Journal for Immunotherapy of Cancer最新文献

The discovery of cancer's hidden antigen landscape-comprising non-canonical 'dark matter' antigens-has unveiled a vast, untapped reservoir of immune targets for next-generation cancer immunotherapy. While most cancer vaccine strategies of the past decade have focused on mutation-derived neoantigens, studies applying sensitive mass spectrometry methods fail to identify the majority of predicted neoepitopes being presented by tumor human leukocyte antigen (HLA) molecules, potentially explaining negative results of several recent neoantigen vaccine trials. By contrast, peptides from non-canonical open reading frames, aberrant splice products, and non-coding RNAs that derive from short-lived proteins (SLiPs) are readily stabilized in class I HLA, and as a consequence of frequently being undetected in the thymus, have demonstrated strong immunogenicity. Early reports suggest some non-canonical immunopeptides are shared within and sometimes across multiple cancer histologies, with early evidence that some have tumor-promoting functions. Because these SLiPs are degraded so quickly and are stabilized in HLA-I, the intact proteins are postulated to not be accessible to antigen-presenting cells and are not efficiently processed and cross-presented-positioning this 'junk DNA'-derived antigen class as an attractive foundation for off-the-shelf vaccines. Here, we trace four phases of cancer vaccine evolution, review the technological advances that enabled the discovery of the dark immunopeptidome and discuss how these findings challenge established paradigms and reinvigorate interest in shared tumor antigens. By embracing this expanded antigenic universe, the field is poised to overcome key limitations of neoantigen-focused immunotherapy and move toward more universally effective cancer vaccines.

Breast cancer brain metastasis (BCBrM) remains one of the most lethal manifestations of breast cancer. Its response to immunotherapy is severely limited by the blood-brain barrier, which restricts immune cell infiltration and antigen presentation, thereby creating an immunosuppressive microenvironment. To overcome these barriers, recent studies have focused on novel immune checkpoints, including the Lymphocyte-Activated Gene 3-Galectin 3 (LAG3-LGALS3) and T-Cell Immunoreceptor with Ig and ITIM Domains-Nectin Cell Adhesion Molecule 2 (TIGIT-NECTIN2) axes, as well as on the reprogrammed metastatic ecosystem driven by immunosuppressive cells such as Forkhead Box P3-positive (FOXP3⁺) Regulatory T (Treg) cells, Lysosomal-Associated Membrane Protein 3-positive (LAMP3⁺) tolerogenic dendritic cells (DCs), C-C Motif Chemokine Ligand 18-positive (CCL18⁺) M2-like macrophages, Regulator of G-Protein Signaling 5-positive (RGS5⁺) cancer-associated fibroblasts (CAFs), Galectin 1-positive (LGALS1⁺) and TANK-Binding Kinase 1-positive (TBK1⁺) microglia, and phosphorylated Signal Transducer and Activator of Transcription 3-positive (pSTAT3⁺) reactive astrocytes. In addition, targeted inhibition of tumor-derived N-acetyltransferase 8-like (NAT8L) and metabolites N-Acetylaspartate (NAA), suppression of the N-Methyl-D-Aspartate Receptor (NMDAR) signaling pathway in tumor cells, and interventions against γ-Aminobutyric Acid (GABA)ergic reprogramming in BCBrM cells. Moreover, targeted interventions against distinct immune escape pathways-such as the Ubiquitin-Conjugating Enzyme E2T (UBE2T)/Cell Division Cycle 42 (CDC42)/Cluster of Differentiation 276 (CD276) and C-C Motif Chemokine Ligand 2-C-C Motif Chemokine Receptor 2/C-C Motif Chemokine Receptor 4 (CCL2-CCR2/CCR4) axes-have shown promise in reshaping the immune microenvironment and enhancing the efficacy of conventional immunotherapy. Collectively, this perspective outlines evolving strategies in immune checkpoint modulation, cellular ecosystem reprogramming, and neuroimmune intervention, providing a forward-looking framework to enhance the efficacy of immunotherapy in BCBrM.

Background: Peritoneal metastasis (PM) from gastric cancer (GC) is associated with poor prognosis and limited treatment options. We investigated a novel peritoneal-targeted therapy using CF33-human sodium iodide symporter (hNIS), a chimeric orthopoxvirus, combined with anti-PD-L1 immune checkpoint blockade in an immunocompetent mouse model of GCPM.

Methods: We evaluated replication and cytotoxicity of CF33-hNIS in human and murine GC cell lines. We assessed a syngeneic mouse model of PM using the transgenic mouse GC ACKPY3944 cells to test the efficacy of intraperitoneal (IP) CF33-hNIS alone and combined with intravenous or IP anti-PD-L1. Next, we performed flow cytometry and immunohistochemistry to analyze immune cell populations of CD3⁺ T cell subsets in the peritoneal cavity and tumor microenvironment. Mice that showed complete tumor regression (CTR) were rechallenged with ACKPY3944 cells to assess memory T cell responses.

Results: CF33-hNIS efficiently infected and killed GC cells. In vivo, IP CF33-hNIS alone significantly prolonged survival and increased infiltration of CD3⁺ and CD8⁺ T cells within the peritoneal cavity and solid tumor. The most pronounced therapeutic effect was observed with a single high-dose of CF33-hNIS (10⁸ plaque-forming units) combined with IP anti-PD-L1 (Combo 2) with 75% CTR. Notably, mice with CTR rejected tumor rechallenge, exhibiting significantly elevated effector and central memory T cell populations in the peritoneal cavity and spleen. IP CF33-hNIS demonstrates robust anti-tumor efficacy against GCPM, particularly when combined with IP anti-PD-L1 in the high-dose Combo 2 regimen.

Conclusions: These findings support a simplified, high-dose IP treatment strategy to overcome immune resistance in GCPM and provide a strong rationale for future clinical evaluation.

Over the past few years, outcomes and findings from several phase 1 clinical trials evaluating chimeric antigen receptor (CAR)-T cell therapies for glioblastoma (GBM) have been reported. For CAR-T cell therapy to succeed in GBM, several challenges must be overcome, including the immunosuppressive microenvironment (immunosuppressive cells, hypoxia, and metabolic constraints), antigen heterogeneity, and the anatomically isolated environment. To address these challenges, next-generation CAR-T cells-those engineered with additional functionalities-have been developed, and phase 1 clinical trials evaluating these next-generation CAR-T therapies for GBM have now been initiated. The development of CAR-T therapy for GBM has entered a new chapter. To date, the antitumor efficacy of CAR-T therapies still needs to be improved, and the high frequency of neurotoxicity remains a major issue that needs to be addressed; however, there is a growing anticipation that next-generation CAR-T therapies may provide clinical benefit to patients.

Background: Immune checkpoint inhibitors have improved outcomes for several malignancies; however, there remains a lack of accurate, non-invasive methods to assess tumor PD-L1 expression levels and guide immunotherapy. This study aimed to evaluate the role of PD-L1-targeted positron emission tomography (PET) imaging in predicting immunotherapy response and prognosis in lung cancer.

Methods: Four healthy volunteers and 22 treatment-naïve lung cancer patients were prospectively enrolled and underwent [⁶⁸Ga]Ga-PDL1p PET imaging. All patients additionally completed paired baseline [¹⁸F]fluorodeoxyglucose ([¹⁸F]FDG) PET scans. Of the 22 patients, 17 received ≥3 cycles of immunotherapy combined with chemotherapy and underwent follow-up [¹⁸F]FDG PET or CT examinations. The correlations of baseline [⁶⁸Ga]Ga-PDL1p and [¹⁸F]FDG uptake with tumor PD-L1 expression were evaluated. Furthermore, the associations of tumor [⁶⁸Ga]Ga-PDL1p uptake, [¹⁸F]FDG uptake, and PD-L1 expression with immunotherapy response were analyzed, along with their predictive values for immunotherapy efficacy and outcomes.

Results: Lesions with high PD-L1 expression exhibited significantly higher [⁶⁸Ga]Ga-PDL1p uptake than those with low expression (p=0.007), whereas [¹⁸F]FDG uptake showed no significant difference (p=0.499). At baseline, [⁶⁸Ga]Ga-PDL1p uptake was significantly higher in responders than in non-responders (p=0.008), with an area under the receiver operating characteristic curve of 0.886. In contrast, neither [¹⁸F]FDG uptake nor PD-L1 expression levels differed significantly between the two groups. Disease progression occurred in 23.5% of patients (4/17) by the final follow-up. Patients with higher [⁶⁸Ga]Ga-PDL1p uptake or higher [¹⁸F]FDG uptake demonstrated significantly longer progression-free survival (PFS) than those with lower uptake (p=0.033 and p<0.001, respectively). However, no significant difference in PFS was observed between patients with high and low PD-L1 expression, using either a 50% (p=0.487) or 1% (p=0.100) cut-off.

Conclusions: [⁶⁸Ga]Ga-PDL1p PET outperforms conventional [¹⁸F]FDG PET and immunohistochemistry-based PD-L1 assessment in predicting immunotherapy response and prognosis. These findings offer new insights for evaluating immunotherapy efficacy and guiding individualized tumor treatment.

Background: Insufficient T-cell infiltration limits the effectiveness of immunotherapy in sarcoma, yet the tumor-intrinsic mechanisms that govern immune exclusion remain poorly defined.

Methods: By integrating patient-derived ex vivo sarcoma spheroids with autologous expanded tumor-infiltrating lymphocytes and an in vivo metastatic osteosarcoma model, antitumor immune regulation by histone modifications was examined.

Results: Histone H3 lysine 27 acetylation (H3K27ac) was identified as a key regulator of CD8⁺ T-cell infiltration in osteosarcoma and other bone and soft-tissue sarcomas. Pharmacological elevation of H3K27ac by the histone deacetylase 1/3 inhibitor entinostat promotes CD8⁺ T-cell activation, cytotoxicity, and the recruitment of CD8⁺CD103⁺ tissue-resident memory T cells. Mechanistically, these immune-boosting effects are triggered by a Hippo pathway switch, in which yes-associated protein 1 (YAP1) is suppressed, and vestigial-like family member 3 (VGLL3) is induced, thereby modulating transcription towards an immune-responsive state. Furthermore, we identified that VGLL3/CD103 signatures predict a response to anti-programmed cell death protein-1 (PD-1) treatment in patients with sarcoma, and that combining H3K27ac induction with anti-PD-1 further augments T cell-mediated killing in ex vivo autologous patient-derived spheroid models.

Conclusions: Our findings reveal an epigenetic-Hippo-immunomodulatory axis in osteosarcoma that also extends to other sarcomas, providing a rationale for incorporating epigenetic preconditioning with immunotherapy to improve patient outcomes and pointing towards novel biomarkers for treatment guidance.

Background: Non-steroidal anti-inflammatory drugs (NSAIDs) are frequently used medications in non-small cell lung cancer (NSCLC). We comprehensively evaluated the effect of NSAID exposure patterns on immunotherapy outcomes, prostaglandin E2 (PGE2) and immune cells.

Methods: This multicenter study employed both prospective and retrospective approaches for patient enrollment. Patients were included if they had advanced NSCLC, and were first-time users of immune checkpoint inhibitor (ICI). Endpoints included progression-free survival (PFS) and overall survival (OS). Serum and tumor tissue were collected in the prospective subset for PGE2 and immune cell assay.

Results: A total of 1,748 patients were included, with 232 patients prospectively enrolled. After multivariable adjustment, NSAID use was independently associated with improved PFS (HR 0.72, 95% CI 0.58 to 0.91, p=0.0053) and OS (HR=0.76, 95% CI 0.58 to 0.98, p=0.0364). Benefit was observed for both before-ICI (HR 0.75, 95 % CI 0.57 to 0.99, p=0.0383) and after-ICI initiation (HR 0.63, 95% CI 0.43 to 0.92, p=0.0159). For aspirin, low-dose aspirin users had better PFS than non-users (HR 0.69, 95% CI 0.51 to 0.94, p=0.0191), especially when used at least 30 days (HR=0.69, 95% CI 0.48 to 0.99, p=0.0464). Among non-selective NSAIDs, only after-ICI initiation improved OS (HR=0.49, 95% CI 0.24 to 0.99, p=0.0458). Any-grade (57.8% vs 53.4%) and grade ≥3 adverse events (9.2% vs 9.8%) were comparable among NSAID users and non-users. Serum sample indicated that aspirin was associated with less PGE2, which was related to better outcomes. On immune cells, RNA-seq revealed a correlation between the cyclooxygenase-2 pathway and neutrophils. Biosample analysis further demonstrated that aspirin use was associated with decreased neutrophils in both circulation and tumor.

Conclusions: Long-term low-dose aspirin enhanced immunotherapy efficacy in patients with advanced NSCLC. PGE2 and neutrophils represented potential biomarkers guiding NSAID-immunotherapy integration.

Trial registration number: NCT05754983.

Background: Immune checkpoint inhibitors (ICIs) therapy targeting programmed cell death protein 1 (PD-1)/programmed cell death-ligand 1 (PD-L1) shows promising clinical benefits in non-small cell lung cancer (NSCLC). However, the relatively low response rate highlights the need to elucidate the regulatory mechanism of PD-L1 expression, and develop an alternative strategy to target PD-1/PD-L1 immune checkpoint pathway. Our study focuses on the role and mechanism of ubiquitin-specific protease 15 (USP15) and its derived peptide U10 on NSCLC immune evasion.

Methods: USP15 as PD-L1's deubiquitinase was identified by screening a human USP complementary DNA (cDNA) library. The role and mechanism of USP15 and its derived peptide U10 on PD-L1 stability in NSCLC cells were analyzed. T cell-mediated tumor cell killing activity and a syngeneic mouse NSCLC model were used to assess the influence of USP15 and U10 on NSCLC immune evasion. The antitumor effect of U10 in combination with PD-1 monoclonal antibody (mAb) via suppressing NSCLC immune evasion was also evaluated in mice. The expression and clinicopathological significance of USP15 and PD-L1 in cancer tissues were evaluated by immunohistochemistry.

Results: We identify USP15 as a novel deubiquitinase of PD-L1. Mechanistically, USP15 binds and stabilizes PD-L1 in NSCLC cells by inhibiting its ubiquitination and degradation. Functionally, USP15 inhibits T cell ability of killing NSCLC cells in vitro, and promotes NSCLC immune evasion in mice via decreasing the population and activation of CD8⁺ T cells in the tumor microenvironment. Based on the interacting regions of USP15 and PD-L1, we develop a 10 amino acid-long USP15-derived peptide U10, which successfully degrades PD-L1 via disrupting USP15 and PD-L1 interaction, dramatically suppresses NSCLC immune escape in vitro and in mice, and enhances the anti-NSCLC effect of PD-1 mAb in mice. Moreover, the expression levels of USP15 and PD-L1 are significantly higher in NSCLC than those in normal lung tissues and are positively correlated. The combination of USP15 and PD-L1 proteins was superior to individual proteins for predicting the efficacy of PD-1 mAb immunotherapy and patient prognosis in NSCLC.

Conclusion: Our findings reveal a critical role for USP15 in PD-L1 stability regulation and NSCLC immune escape and develop a novel peptide as an alternative strategy for ICIs therapy of NSCLC.

Tumor mutational burden (TMB) and PD-L1 are established biomarkers for guiding immune checkpoint inhibitor (ICI) therapy in advanced non-small cell lung cancer (NSCLC). As ICI use expands into early-stage disease, we explored the feasibility of using TMB, which can be determined via a comprehensive genomic profiling assay along with EGFR and ALK genomic alterations, as a biomarker for outcomes in both neoadjuvant and adjuvant settings. TMB-high status (≥10 mut/Mb) showed a numerically higher, but not statistically significant, rate of pathological complete response among patients receiving neoadjuvant ICI and significantly associated with more favorable time to recurrence in patients receiving adjuvant ICI, particularly among patients with PD-L1 expression <50%. TMB should be considered in future early-stage NSCLC ICI clinical trials to further validate these results.

Background: Oncolytic virotherapy represents a promising anticancer strategy by combining direct tumor lysis with in situ immune activation. However, its efficacy remains limited in immune-cold tumors, which are characterized by poor T-cell infiltration and an immunosuppressive microenvironment. Although engineering oncolytic viruses to deliver chemokines has been explored to modulate immune recruitment, most existing approaches activate only a single immune axis. Senecavirus A (SVA), a tumor-selective RNA virus with high genetic stability and flexible transgene capacity, offers an attractive platform for coordinated intratumoral delivery of immune payloads.

Methods: We engineered recombinant SVA vectors to achieve co-expression of CXCL11 and vXCL1 within tumor, aiming to synergistically recruit both T cells and cross-presenting dendritic cells and remodel the tumor immune landscape. In vitro and in vivo assays were conducted to assess viral properties (stability, replication kinetics, gene expression) and cytotoxicity against B16-F10 melanoma cells in different stages, with tumor burden, immune cell infiltration, and mouse survival in C57BL/6 mice analyzed to evaluate the overall therapeutic efficacy.

Results: Signal-peptide deletion significantly improved the genetic stability of transgenes and enhanced intratumoral payload retention, without compromising viral fitness. In murine models, CXCL11-expressing virus monotherapy achieved enhanced tumor control, prolonged survival, and provided preliminary evidence suggestive of protection on tumor rechallenge. Furthermore, the combination of CXCL11 and vXCL1 further augmented dendritic cell activation, promoted CD8⁺ T cell infiltration, and shifted macrophages toward an M1-polarized phenotype, while sustaining higher intratumoral viral loads. These coordinated immune changes led to deeper and more durable tumor regression.

Conclusions: Our findings validate the combination of CXCL11- and vXCL1-armed SVA as a potent immunovirotherapy strategy and propose a design principle for multistage, multigene intervention: concurrently targeting leukocyte recruitment, antigen presentation, and effector activation provides complementary mechanisms that synergistically amplify therapeutic efficacy.