[This corrects the article DOI: 10.3389/fbioe.2023.1156951.].

Figure S1 shows the metabolic alterations induced by anti-PD-1 treatment in mice. Figure S2 presents the effects of itaconate on anti-PD-1 therapy in vivo. Figure S3 shows the impact of Acod1 deletion on the tumor immune microenvironment. Figure S4 presents the oxidative stress levels in DCs and TAMs within the tumor immune microenvironment. Figure S5 shows the efficiency of immune cell depletion in vivo. Figure S6 demonstrates that the impact of Acod1 deletion on anti-PD-1 therapy is dependent on CD8+ T cells. Figure S7 presents ACOD1 expression patterns in the tumor immune microenvironment. t-SNE projections and violin plots illustrate ACOD1 expression across myeloid and immune cell clusters in lung adenocarcinoma samples. Figure S8 shows the transcriptional regulation of Acod1 via the IFN-γ-JAK/STAT1 axis. Figure S9 presents the inhibitory effects of itaconate on DC antigen cross-presentation. Figure S10 shows the effects of itaconate on DC metabolism and signaling pathways.

Radiotherapy is known to trigger immunogenic cell death and activate local anti-tumor immune responses. However, its systemic immunomodulatory effects remain poorly understood. Here, we discovered that irradiated tumor cell-derived microparticles (RT-MPs) are released into the circulation and subsequently taken up by neutrophils in the spleen. The mitochondrial DNA contained within RT-MPs promotes the hyperactivation of neutrophils, leading to the secretion of interleukin-1beta (IL-1β) via the STING/NLRP3/GSDMD axis. IL-1β, in turn, enhances the antigen-presenting capacity of dendritic cells (DCs), which facilitates the formation of cytotoxic T lymphocytes (CTLs) in the spleen. These CTLs then contribute to the destruction of distant, non-irradiated tumors. Our findings provide valuable insights into the mechanisms by which radiotherapy can directly modulate systemic anti-tumor immunity, highlighting the potential for leveraging these effects to improve the efficacy of cancer treatment.

6102 Background: Neoadjuvant chemoimmunotherapy has shown high rates of pathological response in locally advanced head and neck squamous cell carcinoma (LA-HNSCC). However, its long-term survival outcomes, the predictive value of pathological complete response (pCR), and patterns of late failure remain unclear. Methods: In this single-arm phase II trial, 30 patients with resectable LA-HNSCC received three cycles of nab-paclitaxel (or docetaxel) plus cisplatin and camrelizumab, followed by surgery and adjuvant radiotherapy. The primary endpoint was pCR rate. With follow-up until January 2026, the median follow-up was 60 months. Analyses included overall survival (OS), recurrence patterns, and incidence of second primary tumors. Results: The pCR rate was 37.0% (10/27). Key Finding 1: This study represents the first report of 5-year long-term survival data for this regimen, with a 5-year OS rate of 76.7%. Key Finding 2: None of the patients who achieved pCR and completed adjuvant radiotherapy experienced local recurrence (0/7), whereas the recurrence rate was 33.3% (1/3) among those who did not receive radiotherapy, underscoring the role of radiotherapy even in pCR patients. Key Finding 3: Six patients (20%) developed second or third primary tumors during long-term follow-up (sites included larynx, esophagus, and lung), with incidence increasing over time. Key Finding 4: Two patients developed osteoradionecrosis of the jaw more than two years after radiotherapy, leading to malnutrition and subsequent death, highlighting the importance of long-term toxicity management. Conclusions: Neoadjuvant chemoimmunotherapy provides durable survival benefit in LA-HNSCC. pCR is associated with minimal local recurrence. However, with prolonged survival, the risk of second primary tumors becomes significant, and late radiation toxicities can impact outcomes. These findings emphasize the need for establishing lifelong surveillance protocols for multiple primary cancers and systematic management of late toxicities in this population. Clinical trial information: ChiCTR1900025303.

Responsive nanopore-based sensors have evolved to be powerful analytical tools in biosensing territory. Although taking advantages including label-free, low-cost, and analytical universality, the detection sensitivity is hampered by recognition efficiency of the responsive element in the nanoconfinement. Here, we show that improving the sensing performance for ATP molecules can be realized by a dual-site recognition strategy. We engineered a functionalized glass nanopipette using DNA grafted zeolitic iminazolate framework-90 (ZIF-90) as a dual-recognition nanoprobe. By virtue of the rigidity of MOF materials, ZIF-90 was shown to produce a remarkable pore-blockage effect and ion gating function. Particularly, the skeleton of ZIF-90 can be specifically induced to collapse by ATP molecules, resulting in remarkable signal-on ionic-current modulation. Collaborated with the aptamer units, the nanopipette allowed dual-site binding of ATP, thus greatly enhancing the sensing efficiency, with the linear response down to 0.1 nM ATP, compared to traditional single-site recognition mode. This method enabled us to directly probe the ATP concentration level in cells without the need of time-consuming and labor-intensive signal amplification strategies currently widely used. Reasonably, this nanopipette sensor should be promising to be a generalized sensing platform, provided that feasible responsive elements are used.

PURPOSE:We assessed the feasibility and efficacy of a response-adapted surgical strategy in patients with locally advanced head and neck squamous cell carcinoma (LA-HNSCC) achieving major tumor regression after neoadjuvant immunochemotherapy (NAICT). PATIENTS AND METHODS:This phase II, single-arm, prospective trial enrolled patients with resectable, human papillomavirus-negative stage III to IVA LA-HNSCC. All received three cycles of NAICT (penpulimab, nab-paclitaxel, and cisplatin). Pre-cycle 3 imaging and laryngoscopy guided surgery: Patients with ≥50% primary tumor regression underwent reduced-volume surgery (RVS); those with <50% regression received standard-volume surgery (SVS). All received risk-adapted adjuvant radiotherapy. The primary endpoint was 2-year disease-free survival (DFS). RESULTS:Fifty patients completed NAICT, with an objective response rate of 94% [95% confidence interval (CI), 84%-99%]. At a median follow-up of 34.0 months (IQR, 30.1-39.5), the 2-year DFS for the per-protocol cohort was 83% (95% CI, 69%-91%). The RVS group (n = 42) achieved a 2-year DFS of 90% (95% CI, 77%-96%) with 100% laryngeal preservation in laryngeal/hypopharyngeal cancers, whereas the SVS group (n = 5) had a 2-year DFS of 20% (95% CI, 0.8%-58%). Grade 3 adverse events (AE) occurred in 16% of patients related to NAICT and in 15% related to surgery; there were no grade 4 or 5 AEs. At 9 months after radiotherapy, most quality-of-life scores improved or stabilized compared with baseline. CONCLUSIONS:The REMATCH trial provides the first prospective evidence that a response-adapted surgical strategy after NAICT is feasible. Patients achieving ≥50% primary tumor regression derived promising survival and universal laryngeal preservation. These findings support the initiation of phase III randomized trials to validate this de-escalation strategy.

Elderly patients are underrepresented in clinical studies supporting platinum-based concurrent chemoradiotherapy (CCRT) as the standard therapy (ST) for locoregionally advanced nasopharyngeal carcinoma (LA-NPC). Many such patients in real-world settings cannot tolerate CCRT and instead receive individualized therapy (IT). This study aimed to compare the efficacy and safety of ST versus IT in elderly NPC patients. A retrospective analysis was conducted on 393 elderly (≥ 65 years) LA-NPC patients from two centers in China between January 2013 and December 2020. Patients were categorized into ST (platinum-based CCRT with or without induction/adjuvant therapy) or IT (radiotherapy alone, or concurrent radiotherapy with non-chemotherapy agents, with or without induction/adjuvant therapy) groups. Propensity score matching (PSM) generated 141 patient pairs. Survival outcomes were analyzed using the Kaplan-Meier method and Cox proportional hazards models. Toxicities were compared with chi-square or Fisher’s exact tests. In the PSM cohort, no significant differences were observed between the ST and IT groups in overall survival, cancer-specific survival, progression-free survival, locoregional relapse-free survival, and distant metastasis-free survival (all P > 0.05). Multivariable analysis confirmed that treatment group (ST vs. IT) was not a significant predictor for any survival endpoint. In exploratory stratified analysis, radiotherapy alone was associated with inferior OS, CSS, and PFS, whereas radiotherapy combined with systemic therapy showed survival outcomes similar to those of ST. Additionally, compared with the ST group, the IT group had significantly lower rates of grade 3 or higher hematologic toxicities, including leukopenia (8.5

BACKGROUND:Patients with unmethylated O6-methylguanine-DNA methyltransferase (MGMT) glioblastoma derive minimal benefit from the standard temozolomide chemoradiotherapy, leaving an urgent need for more effective therapies. The objective of this phase 2 study was to evaluate the efficacy of anlotinib plus radiotherapy in this disease. METHODS:In this single-arm, phase 2 study, patients with newly diagnosed, unmethylated MGMT glioblastoma were recruited after surgical resection. Patients received standard radiotherapy (60.0 grays) and concomitant with anlotinib (12 mg once daily, 2 weeks on/1 week off), followed by six cycles of anlotinib maintenance. The primary end point was overall survival (OS). Secondary end points were progression-free survival (PFS), OS rates at 12 and 24 months, PFS rates at 6 and 12 months, and safety. RESULTS:Between October 12, 2020, and August 1, 2022, 32 patients were enrolled and received protocol therapy, and all were evaluable for efficacy and safety analyses. At a median follow-up of 18.6 months (range, 8.3-50.3 months), the median OS was 19.1 months (95% confidence interval, 14.4-22.4 months), with 12-month and 24-month OS rates of 90.6% and 30.0%, respectively. The median OS did not reach the prespecified threshold of 20.1 months. The median PFS was 11.1 months (95% confidence interval, 9.6-12.9 months), with 6-month and 12-month PFS rates of 96.9% and 37.5%, respectively. Four patients (12.5%) experienced grade 3 or worse treatment-related adverse events, two of which (6.3%) were serious. No treatment-related deaths occurred. CONCLUSIONS:Postoperative anlotinib plus radiotherapy demonstrated clinically meaningful activity with a manageable safety profile in patients with unmethylated MGMT glioblastoma (Chinese Clinical Trials Registry identifier: ChiCTR2000040116).

Table S1 presents the sequences of primers used for RT-qPCR analysis. Table S2 lists the sequences of primers used for ChIP-PCR. Table S3 contains the differentially expressed metabolites identified in tumor tissues of Lewis tumor-bearing mice treated with IgG2 (Ctrl) or anti-PD-1 monoclonal antibody. Table S4 summarizes the differentially expressed genes in untreated and IFN-γ-treated bone marrow-derived macrophages (BMDMs). Table S5 provides the differentially expressed genes in bone marrow-derived dendritic cells (BMDCs) under IFN-γ versus IFN-γ + 4-OI treatment conditions.

BACKGROUND AND PURPOSE:To explore the efficacy and safety of neoadjuvant chemoradiotherapy (NCRT) plus sequential tislelizumab followed by surgery for esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS:This single-arm, bicentric, phase 2 trial enrolled patients with resectable or potentially resectable thoracic ESCC to receive neoadjuvant radiotherapy (41.4 Gy in 23 fractions) with concurrent chemotherapy (albumin-bound paclitaxel, 50-100 mg/m2, and carboplatin, area under the curve of 2 mg/ml/min, once weekly, five times) plus sequential tislelizumab (200 mg Q3W, three cycles) followed by surgery. The primary endpoint was pathologic complete response (pCR) rate. The secondary endpoints included safety, R0 resection rate, major pathologic response (MPR) rate, disease-free survival (DFS), and overall survival (OS). RESULTS:Of the 30 patients enrolled from January 2021 to October 2022, 24 (80.0 %) completed planned surgery and gained R0 resection (100 %). Among the 24 patients, nine (37.5 %) achieved pCR and 21 (87.5 %) achieved MPR. Ten patients (35.7 %) developed grade 3-4 toxicities during tislelizumab therapy, including lymphopenia (32.1 %), neutropenia (3.6 %), and thrombocytopenia (3.6 %). Grade 5 hematemesis occurred in two patients and both were attributed to aortic invasion. Three patients (12.5 %) developed grade 3 postoperative complications, including pulmonary infection (8.3 %) and hoarseness (4.2 %). After a median follow-up of 35.4 months, the 2-year OS and DFS rates were 83.3 % and 79.2 %, respectively. CONCLUSION:Sequential tislelizumab after NCRT in ESCC is safe and feasible. Further study is warranted to validate the efficacy of this combination mode.

Radioresistance is one of the important reasons for local recurrence and distant metastasis in non-small cell lung cancer (NSCLC). Itaconate primarily functions as an anti-inflammatory metabolite in macrophages, however, its role in radiotherapy remains to be explored. In this study, we demonstrated that radiation significantly increases itaconate in the tumor microenvironment (TME), which is produced by macrophages. Mechanistically, the NF-κB signaling pathway is rapidly activated in macrophages, which enhances the binding of P65 to the Acod1 promoter region, leading to significantly increased secretion of itaconate. Excessive itaconate alleviates oxidative stress of NSCLC cell lines by stabilizing NRF2 protein. Notably, specifically knocking out Acod1 on myeloid cells enhances the activation of the tumor immune microenvironment in response to radiotherapy, particularly increasing the infiltration and activation of CD8+ T cells. Therefore, we propose that targeting Acod1 could be an effective strategy to improve radiosensitivity in NSCLC.

Rationale: mRNA cancer vaccines show great promise for tumor therapy, but the therapeutic efficacy is limited. Metabolites play critical roles in immunomodulation. However, their role in mRNA cancer vaccines remains poorly understood. Methods: Metabolome analysis and single-cell RNA sequence were performed to explore the most important metabolite and its source cell. B16-F10-OVA-bearing wide-type and Irg1-depleted C57BL/6 mice were treated with OVA-LNP, OVA&si-Irg1-LNP, or anti-PD-1 antibody to evaluate therapeutic efficacy. Flow cytometry analysis was used to examine the immune cells within the lymph nodes, spleens, and the tumor immune environment. Results: We found that macrophage-derived itaconate was increased markedly in activated ipsilateral lymph nodes after ovalbumin-encoding mRNA-lipid nanoparticle (OVA-LNP) injection, compared to homeostatic contralateral lymph nodes. Depleting the immune-responsive gene 1(Irg1), which encodes the itaconate-production enzyme aconitate decarboxylase (ACOD1), in macrophages improved dendritic cell antigen presentation and enhances T cell function. Combining Irg1 knockdown via small interfering RNA (siRNA) with OVA mRNA in LNPs augmented the therapeutic efficacy of mRNA cancer vaccines, both as monotherapy and in combination with an anti-programmed cell death-1 antibody. Conclusions: Our findings reveal a link between itaconate and mRNA cancer vaccines, suggesting that targeting Irg1 via siRNA-LNP could be a promising strategy to improve the therapeutic efficacy of mRNA cancer vaccines.

Adaptive resistance to immunotherapy remains a significant challenge in cancer treatment. The reshaping of the tumor immune microenvironment in response to therapeutic pressures is a crucial factor contributing to this resistance. In this study, by comprehensive metabolic profiling of tumor tissues, we identified elevated itaconate in response to anti-PD-1 therapy as an adaptive resistance mechanism that promoted immune escape and tumor progression. CD8+ T-cell-derived IFNγ induced a significant upregulation of cis-aconitate decarboxylase 1 (ACOD1) in macrophages via the JAK-STAT1 pathway, thereby rewiring the Krebs cycle toward itaconate production. In murine models, macrophage-specific deletion of Acod1 increased the antitumor efficacy of anti-PD-1 therapy and improved survival. Additionally, itaconate and its derivative, 4-octyl itaconate, suppressed the tumor antigen presentation and cross-priming ability of dendritic cells, resulting in the impairment of antigen-specific T-cell antitumor responses. In summary, these findings identify an IFNγ-dependent immunometabolic mechanism of anti-PD-1 resistance, providing a promising strategy for combination therapy. Significance: Elevated itaconate production by macrophages induced by IFNγ is a critical negative feedback immunoregulatory metabolic response to anti-PD-1 immunotherapy that inhibits the cross-priming function of dendritic cells and confers immunotherapy resistance.

Globally, colorectal malignancy ranks among the most prevalent forms of cancer and stands as the third principal cause of cancer-associated mortality. Recent studies indicate that inflammatory processes play a significant role in the initiation and advancement of various malignancies, colorectal cancer included. It explores inflammatory biomarkers, with C-reactive protein (CRP) being a key focus. While CRP’s elevation during inflammation is linked to tumorigenesis, studies on its association with CRC risk are inconsistent, showing gender and methodological differences. Interleukin-6 (IL-6), TNF - α, and their receptors also play roles in CRC development, yet research findings vary. Adiponectin and leptin, secreted by adipocytes, have complex associations with CRC, with gender disparities noted. In terms of screening, non-invasive methods like fecal occult blood tests (FOBTs) are widely used, and combining biomarkers with iFOBT shows potential. Multi-omics techniques, including genomics and microbiomics, offer new avenues for CRC diagnosis. Overall, while evidence highlights the significance of inflammatory biomarkers in CRC risk prediction, larger prospective studies are urgently needed to clarify their roles due to existing inconsistencies and methodological limitations.

We identified antitumor effector B cells which demonstrated significant therapeutic efficacy upon adoptive transfer. We recently found that depletion of B cells prior to anti-PD-L1 administration reduced anti-PD-L1 efficacy and administration of anti-PD-L1recovered the humoral immune response, suggesting the involvement of host B cells in the anti-PD-L1 effect. We detected elevated expression of PD-1 on activated B cells, and higher PD-L1 expression on ALDHhigh cancer stem cells (CSCs). Co-culturing ALDHhigh CSCs with B cells significantly reduced IgG secretion, and such CSC-mediated B cell suppression was rescued by adding anti-PD-L1 mAb in a dose-dependent manner. To investigate if exosomes that express PD-L1 contribute to B cell suppression, we generated Rab27ako D5 cells to delete exosomes. D5-derived exosomes suppressed B cell proliferation and IgG production in vitro, and administration of these exosomes promoted tumor growth and reduced animal survival, revealing the role of tumor-derived exosomes in suppressing B cells. Rab27ako resulted in significantly enhanced host anti-tumor immunity by reducing tumor growth. Furthermore, anti-PD-L1 therapy significantly reduced Rab27ako tumor growth and prolonged animal survival vs. the WT tumor controls. We conclude that PD-L1-expresing CSCs and exosomes suppress antitumor effector B cells; targeting ALDHhigh CSCs and/or Rab27a-relevent exosomal PD-L1 may significantly augment the therapeutic efficacy of anti-tumor B cells. Longenbaugh Foundation Tumor Immunology: Cellular Responses and Tumor Microevironment (TIME)

Terminal exhaustion is a critical barrier to antitumor immunity. By integrating and analyzing single-cell RNA-sequencing and single-cell assay for transposase-accessible chromatin with sequencing data, we found that ETS variant 7 (ETV7) is indispensable for determining CD8+ T cell fate in tumors. ETV7 introduction drives T cell differentiation from memory to terminal exhaustion, limiting antiviral and antitumor efficacy in male mice. Mechanistically, ETV7 acts as a central transcriptional node by binding to specific memory genes and exhaustion genes and functionally skewing these transcriptional programs toward exhaustion. Clinically, ETV7 expression is negatively correlated with progression and responsiveness to immune checkpoint blockade in various human cancers. ETV7 depletion strongly enhances the antitumor efficacy of CD8+ T cells and engineered chimeric antigen receptor T cells in solid tumors. Thus, these findings demonstrate a decisive role for ETV7 in driving CD8+ T cell terminal exhaustion and reveal that ETV7 may be a promising target and biomarker for improving the efficacy of cancer immunotherapy. Jiang and colleagues identify ETV7 as a transcriptional node that skews CD8+ T cell transcriptional profiles toward exhaustion, consequently limiting antiviral and antitumor efficacy, and show that it can be targeted in CAR T cells to enhance efficacy.

mRNA cancer vaccines show great promise for tumor therapy, but their therapeutic efficacy is limited. Metabolites play critical roles in immunomodulation, however, their role in mRNA cancer vaccines remains unclear. Using metabolome analysis combined with single-cell RNA sequencing, we found that macrophage-derived itaconate increased markedly after ovalbumin-encoding mRNA-lipid nanoparticle (OVA-LNP) injection in activated ipsilateral lymph nodes compared to that in homeostatic contralateral lymph nodes. Depleting the immune-responsive gene 1(Irg1) in macrophages, which encodes the itaconate-production enzyme aconitate decarboxylase (ACOD1), improved the antigen presentation of dendritic cells and the functions of T cells. Knockdown of Irg1 using small interfering RNA (siRNA) combined with OVA mRNA in LNPs enhanced the therapeutic efficacy of mRNA vaccines alone or combined with an anti-programmed cell death-1 antibody. Taken together, our results reveal a link between itaconate and mRNA cancer vaccines, and suggest that targeting Irg1 via siRNA-LNP may be a promising strategy for improving the therapeutic efficacy of mRNA cancer vaccines. Chao Wan, Wenwen Wei, Xiao Yang, Yeshan Chen, Mengjie Che, You Qin, Qiao Li, Kunyu Yang. Targeting Irg1 expressed on macrophages in lymph nodes promotes response to mRNA cancer vaccines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 854.

Objective.Due to the magnetic hysteresis effect, it normally takes more time to switch the energy layer upward compared to downward in proton arc therapy (PAT), e.g. 5.5 s for energy layer switching upwards and 0.7 s for energy layer switching downwards. Therefore, previous studies have focused on reducing the number of energy jumps (NEJ) as a main direction. However, it is possible to reduce the energy layer switching time, for instance, by using magnetic field feedback in the energy layer switching system (ELSS) of the proton therapy system (PTS), especially for small energy jumps. Thus, we propose a new energy selection optimization strategy to improve the treatment delivery efficiency based on a fast ELSS, which allows energy jumps within a pre-defined threshold energy layer optimization (ELO-EJT).Approach.The ELO-EJT algorithm is based on two sequential steps: ELO and plan-quality re-optimization. The ELO problem was solved using the proximal forward-backward splitting method. The optimization problem of plan-quality re-optimization was solved by the alternating direction method of multipliers. Eight clinical cases were tested on the open-source treatment planning system matRad (German Cancer Research Center DKFZ), assessing plan quality and treatment delivery efficiency to evaluate the ELO-EJT algorithm.Main results.The ELO-EJT algorithm could achieve similar plan quality compared with the spot-scanning proton arc therapy _seq algorithm, which is based on the energy layer sorting and re-distribution mechanisms. Meanwhile, it reduces the beam delivery time without the need to limit energy jumps. More specifically, it reduces BDT by 23.93 ± 4.12% (from 123.95 ± 11.94 s to 94.14 ± 8.82 s).Significance.A new ELO strategy could effectively reduce the treatment delivery time by minimizing the NEJ exceeding a threshold NEJT, which paves the way for the clinical application of spot-scanning PAT.