Journal for Immunotherapy of Cancer最新文献

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.

Background: Cysteine-cysteine chemokine receptors 2 (CCR2) and 5 (CCR5) contribute to immune suppression in tumor microenvironments. CCR2 and CCR5 antagonists have demonstrated antitumor activity in pancreatic ductal adenocarcinoma (PDAC) and colorectal cancer (CRC), respectively. This phase 1b/2, open-label study evaluated BMS-813160, a CCR2/5 dual antagonist, in combination with chemotherapy±nivolumab in advanced PDAC or metastatic CRC.

Methods: Part 1 included patients with metastatic untreated (first-line (1L)) PDAC, 1L CRC, or previously treated (second or third line (2/3L)) microsatellite stable (MSS) CRC. Patients received 2 weeks of BMS-813160 monotherapy (300 mg two times a day, 600 mg once daily, 300 mg once daily, or 150 mg once daily) and then BMS-813160+chemotherapy (gemcitabine+nab-paclitaxel (gem/nabP; 1L PDAC), 5-fluorouracil+leucovorin+irinotecan (FOLFIRI; 1L CRC)), or nivolumab (2/3L MSS CRC).Part 2 included patients with metastatic 1L PDAC or 2L CRC. Patients received BMS-813160 300 mg two times a day+gem/nabP±nivolumab (1L PDAC), BMS-813160 300 mg two times a day or 150 mg once daily+FOLFIRI (2L CRC), or chemotherapy alone. Primary endpoints were safety and pharmacodynamics (Part 1) and efficacy (Part 2).

Results: In Part 1, 22 of 75 (29%) and 54 of 72 (72%) patients experienced a treatment-related adverse event during monotherapy lead-in and overall, respectively. Two dose-limiting toxicities (rash and pericardial effusion with pericarditis, both grade 3) occurred. In Part 2, patients with 1L PDAC who received BMS-813160 300 mg two times a day+gem/nabP+nivolumab achieved an overall response rate (ORR) of 37% (13/35); the median duration of response (DOR) was 45 weeks (95% CI 26.1 to not evaluable). ORRs with BMS-813160 300 mg two times a day+gem/nabP and gem/nabP alone were 26% (9/35) and 28% (9/32), respectively; median DORs were 121 and 31 weeks, respectively. Progression-free survival rates at 24 weeks were 56% (BMS-813160 300 mg two times a day+gem/nabP+nivolumab), 56% (BMS-813160 300 mg two times a day+gem/nabP), and 50% (gem/nabP). ORRs in 2L CRC were 19% (6/32; BMS-813160 300 mg two times a day+FOLFIRI), 13% (4/32; BMS-813160 150 mg once daily+FOLFIRI), and 27% (7/26; FOLFIRI).

Conclusions: In 1L PDAC, BMS-813160 300 two times a day+gem/nabP±nivolumab demonstrated durable antitumor response and was well tolerated. BMS-813160 combination regimens were tolerable in other cohorts, but clinical efficacy was not demonstrated.

Trial registration number: NCT03184870.

Background: Pancreatic ductal adenocarcinoma (PDAC) remains largely refractory to chimeric antigen receptor (CAR)-T cell therapy. Insufficient T cell infiltration, a highly immunosuppressive microenvironment, and antigen loss pose major challenges for CAR-T cell therapy.

Methods: We investigated therapeutic synergies of synthetic 5'-triphosphate RNA (3p-RNA), an agonist of the cytoplasmic double-stranded RNA sensor Retinoic Acid Inducible Gene I (RIG-I), and CAR-T cell therapy using syngeneic and human xenograft PDAC models. Tumor growth, chemokine secretion, immune-cell composition, CAR-T persistence, and endogenous T cell responses were assessed by flow cytometry, multiplex cytokine arrays, Enzyme-linked Immunospot (ELISpot), and vaccination-challenge.

Results: 3p-RNA provoked rapid type I interferon accompanied with chemokine ligand CCL5 and CXCL9/10/11 secretion, creating chemokine gradients that recruited chemokine receptor CCR5⁺/CXCR3⁺ CAR-T cells into tumors. RIG-I activation enhanced CAR-T cell proliferation, activity, and CAR-T persistence. Combination therapy eradicated established tumors in 60%-70% of mice, whereas either monotherapy was largely ineffective. Cured animals rejected CAR antigen-negative tumor cell rechallenge, demonstrating antigen-spreading and endogenous T cell responses.

Conclusions: Intratumoral RIG-I priming reprograms the PDAC microenvironment, transforming a non-responsive cancer into a CAR-T-permissive one, supporting durable, poly-antigenic immunity. These findings position 3p-RNA as a rapid, clinically tractable co-therapy to extend CAR-T efficacy to solid tumors.

Background: Canine oral malignant melanoma (OMM) is a highly aggressive tumor, with several available treatment options, though few achieve durable response or complete remission. Because of its biological similarity to human mucosal melanoma, canine OMM represents a valuable spontaneous model for translational immunotherapy studies. Anti-programmed cell death protein 1 (PD-1) antibody therapy has shown promise in canine OMM; however, predictive biomarkers for treatment response and survival have not been identified.

Methods: We conducted a multicenter, prospective, investigator-initiated clinical trial to evaluate the safety and efficacy of a caninized anti-canine PD-1 monoclonal antibody (ca-4F12-E6) in 150 dogs with advanced OMM. The dogs were administered 3 mg/kg ca-4F12-E6 intravenously every 2 weeks. Treatment response was assessed using cRECIST V.1.0. Biomarker analyses included peripheral blood parameters, cytokine/chemokines, peripheral lymphocyte subsets, microsatellite instability (MSI), immunohistochemistry for immune cell and mismatch repair protein markers, and RNA sequencing of the tumor tissue. Associations with clinical outcomes were determined by logistic regression and Cox proportional hazards models.

Results: The overall response rate was 14.7%, with a best overall response rate of 16.7%. Treatment-related adverse events occurred in 40.0% of the dogs, which were primarily grade 1-3. Increased baseline white blood cell, neutrophil count, and C reactive protein levels were significantly associated with poor response, shorter progression-free survival, and reduced overall survival (OS). MSI-high tumors were associated with significantly prolonged OS compared with MSI-low/microsatellite stable tumors. Transcriptome analysis revealed differentially expressed genes and enriched immune-related pathways in responders, though limited by sample size.

Conclusion: ca-4F12-E6 exhibited durable antitumor activity with manageable safety profile in dogs with OMM. Baseline systemic inflammatory markers and MSI status may serve as predictive biomarkers for clinical outcomes. The results support the use of canine OMM as a comparative model for human immuno-oncology and biomarker discovery.

Background: Oncolytic viruses are tumor-specific immunotherapeutic agents that exploit inherent features of the tumor microenvironment to replicate, spread, and kill cancer cells. The exchange protein activated by cAMP (EPAC) is a cell signaling protein that regulates pathways important for cell growth, survival, and migration, which are commonly associated with cancer progression, but are also very important for regulation of viral infectivity. EPAC antagonism has been explored as a broad-spectrum antiviral strategy, while selective EPAC activation with cAMP analogs has been found to increase virus replication and enhance therapeutic outcome of oncolytic virotherapy. However, systemic EPAC agonism bears risk of cardiovascular complications and may potentiate cancer progression.

Methods: A constitutively active construct of EPAC was encoded into an oncolytic vaccinia virus (VV) and screened using plaque assays, spheroid infections, and Transwell migration assays for its ability to enhance virus replication and spread. In vivo luminescence imaging, titering and immunohistochemical staining was used to measure virus dissemination in primary injected tumors and to track their spread to distal untreated tumors. The impact of the VV-EPAC virus on the immune landscape of MC38 tumors was investigated by flow cytometry, ELISPOTs and cytokine ELISAs, while its overall therapeutic efficacy was explored in MC38, CT26LacZ, and B16F10 models. Combinational synergy was also tested with capecitabine and oxaliplatin chemotherapy, as well as with partial surgical resection.

Results: The EPAC-expressing virus exhibited an increase in migrative ability both in cell culture and in vivo, due in part to remodeling of the actin cytoskeleton leading to intercellular nanotube-like structures and enhanced syncytia formation. It reduced tumor burden and increased survival in multiple colorectal cancer models and reshaped the tumor microenvironment by inducing angiogenesis and recruiting CD8+T cells. The EPAC-expressing virus also synergized with conventional chemotherapy and exhibited a remarkable therapeutic benefit when used together with surgical resection to treat a metastatic melanoma model. Despite the noted benefits that EPAC offers to virus and cancer growth, no significant increase in off-target replication, cytotoxicity, or disease progression was observed.

Conclusions: Altogether, the encoding of cellular signaling proteins into oncolytic viruses that modulate the intracellular and extracellular environments of tumors to create conditions favorable for virus replication and dissemination appears as a promising strategy to treat tumors and synergize with other conventional cancer therapies.

Background: Elevated levels of SPP1⁺ tumor-associated macrophages (TAMs) are associated with reduced CD8⁺ T cell infiltration and poorer prognosis in cancer patients, but direct evidence demonstrating a causal role for SPP1⁺ TAMs in excluding CD8⁺ T cells is still missing. The precise mechanisms by which SPP1-activated signaling pathways and macrophage-derived factors regulate CD8⁺ T cell trafficking remain poorly understood.

Methods: We established multiple tumor mouse models to study the function of macrophage SPP1 in the tumor environment, especially its role in the relationship between macrophages and CD8 T cells. We combined the single-cell (sc) RNA sequencing data of clinical tumor samples and tumor tissues from Spp1^{fl/fl-Lyz2-Cre} mice to identify the differences in SPP1-related genes and found that SPP1 could regulate the expression of CXCL9 and CXCL10 in macrophages. Through Western blotting, immunofluorescence staining, and flow cytometry analyses, we elucidated the mechanistic basis by which macrophage-specific SPP1 deficiency suppressed tumorigenesis.

Results: This study demonstrated that macrophage-derived SPP1 played a crucial role in suppressing CD8 T cell infiltration, promoting tumor progression, and diminishing the effectiveness of immune checkpoint inhibitor (ICI) therapy. Sc-RNA sequencing analysis revealed a marked increase in CD8 T cell populations within tumor tissues of Spp1^{fl/fl-Lyz2-Cre} mice. Furthermore, a negative correlation was observed between CD8 T cells and SPP1 macrophages in human colorectal cancer specimens. Genetic deletion of SPP1 in macrophages markedly enhanced tumor growth suppression in a manner dependent on CD8 T cell-mediated immunity. Mechanistically, SPP1 deficiency in macrophages led to elevated mitochondrial reactive oxygen species (ROS) production, resulting in the accumulation of cytosolic double-stranded DNA (dsDNA) fragments. This accumulated dsDNA activated the cGAS-STING pathway, leading to subsequent STAT1 phosphorylation. The enhanced STAT1 activity upregulated the expression of chemokines CXCL9 and CXCL10, thereby facilitating CD8 T cell recruitment into the tumor microenvironment.

Conclusions: Deletion of SPP1 in TAMs upregulates CXCL9/10 production by activating the ROS-DNA fragment/cGAS-STING/STAT1 pathway, thereby enhancing CD8 T cell infiltration, inhibiting tumor progression, and improving ICI treatment outcomes in tumors.