Journal for Immunotherapy of Cancer最新文献

Background: Mixed response, where different lesions within the same patient show discordant responses to treatment, remains poorly understood. To better understand the complex effects of mixed response on patient survival, we devised three different definitions of mixed response. This retrospective analysis provides the first evaluation of the association between mixed response and survival outcomes in patients with rare cancers treated with dual checkpoint blockade using ipilimumab plus nivolumab, based on data from 52 baskets in the DART SWOG S1609 trial.

Methods: We included 438 patients with Response Evaluation Criteria in Solid Tumors (RECIST) V.1.1-measurable disease and at least two target lesions, after exclusions for ineligibility, early death, or missing data. Overall survival (OS) and progression-free survival (PFS) were compared using log-rank tests and Cox regression, stratified by basket and using a day 65 landmark. A mixed response was evaluated using three definitions: Method 1-RECIST discordance across lesions; Method 2-presence of ≥1 lesion with >5 mm increase or ≥1 with >5 mm decrease; and Method 3-same as Method 2 but with a 1 mm cut-off.

Results: Mixed response was significantly associated with worse OS and PFS using both Method 1 (OS: HR 1.80; PFS: HR 1.58) and Method 2 (OS: HR 1.55; PFS: HR 1.57) compared with "all SD/stable per lesion". Among patients classified as "SD by RECIST", those who exhibited a mixed per lesion response according to Method 1 had significantly worse OS (median 9.9 months (8.8-12.4)) than those with a non-mixed per lesion response (median 22.7 months (19.2-32.0)). Further stratification showed that any lesion increasing >5 mm was linked to worse outcomes (OS: HR >2.37, p<0.05).

Conclusions: Mixed response was significantly associated with worse survival outcomes in patients treated with dual immune checkpoint inhibitors, even among those with RECIST-defined stable disease. Our findings suggest that the "worst"-responding lesion drives prognosis, underscoring the limitations of RECIST in capturing clinically relevant heterogeneity. This study highlights the need to incorporate lesion-level assessment into immunotherapy decision-making and provides a foundation for guiding earlier transition to the next line therapies or other therapeutic options. Future studies integrating molecular biomarkers are warranted to refine response evaluation criteria and optimize immune checkpoint inhibitor-based strategies.

Background: Prostate cancer (PCa) is widely recognized as an immunologically "cold" tumor, characterized by a paucity of effector T cells and a limited response to immune checkpoint blockade therapy. Although the chemokine receptor CXCR6 has been identified as a marker of highly cytotoxic CD8⁺ T cells in other malignancies, its identity, regulatory mechanisms, and clinical significance in PCa remain poorly understood.

Methods: We integrated newly generated and publicly available single-cell RNA sequencing data encompassing 90 651 cells from nine patients. Multicolor immunofluorescence and flow cytometry were performed on human PCa tissue specimens stratified by Gleason Score (GS). In parallel, syngeneic mouse models, bone marrow chimeras, and in vitro T-cell functional assays were employed to investigate the role, recruitment, and transcriptional regulation of CXCR6⁺CD8⁺ T cells. Mechanistic investigations included bulk RNA-seq, chromatin immunoprecipitation-quantitative real-time PCR, pharmacologic modulation of FOXO1, and detailed analysis of the IL-10-STAT3 signaling pathway.

Results: The CXCR6⁺CD8⁺ T cells represent a transcriptionally distinct subset with high expression of cytotoxic markers (GZMA, GZMB, and PRF1), and their frequency declines significantly with increasing GS. Intratumoral maintenance of these cells is sustained by CXCL16 secreted by IL1B⁺ M1-like macrophages; the loss of this macrophage population in advanced tumors leads to depletion of CXCR6⁺CD8⁺ T cells. Genetic ablation of CXCR6 or depletion of CD8⁺ T cells accelerates tumor growth, demonstrating the essential role of CXCR6⁺CD8⁺ T cells in antitumor immunity. At the molecular level, the FOXO1-KLF2 axis transcriptionally represses CXCR6 expression, and IL-10-mediated activation of STAT3 upregulates FOXO1 and KLF2, thereby suppressing CXCR6 expression and impairing cytotoxic function. Pharmacologic inhibition of FOXO1 enhances the expansion of CXCR6⁺CD8⁺ T cells and acts synergistically with anti-PD-1 therapy to inhibit tumor progression.

Conclusions: The CXCR6⁺CD8⁺ T cells are critical yet progressively diminished effectors in PCa. Their persistence within the tumor microenvironment depends on CXCL16 derived from M1-like macrophages and is counteracted by IL-10-driven FOXO1-KLF2 signaling. Targeting this regulatory axis-through inhibition of FOXO1 or IL-10, for example-represents a rational therapeutic strategy to restore CXCR6⁺CD8⁺ T-cell-mediated immunity and enhance the efficacy of immunotherapy in PCa.

Background: Chimeric antigen receptor (CAR)-T cell therapy targeting CD19 has transformed the treatment of hematologic malignancies. The costimulatory domain (CSD) of CAR constructs plays a crucial role in determining T cell metabolism, persistence, and antitumor function. We previously developed a novel CSD combining CD79A and CD40, which conferred superior proliferation and antitumor efficacy compared with CD28-based or 4-1BB-based CAR-T cells. The metabolic mechanisms underlying these effects remain to be elucidated.

Methods: We compared CD28, 4-1BB, and CD79A/CD40 CAR-T cells using transcriptomic analysis, metabolic flux assays, and metabolomics. Patient samples treated with 4-1BB-based or CD28-based CAR-T therapies were analyzed to assess correlations between serum lipids and CAR-T expansion.

Results: Transcriptomic profiling revealed that CD79A/CD40 CAR-T cells shared gene expression patterns with 4-1BB CAR-T cells, particularly in pathways related to oxidative phosphorylation (OXPHOS) and T cell memory differentiation, but were distinct from CD28 CAR-T cells. Both 4-1BB and CD79A/CD40 CAR-T cells relied on OXPHOS and exhibited greater mitochondrial fitness, as evidenced by higher spare respiratory capacity and mitochondrial mass. Notably, CD79A/CD40 CAR-T cells displayed significantly enhanced glycolysis during the early phase following antigen stimulation, distinguishing them from 4-1BB CAR-T cells and supporting rapid initial expansion. Metabolomic profiling showed upregulation of cholesterol biosynthesis enzymes in both CD79A/CD40 and 4-1BB CAR-T cells, suggesting a shared reliance on cholesterol metabolism. Importantly, in patients treated with 4-1BB-based CAR-T therapy, higher serum low-density lipoprotein cholesterol levels positively correlated with CAR-T expansion in the late phase, particularly within CD4+T cell subsets. This relationship was not observed in patients receiving CD28-based CAR-T therapy. These findings indicate that cholesterol availability may influence CAR-T persistence in vivo, and that the metabolic phenotype of CD79A/CD40 CAR-T cells is optimized for both early proliferation and long-term survival.

Conclusions: CD79A/CD40 CAR-T cells exhibit unique metabolic adaptations, including early glycolytic activation, sustained OXPHOS, and upregulated cholesterol metabolism, which together may underpin their enhanced proliferation and persistence. Targeting cholesterol metabolism may represent a novel strategy to optimize CAR-T cell function and improve therapeutic outcomes.

Background: Ferroptosis plays a critical role in immune regulation and tumor microenvironment remodeling. However, its therapeutic potential in enhancing immune checkpoint inhibitor (ICI) efficacy remains incompletely understood and warrants further investigation.

Methods: To investigate the potential of ferroptosis in improving ICI response, we constructed a machine learning-based predictive model using ferroptosis-related genes and analyzed large-scale single-cell RNA sequencing datasets. Mechanistic studies were performed to examine the role of interferon (IFN)-γ signaling in ferroptosis sensitization, including functional validation in vitro and in vivo. Lipidomic, transcriptomic, chromatin Immunoprecipitation sequencing (ChIP-seq) and Cleavage Under Targets and Tagmentation analyses were employed to dissect downstream pathways, focusing on IRF1 and AGPAT3.

Results: Our model successfully predicted ICI response based on ferroptosis-related gene signatures, identifying IFN-γ as a key enhancer of ferroptosis sensitivity in tumor cells. IFN-γ treatment induced activation of the transcription factor IRF1, which in turn upregulated AGPAT3 expression, driving lipid remodeling and accumulation of polyunsaturated ether phospholipids. This lipid remodeling significantly increased tumor cell susceptibility to ferroptosis and enhanced ICI efficacy. Loss of AGPAT3 impaired IFN-γ-mediated tumor elimination both in vitro and in vivo. Clinically, higher AGPAT3 expression in tumors was associated with increased immune activation and improved overall survival in ICI-treated patients.

Conclusion: The IFN-γ-IRF1-AGPAT3 axis represents an important antitumor mechanism that promotes ferroptosis. Targeting this pathway in combination with our ferroptosis-driver model prediction may improve ICI efficacy and patient outcomes.

This research analyzes the global landscape of clinical trials focusing on therapies targeting the Nectin cell adhesion molecule 4 (Nectin-4) across various malignancies, using data from the Trialtrove database. Analysis of 136 interventional trials reveals a rapidly expanding field dominated by antibody-drug conjugates, particularly in urothelial carcinoma, with significant activity in combination regimens and diverse therapeutic modalities under exploration. These findings emphasize the substantial translational potential of Nectin-4 and highlight key trends shaping its future clinical development.

Gastrointestinal malignancies (GIM) impose a substantial global health burden, accounting for approximately 33% of cancer-related mortality worldwide. Although chimeric antigen receptor (CAR)-based cell therapy has achieved remarkable success in hematological malignancies, its application in solid tumors, particularly GIM, remains in its nascent stages. This comprehensive landscape analysis systematically examined the clinical trial ecosystem of CAR-based cell therapy for GIM by retrieving 179 eligible trials from the Trialtrove database as of October 2025 (hepatobiliary and pancreatic malignancies were excluded). The analysis revealed a predominantly early-phase landscape, with Phase I studies constituting 70.39% of all trials. Geographically, China (70.77%) and the USA (16.41%) dominated trial initiation, while academic institutions sponsored 54.92% of investigations. Claudin 18.2, NKG2D(L), mesothelin, and CEA emerged as the most frequently targeted antigens. Autologous therapy administered via peripheral intravenous infusion represented the predominant therapeutic modality. Combination strategies incorporating chemotherapy and immunotherapy demonstrated promising synergistic potential, suggesting that multimodal approaches may enhance therapeutic efficacy while potentially mitigating resistance mechanisms. The convergence of innovative preclinical developments with increasingly supportive regulatory frameworks portends transformative advances in the field, positioning CAR-based cell therapy as an emerging cornerstone modality in the therapeutic armamentarium against GIM.

Background: The orally available toll-like receptor 7 (TLR7) agonist prodrug RO7119929 is converted to active drug predominantly in the liver, where it is hypothesized to reprogram the local immune microenvironment. We aimed to explore the safety, pharmacokinetics (PK), and preliminary antitumor activity and obtain the proof-of-mechanism for RO7119929 in patients with liver cancer.

Methods: RO7119929 was investigated in mouse tumors and liver tissue from cynomolgus monkeys. In the first-in-human, open-label, dose-escalation and expansion study, patients with histologically confirmed advanced or metastatic primary liver cancers or solid tumors with predominant liver involvement received RO7119929 weekly in 3-week cycles either on a flat-dose (FD) or step-up dose (SUD) schedule.

Results: Preclinical results demonstrated antitumor activity and the proinflammatory proof-of-mechanism in mouse liver tumors and cynomolgus monkey liver tissue. The subsequent first-in-human study enrolled 27 patients in five FD dose-escalation cohorts, 18 patients in a FD dose-expansion cohort, and 9 patients in two SUD dose-escalation cohorts. The most common primary tumor type at study entry was hepatocellular carcinoma (HCC) (31%). PK data showed fast conversion of RO7119929 to the active TLR7 agonist. Treatment-related adverse events (TRAEs) were observed in 50 (91%) patients across all treated patients; the most commonly reported TRAE was cytokine release syndrome (CRS) in 48 (81%) patients. CRS was also identified as a dose-limiting safety risk and had a dose-dependent incidence and severity. Grade 3 CRS occurred in six (13%) patients receiving FD RO7119929, while no grade 3 CRS was reported in SUD patients at comparable target dose levels. TLR7-related transient, dose-dependent gene expression and cytokine induction was observed both with FD and SUD treatment and corresponded with treatment-induced local inflammation of the tumor microenvironment induced by reprogramming of the myeloid cells. Among 17 patients with HCC, one durable complete response was observed, and 10 (59%) patients had stable disease.

Conclusions: SUD appears to reduce the risk of CRS while maintaining mode-of-action-relevant pharmacodynamic effects. The clinical activity of RO7119929 as a single agent was limited, suggesting that combination therapy with a checkpoint inhibitor may be needed to leverage the proinflammatory potential of RO7119929 and further increase antitumor activity.

Trial registration number: NCT04338685.

Background: Programmed death-ligand 1 (PD-L1)-targeted radiopharmaceutical therapy (RPT) combined with immune checkpoint blockade (ICB) represents a promising combinatorial treatment because of the "magic" bystander and abscopal effects of RPT. However, the overall efficacy of this combinatorial approach is often constrained by limited tumor retention and insufficient accumulation of existing radiopharmaceuticals. D-peptides, with their superior metabolic stability, offer a promising modality to overcome these limitations. We previously reported [⁶⁸Ga]/[⁶⁴Cu]DPA, which exhibited favorable in vivo stability. Nevertheless, its inadequate tumor retention hampers therapeutic efficacy and immune activation, highlighting the need for further optimization to achieve sustained target engagement.

Methods: Herein, fluorine modification-directed rational design was used to optimize DPA's target engagement and retention, thereby boosting its immune-activating potential in RPT. Following two rounds of screening, a series of fluorinated D-peptides was synthesized. Their in vivo performance was assessed by positron emission tomography (PET)/CT imaging in tumor-bearing mice and subsequently in patients with cancer. The therapeutic efficacy of the corresponding ¹⁷⁷Lu-labeled fluorinated radiopharmaceuticals was further investigated to verify their potential in enhancing antitumor immunity.

Results: [⁶⁸Ga]D3-6, which incorporates a pentafluorinated phenylalanine residue, exhibited optimal tumor uptake and prolonged retention, achieving 22.43±1.63%ID/g at 4 hours post-injection. A preliminary clinical study further validated its diagnostic efficacy across multiple cancers. Additionally, [¹⁷⁷Lu]D3-6 boosted both local and systemic antitumor immunity and, in combination with anti-programmed cell death protein-1 therapy, elicited superior tumor inhibition.

Conclusion: This study highlights the potential of fluorinated D-peptide radiopharmaceuticals as powerful PD-L1 targeting radiotheranostics, enabling more effective systemic antitumor efficacy in RPT and ICB combination therapy.

Background: Peritoneal metastasis (PM) is one of the most challenging clinical problems in gastric cancer (GC), largely due to its high recurrence rate and poor response to current therapies. Increasing evidence indicates that remodeling of the extracellular matrix (ECM) plays an important role in therapeutic failure. However, how specific stromal-immune interactions contribute to PM heterogeneity and immunotherapy resistance remains unclear. In this study, we investigated how ECM composition-particularly the accumulation of hyaluronic acid (HA)-influences the immune microenvironment and therapeutic responses in GC-associated PM.

Methods: We combined histopathological assessment, analyses of patient-derived specimens, single-cell transcriptomic profiling, and murine models of PM to delineate ECM remodeling patterns and immune cell dynamics in therapy-sensitive and therapy-resistant lesions. In addition, functional assays and pharmacological approaches were used to examine HA-CD44 signaling and its impact on CD4⁺ T cell differentiation and responsiveness to immune checkpoint blockade.

Results: Therapy-sensitive PM lesions were characterized by enrichment of elastic fibers, whereas therapy-resistant lesions showed collagen accumulation. Notably, HA deposition emerged as a key feature distinguishing these ECM states and was closely associated with differential therapeutic outcomes. Elevated HA levels activated CD44-dependent signaling in CD4⁺ T cells, driving regulatory T cell (Treg) differentiation through a CD44-IQGAP1-RAC1-SMAD3 signaling pathway and thereby establishing an immunosuppressive microenvironment. Importantly, pharmacological inhibition of CD44 reduced Treg expansion and markedly enhanced the antitumor efficacy of anti-PD-1 therapy in murine PM models.

Conclusions: Our findings identify HA-CD44 signaling as a critical link between ECM remodeling and immune evasion in GC PM. Targeting ECM-driven immunosuppressive mechanisms may represent a promising strategy to overcome therapeutic resistance and improve the efficacy of immunotherapy in this aggressive disease.