. Yes 5 No 9 CD7 CAR-T therapy status — No. Yes 6 No 8 EMD: extramedullary disease. CNSL: central nervous system leukemia. CART: Chimeric Antigen Receptor T-Cell. Keywords: Lymphoma, T cell acute lymphoblastic leukemia

T-acute lymphoblastic leukemia/lymphoma (T-ALL/LBL) patients with t(8;14)(q24;q11)/TCRA/D::MYC translocation represent a rare subgroup, with an aggressive course. In our retrospective analysis of 14 patients, all were identified during refractory or relapsed stages (5 primary tumor, 9 relapse). Notably, extramedullary invasion was detected in most patients. Four exhibited STIL::TAL1 translocation, and six demonstrated CDKN2A/B gene loss. The therapeutic outcomes were notably poor for all seven patients who received only chemotherapy or allogeneic hematopoietic stem cell transplantation (HSCT); all eventually succumbed to the disease with a median OS of 3 months. In the application of CD7 CAR-T therapy in six patients, five achieved CR. Of the four patients who underwent HSCT following CAR-T therapy, all have remained disease-free. The prognosis for T-ALL/LBL patients with t(8;14) translocation remains bleak, but interventions involving CD7 CAR-T may offer a potential pathway to CR. HSCT following CAR-T could be a viable strategy for long-term survival.

Objective:To investigate the relationship between the levels of serum cytokines and chemokines and the prognosis of patients with acute B-ALL after receiving chimeric antigen receptor (CAR)-T cell immunotherapy and acute graft-versus-host disease (aGVHD) in patients after bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:According to the case-control principle, Forty-two patients with B-ALL who received CD19-CAR-T cell immunotherapy bridged to allo-HSCT at Heibei Yanda Ludaopei Hospital from September 18, 2019 to May 9, 2022 were enrolled. Mann-Whitney U test was used to compare the changes of aGVHD-related cytokines and chemokine levels between CAR-T cell immunotherapy and bridging transplantation in different patients at the same time. Their plasma levels of cytokines and chemokines related to aGVHD were monitored at the day before CAR-T therapy and after CAR-T treatment at day 4, 7,14,21,28. The receiver operating characteristic curve was drawn to evaluate the predictive value of cytokines and chemokines in predicting the occurrence and the death of aGVHD patients. Kaplan-Meier method and Log-rank tests were used for Overall survival (OS) analysis. Results:Twenty-four of total 42 patients had aGVHD, of which 11 patients died and 31 patients survived. There was no significant difference in cytokines and chemokines between the aGVHD group and the non-aGVHD group on the day before CAR-T cell treatment. According to statistical analysis, the serum Elafin levels of aGVHD group was higher than that of non-aGVHD group at the 21st day [4 482 (2 811, 6 061) ng/L vs 2 466 (1 948, 3 375) ng/L, Z=3.145, P=0.001] and the 28st day [4 391 (2 808, 5594) ng/L vs 2 463 (1 658, 2 830) ng/L, Z=2.038, P=0.048] separately. At the 14th day, serum cytokines and chemokines levels between the two group were as follows,MIP-1 α [21.02 (12.36, 30.35) ng/L vs 5.56 (3.64, 10.79) ng/L], sCD25 [422.47 (257.99, 1 233.78) IU/ml vs 216.11 (133.75,457.39) IU/ml], Elafin [4 101 (2 393, 5 006) ng/L vs 2 155 (1 781, 3 033) ng/L], IL-6 [119.08 (23.97, 183.43) ng/L vs 8.39 (2.91, 17.42) ng/L] and IL-8 [13.56 (12.50, 24.52) ng/L vs 2.83 (1.73,6.87) ng/L] were at higher levels ( Z=2.653, P=0.007; Z=2.176, P=0. 030; Z=2.058, P=0.041; Z=3.329, P<0.001; Z=3.162, P=0.001). The KM survival curve showed that the cumulative survival rates of patients with higher serum levels of MIP-1α, sCD25, Elafin, IL-6 and IL-8 were lower than those with low levels at day 14, and the difference was statistically significant (χ 2=12.353, 4.890, 6.551, 10.563, 20.755, P<0.05). Conclusion:The outcomes of patients treated with CAR-T cell therapy bridged to allo-HSCT was correlated with serum MIP-1α, sCD25, Elafin, IL-6 and IL-8 levels after receiving CAR-T therapy. High concentrations of MIP-1α, sCD25, Elafin, IL-6 and IL-8 suggest poor prognosis and can be used as biomarkers to suggest appropriate clinical selection of therapy.

CD19 chimeric antigen receptor T-cell (CAR-T) therapy is efficacious for refractory/relapsed (R/R) B-cell hematological malignancies, yet relapse due to CD19 antigen escape remains a challenge. Our trial explored simultaneous targeting of multiple B-cell antigens as a therapeutic approach that may reduce the risk of relapse. We tested the safety and efficacy of CAR19/22 T-cell cocktail therapy including murinized and humanized products among patients with R/R aggressive B-cell lymphoma. In the group that received the humanized product, 11/12 (91.7%) patients achieved an objective response, including 9/12 (75%) complete responses (CRs) by day 28. The overall response rate and CR rate in the murinized group was 92.9% (13/14) and 42.9% (6/14), respectively. Nine of 12 (75%) patients in the humanized group maintained CR at month 3 following infusion, compared to 5/14 patients (35.7%) in the murinized group. Progression-free survival (PFS) was more favorable in the humanized compared to the murinized group. Most patients had mild cytokine release syndrome (CRS) (grade 1–2) in both groups. This study demonstrates that CAR19/22 T-cell cocktail therapy is safe and effective for R/R B-cell lymphoma and that patients treated with a humanized CAR-T exhibited better efficacy compared to patients treated with a murinized CAR-T therapy.

Introduction Anti-CD19 chimeric antigen receptor (CAR) T-cells therapy has achieved unprecedented efficacy in refractory or relapsed (R/R) B-cell acute lymphoblastic leukemia (B-ALL) patients, with up to a 90% complete remission (CR) rate. However, a large body of clinical data has shown that the relapse rate post CD19 CAR-T therapy remains high. Our published study of 254 patients with R/R B-ALL treated with CD19 CAR-T showed the median time to relapse following CAR-T cells infusion was 100 days. Patients who received consolidative allogeneic hematopoietic stem cell transplantation (allo-HSCT) post CAR-T therapy demonstrated much higher overall survival (OS) and leukemia-free survival (LFS) compared to those who received CAR-T therapy alone (1-year OS of 77.8% vs. 35.4%, p<0.0001; 1-year LFS of 70.9% vs. 33.3%, p<0.0001, respectively). However, there are still many patients who cannot undergo allo-HSCT, including those who have relapsed following allo-HSCT and elderly patients. Prolonging disease-free survival (DFS) after CAR-T therapy in these patients remains a challenge for physicians. Studies have shown that the histone deacetylase (HDAC) inhibitor chidamide can up-regulate T-cell activity and down-regulate derived suppressor cell (MDSC) expression. Chidamide also promotes the expression of CD22 on the surface of B-cell tumor cells in vitro and in vivo, and enhances CAR-T function. Here we explore the efficacy of chidamide as maintenance therapy post CAR-T in B-ALL. Patients and methods We analyzed 19 patients with R/R B-ALL who received CD19 CAR-T at our center (NCT04546893) between October 2019 and July 2022. All patients achieved CR on day 30 following CD19 CAR-T cells infusion and were unable to bridge to allo-HSCT within 3 months. We administered chidamide as maintenance therapy to all 19 patients between 30 to 90 days after CAR-T infusion. Chidamide was continuously administered until the patient relapsed, transferred to transplant, or had intolerable side effects. Results On Day 30 post CD19 CAR-T, 19/19 (100%) patients achieved CR or CR with incomplete count recovery (CRi)(Figure 1). Sixteen of 18 (88.89%) who had bone marrow involvement achieved minimal residual disease (MRD)-negative CR. The median time from CAR-T cell infusion to start chidamide therapy was 30 days (range: 30-90 days) and the median duration of chidamide administration was 90 days (range: 12-420 days). No patient experienced cytokine release syndrome (CRS) or encephalopathy while taking chidamide. All side effects that occurred were mild. Two patients experienced nausea and vomiting and one patient had mild bone marrow suppression. No other side effects occurred. The median follow-up time was 12 months (range: 1-33 months), the 12-month OS rate was 83.1% and the 12-month DFS rate was 62.4% (Figure 2). Five patients received allo-HSCT following chidamide maintenance and 13 patients who did not undergo transplantation had 12-month OS of 83.3%, 12-month DFS of 62.9%. Six patients were monitored for the proportion of CAR-T cells in T lymphocytes following chidamide therapy. In all six patients, the proportion of CAR-T cells increased but to different degrees. Conclusions For those patients with R/R B-ALL for whom allo-HSCT is not available following CD19 CAR-T therapy, HDAC inhibitor chidamide as maintenance therapy is a viable option to reduce the risk of relapse rate following CD19 CAR-T therapy. This study demonstrated that chidamide could prolong OS and DFS in some patients who achieved CR after CD19 CAR-T therapy. Chidamide was safe, inexpensive and convenient to take orally. The inclusion of more patients and extended observation time is needed for further evaluation of chidamide maintenance therapy following CAR-T cell therapy in patients with R/R B-ALL. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Objective:To investigate the significance of multicolor flow cytometry (MFC) monitoring of minimal residual disease (MRD) in the course of allogeneic hematopoietic stem cell transplantation (allo-HSCT) after CD19-chimeric antigen receptor(CAR)-T cell immunotherapy for patients with refractory, relapsed B-cell acute lymphoblastic leukemia (r/r B-ALL).Methods:37 patients with r/r B-ALL admitted to Hebei Yanda Lu Daopei Hospital from January to July 2019, aged 15 (6, 19) years old, including 24 males and 13 females, were treated with CD19-CAR-T cell immunotherapy bridging allo-HSCT. MFC with cytoplasmic CD79a antibody to set up B-cell gates was used to monitor patients′ bone marrow (BM), cerebrospinal fluid (CSF), and tissue samples on day 0 (prior to the CAR-T cell immunotherapy), day 15, day 28 post CAR-T cell immunotherapy, and post transplantation.The MRD values of these samples were analyzed to evaluate the residual tumor cells and metastasis. The killing effect of the CAR-T cells was evaluated by the recovery of CD19+B cells before transplantation and the period between the timepoint when CD19+B cells was recovered and the timepoint when CAR-T cells were infused. Peripheral blood CAR-T cells were counted at different time points. Statistic analysis was performed by Kaplan-Meie assay and Log-rank test to analyze the difference of univariate cumulative survival.Results:(1)Among the 37 patients, 8 died and 29 survived. 5 patients relapsed after transplantation, of which 4 relapsed patients died and 1 survived. (2)MFC MRD negative remission rate of the death group was lower than that of the survival group at the following time points: post-CAR-T therapy and prior to transplantation (5/8 vs. 28/29, χ 2=7.540, P=0.006); day 15 of the CAR-T cell reinfusion (3/8 vs. 24/29, χ 2=6.512, P=0.011); day 28 of the reinfusion (3/8 vs. 276/29, χ 2=10.065, P=0.002). The probability of extramedullary MFC MRD positive tumor infiltration in the death group was higher than that in the survival group(7/8 vs. 14/29, χ 2=3.931, P=0.047). After CAR-T cell immunotherapy, the recovery period of CD19-positive cells in the death group, or the time for CAR-T cells to kill CD19-positive cells, was shorter than that in the survival group [42.00 days(30.00,49.00) vs. 55.00 days(41.50,73.50), Z=0.022, P=0.020]. Conclusion:The positive results of MRD by MFC at the following timepoints may predict unfavorable outcomes, such as post-CAR-T therapy and prior to transplantation, day 15 and 28 of the CAR-T cell immunotherapy, which may provide some guidance for clinical management.

目的:研究血小板数值与CD19-CAR-T细胞治疗难治/复发急性B淋巴细胞白血病疗效的关系.方法:回顾性分析2017年5月—2020年5月84例接受CD19-CAR-T细胞治疗难治/复发急性B淋巴细胞白血病患者的临床资料.将患者分为CAR-T细胞治疗后微小残留病(MRD)阴性组(46例)和CAR-T细胞治疗后MRD阳性组(38例),比较2组患者CAR-T细胞回输当天及回输后第7天、第14天、第21天和第28天的血小板数值.结果:CAR-T细胞治疗后MRD阴性组在CD19-CAR-T细胞回输时和回输后第7天、第14天、第21天的中位血小板数值分别为109.0×109/L、57.0×109/L、83.7×109/L和35.0×109/L,明显高于MRD阳性组的31.5×109/L、24.0×109/L、18.0X109/L 和 18.0X109/L,2组比较差异有统计学意义(P＜0.05).结论:血小板数值与CAR-T细胞治疗的疗效呈正相关,且CAR-T细胞回输当天的血小板数值可以预测CAR-T细胞的疗效.

Background: CD19-targeted chimeric antigen receptor (CAR) T-cell therapy has demonstrated about 50~60% of response rate in relapsed or refractory (R/R) B-cell non-Hodgkin lymphoma. However, antigen-escape relapse has emerged as a major challenge for long-term disease control after CD19-targeted therapies. In our previous study, we described a dual-targeted of CD19 and CD22 CAR-T cell cocktail therapy, whichyielded 93.3% of complete remission (CR) rate in R/R B-cell acute lymphoblastic leukemia and could reduce the risk of CD19-negative relapse. Aims: A phase I/II clinical trial (NCT05206071) was designed to explore the safety and efficacy of CAR19/22 T-cell cocktail therapy for patients with R/R aggressive B-cell lymphoma. Methods: Eligible R/R aggressive B-cell lymphoma patients were enrolled from July 2020 toDecember 2021. All patients received fludarabine 30 mg/m2/d and cyclophosphamide 250 mg/m2/d for 3 consecutive days (day −5 to day −3) followed by cocktail CAR-T cell infusion. CAR-T-cell-related cytokine release syndrome (CRS) was graded to assess the safety. PET-CT was performed on day 28 and 3rd month to evaluate the response rate. Results: A total of 26 eligible patients received CAR19/22 cocktail infusion with a median age of 46 years old (4-75) and a median of 3 prior lines of therapies (2-5 lines). Of 26 patients, 4 were Burkitt lymphoma, 2 were follicular lymphoma (grade 3a), and the rest were diffuse large B-cell lymphoma. Besides, 7 patients were diagnosed with double-hit lymphoma and 4 patients were relapsed after previous CD19 CAR-T cell therapy. Fourteen patients received the murinized CAR-T cells at a median dose of 2.18×106/kg (1-3×106/kg) CAR19 and 1.75×106/kg (1-3×106/kg) CAR22, while 12 patients received the does of the humanized CAR-T cells consisting of 2×106/kg CAR19 and 5×105/kg CAR22. At the day 28 assessment, 12/12 (100%) patients received the humanized CAR-T cells achieved overall response, including 9/12 (75%) CR and 3/12 (25%) partial remission (PR), while the overall response rate (ORR) and CR rate of the murinized group was 85.71% (12/14) and 42.86% (6/14) respectively. Among of them, all 4 Burkitt lymphoma patients in both groups achieved CR. In 3 months evaluation, 7/10 (70%,2 cases received infusion for less than 3 months) patients in the humanized group maintained in CR, which was higher than the murinized group with CR rate of 5/14 (35.71%). (Figure 1A-B). At a median follow-up of 291 days (range, 31 to 544), patients in the humanized group had a favorable progression-free survival (PFS) than those in the murinized (1-year PFS of 67.9% vs. 26.8%), although there was no statistical significance (p=0.08) due to the limited number of patients enrolled (Figure 1E-F). Achieving CR on day 28 (HR: 0.21, 95% CI: 0.06-0.72; P=0.012) and maintaining CR till the 3rd month (HR: 0.18, 95% CI: 0.06-0.59; P=0.004) were found as independent prognostic factors associated with favorable PFS. Maintaining CR till the 3rd month (HR: 0.10, 95% CI: 0.01-0.61; P=0.012) and non-double hit (HR: 0.11, 95% CI: 0.02-0.76, P=0.024) predicted a longer overall survival (OS) (Figure 1G-L). Despite impressive CR rates wereachieved, majority of patients had mild CRS (grade 1-2) in both group. Only 2patients experienced grade 3 CRS, and 2 patients developed grade 3 neurotoxicity. Image:Summary/Conclusion: This clinical trial demonstrates promising efficacy and safety of CD19/CD22 CAR-T cocktail therapy for R/R aggressive B-cell lymphoma, and patients in the humanized group showed better results than those with murinized, although this not a randomized trial.

Objective To study the effect of peripheral blood cell paremeters in the patients with refractory/relapsed acute B-lymphoblastic leukemia treated with CD19 chimeric antigen receptor T cells (CD19-CAR-T). Methods We retrospectively analyzed the clinical data of patients with refractory/relapsed acute B-lymphoblastic leukemia treated with CD19-CAR-T cells in Beijing Ludaopei hospital from May 2017 to May 2020 . The peripheral blood cell parameters were compared between 46 patients with minimal residual disease (MRD) negative (MRD negative group) and 38 patients with minimal residual disease positive (MRD positive group) after CAR-T cells therapy. Results The mean values of leukocytes, hemoglobin, platelets and neutrophils(LHPN0) in the MRD negative group were signifi cantly higher than those in the MRD positive group on the day of CAR-T cells transfusion, and the diff erencewas statistically signifi cant (P＜0.05). The mean platelet values on the 7th, 14th and 21st day (PLT7, PLT14, PLT21)after CD19-CAR-T cells transfusion were signifi cantly higher in the MRD negative group than those in the MRD positive group, respectively, and the diff erencewas statistically signifi cant (P＜0.05). Conclusion The higher value of LHPN0, PLT7, PLT14, PLT21 may indicate that CD19-CAR-T cell treatment may be eff ective. The hematopoietic recovery is faster after CD19-CAR-T treatment than after chemotherapy.

Chemotherapy, all-trans retinoic acid (ATRA), and arsenic are effective options for acute promyelocytic leukemia (APL). We conducted a 20-year retrospective analysis of newly diagnosed (ND) APL patients treated with arsenic, ATRA and mitoxantrone. After achieving complete remission (CR), patients received 3–5 cycles of chemotherapy followed by AS4S4 maintenance for 3 years. Eighty-eight ND APL patients were treated with either oral AS4S4 (n = 42) or arsenic trioxide (ATO) (n = 46). The 8-year overall survival (OS) rate was 100

目的:研究培门冬酶治疗结外NK/T细胞淋巴瘤的疗效及安全性.方法:2017年3月-2019年1月收治NK/T细胞淋巴瘤(鼻型)患者86例,随机分为两组,各43例.对照组给予左旋门冬酰胺酶治疗,观察组给予培门冬酶治疗,比较两组治疗效果.结果:观察组疾病缓解率高于对照组,差异有统计学意义(P<0.05);观察组消化道反应、高血糖、过敏反应的发生率均低于对照组,差异有统计学意义(P<0.05);观察组3个月、6个月、9个月及1年生存率均高于对照组,差异有统计学意义(P<0.05).结论:相较于左旋门冬酰胺酶,培门冬酶治疗NK/T细胞淋巴瘤的疗效更好,安全性高.

Abstract Background In October 2020, we began the clinical trials of CD7 CAR-T treatment for CD7-positive hematological malignancies at our center. We found that the proliferation profile and evolution of CD7 CAR-T cells within 1-month following infusion into patients were quite different from those of CD19 CAR-T cells. From these data, we reasoned that the time to occurrence of CAR-T-cell-related side effects might also differ between the two cellular therapies. Here, we systematically compared the proliferation and CAR-T-cell-related side effects of CD7 CAR-T cells to these of CD19 CAR-T cells. Patients and Methods From October 2020 to June 2021, a total of 30 patients (24 male, 6 female) including 22 with T-cell acute lymphoblastic leukemia (T-ALL), 3 with T-cell lymphoblastic lymphoma (T-LBL), and 5 with mixed phenotype acute leukemia (MPAL) received autologous CD7 CAR-T cells manufactured by the SenlangBio company (https://clinicaltrials.gov NCT04572308, NCT04796441 and NCT04938115). The median follow-up time was 116 days (range: 15-221days). On Day 30, 25/30 patients (83.3%) achieved complete remission (CR)/CR with incomplete blood recovery (CRi). From December 2017 to June 2021, 45 B-ALL patients (19 male, 26 female) received CD19 CAR-T cells, also manufactured by SenlangBio (NCT04792593 and NCT04546893). The median follow-up time was 351 days (range: 15-1110days). On Day 30, 43/45 patients (95.6%) achieved CR/CRi. The median infused CD7 CAR-T cell dose was 1×10 6/kg (range: 0.5-2×10 6/kg), and the median infused CD19 CAR-T cell dose was 3×10 5/kg (range: 0.2-10×10 5/kg). The CD7 or CD19 CAR-T cell ratio in peripheral blood lymphocytes (PBLC) and the CD7 or CD19 B-lymphocyte percentage in PBLC samples from patients were analyzed on days 0, 4, 7, 10, 14, 21, and 30 following CAR-T cell infusion using flow cytometry. Results The presence of CD7 CAR-T cells in the PBLC samples were gradually detected following CD7 CAR-T cell infusion. The CD7 CAR-T cell ratio in PBLC increased significantly on Day 10. CD7 CAR-T cell peak appeared on Day 21 with a peak of 39.14% (range: 0.04%-74.58%), and was still detectable on Day 30 with a high CD7 CAR-T ratio of 7.5% (1.15%-70.41%). The ratio of CD19 CAR-T cells in patient PBLC samples showed a significant increase on Day 7 following infusion, and the CAR-T cell peak appeared on Day 10 with a peak of 14.71% (range: 0.11%-89.33%), and then quickly decreased to 0.23% (range: 0%-82.88%) on Day 21 (Figure 1). As the CAR-T cells increased, the proportion of target cells decreases significantly (Figure 2). However, the rate of decrease of CD19 cells differed from that of CD7 cells. CAR-T cell proliferation is also associated with CAR-T-cell-related adverse effects including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Observing the adverse effects after CD7 CAR-T infusion, we found that fever (incidence rate of 83.8%) occurred on the first 1-3 days following infusion, with a body temperature among patients of about 38°C. After patients' body temperature dropped to approximately normal levels, fever occurred again on Day 10-21 (incidence rate of 77.4%), and a higher temperature of 38-40°C was observed. The adverse event profile coincided with the proliferation of CD7 CAR-T cells we observed. Among the 30 cases, 5 had Grade 2 CRS, 2 had CRS of Grade ≥3, and 1 patient had Grade 3 ICANS. Fever following CD19 CAR-T infusion consisted mainly on Day 7-14 after the infusion (incidence rate of 86.6%), followed by a gradual drop of body temperature to normal after Day 14. Among the 45 patients, 5 had Grade 2 CRS, 5 had CRS of Grade ≥3 and 7 had Grade ≥3 ICANS. Conclusions In this clinical study, we found that the proliferation and evolution of CD7 CAR-T cells are distinct from that of C19 CAR-T cells. CD7 CAR-T cells began to proliferate significantly later following patient infusion and persisted longer compared to CD19 CAR-T cells. We found that patients experienced two rounds of fever, appearing on Day 1-3 and Day 10-21 following CD7 CAR-T infusion, which required more attention and prevention compared to the fever experienced by patients infused with CD19 CAR-T cells. However, the incidence of CRS and ICANS did not increase following CD7 CAR-T infusion. More patients and long-term observation are needed to confirm these results and to improve clinical management of patients treated with CAR-T cellular therapies. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

报道1例47岁男性急性髓系白血病(AML)患者,化疗期间予其行PICC置管后,患者表现为高热、右上侧胸痛、右手腕部肿胀疼痛,12d内右前臂肌肉组织肿胀面积逐步扩大,伴右手拇指、食指肿胀及麻木.先后给予多种抗生素治疗,效果欠佳,持续高热.通过宏基因组测序(mNGS)技术快速准确辅助临床检测患者外周血、肌肉组织及痰液样本,确诊患者感染的病原体为微小根毛霉,先后予两性霉素B、安必素、泊沙康唑药物对患者进行治疗,感染得到控制.

目的:探讨异基因造血干细胞移植(allo-HSCT)对第一次形态学完全缓解(CR1)急性髓系白血病(AML)患者预后的影响.方法:回顾性分析2016年8月～2017年8月在本院就诊的AML患者62例,根据治疗方法将患者分为巩固化疗组(n=27)和allo-HSCT组(n=35).巩固化疗组在获得CR1后,采用巩固化疗.allo-HSCT组患者获得CR1后先采用Ara-C巩固化疗1～4个疗程后,再采用allo-HSCT治疗.记录患者达到CR1后1年、2年和3年内的生存、死亡和复发情况,流式细胞仪分析外周血CD3+T、CD4+T、CD4+T/CD8+T、CD8+T和CD4+CD25+Treg水平.结果:allo-HSCT组患者1年、2年、3年总生存率和无病生存率分别为88.57％、77.14％、74.29％和80.00％、71.43％、65.71％,明显高于巩固化疗组的62.96％、44.44％、37.04％和48.15％、33.33％、29.63％;allo-HSCT组患者1年、2年和3年复发率分别为8.57％、17.14％和20.00％,明显低于巩固化疗组的40.74％、51.85％和55.56％,且allo-HSCT组复发死亡率明显低于巩固化疗组,移植物抗宿主病(GVHD)死亡率明显高于巩固化疗组.CR1后1年,allo-HSCT组外周血CD4+T和CD4+T/CD8+T水平明显高于巩固化疗组,CD4+CD25+Treg水平明显低于巩固化疗组.结论:allo-HSCT治疗可以改善CR1后AML患者的免疫功能,提高预后生存率,降低复发率.