7079 Background: Despite high initial cure rates, patients with B-cell hematologic malignancies, including B-ALL and B-NHL, experience substantial relapse, with markedly reduced disease-free and overall survival after second-line therapy. GT801 is a first-in-human, in vivo CAR-T therapy that uses antibody-displayed lipid nanoparticle (LNP) to deliver CD19 CAR mRNA directly to T cells, enabling in vivo reprogramming without ex vivo cell manufacturing or lymphodepletion. We report initial safety and efficacy data from an open-label, single-arm phase I study (NCT07205315). Methods: GT801 was evaluated using a modified 3+3 dose-escalation design across three dose levels (0.5, 1.5, and 3.0 mg). Patients received up to 4 treatment cycles, each consisting of a single intravenous administration of GT801 on Day 1 of a 7-day cycle. The primary endpoint was safety, including treatment-emergent adverse events (TEAEs), graded per CTCAE v5.0. Secondary endpoints included 3-month overall response rate (ORR), best overall response (BOR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS), assessed per Lugano 2014 criteria. Results: As of January 13, 2026, 3 patients were enrolled (median age 60 years; median three prior therapies). One patient received 3 doses at 0.5 mg, and two patients received 4 doses at 1.5 mg. All patients had B-NHL. Premedication included dexamethasone, NSAIDs, antihistamines. Most AEs were Grade 1-2. Common TEAEs included cytokine release syndrome (CRS), cytopenia, and transient liver enzyme elevations. Grade ≥3 events were hematologic toxicities and CRS. No neurotoxicity or organ failure was observed. At Week 4, all patients demonstrated a treatment response. Peripheral blood flow cytometry showed rapid CAR-T generation, with CD8 + CAR-T cells detectable within 4 hours post-infusion and peak levels on Day 1. Reprogramming efficiency reached up to 93% and was maintained with repeat dosing. Minimal off-target CAR expression was detected in monocytes. Complete peripheral B-cell depletion occurred within 4 hours after the first dose and was sustained for at least 7 days; after the third dose, B-cell depletion and CAR-T detection were observed in both peripheral blood and bone marrow. Conclusions: GT801 demonstrated a manageable safety profile and early signs of antitumor activity in heavily pretreated B-NHL patients. The ability to generate functional CAR-T cells in vivo without lymphodepletion supports further clinical evaluation of GT801 as a promising therapeutic approach. Clinical trial information: NCT07205315 . Research sponsor: Vivacta Biotechnology (Shanghai) Co., Ltd.

Despite the critical role of succinylation as a posttranslational modification in regulating cellular homeostasis and metabolic pathways, achieving precise control over mitochondrial function and homeostasis to induce apoptosis in breast cancer remains a significant challenge. In this study, we designed a succinylated D-type neuropeptide integrated with an iron-based probe aimed at regulating mitochondrial function and homeostasis through SIRT5-mediated desuccinylation. The probe is selectively delivered to breast cancer cells via Y1 receptor-mediated endocytosis, where it undergoes SIRT5-catalyzed desuccinylation within the mitochondria. This enzymatic modification induces substantial changes in the secondary structure of the D-type neuropeptide, transforming the probe into aggregates that disrupt the mitochondrial membrane potential and electron transport. By interacting competitively with cellular desuccinylation pathways, this probe further disturbs cellular homeostasis, ultimately leading to apoptosis in tumor cells. Additionally, we observe the upregulation of key genes associated with inflammation and ferroptosis, which may contribute to the establishment of an inflammatory microenvironment, thereby enhancing the antitumor effect. Notably, the aggregation process shifts the magnetic resonance imaging (MRI) contrast from T1 to T2, enabling the real-time visualization of mitochondrial perturbations. This work establishes a versatile platform for designing neuropeptide-based nanoprobes with diagnostic and therapeutic potential for the treatment of breast cancer.

7078 Background: GT719 is an off-the-shelf CAR-NKT therapy derived from umbilical cord blood hematopoietic stem cells and genetically engineered to co-express an anti-CD19 CAR, an invariant natural killer T (NKT)-specific TCR, and secreted interleukin-15 (IL-15). This design enables CD19-targeted tumor killing, avoids risk of graft-versus-host disease (GVHD), and enhanced cellular expansion, persistence, and immune function through IL-15 signaling. We report clinical data from 7 adults with relapsed or refractory (R/R) CD19-positive B-cell hematologic malignancies enrolled in two open-label, single-arm studies (NCT06948981, NCT07131254) evaluating the safety and preliminary efficacy. Methods: The primary endpoint was safety, including treatment-emergent adverse events (TEAEs), graded per CTCAE v5.0. Secondary endpoints included 3-month overall response rate (ORR), best overall response (BOR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS). Efficacy was assessed using Lugano 2014 criteria and the 2024 Chinese guidelines for adult acute lymphoblastic leukemia. Results: As of December 31, 2025, 7 patients were treated (median age 51 years; median two prior therapies), including 3 with B-ALL and 4 with B-NHL. Following FC lymphodepletion, all patients received a single GT719 infusion (5×10⁷ to 1×10⁹ viable cells). Most adverse events (AEs) were Grade 1-2. Grade ≥3 AEs were limited to lymphodepletion-related cytopenia and resolved or improved to Grade ≤2 within 14 days, excluding disease-related cytopenia. One patient experienced Grade 1 cytokine release syndrome that resolved within 1 day. No ICANS, neurotoxicity, or GVHD, severe infections, or serious TEAEs were observed. Efficacy was evaluable in all patients across B-ALL, follicular lymphoma (FL, grade 3a), and diffuse large B-cell lymphoma (DLBCL). The 3-month ORR was 50% (3/6), with all responders achieving CR; one patient was excluded due to insufficient follow-up. The disease control rate (DCR) was 71.4% (5/7). Two of 3 FL patients achieved CR, while one had a PR and remained under follow-up. One of three B-ALL patients achieved stringent complete remission, maintained through 24 weeks of relapse-free follow-up. CAR transgene analysis demonstrated robust in vivo expansion, with peak levels occurring between Days 7-10 in B-ALL and within two months in FL. GT719 persistence was observed for up to six months post-infusion. Conclusions: GT719 demonstrated a favorable safety profile and encouraging antitumor activity in adults with R/R CD19-positive B-cell malignancies. The absence of severe GT719-related toxicities and evidence of durable cellular persistence support further clinical investigation of this off-the-shelf CAR-NKT therapy. Clinical trial information: NCT06948981 , NCT07131254 . Research Sponsor: Grit Biotechnology.

6049 Background: Immune checkpoint inhibitors (ICIs) are the standard treatment for recurrent and/or metastatic (R/M) squamous cell carcinoma of the head and neck (HNC) after failure of platinum-based chemotherapy; however, the clinical benefit remains limited, with reported overall response rate (ORR) of approximately 13% and median progression-free survival (PFS) of about 2 months in the second line setting. GT201 is an autologous TIL therapy engineered to express membrane-bound IL-15 (mbIL-15), which may enhance immune activation in the tumor microenvironment and promote durable response. We report preliminary safety and efficacy results from an open-label, single-arm study evaluating GT201 in combination with the PD-1 inhibitor toripalimab in patients with R/M HNC (NCT06190275). Methods: The primary endpoint was safety, including treatment-emergent adverse events (TEAEs) graded per CTCAE v5.0. Secondary endpoints included ORR, disease control rate (DCR), PFS, duration of response (DOR), and overall survival (OS), assessed per RECIST v1.1. Results: As of November 30, 2025, 6 patients with R/M HNC were treated (median age of 57 years; median 1 prior line of therapy). Histology indicated 5 squamous cell carcinoma and 1 lymphoepithelial carcinoma. All patients received 1–2 cycles of bridge therapy, followed by lymphodepletion (low-dose in 5 patients; intermediated-dose in 1 patient), GT201 infusion (5×10 9 -5×10 10 viable cells), and high-dose IL-2 (600,000 IU/Kg; 4-6 doses). Five patients subsequently received toripalimab; one patient progressed prior to PD-1 inhibitor treatment. Maximum follow-up was 15.5 months. Most of AEs were Grade 1-2. Grade ≥ 3 AEs were primarily related to lymphodepletion and IL-2, and included cytopenia, neutropenia, lymphocytopenia, monocytopenia, hypokalemia, rash, and increased bilirubin; all resolved or improved to Grade ≤ 2 within 14 days. The ORR was 66.7% (4/6), including 2 complete response (CR) and 2 partial response(PR); DCR was 83.3% (5/6). One patient with CR remains progress-free exceeding 12 months. Median PFS and OS have not yet been reached. GT201 cells expanded robustly and persisted in peripheral blood for at least 6 months post-infusion. Conclusions: GT201 combined with toripalimab demonstrated a manageable safety profile and encouraging antitumor activity in heavily pretreated R/M HNC, supporting further clinical development of this combination. Clinical trial information: NCT06190275 . Research Sponsor: Grit Biotechnology.

5532 Background: Adoptive cell therapy with autologous TILs has shown durable clinical benefit in selected advanced solid tumors after progression on standard therapies. We report results from a phase I, open-label, single-arm, multicenter trial (NCT05430373) evaluating GT101 in patients with recurrent or metastatic cervical cancer. Primary objectives were safety; second objectives included preliminary efficacy and PK. Methods: Eleven patients with recurrent or metastatic cervical cancer received a lymphodepletion regimen followed by GT101 infusion and high-dose IL-2. The primary endpoint was safety, including treatment-emergent adverse events (TEAEs), serious adverse events (SAEs) and adverse events (AEs), graded per CTCAE v5.0. Secondary endpoints included objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS), duration of response (DoR), and overall survival (OS), assessed by RECIST v1.1. Flow cytometry of immune cells and T cell receptor (TCR) sequencing were performed on peripheral blood samples collected from patients before and after treatment. Results: As of September, 2025, 11 patients were treated (median age was 48 years; median two prior lines of therapy). Following FC lymphodepletion, patients received GT101 at doses ≥5×10⁹ viable cells (median 4.1×10¹⁰), followed by high-dose IL-2 (600,000 IU/kg, up to 6 doses). Most AEs were Grade 1-2. Grade ≥3 AEs were primarily related to lymphodepletion and IL-2, and included lymphopenia, leukopenia, neutropenia, anemia, pyrexia, and thrombocytopenia; most resolved or improved to Grade ≤2 within 14 days. The ORR was 45.5% (5/11), with DCR of 90.9% (10/11). Median DoR was 6.4 month. Four patients (36.4%) achieved confirmed PR, one patient (9.1%) achieved a CR, and five patients (45.5%) had stable disease (SD). Median PFS was 4.83 months; OS follow-up is ongoing. One patient with stage IIIC2 cervical squamous cell carcinoma achieved a confirmed CR lasting 14.5 months, with tumor burden reduced from a baseline sum of diameters of 71.57 mm to complete resolution by Week 12. Pharmacokinetics and Correlation Analyses: Peripheral blood analyses demonstrated peak circulating T cells 5-7 days post-infusion, followed by contraction to near-baseline by two weeks. Responders exhibited lower proportions of CD39 + PD-1 + T cells. TIL infusion increased T-cell clonal diversity in all patients, with greater expansion observed in non-responders who has lower baseline diversity; persistence of TIL-derived clones did not differ consistently between groups. Conclusions: GT101 demonstrated a manageable safety profile with no treatment-related SAEs or dose-limiting toxicities and showed clinically meaningful and durable antitumor activity in recurrent or metastatic cervical cancer. These findings support further clinical development, and a pivotal phase II study is ongoing. Clinical trial information: NCT05430373 .

Multiple sclerosis (MS) is a chronic autoimmune disorder characterized by the immune-mediated demyelination and neurodegeneration of the central nervous system. Phagocyte mediated myelin debris clearance is required for remyelination. TIM-3 is highly expressed on mononuclear macrophages and promotes the phagocytosis of apoptotic cells. Here, we report that TIM-3 enhances the clearance of myelin debris in experimental autoimmune encephalomyelitis (EAE), a model of MS. Tim-3 knockout (KO) exacerbated EAE severity, neuroinflammation, and demyelination by regulating mononuclear macrophages. TIM-3 promoted the phagocytosis and degradation of myelin debris by macrophages. Mechanistically, Tim-3 deficiency impaired lysosomal biogenesis and function, leading to lysosomal membrane permeabilization and disrupted lysosomal acidification, which further exacerbated neuroinflammation and demyelination. Notably, TIM-3 blocked the interaction of mTOR-TFEB to inhibit TFEB phosphorylation and facilitate its nuclear translocation, followed by increased expression of lysosomal genes critical for myelin degradation. Importantly, the IgV domain is necessary in TIM-3-mediated lysosomal regulation and myelin degradation. These findings highlight TIM-3 as a key regulator of lysosomal homeostasis and the clearance of myelin debris, suggesting that the IgV domain has promise as a therapeutic agent for treating demyelinating diseases such as MS.

Abstract Background: Tumor-infiltrating lymphocyte (TIL) therapy has demonstrated activity in solid tumors but remains constrained by T-cell exhaustion and an immunosuppressive tumor microenvironment. GT201 is an engineered, cytokine-armored TIL product expressing membrane-bound IL-15 (mbIL-15) designed to enhance TIL persistence and antitumor function. A multi-center, open-label, single-arm exploratory clinical study was initiated to evaluate the safety, tolerability, and preliminary efficacy of GT201 in patients with advanced solid tumor. Method: The primary endpoint was to evaluate the tolerance and safety profile according to CTCAE v5.0. Secondary endpoints included preliminary efficacy measures such as overall response rate (ORR), disease control rate (DCR), assessed per RECIST v1.1, as well as pharmacokinetics of GT201. Exploratory endpoint included longitudinal tumor biopsies and peripheral blood sampling, analyzed by single-cell RNA/TCR sequencing and bulk TCR sequencing to track clonal dynamics and identify tumor-specific TCRs. Result: As of November 4, 2025, twelve patients were enrolled (median age 52.5 years; median of two prior lines of therapy). GT201 demonstrated a favorable safety profile: most AEs were Grade 1-2, while Grade ≥3 events were attributable to lymphodepleting chemotherapy or IL-2 support and resolved within 14 days. Preliminary efficacy signals were encouraging, with an ORR of 58.3% (7/12) and a DCR of 83.3% (10/12), including two complete responses (16.7%) and five partial responses (41.7%). Correlative multi-omics profiling revealed that mbIL-15-expressing cells were enriched in post-infusion tumor samples, and their corresponding TCRs underwent marked clonal expansion. A putative tumor-specific T-cell subset (4-1BB+CD8+ with high effector/exhaustion and low stemness signatures in tumor samples pre- and post-infusion) was preferentially expanded during manufacturing and showed a pronounced expansion peak in peripheral blood between Days 7-28 in responders. This subset was scarce at baseline and exhibited minimal expansion in non-responders. In contrast, “TIL-only” TCRs, which were clonotypes present in the infused product but absent pre-infusion, did not expand in peripheral blood in any patient. Conclusion: GT201 exhibited a manageable safety profile and encouraging early efficacy in patients with advanced solid tumor, with ongoing enrollment to further inform clinical outcomes. Integrated multi-omics analyses indicate that expansion of tumor-specific TCRs, during both manufacturing and post-infusion in responding patients, correlates more strongly with clinical benefit than expansion of TIL-only TCRs. Continued longitudinal sampling and functional validation of putative tumor-specific clonotypes will be essential to establish their utility as biomarkers of response in GT201 therapy. Citation Format: Pin Wang, Rong Zhou, Yong Han, Zhengxiang Han, Weijia Fang, Kai Chen, Youguo Chen, Liqing Ma, Lili Lu, Derun Shen, Jiahui Jin, Yiyang Tan, Ke Liu, Zhenjiang Liu, Jingman Wang, Zhao Xu, Jingwei Sun, Jun Cui, Jing Yu, Yue He, Yarong Liu. Armored TIL GT201 induces potent tumor-specific TCR expansion and durable antitumor responses in advanced solid tumor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5257.

Abstract Background: CAR-T therapies have revolutionized hematologic malignancies treatment but remain constrained by complex autologous manufacturing, high cost, and lymphodepletion toxicity. In vivo CAR-T approaches aim to overcome these barriers by engineering T cells directly in patients, which requires targeted delivery, durable CAR expression, potent cytotoxicity, and re-dosing potential. Toward this goal, we present the development, preclinical data, and preliminary clinical results for GT801, a novel anti-CD19 in vivo CAR-T candidate. Methods: GT801 uses T cell-targeted lipid nanoparticles (T-LNPs) encapsulating chemically modified linear mRNA encoding an anti-CD19 CAR. T-LNPs were surface-engineered with a VHH antibody using the CLAMP platform, enabling site-specific antibody attachment, controlled ligand density, and efficient, selective T-cell uptake. Formulation and mRNA design were optimized for targeting specificity, robust CAR expression, and functionality. B-cell depletion, PK, and preliminary toxicology of GT801 were assessed in vitro and in humanized NOG mouse models. Results: With optimized mRNA chemistry, the T-LNP platform achieves robust and durable CAR expression in human PBMCs for >14 days in vitro. Targeted delivery at 0.1 mpk reached receptor-saturating levels across multiple lymphoid tissues, while off-target uptake remained <1%. In human PBMC-engrafted NOG mice, a single i.v. dose as low as 0.01 mpk achieved >95% B-cell depletion, and 0.1 mpk achieved near-complete clearance (<0.1%) across multiple lymphoid tissues. CDX models demonstrated potent antitumor activity and enhanced CAR-T expansion upon repeat dosing, suggesting good in vivo fitness post-transfection and expansion driven by B cell depletion. Primary B-cell killing was robust in PBMCs from healthy donors and autoimmune patients at only 0.1 μg within 24 hours. Serial-dose toxicology elicited minimal cytokine release (IL-6, TNF-α) and no organ toxicity in any tested organs, supporting the platform's good safety profile. Preliminary clinical data with B-cell hematological malignancies and autoimmune diseases demonstrated high in vivo CAR expression and confirmed repeat-dosing feasibility. Conclusion: These findings demonstrate that our T-LNP platform enables efficient, targeted, and sustained in vivo CAR expression with a favorable safety profile and scalable manufacturing. The platform's robust preclinical B cell depletion efficacy, validated by preliminary clinical data confirming high in vivo CAR expression and repeat dosing feasibility, successfully validates the system for clinical use. Ongoing patient accrual will further inform clinical outcomes. Citation Format: Jingwei Sun, Xi Zhu, Jiahui Jin, Yiyang Tan, Liang Lin, Zhao Xu, Jingman Wang, Dalang Li, Hong Chen, Jiaming Ren, Jun Cui, Jing Yu, Pin Wang, Yarong Liu. In vivo CAR-T T-LNP system (GT801) drives potent B cell depletion and clinical feasibility in hematologic and autoimmune conditions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 148.

OBJECTIVE:To investigate the rheumatic antibodies on characteristics of neuromyelitis optic spectrum disorders. METHODS:All the patients were tested for AQP4-IgG. There were three groups, AQP4+NMOSD, AQP4-NMOSD and control group. MRI was performed. Clinical characteristics, including sex, age, disease duration, the number and type of relapse, autoantibodies, and MRI, OCT results and the scores obtained from EDSS at the time of attack and remission were prospectively recorded. Patients and controls underwent peripapillary RNFL and macular volume OCT scanning. RESULTS:In AQP4+ NMOSD patients with optic neuritis (ON), significant associations were identified between various antibodies and OCT parameters. Specifically, INF110 was significantly correlated with dsDNA (r = 0.384, p = 0.023), FT was associated with AHA (r = -0.416, p = 0.015), and optic disc area showed a strong negative correlation with AMA M2 (r = -0.562, p < 0.001). For AQP4-NMOSD individuals, TMP 30 showed a significant relationship with PM-Scl (r = -0.410, p = 0.027), nRNP (r = 0.622, p < 0.001), AHA (r = 0.701, p < 0.001), p-ANCA (r = -0.435, p = 0.018), c-ANCA (r = -0.428, p = 0.021), and CCP (r = -0.428, p = 0.021). SUP 28 was correlated with PCNA (r = 0.426, p = 0.021), INF 110 with nRNP (r = 0.630, p < 0.001) and AHA (r = 0.706, p < 0.001), TMV with nRNP (r = -0.650, p < 0.001) and AHA (r = -0.708, p < 0.001), and FT with nRNP (r = -0.378, p = 0.043) and AHA (r = -0.464, p = 0.011). CONCLUSIONS:Rheumatic antibodies may be as biomarker to induce the severe worsening of development of NMOSD patients.

Neurodegenerative dementia (ND) is characterized by progressive cognitive decline. The role of autoantibodies in ND remains controversial, as they may contribute to neurological damage or, under certain pathophysiological conditions, exert protective effects. In this study, we aimed to analyze the clinical characteristics of patients with autoantibody-positive ND and investigate potential molecular mechanisms using proteomics. The study included 13 patients with autoantibody-positive ND (ND +), 13 with autoantibody-negative ND (ND –), and 13 cognitively normal controls. Differentially expressed proteins (DEPs) were identified through proteomic analysis. Bioinformatics approaches were employed to explore potential pathways and mechanisms. Multiple linear regression analysis was performed to examine the effects of DEPs on cognitive function and their interaction with autoantibody status. The ND + and ND – groups displayed similar overall cognitive status, levels of depression or anxiety symptoms, and mental and behavioral abnormalities. The ND + group demonstrated better daily living abilities and superior frontal lobe cognitive function. Proteomic analysis identified 162 DEPs associated with autoantibodies enriched in pathways related to endocytosis, endoplasmic reticulum protein processing, tight junctions, and cellular adhesion. In the ND + group, β-arrestin-1 (ARRB1, β = 3.31, 95

Neuropeptides and their subtype receptors play crucial roles in a wide range of neurophysiological and pathological processes, particularly in the cross-talk between cancer and nervous system. However, the mechanisms by which chiral amino acid mutations regulate neuropeptide receptor subtype selectivity, binding specificity, and enzymatic resistance remain largely unexplored. In this study, a series of peptides targeting neuropeptide Y subtype receptors (Y1R and Y2R) through in silico amino acid mutagenesis and computational screening were designed, which led to the identification of candidate sequences with predicted receptor selectivity. Molecular dynamics simulations further validated the stability and conformational preferences of these ligand-receptor complexes. The stability and conformational preferences of these complexes were confirmed by molecular dynamics simulations, while the safety, specificity and enzymatic stability of the ligands were validated experimentally. These findings clarified the structural basis of receptor selectivity and guided the design of NPY receptor ligands with enhanced efficacy.

Introducing multiple iodine atoms into a heptamethine cyanine scaffold harnesses the heavy-atom effect to promote non-radiative decay, thereby achieving a remarkably high photothermal conversion efficiency (eta = 61.0%) while retaining intense fluorescence in the near-infrared II (NIR-II) window. This structural optimization enables efficient light-to-heat conversion and enhanced photoacoustic contrast, making the system suitable for dual-modality imaging. Upon encapsulation into biocompatible nanomicelles, the resulting probes form stable, tumor-targeting colloids that exhibit concentration-dependent photothermal heating and superior photoacoustic responses compared with conventional cyanine analogues. In vivo studies confirm efficient tumor accumulation, low systemic toxicity, and optimal photothermal ablation. Collectively, these findings highlight heavy-atom-engineered heptamethine cyanines as a quantitatively optimized and mechanistically innovative platform for NIR-II fluorescence/photoacoustic dual imaging and image-guided photothermal cancer therapy.

CRISPR-Cas systems have transformed genome editing, yet the commonly used Streptococcus pyogenes Cas9 (SpCas9) is limited by off-target effects and chromosomal instability. Here, we characterize AaCas12bMAX, an engineered Alicyclobacillus acidiphilus Cas12b variant, as a high-precision editing platform optimized for tumor-infiltrating lymphocyte (TIL) therapy. Using an FDA-compliant safety assessment framework, we systemically compared AaCas12bMAX- and SpCas9-edited TIL products in terms of on-target efficiency, genome-wide off-target activity, and structural variant (SV) formation. AaCas12bMAX achieved >80% on-target editing efficiency with undetectable off-target events and a 3.3-fold reduction in SVs relative to SpCas9. Mechanistic studies revealed different DNA repair kinetics in AaCas12bMAX-edited cells, reducing sustained DNA damage responses and chromosomal instability. Structural modeling suggested a more stable enzyme-sgRNA-DNA ternary complex, enabling stringent PAM specificity and minimal mismatch tolerance. Functionally, AaCas12bMAX-edited TILs exhibited superior therapeutic potential, including enhanced cellular fitness, a 2-fold increase in expansion capacity, and enrichment of stem-like tumor-reactive CD39-CD69-CD8+ subsets. Together, these results establish AaCas12bMAX as a robust, clinically translatable platform that improves the safety and functional limitations of SpCas9, enabling the development of next-generation T cell therapies.

BACKGROUND:Ovarian cancer (OC) poses a significant challenge for conventional chimeric antigen receptor-engineered T (CAR-T) cell therapy, due to frequent recurrence linked to tumor heterogeneity, platinum resistance, immune evasion, and an immunosuppressive tumor microenvironment (TME). METHODS:Here, we analyze primary OC patient samples and identify a unique opportunity for allogeneic CAR-NKT (AlloCAR-NKT) cells to concurrently attack OC tumor cells and their TME. Leveraging stem cell gene engineering and a clinically guided culture method, we achieve robust generation of AlloCAR-NKT cells at high yield and purity. FINDINGS:Compared to conventional CAR-T cells, AlloCAR-NKT cells demonstrate superior anti-OC efficacy, showcasing multiple OC-targeting mechanisms, focused tumor homing, and pronounced TME modulation. AlloCAR-NKT cells also exhibit a high safety profile with reduced cytokine release syndrome. Additionally, these cells do not induce graft-versus-host disease and resist host immune-cell-mediated allorejection. CONCLUSIONS:These findings underscore the unique efficacy and safety advantages, as well as the off-the-shelf potential of AlloCAR-NKT cell therapy for OC. FUNDING:Major funding was provided by the California Institute for Regenerative Medicine (CIRM).

BackgroundAnti-N-methyl-d-aspartate receptor (anti-NMDAR) encephalitis is a nervous autoimmune disorder discovered in the recent more than 15 years. To understand potential involvement of epigenetic mechanisms in pathogenesis of anti-NMDAR encephalitis, we initiated a study to compare the expression profiles of long non-coding RNAs (lncRNAs) and messenger RNA (mRNAs) in patients of anti-NMDAR encephalitis and healthy controls.MethodsEleven patients who were diagnosed with anti-NMDAR encephalitis were enrolled in our observational studies. Total RNA was extracted from patients’ plasma and the expression levels of lncRNAs and mRNAs were determined. Differential expression analysis via RNA sequencing, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, as well as a co-expression network of lncRNA-mRNA were performed in 5 patients and 5 healthy controls to evaluate potential the changes in expression patterns. The expression levels of certain lncRNAs and mRNAs were further validated in 11patients and 11 healthy controls using quantitative real-time polymerase chain reaction (qRT-PCR).ResultsIt was found that a total of 83 lncRNAs and 2,345 mRNAs were differentially expressed in five patients with anti-NMDAR encephalitis compared with five healthy controls. Of those lncRNAs, 63 were upregulated and 20 downregulated, while 1,509 mRNAs were upregulated and 836 downregulated. GO and KEGG pathway analyses showed that a wide range of biological functions were perturbed during acute anti-NMDAR encephalitis. Differentially expressed genes were found to be involved in autoimmune, B cell signaling, neuroinflammation, or synaptic plasticity. qRT-PCR was conducted in 11 patients and 11healthy controls to further confirm the levels of six lncRNAs and four mRNAs, and the results were found to be consistent with those by RNA sequencing. The co-expression networks of lncRNA-mRNA were performed in some meaningful KEGG analyses, including chemokine signaling pathway, Long-term potentiation, B cell receptor signaling pathway and MAPK signaling pathway.ConclusionTaken together, these findings suggest the involvement of a number of molecular pathways in anti-NMDAR encephalitis, some of which could serve as potential biomarkers to assist in diagnosis, treatment and prognosis.

We summarized the clinical manifestations, auxiliary examinations, treatment, and prognostic characteristics of a patient with neurexin-3a IgG-mediated autoimmune encephalitis. On March 2, 2024, a 43-year-old male patient was admitted to the Second Hospital of Shandong University and had prodromal symptoms of infection before the onset of encephalitis. The main manifestations were episodic loss of consciousness, eyes turned upward to the right, clenched teeth, bleeding from tongue bite, and limb twitching. Imaging results showed that the left frontal lobe was characterized by a patchy, slightly longer T1 and T2 signal foci, with a slightly higher signal in the pressurized water image. The CSF virus test was normal; both the serum and CSF were positive for neurexin-3a antibodies using CBA, which were confirmed by TBA. The patient’s symptoms improved after glucocorticosteroid therapy. Neurexin-3a IgG-mediated autoimmune encephalitis is a new type of autoimmune encephalitis, and suspicion of associated disease requires further testing for neurexin-3a IgG for a definitive diagnosis.