Journal for Immunotherapy of Cancer最新文献

Background: NUT carcinoma is a rare but highly lethal solid tumor without an effective standard of care. NUT carcinoma is caused by bromodomain-containing NUTM1 fusion oncogenes, most commonly BRD4::NUTM1. BRD4::NUTM1 recruits p300 to acetylate H3K27 forming expansive stretches of hyperacetylated chromatin called "megadomains" with the overexpression of corresponding oncogenes, including MYC. We hypothesized that transcriptional dysregulation caused by BRD4::NUTM1 would lead to the generation of cancer-specific antigens that could be therapeutically actionable.

Methods: We integrated genomics, computational antigen prediction software, targeted immunopeptidomics using single-labeled and double-labeled peptide standards, and gain/loss-of-function genetic experiments on a panel of cell lines (N=5), a patient-derived xenograft, a tissue microarray (N=77), and patient samples from the Tempus AI Sequencing Database harboring evidence of NUTM1 fusions (N=165). We created an αPRAME₄₂₅ T-cell receptor (TCR) × SP34 αCD3 bispecific molecule modeled after brenetafusp, an αPRAME₄₂₅ TCR bispecific T-cell engager, as well as αPRAME₄₂₅ TCR T-cells based on anzutresgene autoleucel and we applied these products to NUT carcinoma cells in vitro.

Results: We identified PRAME as the most commonly expressed cancer/testis antigen in patient samples harboring the three canonical NUT carcinoma fusions (BRD4::NUTM1, BRD3::NUTM1, and NSD3::NUTM1). Additionally, 56% (43/77) of NUT carcinoma tissue microarray samples stained positive for PRAME. BRD4::NUTM1 expression in HEK 293T cells enhanced PRAME levels and BRD4::NUTM1 knockout in NUT carcinoma cells reduced PRAME levels. Immunopeptidomics detected more PRAME-derived human leukocyte antigen (HLA) ligands (N=9) than all other cancer/testis antigens combined (N=5). Targeted mass spectrometry detected the HLA-A*02:01/SLLQHLIGL (PRAME₄₂₅) epitope in 100% (4/4) of HLA-A*02+, PRAME+ NUT carcinoma samples at higher levels (>0.01 fM) than HLA-A*02:01/RLDQLLRHV (PRAME₃₁₂) or HLA-A*02:01/YLHARLREL (PRAME₄₆₂). The αPRAME₄₂₅ TCR × SP34 αCD3 bispecific molecule and αPRAME₄₂₅ TCR T-cells each exhibited potent, T-cell mediated cytotoxicity against PRAME+ NUT carcinoma cells.

Conclusions: PRAME is highly and frequently expressed in NUT carcinoma, and the most common oncoprotein causing NUT carcinoma, BRD4::NUTM1, contributes to these high PRAME levels. PRAME epitopes presented by HLA class I are a previously unrecognized therapeutic vulnerability for NUT carcinoma that warrants clinical trials testing PRAME-targeted immunotherapies in this neglected patient population.

Background: Colorectal cancer (CRC) is a leading cause of cancer-related death and remains a significant global health challenge. Cancer vaccines have emerged as a promising immunotherapy for long-term tumor control. While Listeria monocytogenes (Lm)-based intravenous vaccines can generate tumor-reactive CD8 T cells, clinical trial success has been limited. Here, we sought to determine whether in vivo targeting of gastrointestinal tissues with foodborne delivery of Lm-based cancer vaccines controlled tumor growth in murine models of CRC.

Methods: The ActA and InlB virulence genes were deleted from a mouse-adapted Lm strain expressing ovalbumin and containing an internalin A mutation (InlA^M Lm-ova) that allows epithelial cell invasion of mice to generate an oral vaccine administered via consumption of inoculated bread. Immunogenicity and safety were tested in C57Bl/6 mice. Vaccine efficacy was evaluated with CRC tumors delivered by colonoscopy-guided orthotopic transplantation into the colon submucosa. Microsatellite instability high MC38 cell line expressing ovalbumin or genetically engineered microsatellite stable AKPS (Apc ^KO Kras ^G12D Trp53 ^KO Smad4 ^KO) organoids expressing low levels of ovalbumin (lo^SIIN) were used. Vaccines were tested in prophylactic and therapeutic settings and in the context of immune checkpoint inhibitors (ICI).

Results: Oral immunization induced a robust CD8 T cell response that was similar in magnitude and phenotype to the fully virulent Lm. Immunized mice did not lose weight, and Lm was contained to intestinal tissues. Mice prophylactically immunized with the vaccine were protected from CRC tumors. Therapeutic immunization of mice bearing lo^SIIN AKPS tumors revealed curtailed growth of the local tumor but did not improve survival. Immunization with anti-programmed cell death protein-1 and anti-cytotoxic T-lymphocyte-associated protein 4 controlled tumors when coupled with therapeutic immunization. Protection correlated with accumulation of ova-specific CD8 T cells within the tumor.

Conclusions: Oral Lm-based cancer vaccines targeting CRC elicit robust, widely disseminated, and persistent tumor-specific immune responses in mice. These vaccines limit CRC development when administered prophylactically and provide tumor control when administered therapeutically with ICI. Thus, oral delivery of Lm-based cancer vaccines coupled with ICI may provide improved control of CRC progression in clinical application.

Background: Glioblastoma, one of the most aggressive brain tumors, has been largely resistant to conventional immunotherapies, underscoring the need for novel treatment approaches. A promising strategy involves simultaneously inhibiting immunosuppressive pathways in the tumor microenvironment, as these pathways play pivotal roles in immune evasion. However, the therapeutic potential of combined targeting of these key immunosuppressive pathways in glioblastoma remains underexplored. We hypothesized that co-targeting the transforming growth factor (TGF)-β and PD-1 pathways could enhance immune responses against glioblastoma.

Methods: Human glioblastoma datasets were interrogated for the expression of PD-L1, TGF-β, and TGF-β target genes. Bintrafusp alfa, a first-in-class bifunctional fusion protein that blocks PD-L1 while sequestering TGF-β in the tumor microenvironment, was used to simultaneously inhibit both pathways. Its effects were assessed in vitro using human and mouse glioma cells and in vivo in immunocompetent, syngeneic mouse glioma models. High-dimensional flow cytometry was used to analyze treatment-induced changes in the tumor microenvironment.

Results: We observed a strong correlation between TGF-β and PD-L1 co-regulation, suggesting interconnected immunosuppressive mechanisms as part of a gene expression network. In vitro, bintrafusp alfa inhibited TGF-β-induced Smad2 phosphorylation, a bona fide response marker of TGF-β pathway activation, and enhanced immune cell-mediated killing of glioma cells. In vivo, combined targeting of both immunosuppressive pathways significantly improved survival of glioma-bearing mice, with long-term survivors exhibiting protection from tumor re-challenge. This survival benefit was not seen in T cell-deficient mice, confirming the necessity of adaptive immunity. High-dimensional flow cytometry of single-cell suspensions from tumor-bearing hemispheres revealed a distinct remodeling of immune subsets in the bintrafusp alfa-treated group compared with control-treated mice.

Conclusions: Our findings provide strong support for the combined targeting of TGF-β and PD-L1 as a promising immunotherapeutic strategy to overcome immunosuppressive barriers in glioblastoma and induce potent antitumor responses.

Neoadjuvant chemoimmunotherapy (chemo-IO) has fundamentally reshaped the treatment paradigm for resectable non-small cell lung cancer (NSCLC), challenging long-held surgical boundaries and redefining what constitutes "resectable" disease. Trials such as CheckMate-816, KEYNOTE-671, and AEGEAN have demonstrated that integrating immune checkpoint blockade with chemotherapy yields unprecedented rates of pathological response and event-free survival, positioning chemo-IO as the new global standard for stage IB-IIIA NSCLC. Yet these advances bring new complexities; how do we define resectability in an era of immunotherapeutic downstaging, and how should multidisciplinary teams adapt to evolving biology? Traditional radiological and anatomic criteria now sit alongside immune-mediated regression and circulating tumor DNA (ctDNA) kinetics as measures of treatment success. ctDNA clearance and pathological response serve as powerful surrogates for long-term survival, with ongoing studies such as MERMAID-1/2 exploring their potential to guide adjuvant therapy and spare overtreatment. The modern challenge lies in integrating these biomarkers into surgical decision-making and developing standardized, biology-informed resectability frameworks. Future progress will depend on close collaboration between surgeons, oncologists, and translational scientists to expand surgical candidacy safely and define the next generation of curative strategies in lung cancer.

Background: The CDKN2A gene encodes two canonical tumor suppressors, p16INK4A and p14ARF, which safeguard cells from malignant transformation by inducing cell cycle arrest and apoptosis in response to aberrant growth signals. Paradoxically, many cancers overexpress these proteins when downstream effectors that enforce negative feedback regulation are lost or inactivated. For example, p14ARF, which regulates p53 activation, is aberrantly expressed in more than 50% of tumors with inactivating p53 mutations. Here, we evaluated the feasibility of targeting dysregulated p16INK4A and p14ARF expression using TCR-T cell therapeutics.

Methods: We analyzed a panel of p16INK4A- and p14ARF-derived peptides for HLA-A*02:01-associated presentation and recognition by CD8⁺ T cells. Antigen-specific T cell receptors were isolated from healthy donor repertoires and expressed in primary T cells to assess specificity, functional avidity, tumor recognition, and safety using in vitro T cell functional assays, in vivo tumor models, and an in vivo safety model.

Results: We identified a unique and well-presented p14ARF epitope that was consistently detected in the HLA-A*02:01-associated immunopeptidome of cancer biopsies but not in normal tissues. High-avidity ARF-specific TCRs were isolated from the peripheral repertoire of healthy donors, and TCR-transduced T cells mediated potent tumor cell killing in vitro and in vivo in preclinical models. Furthermore, targeting p14ARF-expressing cells did not result in detectable on-target toxicity in an in vivo safety model.

Conclusions: These findings demonstrate the feasibility of targeting dysregulated tumor suppressor proteins with TCR-T cell therapeutics and identify p14ARF as a promising target for future therapies.

Background: Immunotherapy has had limited success in pancreatic cancer, largely due to a low mutational burden and immunosuppressive microenvironment. Here we hypothesized that systemic delivery of viral antigens can redirect pre-existing antiviral immunity against pancreatic tumors.

Methods: Cytomegalovirus (CMV, a β-herpesvirus) was chosen, as the majority of the population is infected and it induces an extremely large/broad memory T-cell response. Mice latently infected with murine CMV (MCMV) were orthotopically implanted with pancreatic cancer cells and treated with systemic injections of MCMV T-cell epitopes. Tumor growth was monitored by ultrasound two times a week, and immune cell infiltration was analyzed by histology, flow cytometry and single-cell RNA sequencing (scRNA-seq). Statistical analysis was performed by two-way analysis of variance with Sidak correction.

Results: MCMV peptide-epitope therapy (MCMVp) promoted preferential accumulation of MCMV-specific T cells within pancreatic tumors, delaying tumor growth and increasing survival. Immunophenotyping and scRNA-seq analyses showed these T cells were highly activated and cytotoxic, leading to increased tumor necrosis and caspase-3 activation. Depletion of CD4 and CD8 T cells abolished the impact of MCMVp therapy, indicating the antitumor response is T-cell dependent. Together, these results show that CMV-specific T cells can be repurposed to combat pancreatic cancer.

Conclusions: Our studies reveal that CMV-specific viral memory T cells can be re-directed to control a solid tumor normally refractory to immunotherapy via a simple, intravenous injection of T-cell peptide epitopes. This mutation-agnostic approach has significant potential for the development of "off-the-shelf" therapeutics by stimulating pre-existing antiviral memory, and it is widely applicable due to the high prevalence of CMV.

Background: The rapidly advancing field of cancer therapy has sparked growing interest in the potential synergy between anticoagulation and immune checkpoint inhibitor (ICI) therapy. Recent research highlights that anticoagulants, traditionally used for thromboprophylaxis and managing thromboembolic events, may also exhibit immunomodulatory properties. These properties can influence the tumor microenvironment by promoting immune cell infiltration, enhancing antitumor immune responses, and potentially reducing metastasis. This emerging evidence underscores the complex interplay between coagulation pathways and immune regulation, paving the way for further exploration of the clinical benefits of combining anticoagulation with ICI therapy.

Methods: A systematic review was conducted to synthesize the current evidence on the interplay between anticoagulation and ICI. Relevant studies examining their mechanisms of action, clinical outcomes, and potential interactions were identified and analyzed. Comprehensive database searches were performed to ensure a thorough and inclusive review of the literature.

Results: Preclinical studies consistently show that combining ICIs with anticoagulants can enhance cancer treatment by inhibiting tumor growth and metastasis. In particular, low molecular weight heparin, oral factor Xa (FXa) inhibitors, and platelet inhibitors have demonstrated synergistic effects with ICI. However, these findings have not been consistently replicated in clinical settings. While two retrospective studies reported no significant impact of anticoagulants on ICI efficacy, one retrospective study found improved outcomes in advanced melanoma patients treated with ICI and FXa inhibitors. Additionally, another retrospective study revealed a significant association between platelet aggregation inhibition and extended progression-free survival.

Conclusions: Our literature review underscores the intricate relationship between anticoagulation and ICIs in cancer therapy. Future studies should prioritize exploring the interactions between ICI, FXa inhibitors, and antiplatelet agents.

Background: Acquired resistance to KRAS G12C inhibitor sotorasib remains a critical challenge in non-small cell lung cancer treatment. A deeper, rational understanding of resistance mechanisms can enable the development of therapeutic strategies to overcome resistance.

Methods: We established a syngeneic resistant model after prolonged AMG-510 treatment in C57BL/6 mice. In addition, the in vitro co-culture model and multiple methods including flow cytometry and western blot were used to assess the changes of immune microenvironment during resistance. Finally, a serial combinatorial therapy strategy was applied in the resistant mouse model to evaluate its ability to reverse resistance.

Results: Upregulation of PD-L1 in KRAS G12C tumors drives an immunosuppressive tumor microenvironment and promotes acquired resistance characterized by reduced infiltration of cytotoxic CD8⁺ T cells and a marked expansion of myeloid-derived suppressor cells through JAK2/STAT3/IL-6 Pathway. These mechanisms promote tumor immune evasion and protection from cell apoptosis, thereby establishing a microenvironment that sustains acquired resistance to sotorasib. Critically, sequential administration of a PD-L1 inhibitor (PD-L1i) effectively reprogrammed the immunosuppressive microenvironment, restoring antitumor immunity and re-sensitizing resistant tumors to sotorasib treatment.

Conclusions: These results identify the PD-L1-driven immunosuppressive microenvironment as a key mediator of sotorasib resistance and propose PD-L1i as a synergistic strategy to overcome resistance, which warrants clinical exploration of sequential or combinatorial regimens.

Background: Crosstalk between inflammation and the immune system plays an important role in tumor malignant progression, immune evasion, and immunotherapy efficacy. This study aims to explore the significance of inflammation-associated gene ribosomal-binding protein 1 (RRBP1) in modulating tumor malignant progression and immune escape.

Methods: This study was used transcriptome, proteomic and in vivo anti-programmed death-ligand 1 (PD-L1) antibody CRISPR Cas9 screening data to identify RRBP1 as an inflammation-immune-associated gene in bladder cancer (BC). Immunohistochemistry, single-cell RNA sequencing, multiplex immunofluorescence, flow cytometry, RNA sequencing, and animal experiments were used to study the role of RRBP1 in regulating tumor malignant progression and immunotherapy efficacy.

Results: RRBP1 overexpression promoted the proliferation and metastasis of BC both in vitro and in vivo. RNA sequencing and single-cell RNA sequencing revealed that RRBP1 inhibition activated immune-associated pathways and reshaped the tumor immune microenvironment by altering the infiltration of CD8⁺ T-cell subpopulations, thereby enhancing antitumor immunity. Mechanistically, RRBP1 inhibition enhances the secretion of CXCL10 by cancer cells, which binds to CXCR3 on CD8⁺ T cells to promote interferon-γ and Granzyme B expression. Furthermore, genetic and pharmacological inhibition of RRBP1 sensitizes tumors to anti-PD-L1 therapy.

Conclusions: Our findings highlight RRBP1 as an inflammation-immune-associated gene that inhibits tumor progression and improves immunotherapy efficacy by regulating the CXCL10-CXCR3 axis in the tumor microenvironment.