Journal for Immunotherapy of Cancer最新文献

Background: Insufficient infiltration of CD8⁺ T cells in the tumor microenvironment (TME) critically restricts antitumor immunity and cancer immunotherapy efficacy. The purpose of this study was to identify novel tumor cell-intrinsic regulators of T-cell infiltration and to elucidate their mechanisms of action.

Methods: We performed a genome-wide Sleeping Beauty transposon mutagenesis screen in murine breast cancer models. Protein-protein interactions were identified by mass spectrometry and validated by co-immunoprecipitation. Gene and protein expression levels were assessed by reverse transcription and quantitative PCR and western blotting. T-cell infiltration and function were evaluated using flow cytometry, immunohistochemistry (IHC), multiplex IHC, and by analyzing bulk and single-cell RNA sequencing data complemented by bioinformatic analysis. The specific dephosphorylation sites on LGALS1 were confirmed through phosphomimetic mutant experiments. T-cell infiltration was further validated using an in vitro T-cell transendothelial migration assay and in vivo mouse models.

Results: Our screening identified 39 candidate genes, with tumor cell-intrinsic dual-specificity phosphatase 22 (DUSP22) expression correlating with enhanced CD8⁺ T-cell accumulation and suppressed tumor progression. Overexpression of DUSP22 resulted in increased CD8⁺ T-cell infiltration and enhanced T-cell function. Mechanistically, DUSP22 binds to LGALS1 and dephosphorylates it at the Ser8 and Thr58 residues, leading to LGALS1 degradation and subsequent alleviation of LGALS1-mediated immunosuppression. In human breast cancer samples, LGALS1 expression was negatively correlated with both DUSP22 levels and CD8⁺ T-cell infiltration. Therapeutic targeting of the DUSP22-LGALS1 axis significantly enhanced CD8⁺ T-cell infiltration and synergized with anti-programmed cell death protein-1 therapy to boost antitumor responses.

Conclusions: Our findings unveil a novel phosphorylation-dependent DUSP22-LGALS1 axis that reprograms the immunosuppressive TME. This work thus proposes a promising therapeutic strategy to overcome immune checkpoint blockade resistance in breast cancer.

Background: Nanosecond pulsed electric field (nsPEF) ablation has demonstrated limited and transient efficacy in suppressing tumor progression. Oncolytic peptide LTX-315 is known to elicit a strong antitumor immune response and durable immune memory. This study aimed to investigate whether LTX-315 could enhance nsPEF-induced antitumor immunity in liver cancer.

Methods: Both cell assays and mouse models were used to evaluate the therapeutic efficacy of nsPEF, LTX-315, and combination therapy. Flow cytometry and immunofluorescence were performed to assess the tumor immune microenvironment. Co-culture models were established to evaluate the functional modulation of immune cells.

Results: nsPEF upregulated the programmed cell death 1 ligand 1 (PD-L1) expression in liver cancer cells, leading to CD8⁺ T-cell dysfunction. LTX-315 reduced the nsPEF-mediated elevated PD-L1 level and restored the cytotoxicity of CD8+ T cells. Furthermore, LTX-315 acted with nsPEF to induce enhanced immunogenic cell death for the activation of dendritic cells and CD8+ T cells. In addition, LTX-315 improved antigen processing and presentation in nsPEF-treated liver cancer cells. Notably, the combination of nsPEF and LTX-315 achieved durable tumor control and prolonged survival of the tumor-bearing mice, by promoting the migration of dendritic cells to tumor-draining lymph nodes, the infiltration of immune cells within the tumor and potential immune memory to prevent tumor metastasis.

Conclusions: LTX-315 functions as an immune stimulant to improve the antitumor efficacy of nsPEF. The combination of nsPEF and LTX-315 represents a promising interventional immunotherapy strategy for liver cancer.

Key points: LTX-315; Antitumor immune response; nsPEF; Liver cancer.

Background: Tumor-associated macrophages (TAMs) are key drivers of the immunosuppressive tumor microenvironment (TME), thereby limiting the efficacy of immune checkpoint inhibitors (ICIs). However, the underlying mechanisms remain unclear.

Methods: Both genetic (Akr1b3 knockout) and pharmacologic (epalrestat) approaches were employed to examine the impact of Aldo-keto reductase family 1 member B1 (AKR1B1) inhibition on TAMs and T-cell function in vitro and in vivo. Mechanistic insights were obtained through RNA sequencing, flow cytometry, immunofluorescence staining, and co-culture assays. To assess therapeutic relevance, 4T1 breast cancer and LLC lung carcinoma mouse models were used to evaluate the effects of epalrestat on tumor growth, immune infiltration, and T-cell responses. Clinical relevance was validated in patient cohorts with triple-negative breast cancer (TNBC) and lung adenocarcinoma (LUAD).

Results: AKR1B1 is highly expressed in TAMs and correlates with CD8⁺ T-cell dysfunction. Targeting AKR1B1 enhances antitumor immunity by reprogramming TAMs. Mechanistically, AKR1B1 modulates macrophage metabolism via the glutathione/reactive oxygen species axis, suppressing nuclear factor κB activation and downregulating C-C motif chemokine ligand 5 (CCL5) production, thereby inducing CD8⁺ T-cell dysfunction and establishing an immunosuppressive TME. Inhibition of AKR1B1, either by gene knockout or selective pharmacologic blockade, reprograms TAMs toward an immunostimulatory phenotype, increases CCL5-CCR5 (C-C motif chemokine receptor 5) signaling, restores CD8⁺T cell effector function, and strengthens antitumor immunity. Clinically, high AKR1B1 expression is associated with poor prognosis and immune suppression in TNBC and LUAD. Notably, targeting AKR1B1 improves responses to ICIs in both breast and lung cancer models.

Conclusions: AKR1B1 as a critical regulator of TAM-mediated immunosuppression and highlight its therapeutic potential to enhance the efficacy of ICIs.

Introduction: An adaptive immune response to cancer requires three main signals: antigen presentation and recognition ("signal 1"), costimulation ("signal 2"), and secreted immunostimulatory cytokines ("signal 3"). Expression of these signals in tumors via non-viral gene delivery represents a promising strategy to reprogram the tumor microenvironment (TME) and prime antitumor immunity.

Methods: We used modular polymeric poly(beta-amino ester)-based nanoparticles (NPs) to investigate codelivery of plasmids encoding signal 3s (interleukin (IL)-2, IL-12, IL-15, IL-23, IL-35, or granulocyte-macrophage colony-stimulating factor) and signal 2s (4-1BBL, CD80, CD86, or OX40L) to B16F10 melanoma tumors in vivo. Downstream immune responses and impact on the TME were assessed via flow cytometry and spatial proteomics using a 27-marker PhenoCycler panel.

Results: Ex vivo flow cytometry and PhenoCycler tissue analysis revealed that multiple signal 2/3 NP combinations led to decreased tumor growth, increased immune-cell infiltration, and skewing of adaptive and innate populations towards immunostimulatory phenotypes with increased CD8⁺ T-cell and M1 macrophage infiltration. Signal 2/3 NPs also drove antigen presentation on tumor and antigen-presenting cells (APCs) (macrophages, dendritic cells), and led to induction of major histocompatibility complex class I and class II on melanoma cells. Evaluation of putative biomarkers of treatment response to signal 2/3 NP delivery demonstrated that multiple markers of an inflamed TME correlated negatively with tumor growth, with antigen presentation induction highly correlated with tumor size reduction. Spatial analysis on top-performing NP formulations demonstrated that immune cell populations entered the TME via both the tumor-stroma interface and intratumoral vessels; CD8⁺ T cells and proinflammatory M1 macrophages were brought into proximity. Furthermore, top NP formulations increased the density of intratumoral CD8⁺-CD4⁺-proinflammatory APC triads, which have been identified as key structures for immunotherapy-mediated clearance of solid tumors.

Conclusions: These results demonstrate the utility of a modular NP design for systematic screening of signal 2/3 delivery to melanoma in vivo and highlight the benefit of in-depth profiling via spatial proteomics to evaluate local antitumor immune responses. The results provide insight into the mechanisms underpinning this therapeutic reprogramming strategy, emphasizing the relationship between signal 2/3 NPs and their ability to drive signal 1 for productive antitumor responses in vivo.

Background: Immune checkpoint inhibitors have transformed melanoma therapy but frequently cause immune-related adverse events (irAEs), including colitis, that limit treatment. Reliable biomarkers predicting toxicity remain lacking.

Methods: In this retrospective, multicenter study, we analyzed pretreatment serum samples from 331 patients with metastatic melanoma treated with anti-CTLA-4 (ipilimumab), anti-PD-1 (pembrolizumab or nivolumab), or combination ipilimumab/nivolumab. IgG autoantibody reactivity against 832 human protein antigens, including autoimmune targets, cytokines, tumor-associated antigens, and cancer pathway proteins, was profiled using multiplex bead-based arrays. Statistical analysis (Significance Analysis of Microarrays and Cox regression) identified autoantibody signatures associated with subsequent irAEs and immune-related colitis (ir-colitis).

Results: We detected 47 autoantibodies predictive of irAEs, with KRT7, RPLP2, UBE2Z, and GPHN emerging as the strongest markers. Anti-KRT7 and anti-GPHN were specifically predictive in patients receiving PD-1 monotherapy, whereas anti-RPLP2 was associated with irAEs in ipilimumab/nivolumab combination therapy. For ir-colitis, 38 autoantibodies were identified, with five (PIAS3, RPLP0, UBE2Z, KRT7, and SDCBP) showing consistent predictive value across treatment groups. Anti-PIAS3 and anti-RPLP0 increased ir-colitis risk, while anti-SDCBP conferred protection. Notably, predictive profiles differed between PD-1-based and CTLA-4-based regimens, underscoring divergent mechanisms of toxicity. Several autoantibodies predictive of irAEs or ir-colitis also correlated with clinical outcome. ATG4D, MAGEB4, and IL4R were associated with prolonged progression-free and overall survival, whereas FGFR1 predicted both reduced irAE risk and inferior survival, consistent with the link between heightened immune activation, toxicity, and therapeutic benefit.

Conclusions: This study, to our knowledge, is the largest pretreatment autoantibody screen in melanoma immunotherapy, demonstrates that serum autoantibody profiles can stratify patients at risk for irAEs and ir-colitis. The identified signatures connect tumor-related and immunity-related antigens, stress-response pathways, and autoimmune mechanisms. Pretreatment autoantibody profiling offers a promising biomarker-driven approach for individualizing risk assessment, improving patient selection, and guiding early intervention strategies to enhance the safety of immune checkpoint blockade in melanoma. Beyond toxicity prediction, our findings also suggest that specific autoantibodies may reflect underlying immune activation states linked to therapeutic response.

Background: Hepatocellular carcinoma (HCC) is a global health challenge with high mortality rates, particularly in patients with advanced disease and lung metastasis. T-cell receptor (TCR)-T cell therapy based on specific neoantigens, is an emerging treatment with potential for HCC. However, the prognosis of patients remains poor, underscoring the need for novel targets and strategies.

Methods: We conducted a comprehensive study to investigate the role of C7orf50 and its neoantigens in HCC. We evaluated the functional impact on HCC progression and metastasis in vitro and in vivo, and further explored the mechanism by which C7orf50 promotes cancer metastasis and remodels tumor immune environment. Using exome and transcriptome sequencing, we identified neoantigens associated with C7orf50 and assessed their potential in TCR-T therapy.

Results: Our in vitro experiments revealed that C7orf50 overexpression enhances HCC cell proliferation, migration, and invasion, while knockdown inhibits these processes. In vivo, C7orf50 promoted tumor growth and lung metastasis, with a significant correlation between C7orf50 expression and poor clinical outcomes in patients with HCC. We further demonstrated that C7orf50 activates the NF-κB/PAI-1 pathway by binding to AEG-1 and facilitating its nuclear translocation, thereby promoting tumor-associated macrophage recruitment. Meanwhile, we found that TCR-T from C7orf50-related neoantigen could obviously realize the killing effect on HCC cells, revealing its great role in cell therapy.

Conclusion: C7orf50 is a critical mediator of HCC progression and lung metastasis, acting through the NF-κB/PAI-1 pathway and AEG-1. Its expression levels, along with those of PAI-1 and CD68, serve as independent prognostic markers. And C7orf50-related neoantigen shows great application potential in TCR-T therapy. These findings provide a foundation for developing C7orf50-targeted therapies and highlight its potential in precision medicine and immunotherapy for HCC.

Background: Cancer-associated fibroblasts (CAFs) significantly impact cancer progression and CAF subtypes are key determinants of response to immune checkpoint therapy (ICT). The transforming growth factor-β (TGF-β) signaling is a main pathway in protumorigenic activity of CAFs and resistance to ICT. The actin cytoskeleton regulator hMENA plays crucial roles in epithelial-mesenchymal transition (EMT) and regulates pathways critical to antitumor immune response, such as interferon-I and Axl-GAS6.

Methods: We constructed a single-cell atlas of CAFs using integrated public dataset. Experimental data were obtained by biochemical and molecular approaches in CAFs from freshly explanted non-small cell lung cancer (NSCLC) tissues hMENA silenced and in tumor cell lines or peripheral blood mononuclear cells treated with CAF conditioned medium. Multiparametric flow cytometry was used to characterize T cells. A gene signature indicative of ICT response was developed by a machine learning model.

Results: Computational analysis indicates that hMENA is primarily overexpressed in a myofibroblast cluster enriched for a TGF-β-activated CAF signature. Experimentally, we showed that TGF-β1 treatment increases hMENA expression and, reciprocally, hMENA/hMENAΔv6 modulate TGF-β1/2 production and secretion and transforming growth factor-β type II receptor expression in CAFs. Functionally, hMENA contributes to TGF-β1-driven CAF phenotype, programmed death-ligand 1 (PD-L1) upregulation, extracellular matrix remodeling and secretion of immunosuppressive cytokines/chemokines. The hMENA-driven TGF-β secretion in CAFs promotes PD-L1 expression and EMT in cancer cells by activating TGF-β signaling. On the tumor cell side, hMENA expression sustains the TGF-β signaling and EMT mediated by hMENA-driven CAFs secretome. This immunosuppressive secretome favors regulatory T cell (Treg) abundance and reduces CD8+ and CD4+ T cell functionality. Finally, based on the hMENA and TGF-β enriched CAF subtype, we developed a 9-gene signature, which in combination with hMENA/hMENAΔv6 correlates with increased Treg abundance and poor prognosis in the Cancer Genome Atlas NSCLC and associates with ICT resistance in Stand Up To Cancer (SU2C) and in phase III clinical trial (OAK) (NCT02008227) datasets.

Conclusions: Our findings indicate that hMENA overexpression in CAFs defines a myofibroblast-like subset predominantly driven by TGF-β signaling, which sustains TGF-β1-mediated crosstalk between cancer cells and CAFs and impairs T-cell functionality. In NSCLC tissues, hMENA^high CAFs associate with TGF-β and regulatory T-cell signatures and correlate with poor patient prognosis and resistance to immune checkpoint therapies, supporting their role as key contributors to an immunosuppressive, ICT-refractory tumor microenvironment.

Background: Current second-generation CAR T cell products rely on CD28 or 4-1BB costimulatory domains, additions that respectively favor rapid cytolysis or long-term persistence, but rarely both. Preclinical modeling and retrospective analysis have linked CD2-CD58 engagement to superior preclinical and clinical responses, yet the direct contribution of CD2 intracellular signaling remains undefined.

Methods: We replaced the costimulatory domain of anti-mesothelin (SS1) and anti-TnMUC1 (5E5) CARs with the human CD2 cytoplasmic tail (CD2z) and benchmarked them against 28z and BBz formats. Transient mRNA expression was used to profile proximal Ca²⁺ flux and degranulation free of tonic viral signals; durable functional assays employed lentiviral CARs. Cytokine release, genome-wide transcriptional programs, and anti-tumor activity were assessed in vitro and in NSG xenograft models.

Results: CD2z CAR T cells degranulated as efficiently as other z-containing CARs and generated a Ca²⁺ flux signal intermediate to 28z and BBz CARs. Lentiviral CD2z CARs released a Th1-skewed cytokine panel and matched 28z cytolysis despite a lower acute cytokine release. Transcriptomic analysis characterized CD2z cells in an early effector-memory state: glycolytic, mTORC1, and TNFa-NF-κB hallmarks were upregulated, whereas exhaustion-up signatures were selectively depleted vs 28z. In vivo, a single CD2z infusion induced deep and durable tumor regressions over the 60-day observation period in subcutaneous mesothelin-positive mesothelioma and orthotopic TnMUC1-positive pancreatic tumor models, achieving tumor control comparable to 28z and more rapid early tumor clearance than BBz, while supporting peripheral T cell persistence similar to BBz.

Conclusions: The CD2 cytoplasmic tail, in combination with CD3z, delivers balanced costimulation that couples brisk tumor debulking to T cell persistence. CD2z therefore may provide a simple, versatile alternative to canonical CD28 and 4-1BB modules for next-generation CAR T therapies.

The phase III ATOMIC trial recently showed that adding atezolizumab to standard adjuvant chemotherapy improved disease-free survival (DFS) in patients with mismatch-repair-deficient (dMMR) stage III colon cancer; however, its efficacy in mismatch-repair-proficient (pMMR) disease remains unknown. To address this issue, we reconstructed individual patient-level survival data (IPD) for the centrally confirmed dMMR subgroup, as well as those without centrally confirmed dMMR status (an approximation of the pMMR subgroup) in ATOMIC. The analysis of reconstructed IPD faithfully reproduced the trial's reported DFS gain by adding atezolizumab to adjuvant chemotherapy in the dMMR subgroup (3-year DFS, 86.2% vs 77.0%; HR, 0.52 (95% CI 0.36 to 0.77)). Intriguingly, DFS was also in favor of the chemo-immunotherapy arm in the approximated pMMR subgroup (3-year DFS, 87.1% vs 77.4%; HR, 0.45 (95% CI 0.16 to 1.27)). The interaction test further demonstrated that treatment effect did not differ by MMR status (P_interaction=0.808). These findings raise the possibility that adjuvant chemo-immunotherapy may confer benefit even in pMMR stage III colon cancer, which underscores the need for prospective validation in this population.

The recent study by Di Federico et al provides valuable insights into the intrapatient heterogeneity of programmed death-ligand 1 (PD-L1) expression and tumor mutational burden (TMB) in non-small cell lung cancer (NSCLC), and its potential impact on responses to immune checkpoint inhibitors. This commentary examines several biological factors that may contribute to such variability, including cytokine signaling, metabolic changes within the tumor microenvironment, and intrinsic tumor heterogeneity. We also consider possible interactions between PD-L1 and TMB in the context of immune evasion. Furthermore, we highlight the need for more rigorous patient stratification in future studies and suggest that dynamic monitoring tools like liquid biopsy could enhance clinical decision-making. A deeper understanding of biomarker variability mechanisms may ultimately support more precise and effective personalized immunotherapy strategies for NSCLC.