bims-tuinly 2026-02-01 papers

bims-tuinly

Biomed News

on Tumor-infiltrating lymphocytes therapy

Issue of 2026–02–01
twenty-two papers selected by
Pierpaolo Ginefra, Ludwig Institute for Cancer Research

Non-Genetically Modified Adoptive Cell Therapies for Solid Tumors: Current Landscape and Future Challenges.
Tanshinone IIA&Icaritin -MPs regulated vascular normalization and restored tumor-infiltrating T lymphocyte function to boost anti-PD-1 therapy in melanoma lung metastasis.
Discrepant CD3+ TILs in PD-L1-Negative NSCLC: Favorable Outcome in an Elderly Patient Treated With Nivolumab, Ipilimumab, and Chemotherapy.
FOSL2 regulates PD-L1 and modulates hormone therapy response heterogeneity.
Bispecific T-Cell Engagers, Cell Therapies, and Other Non-Checkpoint Immunotherapies for Metastatic Uveal Melanoma: A Narrative Review.
Terminally exhausted CD8+ T cells in solid tumors: biology, biomarker potential and translational tools for precision oncology.
Spatial Profiling and Prognostic Role of Tumor-Infiltrating CD8+ T and CD20+ B Cells in Metastatic Clear Cell Renal Cell Carcinoma Treated with Sequential Tyrosine Kinase Inhibitors and Nivolumab.
CRISPR screens identify targets to rescue age-related T cell dysfunction in cancer.
CAR-T cell immunotherapy of malignant melanoma.
Decorin facilitates T cell-mediated antitumor immunity and augments the efficacy of anti-PD1 immunotherapy.
CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma.
Prediction of tumor-infiltrating lymphocytes through habitat radiomics and exploration of response mechanisms in neoadjuvant immunochemotherapy-treated lung cancer.
Autocrine activity of engineered IL-33 mRNA enhances adoptive T-cell therapy for peritoneal carcinomatosis and synergizes with IL-12 mRNA.
Integration of the GRIm Score with Pathologic Immune and Stromal Markers to Develop a Combined Prognostic Model in Gastric Cancer: A Retrospective Single-Center Study.
Neoadjuvant ipilimumab plus nivolumab in melanoma: 5-year survival and biomarker analysis from the phase 2 PRADO-trial.
Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects.
DUSP22 dephosphorylates LGALS1 to enhance T cell-driven antitumor immunity.
GM-CSF Armed Oncolytic Adenovirus Enhances T-Cell Infiltration and Suppresses Local and Distal Tumor Growth.
Enhancing oncolytic virotherapy with a tri-specific T-cell engager targeting CD3ε, EpCaM, and 4-1BB: preclinical evaluation and implications for cancer immunotherapy.
Association of PD-L1 and PD-1 Expression With Clinicopathological Characteristics and Prognosis in Upper Tract Urothelial Carcinoma.
In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab.
A ligand-centered framework for γδ T cell activation in colorectal cancer revealed by single-cell and transformer-based perturbation.

Cancer Immunol Immunother. 2026 Jan 27. 75(2): 40

Non-Genetically Modified Adoptive Cell Therapies for Solid Tumors: Current Landscape and Future Challenges.

Qinghuizi Luo, Wendi Jiang, Yiyang Xie, Zhe Dai, Jialong Zhu, Yuanyang Du, Xiaoyue Tao, Zengjie Lei, Xiaoyuan Chu, Bingji Wang, Gongbo Fu.

  Non-genetically modified adoptive cell therapies (ACTs) represent a rapidly advancing frontier in solid tumor immunotherapy, offering a safe and adaptable alternative to genetically engineered approaches by capitalizing on the intrinsic plasticity of immune cells. Genetic engineering strategies, including CAR-T cells, encounter significant obstacles in solid tumors, including on-target off-tumor toxicity, an immunosuppressive tumor microenvironment, and drug resistance. Although non-genetically modified ACTs-including tumor-infiltrating lymphocytes (TILs), cytokine-induced killer (CIK) cells, natural killer (NK) cells, and γδ T cells-offer unique advantages, their clinical application remains underexplored. This review consolidates the mechanistic basis, clinical progress, and limitations of non-genetically modified ACTs, proposing a paradigm shift toward combinatorial strategies. We systematically assessed how TILs overcome tumor microenvironment (TME) inhibition through lymphodepletion and cytokine assistance, compared the histocompatibility complex-unrestricted cytotoxicity of CIK cells with their functional diversity, and emphasized the innate flexibility of NK/γδ T cells against antigen-loss variants. By integrating preclinical and clinical data, we identify critical challenges: in vitro expansion inefficiency, absence of standardized protocols, and dynamic TME interactions. Furthermore, we advocated patient stratification by biomarkers, the addition of optimized cytokines, and rational combinations with checkpoint inhibitors or metabolic modulators to enhance efficacy. This review outlines the current landscape and proposes actionable solutions to reconcile the disparity between experimental potential and clinical applicability in non-genetically modified ACT.

Keywords:  Adoptive cell therapy; Non-genetically modified; Solid tumor treatment; Tumor microenvironment; Tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.1007/s00262-025-04274-y
J Nanobiotechnology. 2026 Jan 27.

Tanshinone IIA&Icaritin -MPs regulated vascular normalization and restored tumor-infiltrating T lymphocyte function to boost anti-PD-1 therapy in melanoma lung metastasis.

Xiaoyu Che, Yaqin Li, Wenjing Chen, Xinyu Yang, Hong Wang, Shan Deng, Xiaoqi Li, Xiaoying Qin, Yan Chen, Yuping Liu.

   BACKGROUND: PD-1 inhibitors are a promising treatment for melanoma, but over 50% of patients with metastatic melanoma do not respond well. This limited efficacy is partly due to the aberrant vascular structure and immunosuppressive microenvironment in metastatic lung tissue.
METHODS: We developed an extracellular vesicle-based delivery system Tanshinone IIA & Icaritin -MPs (TSA&ICT-MPs) that targets lung metastases. In vivo in vitro models, cell experiments, immunofluorescence, immunohistochemistry, flow cytometry, and mass spectrometry flow cytometry were used to validate the efficacy of TSA&ICT-MPs in promoting vascular normalization, enhancing the activity of tumor-infiltrating lymphocytes (TILs), and reducing myeloid-derived inhibitory cell (MDSC) infiltration by modulating the adenosine metabolic pathway.
RESULTS: TSA&ICT-MP contributes to vascular normalization by modulating ELTD1, thereby enhancing TIL infiltration, and reduces adenosine release by targeting ENPP1, thus enhancing anti-tumor immunity. Combining TSA&ICT-MP with α-PD-1 achieved a 70.33% suppression rate of lung metastasis and prolonged survival in murine models. This approach offers a promising strategy to enhance the efficacy of melanoma immunotherapy.

Keywords:  Combination therapy; Immunosuppressive microenvironment; Metastasis; Microparticles; PD-1 inhibitors; Vascular normalization

DOI:  https://doi.org/10.1186/s12951-026-04042-9
Am J Case Rep. 2026 Jan 27. 27 e951075

Discrepant CD3+ TILs in PD-L1-Negative NSCLC: Favorable Outcome in an Elderly Patient Treated With Nivolumab, Ipilimumab, and Chemotherapy.

Mataichi Sekiya, Munehide Nakatsugawa, Nobuyuki Koyama, Naohiro Kajiwara, Takuya Aoki.

BACKGROUND Immune-checkpoint inhibitors (ICIs) targeting programmed cell death-1 (PD-1), its ligand PD-L1, and cytotoxic T lymphocyte antigen-4 (CTLA-4) have revolutionized the treatment landscape of non-small cell lung cancer (NSCLC). These agents restore antitumor immunity by reactivating suppressed T cells. Although PD-L1 expression is widely used as a predictive biomarker, responses to ICIs can occur even in tumors lacking PD-L1 expression, underscoring the complexity of the tumor immune microenvironment. Ongoing research on the tumor microenvironment aims to achieve a better understanding of cancer progression mechanisms and to improve the assessment of therapeutic efficacy. CASE REPORT We present an 80-year-old man with advanced NSCLC, without any remarkable past medical history, clinically staged as IVB (cT4N3M1c), and demonstrating a PD-L1 tumor proportion score (TPS) of less than 1%. Despite this, he exhibited an excellent response to combination therapy with anti-PD-1, anti-CTLA-4 monoclonal antibodies, and cytotoxic chemotherapy during hospitalization, with manageable adverse events. Notably, pathological analysis revealed marked infiltration of CD3-positive tumor-infiltrating lymphocytes (TILs), averaging 1100/mm². CD4- and CD8-positive TILs were present in equal numbers, suggesting a balanced population of helper and cytotoxic T cells. The patient received a total of 24 cycles of immunotherapy before disease progression was confirmed. CONCLUSIONS This case highlights a striking dissociation between TIL density and PD-L1 expression, suggesting that CD3-positive TILs may reflect underlying immune activity not captured by PD-L1 status alone. Our findings emphasize the need to further explore TIL profiling as a complementary biomarker, particularly in patients treated with anti-PD-1/anti-CTLA-4-containing regimens.

DOI: https://doi.org/10.12659/AJCR.951075
Mol Cancer Res. 2026 Jan 30.

FOSL2 regulates PD-L1 and modulates hormone therapy response heterogeneity.

Feifei Sun, Baozhen Wang, Shuyu Mei, Lin Zhang, Xinpei Wang, Hui Liu, Wenyao Liu, Jiajia Wang, Qianqian Zhou, Bo Han, Jing Hu, Lin Gao, Xueli Wang.

The response to androgen-targeted therapy in prostate cancer (PCa) is highly heterogeneous. While previous studies have primarily concentrated on tumor cell-intrinsic signaling changes, the tumor microenvironment, particularly the interactions between tumor-infiltrating lymphocytes (TILs) and tumor cells, is equally critical in shaping treatment responses. Building on our previous observations linking TILs to treatment efficacy in the context of neoadjuvant androgen deprivation therapy (NADT), we employed publicly available clinical datasets, in vitro T cell-PCa cell co-culture systems, and murine xenograft models to investigate this interplay. We found treatment-related dynamic change in TILs populations, accompanied by a concordant expression pattern of FOSL2 and PD-L1. Mechanistically, FOSL2 directly bound to the PD-L1 promoter to transcriptionally upregulate PD-L1, thereby modulating T cell infiltration and function. Importantly, in vivo results demonstrated that targeting FOSL2 enhanced the antitumor effect when it combined with hormone therapy and anti-PD-L1 treatment. These findings suggest that FOSL2 may contribute to treatment response heterogeneity by shaping the tumor immune microenvironment, offering novel insights into resistance mechanisms and uncovering potential strategies to enhance the efficacy of hormone therapy in PCa. Implications: Targeting FOSL2-mediated PD-L1 regulation offers a promising strategy to overcome immune microenvironment-mediated resistance and improve the therapeutic efficacy of androgen-targeted therapy in PCa.

DOI: https://doi.org/10.1158/1541-7786.MCR-25-0611
J Clin Med. 2026 Jan 13. pii: 641. [Epub ahead of print]15(2):

Bispecific T-Cell Engagers, Cell Therapies, and Other Non-Checkpoint Immunotherapies for Metastatic Uveal Melanoma: A Narrative Review.

Jakub Kleinrok, Weronika Pająk, Joanna Pec, Kamil Rusztyn, Joanna Dolar-Szczasny, Alicja Forma, Grzegorz Teresiński, Jacek Baj.

  Metastatic uveal melanoma (MUM) remains largely refractory to immune-checkpoint inhibition, so recent research has turned to bispecific T-cell engagers (BTCEs), adoptive-cell therapies (ACTs), and oncolytic viruses (OVs). To summarize the available clinical evidence, we performed a structured literature search across PubMed, Scopus, and Europe PMC for primary studies published between 1 January 2010 and 31 May 2025 that enrolled at least three adults with MUM, treated with one of these modalities, and that reported efficacy or grade-3+ safety outcomes; two reviewers independently performed screening, data extraction, and risk-of-bias assessment, and because of notable heterogeneity, we synthesized the findings narratively. Twenty-two studies met the criteria-thirteen phase I-III trials, eight observational cohorts, and one case series-covering fifteen BTCE cohorts, four ACT cohorts, and three OV cohorts. Tebentafusp, the dominant BTCE evaluated in roughly 1150 HLA-A*02:01-positive patients, extended median overall survival from 16.0 to 21.7 months (hazard ratio 0.51, with three-year follow-up HR 0.68) in its pivotal phase-III trial despite objective response rates of only 5-12%, with early skin rash and week-12 circulating-tumor-DNA clearance emerging as consistent markers of benefit. Tumor-infiltrating lymphocyte therapy, administered to about thirty patients, produced objective responses in 11-35% and occasional durable complete remissions, although median progression-free survival remained 2-6 months and severe cytopenias were universal. Three early-phase OV studies, totaling twenty-nine patients, yielded no radiographic responses but showed tumor-specific T-cell expansion and transient disease stabilization. Safety profiles reflected the mechanism of action: tebentafusp most often caused rash, pyrexia, and usually manageable cytokine-release syndrome with grade-3+ events in 40-70% yet discontinuation in roughly 2%; TIL therapy toxicity was driven by lymphodepleting chemotherapy and high-dose interleukin-2 with one treatment-related death; and OVs were generally well tolerated with no more than 20% grade-3 events.

Keywords:  HLA-A*02:01; adoptive cell therapy; bispecific T-cell engager; immunotherapy; metastatic uveal melanoma; oncolytic virus; tebentafusp; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3390/jcm15020641
Front Immunol. 2025 ;16 1709852

Terminally exhausted CD8+ T cells in solid tumors: biology, biomarker potential and translational tools for precision oncology.

Xuejun Guo, Shuhan Ma, Jingwen Wang, Yilin Fu, Wenxue Ma.

  Terminally exhausted CD8+ T cells (Ttex) are emerging as clinically relevant immune subsets across solid tumors, marked by sustained inhibitory receptor expression, loss of TCF1, and limited proliferative capacity. Once considered functionally inert, Ttex are now recognized for their residual cytotoxic potential and strong associations with tumor immunogenicity, including microsatellite instability (MSI), high tumor mutational burden (TMB), and neoantigen load. Importantly, the prognostic significance of Ttex is highly tumor-context-dependent, shaped by stromal architecture, mutational burden, and progenitor Tpex availability. This review examines the biology, spatial localization, and prognostic value of Ttex, highlighting the Ttex/CD8+ ratio as a promising biomarker in cancers such as colorectal, lung, and esophageal carcinoma. We summarize recent advances in multiplex imaging, digital pathology, and AI-driven quantification that support the clinical integration of Ttex assessment. In addition, we discuss emerging therapeutic strategies targeting Ttex through immune checkpoint combinations, thymocyte selection-associated high mobility group box protein (TOX) and circRNA-mediated reprogramming, and exhaustion-resistant T cell engineering. Finally, we outline translational priorities including assay harmonization, functional validation, and longitudinal profiling to advance Ttex-based precision oncology.

Keywords:  T cell exhaustion; digital pathology; immunotherapy biomarkers; precision oncology; terminally exhausted CD8⁺ T cells; tumor microenvironment

DOI:  https://doi.org/10.3389/fimmu.2025.1709852
J Cancer. 2026 ;17(2): 372-381

Spatial Profiling and Prognostic Role of Tumor-Infiltrating CD8+ T and CD20+ B Cells in Metastatic Clear Cell Renal Cell Carcinoma Treated with Sequential Tyrosine Kinase Inhibitors and Nivolumab.

Andriy Trailin, Lenka Červenková, Petr Hošek, Kristýna Pivovarčíková, Michaela Tkadlecová, Petr Stránský, Kari Hemminki, Ondřej Fiala.

  Background: Tumor-infiltrating lymphocytes (TILs) are known to influence disease progression and treatment response in clear cell renal cell carcinoma. This study aimed at evaluating the prognostic and predictive relevance of T and B cell infiltration patterns in patients with metastatic clear cell renal cell carcinoma (mRCC-cc) treated sequentially with tyrosine kinase inhibitors (TKIs) and the immune checkpoint inhibitor nivolumab. Methods: In this retrospective cohort study, immune cell densities (CD3+, CD8+ T cells and CD20+ B cells) were analyzed by immunohistochemistry and quantified using digital image analysis software QuPath in distinct tumor regions of primary tumor: tumor center (TC), inner margin (IM), outer margin (OM), and peritumoral (PT) region. Samples were obtained from 36 patients with mRCC-cc treated with TKIs in the first line and sequentially with nivolumab in the second or third-line setting. Associations between immune cell densities, clinicopathological features, and survival outcomes were assessed using univariable and multivariable Cox regression models. Progression-free survival (PFS), overall survival (OS), and objective response rate (ORR) were evaluated. Results: Densities of all immune cells were significantly higher in the OM and PT regions than in the TC and IM. Older age correlated with lower CD8+ T cell and CD20+ B cell densities, whereas higher tumor grade was associated with increased CD20+ B cell infiltration in IM. High CD20+ B cell density in IM and OM was significantly associated with shorter PFS during first-line TKI therapy (hazard ratio (HR) = 3.30, P = 0.015 and HR = 3.25, P = 0.016, respectively). In contrast, an intermediate CD8+ T cell density in the PT region was associated with longer PFS during sequential nivolumab treatment (HR = 0.26, P = 0.007). No significant associations between immune cell densities and ORR or OS were observed. Conclusions: Our findings suggest that spatial localization and density of tumor-infiltrating CD20+ B cells are potential predictors of poor PFS on TKIs, whereas higher CD8+ T cell infiltration in peritumoral areas may be a potential predictor of prolonged PFS on nivolumab. These immune-cell-based parameters may refine prognostic models and help guide treatment selection in mRCC-cc.

Keywords:  immune checkpoint inhibitors; metastatic renal cell carcinoma; progression-free survival; tumor-infiltrating lymphocytes; tyrosine kinase inhibitors

DOI:  https://doi.org/10.7150/jca.125509
bioRxiv. 2026 Jan 24. pii: 2026.01.22.701075. [Epub ahead of print]

CRISPR screens identify targets to rescue age-related T cell dysfunction in cancer.

Alex C Y Chen, Keely Y Ji, Cansu Yerinde, Nelson H Knudsen, Kevin Bi, Daniela Martinez, Thomas J Carmona-LaSalle, Kazuhiro Taguchi, Katherine H Xu, Elizabeth M Seider, Marc A Schwartz, Maria Zschummel, Linda T Nieman, Kathleen B Yates, Thorsten R Mempel, Robert T Manguso, Nir Hacohen, Debattama R Sen.

Immune aging is being increasingly recognized as a critical barrier to effective cancer immunotherapy, as the aged tumor microenvironment (TME) drives T cell dysfunction and impairs immune control of cancer. However, the key molecular drivers of this process as well as potential targets to rescue T cell dysfunction in aged tumors remain incompletely understood. Therefore, we performed in vivo single-cell CRISPR screens in CD8 + T cells within aged tumors and tumor-draining lymph nodes (tdLNs). We identified Dusp5 and Zfp219 as key regulators of T cell persistence and effector differentiation in aged hosts. Loss of Dusp5 , a negative regulator of ERK signaling, increased ERK1/2 phosphorylation and enhanced T cell proliferation in both young and aged tumors. In contrast, loss of Zfp219 , a transcriptional repressor, induced epigenetic reprogramming of cytotoxic gene programs, thereby increasing granzyme secretion and enhancing antitumor immunity. Moreover, expression of the human ortholog gene ZNF219 is increased within intratumoral CD8 + T cells in older cancer patients. High ZNF219 expression correlates with poorer survival following immune checkpoint blockade (ICB) and reduces persistence of human intratumoral T cells. Notably, Zfp219 ablation synergized with anti-PD-1 blockade in mice to expand effector-like CD8 + T cells, leading to significantly enhanced anti-tumor immunity and tumor clearance in aged hosts. Together, these findings highlight Dusp5 and Zfp219 as critical drivers of age-related T cell dysfunction and as potential therapeutic targets to rejuvenate T cell antitumor immunity in older cancer patients.

DOI: https://doi.org/10.64898/2026.01.22.701075
Expert Rev Clin Immunol. 2026 Jan 27.

CAR-T cell immunotherapy of malignant melanoma.

Olha Kharasakhal, John Maher.

   INTRODUCTION: Management of inoperable and advanced malignant melanoma has been transformed in recent years by the advent of a number of approaches, including immune checkpoint blockade, small molecule inhibitors and adoptive immunotherapy with ex vivo expanded tumor-infiltrating lymphocytes.
AREAS COVERED: In this review, we describe efforts made to develop an alternative immunotherapeutic approach for this disease using chimeric antigen receptor (CAR) engineered T-cells. Literature was reviewed in the PubMed database and clinicaltrials.gov website (1998-2025).
EXPERT OPINION: CAR T-cell immunotherapy has proven transformative in the treatment of selected hematological malignancies. However, solid tumors such as melanoma remain much more challenging to treat using this emerging modality. Here we consider issues surrounding target selection, encompassing both tumor cells and accompanying stroma, in addition to armoring approaches that may potentiate delivery to or efficacy within the tumor microenvironment. We also consider a number of advanced CAR-based architectures to enable multi-antigen or universal antigen targeting and combination-based approaches.

Keywords:  CAR armoring; CAR combination therapies; CAR macrophages; CAR manufacture; Chimeric antigen receptor (CAR); NK cells; NKT-cells; cancer immunotherapy; dual targeting; gd T-cells; malignant melanoma; target selection; tumor microenvironment (TME); universal CAR

DOI:  https://doi.org/10.1080/1744666X.2026.2621811
Cancer Immunol Immunother. 2026 Jan 27. 75(2): 45

Decorin facilitates T cell-mediated antitumor immunity and augments the efficacy of anti-PD1 immunotherapy.

Ningqian Zheng, Lvzhu Xiang, Guiqin Xu, Yun Liu, Chen Xu, Li Zhang, You Zuo, Zhiqian Ye, Yongzhong Liu, Zhaojuan Yang.

   BACKGROUND: Decorin (DCN) predominantly produced by fibroblasts is a small leucine-rich proteoglycan with tumor-suppressive property. However, whether DCN has a role in shaping the tumor immune microenvironment remains elusive.
METHODS: The TCGA and GEO databases were analyzed to identify fibroblast-specific secretory proteins that are downregulated in most types of human tumors, positively correlate with CD8⁺ T cell infiltration, and associate with improved response to immune checkpoint blockade (ICB) therapy. The function of DCN in vivo was assessed using cell lines with stable DCN overexpression in both immunocompetent and immunodeficient mice. The changes in the composition and function of immune cell subpopulations in tumors were analyzed by flow cytometry analysis (FCM) and immunofluorescence staining. The role of CD8⁺ T cells in the DCN-mediated tumor suppression was further elucidated by utilizing B2m-knockout tumor cells and CD8⁺ T cell depletion assays. The in vitro co-culture system of tumor cells and T cells was applied to dissect the effects of DCN on CD8+ T lymphocyte activation and functions. Finally, the therapeutic efficacy of DCN in combination with anti-PD1 antibody was evaluated in mouse tumor model.
RESULTS: DCN-mediated tumor suppression was present in immunocompetent mice, wherein either depletion of CD8⁺ T cells in mice or ablation of β2M in tumor cells abrogated the tumor-inhibitory effects mediated by DCN, indicating the importance of CD8⁺ T cells in the DCN-antitumor activities. DCN overexpression promoted CD8⁺ T cell infiltration into tumors and increased the production of TNF-α, IFN-γ, and perforin in infiltrating T cells, and DCN expression substantially enhanced the tumor-suppressive efficacy of anti-PD1 therapy. More importantly, integrative analyses of clinical data indicated that DCN expression is downregulated in multiple types of human tumors and positively associated with the presence of CD8⁺ T cells and their expression of cytotoxic genes. Furthermore, high DCN levels were correlated with favorable prognosis in certain types of cancer patients with ICB therapy.
CONCLUSIONS: Our study reveals that DCN functions as a tumor-suppressive factor by enhancing T cell response, and proposes the potential of exogenous DCN as a supplement to cancer immunotherapy.

Keywords:  Antitumor; CD8+ T cells; Decorin; Immunotherapy; Tumor microenvironment

DOI:  https://doi.org/10.1007/s00262-025-04290-y
J Nanobiotechnology. 2026 Jan 29.

CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes delivering C646 reverse CD8⁺ T Cell exhaustion via H3K18la histone delactylation in gastric cardia adenocarcinoma.

Zheng Xiang, Xinxin Zhang, Xinlei Liu, Xiaohu Lv, Yubo Hu, Chi Zhang, Chunlei Zou, Anqi Wang, Bo Zhang, Wei Wang, Guodong Cao, Jianguang Jia.

  The functional exhaustion of CD8+ T cells in the tumor microenvironment (TME) severely limits anti-tumor immunity in gastric cardia adenocarcinoma (GCA). Here, we developed CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes (CD8a-NVEs) encapsulating the p300 inhibitor C646 to selectively target and reprogram exhausted CD8+ T cells. Single-cell RNA sequencing of human GCA tissues revealed lactate-driven epigenetic remodeling, characterized by elevated H3K18 lactylation (H3K18la) at the PDCD1 promoter, which correlated with impaired CD8⁺ T cell function. In vitro, C646 effectively reduced H3K18la, suppressed PDCD1 transcription, and restored effector molecule expression, including IFN-γ and GZMB. CD8a-NVEs@C646 exhibited superior targeting specificity, biocompatibility, and functional efficacy, markedly enhancing CD8⁺ T cell proliferation and cytotoxicity compared with free C646. In a humanized orthotopic GCA model, CD8a-NVEs@C646 significantly inhibited tumor growth, and its combination with anti-PD-1 therapy further enhanced T cell infiltration and tumor apoptosis. This biomimetic nanoplatform enables precise epigenetic reprogramming of tumor-infiltrating CD8⁺ T cells, overcoming lactate-induced histone modifications and reversing exhaustion. Collectively, these findings present a translational nanobiotechnology-based strategy to potentiate immunotherapy efficacy in GCA and potentially other malignancies driven by T cell dysfunction.

Keywords:  C646; Gastric cardia adenocarcinoma; H3K18la; Programmed cell death protein 1; Red blood cell membrane-derived ectosomes

DOI:  https://doi.org/10.1186/s12951-025-03957-z
Cancer Immunol Immunother. 2026 Jan 27. 75(2): 42

Prediction of tumor-infiltrating lymphocytes through habitat radiomics and exploration of response mechanisms in neoadjuvant immunochemotherapy-treated lung cancer.

Zuhan Geng, Yihong Hu, Shunchen Zhou, Guangyao Wu, Zhenyu Gong, Ruiping Feng, Zhangfeng Huang, Peiyuan Mei, Kuo Li, Guanchao Ye, Yongde Liao.

   BACKGROUND: Neoadjuvant immunochemotherapy (NAIC) induces tumor microenvironment remodeling in non-small cell lung cancer (NSCLC), presenting challenges for treatment response assessment. This study developed and validated a habitat radiomics approach for non-invasive prediction of tumor-infiltrating lymphocyte (TIL) status to evaluate NAIC response in NSCLC.
METHODS: This retrospective study enrolled 238 NSCLC patients following NAIC for clinical analysis, of which 201 patients met criteria for radiomics analysis. Patients were classified into TIL-positive and TIL-negative groups based on pathological assessment. Post-treatment computed tomography (CT) images were analyzed using K-means clustering to identify tumor habitat sub-regions for radiomic feature extraction. Seven machine learning algorithms were evaluated for TIL status prediction. Model interpretability was assessed through SHapley Additive exPlanations (SHAP) analysis. Single-cell RNA sequencing (scRNA-seq) data were analyzed to compare major pathological response (MPR) and non-MPR tumor microenvironments through cell type annotation, differentiation trajectory analysis, and intercellular communication network analysis.
RESULTS: Pre-treatment neutrophil-to-lymphocyte ratio (NLR) showed association with pathological response in multivariable analysis. The radiomics cohort was randomly divided 7:3 into training (n = 140) and test (n = 61) sets. The Random Forest model achieved an area under the receiver operating characteristic curve (AUC) of 0.823 (95% CI: 0.694-0.932) in the test set, and the habitat radiomics model stratified patients into high and low recurrence risk groups. Single-cell analysis identified immunosuppressive features in non-responding tumors, characterized by expansion of SERPINB9 + regulatory T cells (Tregs) that regulated suppressive intercellular communication networks.
CONCLUSIONS: This study establishes a habitat radiomics model for non-invasive assessment of TIL status following neoadjuvant immunochemotherapy in NSCLC. The model shows reliable predictive performance and prognostic stratification capability, offering potential clinical utility for treatment response evaluation and patient selection.

Keywords:  Habitat radiomics; Neoadjuvant immunochemotherapy; Non-small cell lung cancer; Tumor microenvironment; Tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.1007/s00262-025-04291-x
Theranostics. 2026 ;16(7): 3541-3555

Autocrine activity of engineered IL-33 mRNA enhances adoptive T-cell therapy for peritoneal carcinomatosis and synergizes with IL-12 mRNA.

Leire Arrizabalaga, Claudia Augusta Di Trani, Celia Gomar, Daniel Moreno-Luqui, Nuria Ardaiz, Virginia Belsue, José González-Gomariz, David Ruiz-Guillamon, Aline Risson, Yufei Zheng, Eduardo Huarte, Ignacio Matos, Ignacio Melero, Pedro Berraondo, Fernando Aranda.

  Rationale: Peritoneal carcinomatosis (PC) remains a major clinical challenge with limited therapeutic options across tumor types. Adoptive cell therapy (ACT) with tumor-specific T cells offers promise, but its efficacy is often impaired by the immunosuppressive tumor microenvironment (TME). Intraperitoneal ACT is under investigation to improve its effectiveness against metastases within the peritoneal cavity. IL-33, a cytokine of the IL-1 family, plays dual roles in immunity and inflammation and may enhance antitumor responses. We evaluated whether IL-33 mRNA-engineered T cells improve ACT efficacy in murine PC models and assessed potential synergy with IL-12 mRNA. Methods: OT.I, PMEL-1, and CEA-specific CAR T cells were electroporated with mRNA encoding IL-33, IL-12, or an IL-33 mutein. In vitro assays measured cytokine production and cytotoxicity. RNA-seq was performed to analyze transcriptomic changes following IL-33 mRNA electroporation. ST2-/- T cells were used to evaluate the role of IL-33 receptor expression on transferred T cells versus host cells. In vivo studies in murine PC models assessed survival and immune responses using ELISA, ELISpot, and flow cytometry. Results: IL-33 mRNA-electroporated OT.I T cells exhibited enhanced IFN-γ expression in a ST2-dependent, T cell-intrinsic manner. In vivo, IL-33-engineered T cells significantly improved survival in PC models. IL-33 reshaped the TME by increasing infiltration of innate lymphoid cells and eosinophils while reducing neutrophils. Engineering T cells with a stabilized IL-33 mutein further enhanced antitumor activity. Co-electroporation of IL-33 mutein and IL-12 mRNA in PMEL-1 T cells led to synergistic increases in IFN-γ production, cytotoxicity, and long-term memory, resulting in superior tumor control and protection upon rechallenge. These findings were confirmed using IL-33 mutein/IL-12 mRNA-electroporated CEA CAR T cells in peritoneal tumor models. Conclusions: IL-33 enhances ACT efficacy by promoting IFN-γ expression via autocrine ST2 signaling and by modulating the TME. The IL-33 mutein improves cytokine stability and antitumor activity, while combination with IL-12 yields synergistic effects. This strategy holds promise for enhancing ACT in peritoneal carcinomatosis.

Keywords:  RNA electroporation; T cell therapy; cytokine engineering; immune modulation; solid tumor immunotherapy; tumor microenvironment

DOI:  https://doi.org/10.7150/thno.122132
Medicina (Kaunas). 2026 Jan 16. pii: 192. [Epub ahead of print]62(1):

Integration of the GRIm Score with Pathologic Immune and Stromal Markers to Develop a Combined Prognostic Model in Gastric Cancer: A Retrospective Single-Center Study.

Gökhan Öztürk, Ebru Taştekin, Canberk Topuz, Aysun Fatma Akkuş, Tayyip İlker Aydın, Sernaz Topaloğlu, Bülent Erdoğan, Muhammet Bekir Hacıoğlu, Ahmet Küçükarda.

  Background and Objectives: The Gustave Roussy Immune (GRIm) score, reflecting systemic inflammation and nutritional status, has emerged as a simple and reproducible prognostic biomarker in various malignancies. However, its prognostic interaction with tumor microenvironmental factors remains unclear in gastric cancer. The primary aim of this study was to evaluate the prognostic value of the GRIm score in patients with resectable gastric adenocarcinoma, while the secondary aim was to determine whether integrating the GRIm score with tumor microenvironment-related pathological markers could improve prognostic stratification. Materials and Methods: This retrospective study analyzed 188 patients with resectable gastric adenocarcinoma treated at the Trakya University Faculty of Medicine between 2007 and 2018. GRIm scores were calculated from preoperative lactate dehydrogenase (LDH), albumin, and neutrophil-to-lymphocyte ratio (NLR) values. Pathologic parameters, including programmed death-ligand 1 (PD-L1) expression (combined positive score [CPS] ≥ 1 vs. <1), tumor-stroma ratio (TSR; stromal component ≥ 50% vs. <50%), and tumor-infiltrating lymphocyte (TIL) density (CD8+ ≥ 10% vs. <10%), were evaluated on surgical specimens. Survival outcomes were assessed using Kaplan-Meier and multivariate Cox analyses. Results: The study population had a mean age of 61.8 years and was predominantly male (72.3%). Patients with low GRIm scores had significantly longer disease-free survival (DFS; 24 vs. 12 months; p = 0.004) and overall survival (OS; 32 vs. 19 months; p = 0.006). In multivariate analysis, the GRIm score remained an independent predictor for both disease-free survival (p = 0.035) and overall survival (p = 0.044). Among combined models, the GRIm-TSR classification provided the most pronounced stratification (median DFS = 35 vs. 12 months; OS = 45 vs. 19 months; p = 0.014 and 0.001, respectively), retaining independent prognostic significance (hazard ratio [HR] = 1.23; p = 0.005). Integrating GRIm with PD-L1 and TIL density also improved prognostic discrimination. Conclusions: The GRIm score is a robust and cost-effective biomarker that independently predicts disease-free survival and overall survival in resectable gastric adenocarcinoma. Its combination with microenvironmental markers-PD-L1, TIL, and TSR-captures complementary biological dimensions of tumor aggressiveness, offering an integrative and clinically feasible framework for individualized risk assessment and postoperative management. Prospective multicenter validation is warranted.

Keywords:  GRIm score; gastric cancer; prognostic biomarker; programmed death-ligand 1 (PD-L1); systemic inflammation; tumor microenvironment; tumor-infiltrating lymphocytes (TIL); tumor-stroma ratio (TSR)

DOI:  https://doi.org/10.3390/medicina62010192
Nat Med. 2026 Jan 28.

Neoadjuvant ipilimumab plus nivolumab in melanoma: 5-year survival and biomarker analysis from the phase 2 PRADO-trial.

Lotte L Hoeijmakers, Petros Dimitriadis, Steven C M A Wijnen, Irene L M Reijers, Marta Lopez-Yurda, Alexander M Menzies, Annegien Broeks, Sten Cornelissen, Alejandro Torres Acosta, Anja van der Wal, Robyn P M Saw, Judith M Versluis, Winan J van Houdt, Michel W Wouters, Jurriaan Romano, Eliza A Rozeman, Lindsay G Grijpink-Ongering, Ellen Kapiteijn, Astrid A M van der Veldt, Karijn P M Suijkerbuijk, Hanna Eriksson, Geke A P Hospers, Jos A van der Hage, Dirk J Grünhagen, Arjen J Witkamp, Judith M Lijnsvelt, Willem M C Klop, Charlotte L Zuur, Annemarie Bruining, Abrahim Al-Mamgani, Thomas E Pennington, Kerwin F Shannon, Sydney Ch'ng, Andrew J Colebatch, Maria Gonzalez, Andrew J Spillane, John B A G Haanen, Robert V Rawson, Richard A Scolyer, Bart A van de Wiel, Alexander C J van Akkooi, Georgina V Long, Christian U Blank.

Neoadjuvant ipilimumab plus nivolumab has become standard therapy for stage III melanoma based on the NADINA trial, although long-term data are lacking. In the phase 2 PRADO cohort of OpACIN-neo, 99 patients with stage III macroscopic melanoma received this regimen. Here we report first-time 5-year survival data: 71% event-free survival, 74% relapse-free survival, 79% distant metastasis-free survival and 86% overall survival. Ongoing grade 1-2 immune-related adverse events occurred in 69% of patients alive, predominantly vitiligo and hypothyroidism. Major pathologic response (MPR), high tumor mutational burden (TMB), high interferon-gamma (IFNγ) signature and programmed cell death ligand 1 (PD-L1) expression of 1% or higher were associated with favorable outcomes. Combined high TMB, IFNγ and PD-L1 expression yielded 100% MPR and 100% 5-year event-free survival, whereas triple low expression had only 18% MPR and 41% event-free survival. Our findings demonstrate favorable long-term outcomes for patients with an MPR and identify IFNγ and PD-L1 as promising baseline biomarkers. ClinicalTrials.gov identifier: NCT02977052 .

DOI: https://doi.org/10.1038/s41591-025-04158-9
Acta Pharm Sin B. 2026 Jan;16(1): 239-251

Attenuated Salmonella secreting interleukin-21 activates T-cells and induces anti-tumor effects.

Minju Han, Eunji Kim, Minchan Jeong, Solbi Kim, Hyo-Jin Lee, Heung Jin Jeon.

  The tumor microenvironment is characterized by an immunosuppressive state. Although PD-1/PD-L1 blockade therapy activates the immune system against tumors, it has limited long-term efficacy, prompting the development of combination therapies with targeted treatments to improve cancer treatment outcomes. Recent advancements have revitalized interest in using attenuated Salmonella strains as cancer therapeutics that target tumors, induce immune responses, and promote tumor cell death, although complete tumor suppression remains challenging. We aimed to induce antitumor effects by activating the suppressed immune system within the tumor microenvironment using Salmonella-mediated secretion of interleukin-21 (IL-21). We used the tumor-targeting ability of Salmonella and its flagellar type-3 secretion system (FT3SS) to induce the secretion of IL-21 into the tumor microenvironment via the flagellar system and evaluated the local immune response. We also evaluated the efficacy of combining Salmonella-mediated IL-21 delivery and anti-PD-L1 therapy in a mouse model. IL-21 secretion promoted the recruitment of CD4+ and CD8+ T cells and enhanced the expression of cytotoxicity-related molecules. Tumor-bearing mice treated with the combination therapy with anti-PD-L1 antibodies showed improved survival rates and enhanced tumor growth inhibition. This study demonstrates the tumor-targeting capability and in vivo safety of Salmonella, highlighting its potential as a powerful cancer therapy platform.

Keywords:  Anti-tumor effect; Combination therapy; Engineering Salmonella; Flagellar type-3 secretion system; Immune checkpoint inhibitor; Immunotherapy; Interleukin-21; Tumor microenvironment

DOI:  https://doi.org/10.1016/j.apsb.2025.09.025
J Immunother Cancer. 2026 Jan 28. pii: e013142. [Epub ahead of print]14(1):

DUSP22 dephosphorylates LGALS1 to enhance T cell-driven antitumor immunity.

Lijian Wang, Yutong Guo, Yujie Dai, Wangsheng Sun, Xiaoying Huang, Haipeng Lei, Aiping Zhang, Shuwen Chen, Yiting Li, Jiani Pan, Yangjian Hong, Lingchuan Ma, Yangyang Feng, Fangyuan Shao, Jianming Zeng, Peng Luo, Junqi Li, Weiting Chen, Na Zhou, Yang Li, Heng Sun, Xiaoling Xu, Chu-Xia Deng, Kai Miao.

   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.

Keywords:  Breast Cancer; Immunotherapy; Tumor infiltrating lymphocyte - TIL; Tumor microenvironment - TME

DOI:  https://doi.org/10.1136/jitc-2025-013142
Viruses. 2026 Jan 12. pii: 102. [Epub ahead of print]18(1):

GM-CSF Armed Oncolytic Adenovirus Enhances T-Cell Infiltration and Suppresses Local and Distal Tumor Growth.

Hua-Wei Xu, Qing-Wen Wang, Min Zhao, Jie Jun, Ri-Gan Shu, Yu-Sen Shi, Xiang-Lei Peng, Jie-Mei Yu, Yan-Peng Zheng, Yuan-Hui Fu, Jin-Sheng He.

  The limited ability of the immune system to infiltrate solid tumors, attributed to the immunosuppressive tumor microenvironment (TME), remains a significant challenge in cancer therapy oncolytic adenovirus (OAd) that can directly kill tumor cells in addition to inducing both innate and adaptive immune responses. Therefore, the use of OAd to treat tumors is an appealing approach. In this study, we engineered an OAd armed with a human granulocyte-macrophage colony-stimulating factor (GM-CSF), controlled by the E2F promoter, Ad5/3-E2F-d24-GM-CSF (named OAd-Z1). The antitumor activity of OAd was tested in vitro and in vivo. These findings demonstrated that OAd expressed GM-CSF, replicated effectively in tumor cells, inhibited tumor growth, activated the de novo antitumor response, promoted apoptosis and immunogenic cell death in tumor cells, and increased cytokine and chemokine production both in vitro and in vivo. Additionally, OAd demonstrated an abscopal effect and stimulated T lymphocyte infiltration in vivo. Our findings demonstrate that OAd-Z1 represents promising immunotherapeutic candidates for lung cancer, with the potential to enhance systemic antitumor immunity.

Keywords:  GM-CSF; abscopal effect; oncolytic adenoviruses; tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.3390/v18010102
BMC Med. 2026 Jan 30.

Enhancing oncolytic virotherapy with a tri-specific T-cell engager targeting CD3ε, EpCaM, and 4-1BB: preclinical evaluation and implications for cancer immunotherapy.

Zhongyi Dong, Haoyu Zhang, Hailin Zhang, Long Bai, Qianyu Guo, Guy Robinson, Aya Elalfy, Tao Liu, Muerzhate Aimaiti, Xiang Xia, Shuguang Zuo, Zizhen Zhang.

   BACKGROUND: Cytotoxic T-cell-mediated tumor lysis is a key mechanism of oncolytic virotherapy. While oncolytic viruses expressing bispecific T-cell engagers (BiTE) enhance tumor targeting, they lack costimulatory signals, leading to T-cell exhaustion. We developed an oncolytic adenovirus expressing a trispecific T-cell engager (TriTE) to improve antitumor responses in colorectal carcinoma models.
METHODS: An oncolytic adenovirus (Ad5-TriTE) was engineered to express a TriTE molecule targeting EpCAM (tumor antigen), CD3ε (T-cell activation), and 4-1BB (co-stimulation). For comparison, a BiTE-expressing virus (Ad5-BiTE) lacking 4-1BB was used. BiTE (αCD3ε-αEpCAM) facilitates T-cell redirection, whereas TriTE (αCD3ε-α4-1BBL-αEpCAM) adds co-stimulation for enhanced T-cell activation. Antitumor efficacy was evaluated in syngeneic and humanized colorectal carcinoma mouse models.
RESULTS: Ad5-TriTE demonstrated efficient tumor infection and TriTE secretion, leading to superior tumor control compared to Ad5-BiTE. Enhanced CD8 + T-cell infiltration and activation correlated with improved antitumor effects in both subcutaneous and peritoneal metastasis models. The humanized version, Ad5-hTriTE, exhibited potent activity in a humanized colon cancer model.
CONCLUSIONS: Oncolytic adenovirus armed with TriTE enhances antitumor immunity by integrating costimulatory signaling. Incorporating costimulatory molecules into oncolytic virotherapy may offer more effective cancer immunotherapy strategies.

Keywords:  Colon cancer; Immunotherapy; Oncolytic virotherapy

DOI:  https://doi.org/10.1186/s12916-026-04661-x
Clin Genitourin Cancer. 2026 Jan 01. pii: S1558-7673(25)00197-1. [Epub ahead of print]24(2): 102497

Association of PD-L1 and PD-1 Expression With Clinicopathological Characteristics and Prognosis in Upper Tract Urothelial Carcinoma.

Peng Li, Xiaolei Zhang, Xiao Yang, Hai Li, Guoxin Song, Qiang Lu, Li Pengchao.

   INTRODUCTION: To investigate the expression of PD-L1/PD-1 in tumor cells and tumor-infiltrating immune cells (TIICs) and their association with clinicopathologic characteristics and prognosis in UTUC.
PATIENTS AND METHODS: Immunohistochemistry (IHC) was performed to detect the expression of PD-L1 and PD-1 in 124 formalin-fixed paraffin embedded tumor specimens in UTUC. Clinicopathological variables and overall survival (OS) were documented. Association between PD-L1, PD1 with clinicopathologic characteristics and OS were evaluated using binary logistic regression or Cox regression. Statistical significance was considered at 0.05.
RESULTS: Of 113 eligible patients (11 excluded due to insufficient tissue), 31.9% (36/113) showed PD-L1⁺ tumor cells, and 28.8% (29/101) had PD-L1⁺ TIICs. PD-1⁺ TIICs were observed in 58.4% (59/101). PD-L1 expression in tumor cells correlated significantly with high tumor grade (P = .029). PD-L1⁺ tumor cells predicted reduced OS in univariate (HR = 1.83, P = .025) and multivariable analyses (HR = 2.15, P = .007). PD-1⁺ TIICs associated with early-stage disease (P = .005) but not OS (P = .166). PD-L1⁺ TIICs also showed no OS association (P = .644).
CONCLUSIONS: PD-L1 is expressed in 31.9% of UTUC tumors and 28.8% of TIICs, while PD-1 is detected in 58.4% of TIICs. Tumor cell PD-L1 positivity independently predicts poor survival and may drive UTUC progression. PD-1⁺ TIICs correlate with early-stage disease but lack prognostic value for OS. These findings highlight PD-L1 as a potential biomarker for risk stratification in UTUC.

Keywords:  Clinicopathological features; Immune checkpoint; Immunotherapy; Tumor infiltrating immune cells; Tumor prognosis

DOI:  https://doi.org/10.1016/j.clgc.2025.102497
Front Oncol. 2025 ;15 1747065

In vivo detection of HER2-positive breast cancer and sentinel lymph node metastasis using activatable ICG-conjugated Trastuzumab.

Yunpeng Liu, Mahendra Isa, Takahito Nakajima.

  Breast cancer continues to pose a significant health threat to women globally, with sentinel lymph node (SLN) metastasis playing a crucial role in treatment planning and prognosis. Conventional diagnostic techniques are invasive, lack dynamic capability, and have limited specificity. Moreover, detecting intraductal spread or microinvasion remains a challenge. Here, we utilized an activatable (quenched) fluorescence probe by conjugating indocyanine green (ICG) with the anti-HER2 antibody Trastuzumab for non-invasive detection of HER2-positive breast cancer and its sentinel lymph node metastasis. Specifically, we conjugated ICG-sulfo-EG4-OSu to Trastuzumab and purified it to obtain Trastuzumab-ICG. In vitro live-cell imaging and flow cytometry demonstrated that the conjugate bound specifically to HCC-1419-Luc (HER2-positive) cells and emitted fluorescence, whereas no fluorescence was observed in BT-20-Luc (HER2-negative) cells. Orthotopic tumor models and lymph node metastasis models further confirmed its in vivo specificity. These results indicate the potential of Trastuzumab-ICG for high-specificity detection of HER2-positive breast cancer and its lymph node metastases, offering a promising tool for high-contrast tumor visualization and personalized treatment guidance.

Keywords:  HER2; Trastuzumab; breast cancer; indocyanine green; metastasis detection; near-infrared fluorescence; sentinel lymph node

DOI:  https://doi.org/10.3389/fonc.2025.1747065
Front Immunol. 2025 ;16 1715827

A ligand-centered framework for γδ T cell activation in colorectal cancer revealed by single-cell and transformer-based perturbation.

Ran Ran, Douglas K Brubaker.

  Understanding the activation mechanisms of γδ T cells in colorectal cancer (CRC) is critical for harnessing their therapeutic potential. Here, using an atlas of human CRC-infiltrating γδ T cells that we built by integrating multiple single-cell RNA-seq datasets, we developed a γδ T cell-refined ligand inference pipeline by combining differential gene expression, gene regulatory network prediction, ligand inference, and in silico perturbation analysis. This approach identified ligands, including IL-15 and TNFSF9 (4-1BBL), as candidates promoting γδ T cell effector function and highlighted NCR2 and KLRC3 (NKG2E), whose in silico overexpression was associated with γδ T cell activation. Ligand enrichment analyses further indicated that monocytes and dendritic cells are key contributors to γδ T cell activation in the tumor microenvironment. Our results also highlighted transcription factors IKZF1, FOSL2, and FOXO1 in the less activated γδ T cells and IRF1, KLF2, and BHLHE40 in the effector γδ T cells that plausibly regulated the differential activation state. Together, our results offer a systems-level view of the signaling and transcriptional programs governing γδ T cell phenotypes in CRC and provide a foundation for γδ T cell-based immunotherapies with enhanced antitumor functions.

Keywords:  cancer colorectal; deep learn ing; gamma delta (γδ) T cells; ligand; perturbation prediction; transformer

DOI:  https://doi.org/10.3389/fimmu.2025.1715827