Protease nexin 1 (PN1) is an endogenous serine protease inhibitor (SERPIN), expressed at high levels in the prostate, and capable of inhibiting the proliferation of prostate cancer cells. We previously showed that PN1-uPA complexes inhibited Sonic Hedgehog (SHH) signalling through engagement of the LRP receptor. Here, we describe an alternative anti-proliferative mechanism through which PN1 expression leads to apoptosis. In prostate cancer cells, increased expression of PN1 led to substantial reduction of XIAP levels and apoptosis mediated through the uPAR, but not the LRP receptor. The alterations in XIAP were effected in two ways 1) via alteration in the NF-κB pathway, a pathway known to signal XIAP transcription and 2) by promoting XIAP instability. The AKT pathway is known to phosphorylate XIAP at serine 87 leading to protein stability and PN1 expression is shown to interfere with this process. As a result of both mechanisms, programmed cell death is substantially increased. Consistent with these observations, reduced PN1 protein correlated with elevated p65/XIAP expression and with higher Gleason scores in human prostate tissue arrays. Thus, PN1 expression appears to differentially down-regulate distinct oncogenic pathways depending upon the cell surface receptor engaged by its complexes and demonstrates a novel molecular mechanism by which the protein can promote tumor cell apoptosis.

Abstract Purpose: This report presents long-term outcomes of third-generation anti-CD30 chimeric antigen receptor (CAR) T-cell therapy in patients with relapsed/refractory (r/r) CD30+ lymphoma. Patients and Methods: In this single-arm, multicenter, phase I/II trial, patients received a lymphodepletion regimen comprising fludarabine and cyclophosphamide, followed by infusion of anti-CD30 CAR T cells. Primary endpoints included safety and overall response rate (ORR), whereas secondary endpoints were progression-free survival (PFS) and overall survival (OS). Results: Forty-four patients were enrolled, including 33 cases of Hodgkin lymphoma. Of 44 patients, 23 achieved complete response (CR) to CAR T, and 19 achieved PR, resulting in a CR rate of 52.3% and an ORR of 95.5%. The most frequent toxicities were hematologic adverse events of grade 3 or higher (68.2% of neutropenia). Cytokine release syndrome occurred in 18 (40.9%) patients, with 2 (4.5%) cases being ≥ grade 3. In the follow-up period, 24 patients underwent autologous hematopoietic stem cell transplantation (auto-HSCT) after CAR T within a median of 3 months. The best ORR was 95.5%, with 27 (61.4%) patients achieving CR. The best CR rate was higher in patients receiving CAR T followed by auto-HSCT compared with those receiving CAR T alone (75% vs. 45%). Three-year OS and PFS rates for all patients were 79% [95% confidence interval (CI), 66.1%–91.9%] and 74.2% (95% CI, 60.3% –88.1%), respectively. Patients receiving consolidated auto-HSCT following CAR T exhibited significantly longer OS and PFS compared with those treated with CAR T alone. Conclusions: Third-generation anti-CD30 CAR T demonstrates high efficacy and a favorable safety profile in patients with r/r CD30+ lymphoma. Addition of auto-HSCT following CAR T-cell therapy improves depth of remission and potentially enhances OS and PFS.

CD19/CD22 dual-target chimeric antigen receptor (CAR19/22) T cell therapies mark a key advance over single-target options for refractory B-cell malignancies. However, over 20% of patients relapse and underlying mechanisms remain less understood. This study evaluated relapse in 91 B-cell acute lymphoblastic leukemia (B-ALL) patients treated with CAR19/22 T cells a clinical trial (ChiCTR-OPN-16008526). The complete remission rate was 91.9%, and 32.9% (26/79) of responders relapsed at a median of approximately 7 months. Among late relapses (>7 months), 92.3% was CAR T functional insufficiency showing lack of persistence or recovery of CD19⁺ B lymphocytes. Early relapses showed greater heterogeneity with 30.8% attributable to CAR T functional insufficiency and 30.8% to antigen insufficiency. Notably, two early relapses (15.4%) exhibited concurrent CD19/CD22 downregulation and harbored a preexisting PAX5 deletion or frameshift insertion. These PAX5-mutated subclones persisted and expanded at relapse, with one patient acquiring additional CD19 mutations. PAX5 knockout in leukemic cells reduced CD19/CD22 expression and proximal enhancer activities, causing in vitro resistance to CAR19/22 T cells. Collectively, CAR T functional insufficiency and antigen insufficiency are the most (61.5%) and second (15.4%) frequent drivers of relapse, respectively. Disruptive PAX5 mutations define a distinct molecular mechanism of early relapse, guiding rational targeted strategies.

Abstract Introduction CD5 is broadly expressed across hematologic malignancies, notably in ~85% of T-cell tumors and is associated with poorer prognosis in certain B-cell tumors, making it a promising target for CAR-T cell therapy. Two primary clinical studies have reported on CD5-targeted CAR-T therapy: one using autologous CAR-T cells without CD5 editing (NCT03081910) and one using allogeneic CD5-knockout (CD5-KO) CAR-T cells (NCT05032599). To further explore its clinical utility, we conducted a Phase I trial (NCT04767308) of autologous CD5-KO anti-CD5 CAR-T cells (CAR55ko-T) in patients with relapsed and refractory (r/r) CD5⁺ hematologic malignancies. Methods Patients received a single dose of CAR55ko-T cell infusion after a three-day FC lymphodepletion regimen, followed by a 1-day rest. Dose levels evaluated were 1.0 × 10⁶, 2.0 × 10⁶, and 1.0 × 10⁵ cells/kg. Adverse events (AEs) were graded using CTCAE v5.0, and treatment responses were assessed per Lugano 2014 criteria. CAR-T kinetics were measured by ddPCR and flow cytometry. Lymphocyte subset dynamics were analyzed via multicolor flow cytometry. The study was conducted in accordance with the Declaration of Helsinki, and all participants provided informed consent. Results Seven patients (4 males, 3 females) were treated from February 2021 to October 2021, including 1 ALCL, 2 AITL, 1 SPTCL, 2 MCL, and 1 DLBCL. Median age was 47 years (range: 17–67), with a median of 3 prior therapy lines (range: 2–6). Patient 2 had previously received anti-CD19/22 CAR-T, and Patient 3 had undergone autologous HSCT. The overall response rate was 85.7%, with 4 CR and 2 PR. Median PFS and OS were 170 and 205 days, respectively. Patient 1 remained in remission > 902 days post-infusion. Notably, CD5+T cells were absent at the disease relapse or resistance. CAR55ko-T cells expanded robustly and persisted long-term. Median peak copy number was 87,178 copies/μg DNA (range: 31,618–118,000), with a median time to peak of 14 days. During a median follow-up period of 124 days, CAR-T cells remained detectable in all patients, with persistence >809 days in patient 1. CRS and ICANS were generally mild; only one case of grade 3 CRS occurred. The most common ≥ grade 3 AEs were cytopenia: neutropenia (100%), lymphopenia (100%), thrombocytopenia (78%), and anemia (89%). Notably, grade 4 lymphopenia and neutropenia recured after one-month post-infusion. Consistently, EBV (6/7) and CMV (4/7) reactivations were frequent. Four patients had bacterial infections, resulting in to two deaths. Uncommon AEs were observed, including rash (71.4%), skin tingling (71.4%) and autoimmune-associated AEs: xerostomia (28.6%), hypothyroidism (14.3%), and adrenal insufficiency (14.3%). Immune reconstitution process was monitored. Following the transient nadir, CD3+ T cells recovered to pre-lymphodepletion levels within 1 month and maintained stable. Notably, the reconstituted T cells was CD5 negative. The CD4/CD8 ratio decreased rapidly and remained < 1 throughout the entire follow-up. B cells were rapidly depleted and failed to recover during long-term follow-up. NK and NKT cells showed transient decline but rebounded to baseline. Conclusion CD5-KO anti-CD5 CAR-T therapy demonstrates promising efficacy in treating CD5+ hematological malignancies, including both T-cell and B-cell tumors. The knockout of CD5 enhances CAR-T persistence and therapeutic durability. However, sustained CD5+ T cells depletion and expansion of CD5- T cells may impair immune regulation and infection control in patients. These findings highlight the need to modulate the CAR-T persistence duration clinical application and support the consideration of consolidative HSCT therapy in patients achieving remission.

>CD5-positive （CD5+） diffuse large B-cell lymphoma（DLBCL） represents a special subgroup of DLBCL with a more aggressive disease course and is more likely to develop into relapsed/refractory （r/r） DLBCL in response to immunochemotherapy. The incidence of CD5+DLBCL is 5%–10%among DLBCL patients 1 .

KRT17 has no effect on proliferation, migration and invasion of CRC cells.

Abstract Background While CD19-targeting chimeric antigen receptor (CAR) T-cell therapy has demonstrated remarkable efficacy in relapsed/refractory non-Hodgkin lymphoma (r/r NHL), high relapse rates and CD19 antigen escape remain major clinical challenges. Given that CD19 loss represents a well-characterized resistance mechanism, we developed a structure-optimized CD19/CD20 bispecific CAR-T cell (SL1716) to improve clinical outcomes and reduce relapse risk in r/r NHL patients. Objective This study aims to develop and validate a structurally optimized bispecific nanobody-based CD19/CD20 CAR-T construct (SL1716) through comprehensive in vitro functional screening, and evaluate its safety and preliminary efficacy in the patients with r/r NHL. Methods We engineered an optimized bispecific CAR construct and systematically characterized its functional advantages, including dual-target synergy, persistence, and memory phenotype. The lead construct was evaluated in an investigator-initiated trial (IIT) involving heavily pretreated r/r B-NHL patients who were refractory to or relapsed after prior therapies. The primary objective was to assess the incidence and severity of treatment-emergent adverse events (per CTCAE v5.0 criteria), with secondary objectives including overall response rate (ORR; Lugano 2014 criteria). Comprehensive assessments were conducted, including clinical responses, adverse events, and CAR-T cell expansion kinetics. Results Using our alpaca immunization and phage display platform, we identified nanobodies specific to novel epitopes on CD19 and CD20. Through systematic optimization of each CAR module, we developed SL1716 as the lead bispecific CAR-T candidate. Functional characterization demonstrated its superior synergistic efficacy, as evidenced by dual-target synergistic cytotoxicity compared to monospecific CAR-Ts, along with robust antitumor activity across various lymphoma subtypes. Notably, SL1716 indicated sustained cytotoxic potency and an enhanced memory phenotype in multi-cycle antigen rechallenge assays. Between August 2024 and May 2025, 10 eligible patients with r/r B-NHL received SL1716 infusion, including 7 (70%) with DLBCL, 2 (20%) with MCL, and 1 (10%) with PMLBCL. The cohort had a median age of 55 years (range: 19-70), with 5 female (50%) and 5 male (50%) patients. All enrolled patients had disease progression after ≥1 prior line of therapy (median 4.5 lines, range 1-11) with ECOG scores of 1-2. Disease staging revealed 6 (60%) with stage IV, 1 (10%) with stage III, and 3 (30%) with bone marrow involvement, none with CNS infiltration. Among treatment-refractory cases, 3 had prior CAR-T exposure, including 1 case of CD19/CD22 CAR-T and 2 case of CD19 CAR-T therapy, all with subsequent relapse. Following lymphodepletion, all patients received SL1716 (CD19/CD20 bispecific CAR-T) at 4×106 cells/kg, with 100% manufacturing success rate. SL1716 exhibited excellent tolerability profile, with grade Ⅰ cytokine release syndrome (CRS) observed in 9 patients (90%) and no CRS in 1 patient (10%). Importantly, no immune effector cell-associated neurotoxicity syndrome (ICANS) events were documented during the observation period. Pharmacokinetic evaluation demonstrated favorable expansion kinetics, achieving a median peak CAR copy number (Cmax) of 4.15×104 copies/μg DNA (range: 7.23×103-2.05×105) at a median time to peak (Tmax) of 12 days post-infusion (range: 7-14 days). As of the data cut-off date (July 30, 2025), the median follow-up time was 6.5 months (range 2.03–11.7). A best overall response rate (ORR) of 90% was observed, with 6 patients achieving complete remission (CR; 60%) and 3 patients achieving partial remission (PR; 30%). By the 3-month post-infusion assessment, SL1716 achieved an ORR of 87.5% among 8 evaluable patients, with 5 patients (62.5%) achieving CR and 2 patients (25%) achieving PR. One patient experienced progressive disease (PD; 12.5%). Notably, all treated patients exhibited a clinical response to SL1716, with extended follow-up data currently being collected. Conclusion SL1716, a nanobody-based CD19/CD20 bispecific CAR-T cell therapy, demonstrates a favorable safety profile and durable clinical efficacy in patients with r/r NHL. These promising results support its potential as an effective treatment option for this high-risk patient population.

Importance Previous studies have demonstrated the advantages of short-term histopathological outcomes and complications associated with transanal total mesorectal excision (TME) compared with laparoscopic TME. However, the long-term oncological outcomes of transanal TME remain ambiguous. This study aims to compare 3-year disease-free survival of transanal TME with laparoscopic TME. Objective To evaluate 3-year disease-free survival between transanal TME and laparoscopic TME in patients with rectal cancer. Design, Setting, and Participants This randomized, open-label, noninferiority, phase 3 clinical trial was performed in 16 different centers in China. Between April 2016 and June 2021, a total of 1115 patients with clinical stage I to III mid-low rectal cancer were enrolled. The last date of participant follow-up was in June 2024. Interventions Participants were randomly assigned in a 1:1 ratio before their surgical procedure to undergo either transanal TME (n = 558) or laparoscopic TME (n = 557). Main Outcomes and Measures The primary end point was 3-year disease-free survival, with a noninferiority margin of -10% for the comparison between transanal TME and laparoscopic TME. Secondary outcomes included 3-year overall survival and 3-year local recurrence. Results In the primary analysis set, the median patient age was 60 years. A total of 692 male and 397 female patients were included in the analysis. Three-year disease-free survival was 82.1% (97.5% CI, 78.4%-85.8%) for the transanal TME group and 79.4% (97.5% CI, 75.6%-83.4%) for the laparoscopic TME group, with a difference of 2.7% (97.5% CI, -3.0% to 8.1%). The lower tail of a 2-tailed 97.5% CI for the group difference in 3-year disease-free survival was above the noninferiority margin of -10 percentage points. Furthermore, the 3-year local recurrence was 3.6% (95% CI, 2.0%-5.1%) for transanal TME and 4.4% (95% CI, 2.6%-6.1%) for laparoscopic TME. Three-year overall survival was 92.6% (95% CI, 90.4%-94.8%) for transanal TME and 90.7% (95% CI, 88.3%-93.2%) for laparoscopic TME. Conclusions and Relevance In patients with mid-low rectal cancer, 3-year disease-free survival for transanal TME was noninferior to that of laparoscopic TME.

Background:Although the overall survival of multiple myeloma (MM) has improved significantly, patients with ultra-high-risk features (UHR-MM) had dismal outcomes. New therapies to address this unmet medical need are warranted. Equecabtagene autoleucel (eque-cel) has been approved for patients who have received at least three previous lines of therapy by the Chinese National Medical Products Administration. With good efficacy and safety profile in these patients, eque-cel is being explored in early relapse or newly diagnosed UHR-MM patients, Hereby, we report the primary real world data of eque-cel followed ASCT in UHR-MM patients. Methods:We conducted a retrospective chart review on UHR-MM patients who received eque-cel followed ASCT. UHR-MM are defined as: 1) Genetic ultra-high risk: del(17p)≥60%; or ≥2 high-risk cytogenetic abnormalities including TP53 mutation, del(17p)/P53 deletion, t(4;14), t(14;16), t(14;20), 1q21 gain/ amplification; 2) Primary refractory: Results:From August 2023 to April 2025, 12 UHR-MM patients completed ASCT followed by eque-cel infusion. Six patients received melphalan, five received bendamustine combining melphalan and one patient received fludarabine combining melphalan conditioning. Peripheral stem cell was administrated at (2.3-5.5) × 106 cells/kg, and eque-cel was administrated at 1 × 106 cells/kg for all 12 patients. The median age was 53 years (range: 36-67) and 11 (91.7%) were male. Eight patients (72.7%) had genetic ultra-high risk features, one patients (8.3%) were early progression, and two patients (16.7%) had primary PCL history. Two (16.7%) patients had extramedullary disease. With median 2 (range: 1-4) previous line of therapy, the median disease course is 10.5 months before ASCT. Eleven (91.7%) patients had received daratumumab based triplet or quadruplet therapy. All patients received bridging and 11 received maintenance therapy. Two patients, who had received eque-cel infusion within the preceding 6 months but did not achieve a complete response (CR), subsequently underwent consolidation therapy with eque-cel followed ASCT. With a median follow-up of 196 days (from ASCT date), seven patients (58.3%) experienced grade 1 and three patients (25.0%) experienced grade 2 CRS and fully recovered. Four patients were treated with glucocorticoids. No ICANS event was reported. As expected, AEs are dominated by hematological toxicities. All 12 patients achieved hematopoietic reconstitution within 1 months after ASCT, with a median time of 15 days for ANC reconstitution (≥0.5×109/L) and 11.5 days for PLT reconstitution (≥20×109/L) post ASCT. By July 1st, 2025, the ORR was 100%, with all 12 patients reached CR. All patients are MRD negative. Two patients received first eque-cel 3 month before ASCT, which achieved both VGPR. After ASCT and second eque-cel infusion, achieved sCR and MRD negative status on day 23 and day 190 post-ASCT, respectively. The early progression patient relapsed at 55 days post ASCT, all the other patients are keeping their response and under follow-up. The median DOR, PFS and OS were not reached by cutoff date. Robust CAR T-cell expansion was observed, with a median Tmax of 11 days (range 7~21). The median Cmax was 665.04 cells/μL. The pharmacokinetic profile is similar to that of eque-cel in R/RMM patients. Conclusion:Eque-cel followed ASCT demonstrated promising deep and durable response and was well tolerated in UHR-MM patients. CRS events are slight, hematopoietic reconstruction rate was 100%. We are looking forward to more patients gaining long-term benefit from this new treatment.

BACKGROUND:Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of aggressive non-Hodgkin's lymphoma with distinct clinical and molecular heterogeneity. DLBCL that arises in extranodal organs is particularly linked to poor prognosis. This study aimed to determine the clinical and molecular characteristics of extranodal involvement (ENI) in DLBCL and assess the actual survival status of the patients. METHODS:In this population-based cohort study, we investigated the clinical features of 5,023 patients newly diagnosed with DLBCL. Their clinical conditions, eligibility criteria, and sociodemographic details were recorded and analyzed. Gene panel sequencing was performed on 1,050 patients to discern molecular patterns according to ENI. RESULTS:The 2-year overall survival (OS) rate was 76.2% [95% confidence interval (CI), 74.0%-78.2%], and the 5-year OS rate was 67.9% (95% CI, 65.2%-70.4%). The primary treatment was immunochemotherapy with rituximab. Specific lymphoma involvement sites, especially the bones, bone marrow, and central nervous system, were identified as independent adverse prognostic factors. A high prevalence of non-germinal center B-cell (non-GCB) phenotype and myeloid differentiation primary response 88 (MYD88)/CD79B mutations were noted in lymphomas affecting the breasts, skin, uterus, and immune-privileged sites. Conversely, the thyroid and gastrointestinal tract showed a low occurrence of non-GCB phenotype. Remarkably, patients with multiple ENIs exhibited a high frequency of MYD88, tet methylcytosine dioxygenase 2 (TET2), CREB binding protein (CREBBP) mutations, increased MYD88L265P and CD79B mutation (MCD)-like subtypes, and poor prognosis. Genetic subtype-guided immunochemotherapy showed good efficacy in subgroup analyses after propensity score matching with 5-year OS and progression-free survival rates of 85.0% (95% CI, 80.6%-89.5%) and 72.1% (95% CI, 67.3%-76.7%). CONCLUSIONS:In the rituximab era, this large-scale retrospective analysis from Asia confirmed the poor prognosis of DLBCL with multiple ENIs and underscored the efficacy of genetic subtype-guided immunochemotherapy in treating extranodal DLBCL.

The urban agglomeration in central Guizhou is located in a crustal deformation area caused by tectonic uplift between the Mesozoic orogenic belt of East Asia and the Alpine-Tethys Cenozoic orogenic belt, with high mountains, steep slopes, fractured rock masses and a fragile ecological environment; this area is the most affected by landslides in Guizhou Province, China. In the past decade, there were a total of 613 medium and large landslide disasters, resulting in 137 deaths and a direct economic loss of 1.032 billion yuan. Therefore, this study selected 12 indicators from the topography, geological structure, and external inducing factors, and conducted factor collinearity analysis using the variance expansion coefficient to construct a landslide hazard assessment index system. The statistical analysis model was combined with a variety of machine learning models, and the selection of negative sample points was restricted in various ways to improve training data accuracy and enable machine learning model predictions with sufficiently supervised prerequisites. The accuracy of the model was validated by ROC curve analysis. The AUC values of the SVM, DNN, and bagging models were all greater than 0.85, indicating that the results were credible. However, the overall accuracy was SVM > DNN > Bagging; that is, SVM was more suitable for landslide hazard assessment of the urban agglomeration in central Guizhou. Finally, field surveys were used to validate multiple sites with historical landslides in extremely high-hazard areas and analyse their development characteristics. The evaluation results can provide strong guidance for engineering design, construction and disaster prevention decision-making of urban agglomeration in central Guizhou.

Background Polatuzumab vedotin (Pola), an anti-CD79b antibody-drug conjugate delivering monomethyl auristatin E (MMAE), is approved for diffuse large B-cell lymphoma (DLBCL). CD79B mutations are recurrent in DLBCL, particularly in patients with multiple extranodal diseases and MCD subtype, and associate with inferior prognosis following standard immunochemotherapy. Whether these mutations confer resistance to Pola remains clinically significant yet undetermined. Methods In vitro, CD79B-null K562 and 293T cell lines were transduced to express CD79B Y196H mutant variant, while endogenous CD79B in TMD8 cell line was modified to generate homozygous mutations using CRISPR-Cas9 gene editing. Cells were exposed to Pola (5 μg/mL, 72 hours), and cytotoxicity was evaluated by trypan blue exclusion assay. Clinically, 20 treatment-naïve DLBCL patients with CD79B Y196 mutation received either R-CHOP (n=10) or Pola-based regimens (Pola-R-CHP: n=9; Pola-R2: n=1) for six cycles. Treatment responses (ORR, CRR) and progression-free survival (PFS) were compared between cohorts. Results Pola demonstrated potent cytotoxicity against CD79B-mutated cells in vitro. Viability plummeted to 6.39% ± 1.71% (vs. 81.27% ± 2.25% in controls, p<0.001) in mutant K562 cells, 2.29% ± 0.90% (vs. 88.22% ± 1.82%, p<0.001) in 293T transfectants, and 6.57% ± 0.69% (vs. 82.47% ± 1.37%, p<0.001) in homozygous mutant TMD8 cells. Clinically, Pola-based regimens achieved superior best overall response rates (ORR 100% vs. 90%; CRR 80% vs. 60%) and end-of-treatment responses (Cycle 6 ORR 100% vs. 80%; CRR 88.9% vs. 60%) compared to R-CHOP. At median 6-month follow-up, PFS rates were 100% for Pola-based therapy versus 90.0% for R-CHOP (log-rank p=0.705). Conclusions Pola retains potent activity against CD79B-mutant DLBCL models and demonstrates clinically meaningful efficacy in treatment-naïve patients, achieving 100% response rates despite this historically adverse biomarker. These findings position Pola-based regimens as a promising therapeutic strategy for CD79B-mutated DLBCL.

Alzheimer's disease (AD) is a prevalent neurodegenerative disorder characterized by memory decline and cognitive impairments. The clinical treatments for AD have numerous adverse effects; hence, the exploration of natural products for AD therapy is of significant importance. Protocatechuic acid (PA), a natural phenolic acid, has been shown to possess various pharmacological activities, including anti-inflammatory, antioxidant, and antitumor effects. However, the mechanisms underlying its therapeutic potential for AD remain elusive. This study utilized a β-amyloid (Aβ) injection into the hippocampus of mice as an AD model and L-glu-induced HT-22 cell neurotoxicity and lipopolysaccharides (LPS)-induced cellular neuroinflammation models to assess reactive oxygen species (ROS), JC-1, and relevant biochemical markers. This study examined behavioral, pathological, and inflammatory factors and investigated the molecular mechanisms through transcriptomics, western blot, and molecular docking studies. This study's findings reveal that high-dose PA (50 mg/kg) improves symptoms in AD mice through the cholinergic synaptic signaling pathway. This study indicates that PA is a potential candidate for AD treatment targeting the cholinergic synaptic signaling pathway, providing a lead compound for AD therapy.

Chimeric antigen receptor (CAR)-T-cell therapy has achieved remarkable clinical success in the treatment of B-cell malignancies; however, its efficacy can be limited by poor T-cell persistence and insufficient antitumor activity in certain cases. Moreover, interleukin-12 (IL-12) is a prominent agent in cancer immunotherapy, but its clinical application is constrained by severe toxicity associated with systemic exposure. In this study, we developed a novel cytokine delivery platform based on CAR target-modified cell-derived extracellular vesicles (EVs) that preferentially bind CAR-T cells to improve CAR-T-cell function. EVs with surface-displayed CD19 and/or IL-12 were successfully generated from HEK-293T cells. Compared with an equivalent concentration of rhIL-12, IL-12 EVs significantly enhanced the effector function of anti-CD19 CAR-T cells in vitro, resulting in increased Interferon-γ (IFN-γ) and TNF-α secretion, cytolytic activity, and T-cell expansion. Additionally, compared with EVs expressing IL-12 alone, EVs co-expressing IL-12 and CD19 (CD19/IL-12 EVs) exhibited superior binding efficiency to CAR-T cells but not to T cells, as indicated by flow cytometry. In xenograft model mice bearing CD19 + Raji tumors, intratumoral injection of CD19/IL-12 EVs resulted in durable antitumor responses and enhanced the in vivo expansion of CAR-T cells, outperforming CD19 EVs, IL-12 EVs and control EVs, without causing systemic toxicity. RNA sequencing (RNA-seq) analysis of CAR-T cells stimulated with EVs suggested that the increased efficacy was driven by IL-12 signaling. These data demonstrate that CAR-targeted modified EVs may serve as targeted cytokine delivery systems for CAR-T cells, offering a safe and effective strategy to augment CAR-T-cell function.

Abstract Background Acute myeloid leukemia (AML) is a genetically heterogeneous hematological malignancy. Risk stratification in AML is primarily determined by genomic classification and cytogenetic alterations. The International Consensus Classification (ICC)-2022 classifies AML with mutations in ASXL1, BCOR, EZH2, SF3B1, SRSF2, STAG2, U2AF1, ZRSR2, or RUNX1 as AML with myelodysplasia-related gene mutations (AML-MR). According to the European Leukaemia Net-2022 guidelines (ELN-2022), MR-associated mutations are categorized as adverse-risk. Allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains an important curative approach for AML, however, its survival benefit in patients harboring AML-MR mutations with additional co-mutations remains controversial. Methods This is a multicenter, retrospective designed study, based on the transplant database of the TROPHY group. AML patients receiving allo-HSCT from February 2016 to December 2023 were screened. Eligibility criteria included: (1) diagnosis of AML, (2) ELN-2022 adverse-risk classification, (3) presence of ≥1 AML-MR mutation. Induction therapy was administered following the recommendations outlined in the National Comprehensive Cancer Network (NCCN) guidelines. Complete remission (CR) was defined as <5% blasts in bone marrow, absence of circulating blasts or extramedullary leukemia and peripheral blood recovery (absolute neutrophil count ≥1,000/μL and platelet count ≥100,000/μL). If CR was not achieved after the first cycle, a second induction using the same or an intensified cytarabine-based regimen was administered. Selection of consolidation treatment was based on measurable residual disease (MRD) status, donor availability, and the patients’ fitness. Key outcomes included relapse-free survival (RFS), overall survival (OS), cumulative incidence of relapse (CIR), and non-relapse mortality (NRM). The study was approved by the institutional review board of each participated hospital and was conducted in accordance with the Declaration of Helsinki. Results A total of 341 acute myeloid leukemia patients were enrolled. Among the AML-MR defining genes, the most frequently mutated were ASXL1 (32%), RUNX1 (29%), and BCOR (22%). Common co-mutated genes included TP53 (7%) and epigenetic regulator genes such as IDH1/2 (6%), DNMT3A (6%), and TET2 (6%). Overall, 193 patients (56.6%) achieved CR after the first induction. Regimens incorporating hypomethylating agents or venetoclax yielded significantly higher CR rates than conventional idarubicin-cytarabine (IA) regimens (64.4% vs. 43.3%; HR = 1.979, 95% CI: 1.197–3.272, p = 0.008). A total of 322 patients (94.4%) had achieved CR prior to allo-HSCT, the entire cohort demonstrated a 2-year OS of 77.4% and RFS of 70%. Patients achieved CR prior to transplantation had significantly improved RFS (2y-RFS: 71.1% vs. 49.3%, P=0.026) and a trend toward lower CIR (2y-CIR: 13.7% vs. 33.7%, P=0.052), although OS was not significantly different. Achieving CR after first induction was associated with superior RFS (2y-RFS: 81.8% vs. 55,5%, P＜0.001), OS (2y-OS: 85.6% vs. 66.3%, P＜0.001), lower CIR (2y-CIR: 9.5% vs. 18.8%, P=0.025), and reduced NRM (2y-NRM: 8.7% vs. 25.5%, P＜0.001). Multivariate analysis identified post-remission consolidation therapy as an independent predictor of improved OS (HR = 0.56, 95% CI: 0.32–0.97, p = 0.015). Conversely, Grade III-IV aGvHD and TET2 co-mutation were associated with inferior OS (HR= 2.32, 95% CI: 1.35–3.98, p = 0.002; HR= 4.15, 95% CI: 1.90–9.05, p< 0.001, respectively). Additionally, both Grade III-IV aGvHD and pre-transplant MRD positivity predicted inferior RFS (HR= 1.86, 95% CI: 1.14–3.05, p = 0.014; HR= 1.73, 95% CI: 1.10–2.74, p = 0.019, respectively). NPM1 co-mutation (n=9) and TP53 co-mutation (n=25) showed no significant impact on RFS or OS. However, patients harboring epigenetic regulator mutations had significantly reduced OS (2y-OS: 73.0% vs. 83.1%, P=0.044), without affecting other endpoints. Conclusion In patients with AML-MR, induction regimens incorporating hypomethylating agents or venetoclax are recommended due to their efficacy in improving CR rates. Achieving CR after first induction significantly improves post-transplant RFS and OS along with reduced CIR and NRM. Co-mutations in TP53 and NPM1 did not significantly impact survival. Notably, epigenetic regulator mutations were linked to reduced OS, highlighting the prognostic relevance of specific co-mutations in AML-MR.

Hydrogen and Al2O3 supported NiCu alloy bimetallic nanoparticle (NP) catalysts were simultaneously prepared through Al-ethanol reaction with a mixed dispersion of Al powders, ethanol, nickel chloride hexahydrate, copper chloride dihydrate and aluminum chloride hexahydrate. The addition of Cu improved the dispersion of Ni and enhanced antioxidation ability. The Ni80Cu20 NPs demonstrated an outstanding in-situ hydrogen production performance with a hydrogen yield of 97% and a generation rate of 85 mL-H2-min-1-g-Al-1, which was 1.1 and 3.4 times compared to that of monometallic Ni and Cu NPs. Moreover, the ethylene selectivity of Al2O3 supported Ni80Cu20 NPs catalysts synthesized via subsequently freeze-drying the products from the Al-ethanol reaction, was 2 and 8 times higher than that of monometallic counterpart samples. This synergistic effect between the metals altered both the geometric and electronic properties of the catalyst. Furthermore, compared to Pd-based catalysts, the Ni80Cu20/Al2O3 catalyst exhibited comparable ethylene selectivity for the semi-hydrogenation of acetylene.

Clear cell renal cell carcinoma (ccRCC) is the most prevalent type of renal malignancy, and the deubiquitinase USP3 has been implicated as a critical factor in tumor biology. However, the precise mechanisms by which USP3 contributes to ccRCC progression remain unclear. This study investigates the role of USP3 in ccRCC and elucidates its underlying molecular mechanisms. Data from TCGA and GTEx databases showed elevated USP3 expression in ccRCC tissues and cell lines compared to normal renal tissues. Further analysis using qPCR and Western blot confirmed this upregulation in ccRCC cell lines. Functional assays revealed that silencing USP3 significantly impaired cell proliferation, migration, and invasion, while promoting apoptosis. Additionally, co-immunoprecipitation assays demonstrated an interaction between USP3 and MYC, with subsequent ubiquitination assays showing that USP3 regulates MYC stability. USP3 depletion also led to alterations in glycolysis-related gene expression, which could be partially reversed by MYC overexpression. These findings suggest that USP3 modulates ccRCC progression by stabilizing MYC, highlighting its potential as a therapeutic target in ccRCC treatment.

Abstract Background: Preliminary studies, including ours, have demonstrated that CAR-T (CAR19) therapy confers suboptimal therapeutic efficacy in high-risk B-cell acute lymphoblastic leukemia (B-ALL), particularly in cases with IKZF1 deletion (IKZF1DEL) and Ph-like phenotype (2023 EHA oral presentation, 2024 Hemasphere). Yet such findings in the Ph-like ALL cohort lack validation in large cohorts. Here we seek to explore the efficacy of CAR19 therapy in high-risk Ph-like ALL. Method: This multicenter, retrospective cohort study included 75 patients with Ph-like ALL in five centers of China from 2017 to 2023. We collected and analyzed the data of clinical characteristics. The event-free survival (EFS) and overall survival (OS) were investigated. Result: A total of 75 patients with Ph-like ALL receiving CAR19 therapy were enrolled. Patients were aged from 6 to 64 years, with the median follow-up time of 445 days (95%CI: 330-559). This cohort was grouped as ABL-class fusions (11/75, 14.6%; ABL1, ABL2, CSF1R, PDGFRB), CRLF2 positive (16/75, 21.3%; rearrangement or high-expression), EPOR or JAK2 rearrangement (8/75, 10.6%), JAK-STAT pathway mutations (16/75, 21.3%; IL7R, SH2B3, JAK1, JAK3, FLT3), Ras pathway mutations only (24/75, 32%; KRAS, NRAS, PTPN11, NF1). Consistent with our previous study, patients with IKZF1 deletion (24/75, 32%) conferred inferior outcome compared with those without (51/75, 68%), with 1y-EFS 14.1% vs. 71.5%, (P=0.002), and 1y-OS 55.7% vs. 80.8% (P=0.22), respectively. Furthermore, within the IKZF1WT group, CAR19 therapy showed different efficacy among various Ph-like subgroups. Synthesized the above findings, patients were further divided into three groups following the survival difference: Standard Risk (SR, IKZF1WTwith ABL1-class fusion or CRLF2 positive, n=18); Medium Risk (MR, IKZF1WT with JAK-STAT pathway mutation or Ras pathway mutation, n=28), and High Risk (HR, IKZF1WT with EPOR or JAK2 rearrangement and IKZF1DEL, n=29). Results demonstrated that HR group showed significantly worse outcomes compared to the other two groups (1y-EFS 79.6% in SR, 73.2% in MR, 12.7% in HR, respectively, P=0.02; 1y-OS 100% in SR, 73.6% in MR, 54.6% in HR, respectively, P=0.018). Conclusion: This study isolated the highest-risk subgroup of Ph-like ALL for CAR19 therapy, which serves as a valuable prognostic feature in Ph-like ALL.

Abstract Background Equecabtagene autoleucel (eque-cel), a fully human BCMA-targeted CAR-T therapy, has exhibited unprecedented efficacy in relapsed/refractory multiple myeloma (RRMM) patients with ≥3 prior lines of therapy (LOT), achieving 96% overall response rates (ORR) in clinical trials. Despite its 2023 approval in China, critical knowledge gaps persist regarding its real-world applicability across heterogeneous patient populations. Method This is a single-center retrospective study that evaluates the efficacy, safety, and durability of eque-cel in Chinese RRMM patients. The prior lines of treatment (LOT), baseline characteristics before eque-cel infusion, as well as the safety and efficacy of eque-cel were recorded. Categorical variables were analyzed using Fisher's exact test. Survival outcomes including progression-free survival (PFS) and overall survival (OS) were evaluated by Kaplan-Meier methodology and significance testing was conducted using the Log-rank method. We also performed Cox regression analyses for PFS as univariate analyses. Results From June 2023 to February 2025, totally 45 patients were enrolled, 22 (48.9%) were male, with a median age of 61 years (range, 30-75 years). High-risk cytogenetic abnormalities included G(1q21) in 46.7% (21/45), t(4;14) in 15.6% (7/45), and TP53/del(17p) mutation in 33.3% (15/45). Fifteen patients (33%) harbors at least two high-risk cytogenetic abnormalities. The median number of prior LOT was 2 (range, 1-7), with 46.7% (21/45) having received ≥3 lines. The triple-class exposed rate was 68.9% (31/45), and the autologous stem cell transplantation (ASCT) rate was 37.8% (17/45). All patients received FC lymphodepleting regimen and underwent infusion of 1×10⁶/kg CAR-T cells. The median leukapheresis to infusion interval was 57 days (range, 22-203 days). Individualized bridging therapies were used in 100% of patients. Adverse events were manageable. Cytokine release syndrome (CRS) occurred in 84.4% (38/45) of patients, with grade 3 CRS in 4.4% (2/45) who had received 5 and 7 prior LOT, respectively; immune effector cell-associated neurotoxicity syndrome (ICANS) occurred in 6.7% (3/45), comprising two grade 1 events and one grade 3 event. The patient with grade 3 ICANS had received 5 prior LOT. Hematologic toxicities were common, with grade 3-4 neutropenia, anemia, and thrombocytopenia occurring in 60.0% (27/45), 31.1% (14/45), and 24.4% (11/45) of patients, respectively. Non-hematologic toxicities such as nausea (11.1%), diarrhea (20.0%), and headache (4.4%) were less frequent. Infections occurred in 51.1% (23/45) of patients, with grade 3-4 infections in 26.7% (12/45). Before eque-cel infusion, 16 attained complete response (CR), 4 attained VGPR, 12 patients remained with a high M-protein load (≥10 g/L), 2 patients PD, while all the rest were SD. Median follow up 196 days (range, 18-596 days), the overall response rate (ORR) was 95.6%, with 82.2% achieving ≥CR. Furthermore, 93.3% of patients achieved minimal residual disease (MRD) negativity. The median time to response was 28 days (range, 14-85 days). The median time to best response was 60 days (range, 14-231 days). The median PFS and OS were both not reached. For all patients, 1-2 LOT and ≥3 LOT, the 12-month PFS rate were 74.9%, 91.5% and 63.1%, respectively, and the 12-month OS rate were 91.0%, 100% and 82.5%, respectively. Two patients died from infection at 1 month, the other one died at 3 month with unknown reason. Cell kinetic analysis confirmed the presence of CAR-positive T cells in all patients, with median time to peak was 10 days, and the median peak concentration was 423.32 cells/μL. Univariate analysis indicated that 1q21 amplification was significantly negatively correlated with ORR (100% vs 61.9%, p=0.001). Heavily pretreated patients (≥3 lines) and triple-refractory disease showed a trend toward inferior ORR, while patients with t(11;14) translocation showed a trend toward longer PFS (the 12-month PFS rate 100% vs 68.3%, p=0.056). Conclusion This is the first report on the efficacy and safety of eque-cel in real-world patients with RRMM, which confirmed that eque-cel provided early and deep responses in heavily pretreated RRMM patients, with a manageable safety profile. Preliminary findings from the exploration in early-line RRMM patients also support further investigation in this population.