Background: Concurrent chemoradiotherapy (CCRT) combined with induction chemotherapy (IC) is the standard first-line treatment for locally advanced nasopharyngeal carcinoma (LA-NPC). However, treatmentrelated toxicities and the risk of recurrence or metastasis remain clinical challenges, which have prompted the development of strategies that can improve efficacy while minimizing adverse effects. The integration of programmed cell death receptor 1 (PD-1) inhibitors with IC has demonstrated significant survival benefits in pivotal phase III trials. This study aimed to assess the efficacy and safety of PD-1 inhibitors combined with IC consisting of a paclitaxel-platinum (TP) regimen for LA-NPC in a real-world clinical setting. Methods: This retrospective study enrolled patients diagnosed with stage III or IVA NPC who received treatment between January 2021 and December 2024. Clinical records and radiographic data were systematically reviewed. Based on the administration of PD-1 inhibitor immunotherapy during IC, patients were categorized into a control group and an immunotherapy group. Propensity score matching (PSM) was performed sequentially after IC and again after CCRT to minimize confounding factors. Matching was based on clinically relevant covariates, including age, gender, differentiation, tumor-node-metastasis stage, Epstein-Barr virus DNA level and treatment cycle. Between-group differences in treatment response and adverse events were assessed via Chi-squared tests for categorical variables. The primary endpoint is the disease control rate (DCR) after IC. The secondary endpoints include the objective response rate (ORR) after CCRT, the incidence of adverse events, and their severity. Results: A total of 150 patients with LA-NPC were included, among whom 76 received standard IC, and 74 received combined immunotherapy. In the PSM analysis, 54 patients were matched to each group, and the immunotherapy group demonstrated significantly higher disease control during the IC phase (P=0.004), with a partial response (PR) rate of 94.4% compared to 70.4% in the control group. Following CCRT, 49 patients per group matched by PSM showed no significant differences in complete response (CR) or PR rates (P=0.25). The CR rates were 10.2% and 18.4% in the control group and the immunotherapy group, respectively, while the PR rates were 89.8% and 81.6%, respectively. Furthermore, the groups showed no statistical differences in terms of the incidence of radiotherapy-related adverse events, including oral mucositis (P=0.26), skin reactions (P=0.20), and myelosuppression (P=0.33). No grade 3 or higher immune-related adverse events (irAEs) were reported in the immunotherapy group. Conclusions: This real-world study successfully verified that adding PD-1 inhibitors to IC improves early efficacy without increasing toxicity in LA-NPC, providing external validation of key trial results related to routine clinical practice.

Background Leptomeningeal metastasis (LM) represents one of the most severe complications in advanced lung cancer, indicating a poor prognosis. However, the mechanisms underlying LM progression remain incompletely understood, and reliable biomarkers for monitoring disease progression and treatment response are lacking.Methods This retrospective observational study was conducted at the Zhejiang Provincial People's Hospital. Cerebrospinal fluid (CSF) samples from 13 patients with non-small cell lung cancer (NSCLC) and LM were analyzed, including biochemical parameters, tumor markers, differential cell counts, and T-cell subsets assessed by flow cytometry.Results The median overall survival (mOS) after LM diagnosis was 15 months. During LM progression, increased CSF levels of albumin, lactate dehydrogenase (LDH), lactic acid, and carcinoembryonic antigen (CEA) were observed, accompanied by decreased glucose, chloride, lymphocyte proportion, and CD8+ T-cell proportion. In some cases, lymphocyte percentages and CD8+ T-cell proportions increased during subsequent evaluations following treatment adjustment.Conclusions These findings suggest that CSF biochemical and immune parameters may be associated with disease progression in LM. Dynamic changes in CD8+ T-cell proportions may also reflect alterations in the CSF immune microenvironment and could represent potential targets for future immunotherapeutic strategies.

Background:Despite notable advances with immune checkpoint inhibitors (ICIs) in esophageal squamous cell carcinoma (ESCC), their clinical efficacy remains limited, largely due to CD8⁺T cell exhaustion. Among these, progenitor exhausted T cells (Tpex) represent a key subset with stem cell-like features that sustain durable anti-tumor immunity. Methods:We applied multi-color immunohistochemistry (mIHC) to determine the spatial distribution and clinical significance of Tpex cells within the tumor microenvironment (TME) of ESCC. Publicly available single-cell RNA sequencing (scRNA-seq) datasets were further analyzed to characterize Tpex cell phenotypes, differentiation trajectories, and intercellular communication networks. Results:Tpex cells constituted a distinct subset of infiltrating CD8⁺T cells and represented a transitional stage of the exhaustion continuum. A higher degree of Tpex infiltration was significantly associated with improved overall survival in ESCC patients. Moreover, scRNA-seq data from patients treated with PD-1 blockade revealed that responders harbored markedly enriched Tpex populations compared with non-responders. Conclusion:Our findings identify Tpex cells as a critical prognostic and immunologically active CD8⁺T cell subset in ESCC. Their abundance and functional engagement are closely associated with favorable clinical outcomes and response to PD-1 blockade. Furthermore, their stem cell-like properties may be pivotal in shaping durable anti-tumor immunity and could provide novel therapeutic targets to enhance the efficacy of PD-1-based immunotherapy.

Cancer-associated fibroblasts (CAFs) are key regulators in tumor microenvironment and tumor immunity, partly through MHC-II expression that modulates T-cell differentiation. However, the upstream cytokine signals controlling MHC-II expression in fibroblasts still remain poorly defined. We examined MHC-II expression on fibroblasts under stimulation with interferon-γ (IFN-γ) and interleukin-1β (IL-1β) by using flow cytometry, transcriptomic analysis, and qRT-PCR. To dissect transcriptional regulation, we generated CIITA-overexpressing and CIITA-deficient fibroblast lines by lentiviral transduction and CRISPR/Cas9-mediated editing. Public scRNA-seq, ATAC-seq, and ChIP-seq datasets were further analyzed to validate molecular mechanisms. IFN-γ robustly up-regulated MHC-II expression on fibroblasts, while IL-1β selectively suppressed this induction without affecting PD-L1. Mechanistically, IL-1β attenuated IFN-γ–induced CIITA expression at the mRNA level but did not alter STAT1 abundance or phosphorylation. Functional assays confirmed that CIITA was indispensable for IFN-γ–driven MHC-II expression in fibroblasts. Integration of transcriptomic and epigenomic data demonstrated that CIITA directly bound MHC-II gene promoters and regulated chromatin accessibility. Our study identifies an IFN-γ/STAT1/CIITA axis as the central regulator of MHC-II expression in fibroblasts and reveals IL-1β as a potent suppressor of this pathway. These findings highlight a novel cytokine-mediated regulatory mechanism underlying CAF-driven immunosuppression within the tumor microenvironment.

Colon cancer is a prevalent malignancy of the digestive tract, consistently ranking among the top three cancers globally in both incidence and mortality. Current standard treatments for colon cancer primarily include surgical resection and adjuvant chemoradiotherapy, while only a limited subset of patients derive substantial benefits from immunotherapy. Consequently, further investigations into the pathogenesis and immune microenvironment of colon cancer, along with the identification of effective therapeutic targets, are crucial for improving patient prognoses. In this study, we utilized a multi-color immunohistochemistry (mIHC) assay to examine the expression patterns of IL22RA1, IL22, CD155, and IL22BP in a tissue microarray (TMA) comprising 90 colon cancer samples. Additionally, we assessed the immunolocalization, clinical relevance, and prognostic significance of these molecules. The mIHC results, supported by multispectral tissue imaging, revealed that IL22RA1 was predominantly expressed in tumor epithelial cells. Expression levels of IL22RA1, CD155, and IL22BP in colon cancer tissues were markedly higher than those in adjacent normal tissues, with statistically significant differences (t = 6.05, 9.53, 3.91, all P < 0.001). Analysis of TMA and publicly available databases demonstrated that patients with low IL22RA1 expression exhibited significantly improved overall survival (OS) compared to those with high IL22RA1 expression. Furthermore, patients characterized by high co-expression of IL22RA1 and IL22 (IL22RA1highIL22high) had notably poorer OS compared to others. Similar trends were observed for patients with IL22RA1highCD155high or IL22highCD155high expression patterns, all of whom experienced significantly worse OS. Conversely, patients with low IL22RA1 expression and high IL22BP expression (IL22RA1lowIL22BPhigh) exhibited substantially better OS than those with IL22RA1highIL22BPlow (all P < 0.05). Correlation analysis highlighted a strong positive relationship between the infiltration levels of IL22RA1+CK+ cells and CD155+CK+ cells in tumor tissues (r = 0.55, P < 0.001). Cox multivariate regression analysis identified high IL22RA1 expression in tumor cells as an independent risk factor for poor prognosis (HR = 2.726, 95

Liver cancer, specifically hepatocellular carcinoma (HCC), stands out as one of the most formidable solid tumors, characterized by a dauntingly low survival rate. At the forefront of the tumor microenvironment (TME) orchestrating the initiation and advancement of HCC are cancer-associated fibroblasts (CAFs). TGF-β, widely recognized as a potent activator of CAFs, not only regulates their activity but also assumes a pivotal role in the metastatic journey of the tumor. In our recent study, drawing from the GEO database, we identified two fibroblast subtypes in HCC through single-cell RNA sequencing (scRNA-seq) and explore the expression and distribution of TGF-β and its receptors in the TME of HCC. Subsequently, we investigated the interactions between tumor cells expressing high levels (TGFB1high) and low levels (TGFB1low) of TGF-β in the HCC TME and the two subtypes of CAFs. We also employed multi-color immunohistochemistry (mIHC) technology to examine the expressions of FAP, α-SMA, CD4, Foxp3, and TGF-β in HCC tissues within a tissue microarray. Additionally, we analyzed clinical associations, prognostic values, and the correlation of these molecules. These insights advance our understanding of the molecular mechanisms driving HCC progression and underscore the intricate interplay between tumor cells and the stromal components of the TME.

Exhausted CD8+T cells represent a distinct cellular lineage that emerges during both chronic infections and cancers. Recent studies have shown that persistent antigen exposure can drive the differentiation of precursor exhausted CD8+T cells, termed Tpex cells, which are characterized as TCF-1+PD-1+CD8+T cells. Elevated Tpex cell frequencies in the tumor microenvironment (TME) are associated with improved overall survival (OS) in cancer patients and heightened responsiveness to anti-PD-1 therapy. In our present study, we utilized multi-color immunohistochemistry (mIHC) to determine the localization and clinical implications of tumor-infiltrating Tpex cells within the TME of human colorectal cancer (CRC) tissues. We also conducted a multi-omics integrative analysis using single-cell RNA sequencing (scRNA-seq) data derived from both the murine MC38 tumor model and human CRC tissues. This analysis helped delineate the transcriptional and functional attributes of Tpex cells within the CRC TME. Furthermore, we employed spatial transcriptome sequencing data from CRC patients to investigate the interactions between Tpex cells and other immune cell subsets within the TME. In conclusion, our study not only established a method for Tpex cell detection using mIHC technology but also confirmed that assessing Tpex cells within the CRC TME could be indicative of patients’ survival. We further uncovered the transcriptional and functional characteristics of Tpex cells in the TME and ascertained their pivotal role in the efficacy of immunotherapy against CRC.

Background Immune-based therapies are commonly employed to combat hepatocellular carcinoma (HCC). However, the presence of immune-regulating elements, especially regulatory T cells (Tregs), can dramatically impact the treatment efficacy. A deeper examination of the immune-regulation mechanisms linked to these inhibitory factors and their impact on HCC patient outcomes is warranted. Methods We employed multicolor fluorescence immunohistochemistry (mIHC) to stain Foxp3, cytokeratin, and nuclei on an HCC tissue microarray (TMA). Leveraging liver cancer transcriptome data from TCGA, we built a prognostic model focused on Treg-associated gene sets and represented it with a nomogram. We then sourced liver cancer single-cell RNA sequencing data (GSE140228) from the GEO database, selectively focusing on Treg subsets, and conducted further analyses, including cell-to-cell communication and pseudo-time trajectory examination. Results Our mIHC results revealed a more substantial presence of Foxp3 + Tregs in HCC samples than in adjacent normal tissue samples ( P < .001). An increased presence of Foxp3 + Tregs in HCC samples correlated with unfavorable patient outcomes ( HR = 1.722, 95% CI :1.023-2.899, P = .041). The multi-factorial prognosis model we built from TCGA liver cancer data highlighted Tregs as a standalone risk determinant for predicting outcomes ( HR = 3.84, 95% CI: 2.52-5.83, P < .001). Re-analyzing the scRNA-seq dataset (GSE140228) showcased distinctive gene expression patterns in Tregs from varying tissues. Interactions between Tregs and other CD4 + T cell types were predominantly governed by the CXCL13/CXCR3 signaling pathway. Communication pathways between Tregs and macrophages primarily involved MIF-CD74/CXCR4, LGALS9/CD45, and PTPRC/MRC1. Additionally, macrophages could influence Tregs via HLA-class II and CD4 interactions. Conclusion An elevated presence of Tregs in HCC samples correlated with negative patient outcomes. Elucidating the interplay between Tregs and other immune cells in HCC could provide insights into the modulatory role of Tregs within HCC tissues.

Nuclear regulation has potential in cancer therapy, with the nuclear pore complex (NPC) serving as a critical channel between the nucleus and cytoplasm, playing a role in regulating various biological processes and cancer. DNA methylation, an epigenetic modification mediated by DNA methyltransferases (DNMTs), influences gene expression and cell differentiation, and is crucial for the development and progression of tumor cells. Gliomas are the most common primary brain tumors, with glioblastoma being particularly aggressive, characterized by invasiveness, migration capability, and resistance to conventional treatments, resulting in poor prognosis. Our study revealed that the expression level of NUP37 affects the proliferation and invasion of glioma cells, and that the overexpression of DNMT1 can alleviate the adverse effects caused by NUP37 depletion. These findings suggest that NUP37 promotes the proliferation and invasion of glioma cells through its interaction with DNMT1.

HHLA2 (human endogenous retrovirus-H long terminal repeat-associating protein 2) represents a recently identified member of the B7 immune checkpoint family, characterized by limited expression in normal tissues but notable overexpression in various cancer types. Nevertheless, the precise function and interaction with immune cells remain poorly understood, particularly in laryngeal squamous cell carcinoma (LSCC). This investigation endeavored to elucidate the biological significance of HHLA2 within the tumor microenvironment of human LSCC tissues and delineate the clinical relevance and functional roles of HHLA2 in LSCC pathogenesis. Through multiplexed immunohistochemistry analyses conducted on tissue microarrays sourced from LSCC patients (n = 72), the analysis was executed to assess the expression levels of HHLA2, density and spatial patterns of CD68+HLA-DR+CD163− (M1 macrophages), CTLA-4+CD4+FoxP3+ (CTLA-4+Treg cells), CTLA-4+CD4+FoxP3− (CTLA-4+Tcon cells), exhausted CD8+T cells, and terminally exhausted CD8+T cells in LSCC tissues. Survival analysis was conducted to evaluate the prognostic significance of HHLA2 and these immune checkpoints or immune cell populations, employing COX regression analysis to identify independent prognostic factors. Kaplan–Meier (K–M) survival curves revealed a significant association between HHLA2 expression and overall survival (OS) in LSCC. Elevated levels of HHLA2 were linked to reduced patient survival, indicating its potential as a prognostic marker (HR: 3.230, 95

While biomarkers have been shown to enhance the prognosis of patients with colorectal cancer (CRC) compared to conventional treatments, there is a pressing need to discover novel biomarkers that can assist in assessing the prognostic impact of immunotherapy and in formulating individualized treatment plans. The RUNX family, consisting of RUNX1, RUNX2, and RUNX3, has been recognized as crucial regulators in developmental processes, with dysregulation of these genes also being implicated in tumorigenesis and cancer progression. In our present study, we demonstrated a crucial regulatory role of RUNX in CD8+T and CD103+CD8+T cell-mediated anti-tumor response within the tumor microenvironment (TME) of human CRC. Specifically, RUNXs were significantly differentially expressed between tumor and normal tissues in CRC. Patients with a greater proportion of infiltrating CD8+RUNX1+, CD103+CD8+RUNX1+, CD8+RUNX2+, CD103+CD8+RUNX2+, CD8+RUNX3+, or CD103+CD8+RUNX3+ T cells demonstrated improved outcomes compared to those with lower proportions. Additionally, the proportions of infiltrating CD8+RUNX1+T and CD8+RUNX3+T cells may serve as valuable prognostic predictors for CRC patients, independent of other clinicopathological factors. Moreover, further bioinformatic analysis conducted utilizing the TISIDB and TIMER platforms demonstrated significant associations between the members of the RUNX family and immune-infiltrating cells, specifically diverse subpopulations of CD8+TILs. Our study of human colorectal cancer tissue microarray (TMA) also revealed positive and statistically significant correlations between the expressions of RUNX1, RUNX2, and RUNX3 in both CD8+T cells and CD103+CD8+T cells. Our study comprehensively revealed the varied expressions and prognostic importance of the RUNX family in human colorectal cancer tissues. It underscored their potential as vital biomarkers for prognostic evaluation in colorectal cancer patients and as promising targets for immunotherapy in treating this disease.

PD-L1 is expressed on antigen-presenting cells and tumor cells, thus allows tumor cells to escape immune surveillance. Moreover, targeting PD-L1 was also recommended and selected as important immune checkpoint inhibitors (ICIs) strategy in the treatment of advanced cancers due to the safety and activity. However, the detailed alteration of tumor microenvironment (TME) upon anti-PD-L1 therapy in lung cancer tumor model still needs to be resolved. In our present study, first, we characterized PD-L1 expression in human lung adenocarcinoma tissues by using public data, then we established the subcutaneous tumor-bearing model by using murine lung cancer cell line 3LL to perform the anti-PD-L1 therapy and the single-cell RNA sequencing (scRNA-seq) to reveal the remodeling of TME. We confirmed that PD-L1 blockade significantly inhibited tumor progression in 3LL mouse lung cancer model. The scRNA-seq depicted the detailed TME landscape of 3LL tumor model upon anti-PD-L1 treatment. Five major populations according to the marker genes were identified, including tumor cells, stromal cells, myeloid cells, T cells, and NK cells. In addition, we found that anti-PD-L1 treatment enhanced tumor immunogenicity and promoted inflammation in TME and promoted cancer-associated fibroblasts (CAFs)-mediated T-cell migration and infiltration. We also found that anti-PD-L1 treatment can increase dendritic cells (DCs) population and enhance the antigen-presenting ability to CD8 + T cells and promote the transition of monocytes to macrophages and tumor-associated macrophages 2 (TAM2) to TAM1. We also revealed that Nfatc1 was up-regulated in the anti-PD-L1 treatment group, the frequencies of effector CD8 + T cells, exhausted CD8 + T cells, cycling T cells, and NKT were increased, and the frequencies of conventional CD4 + T cells, Treg, IFN-induced T cells, and γδT cells were decreased. Therefore, our scRNA-seq data of the lung cancer tumor model upon anti-PD-L1 treatment made a comprehensive presentation and description about the remodeling of TME and will benefit us to understand the underlying mechanisms and to design combinational therapeutic strategies based on anti-PD-L1 therapy against lung cancer.

Purpose The objective of this study was to determine whether nucleoporin 37 (NUP37) could control the proliferation and invasion of glioma cells through DNA methyltransferase 1 (DNMT1), thus contributing to the onset and progression of glioma. Methods TCGA and GTEx public databases were employed to examine the expression of NUP37 and DNMT1 in glioma. The correlation between NUP37 and DNMT1 expression levels and clinical features, such as prognosis, World Health Organization (WHO), and histopathological types of glioma patients, was analyzed based on the TCGA database. qRT-PCR and Western blotting analysis were used to detect the expression levels of NUP37 and DNMT1 in glioma tissues, cell lines, and post-lentivirus transfection cells. Assays, such as MTT assay, CCK-8 assay, Transwell assay, flow cytometry, scratch test, and cell counting assay, were employed to identify the regulatory effects of NUP37 depletion on the proliferation, apoptosis, invasion, and cell cycle of glioma cells. Transcriptome sequencing combined with proteomic was utilized to examine the changes in genes, proteins, and signaling pathways post-NUP37 knockdown in glioma cells to uncover the effects of changes in target molecules upstream and downstream of NUP37 on glioma cell biological functions. The co-immunoprecipitation (Co-IP) assay was used to investigate the interaction between NUP37 and DNMT1. Lastly, the rescue assay was used to assess whether NUP37 regulated the proliferation and invasion of glioma cells via DNMT1. Results Bioinformatic analysis revealed that NUP37 and DNMT1 were overexpressed in glioma and closely correlated with clinical features, such as prognosis and WHO grades of glioma. The expression of NUP37 and DNMT1 in glioma tissues was significantly higher than in normal brain tissue respectively. NUP37 depletion could suppress the proliferation and invasion of U87 and U251 glioma cells, induce apoptosis, and cause cell cycle arrest. Co-IP experiments indicated that NUP37 could bind DNMT1. Transcriptome sequencing combined with proteomic sequencing showed a decrease in the expression of certain genes, proteins (including DNMT1), and some signaling pathways following NUP37 depletion in glioma cells. Western blotting analysis indicated a decrease in the expression of DNMT1 at the protein level upon NUP37 depletion. The recovery experiment demonstrated that DNMT1 overexpression could restore the proliferation and invasion capacity of glioma cells and reduce the apoptosis rate of these cells. Conclusion These findings suggested that high expression of NUP37 regulated the proliferation and invasion of glioma cells by binding DNMT1.

It is well-known that CD226 serves as a critical activating receptor on various immune cells, such as lymphocytes and monocytes, and it is suggested to promote anti-tumor immunity in the tumor microenvironment (TME). Herein, we showed a crucial regulatory role of CD226 in CD8 + T cell-mediated anti-tumor response in TME of human gastric cancer (GC). Specifically, the increased CD226 expression in cancer tissues was significantly associated with better clinical outcomes in GC patients. Moreover, the increased infiltrating CD226 + CD8 + T cells and the increased ratio of infiltrating CD226 + CD8 + T cells in CD8 + T subpopulation within cancer tissues could also be valuable prognostic predictors for GC patients. Mechanically, the assay for transposase-accessible chromatin using sequencing (ATAC-seq) analysis revealed that the chromatin accessibility of CD226 in CD4 + and CD8 + TILs was significantly higher than that in CD8 + T cells in normal tissues. Further analysis showed that CD8 + TILs highly expressed immune checkpoint molecules, such as TIGIT , LAG3 , and HAVCR2 , which means CD8 + TILs are more exhausted. In addition, our multi-color immunohistochemical staining (mIHC) revealed that GC patients with higher frequency of IFN-γ + CD226 + CD8 + TILs showed poorer prognosis. Combined with the single-cell transcriptome sequencing (scRNA-seq) data analysis, we found that the expressions of IFN-γ and TIGIT in CD8 + TILs were significantly and positively correlated. The expression of TIGIT in IFN-γ + CD226 + CD8 + TILs was higher, while that in IFN-γ - CD226 + CD8 + TILs was significantly lower. The correlation analysis showed that the expression of CD226 was positively correlated with the score of effector T cells but negatively correlated with that of immunosuppressive factors, such as Tregs and tumor-associated macrophages (TAMs). Collectively, we showed that the frequency of CD226 + CD8 + TILs was an excellent prognostic predictor for GC patients. Our findings provided insights into the interaction pattern between co-stimulatory receptor CD226 and tumor cells as well as the infiltrating immune cells in the TME in GC.

BackgroundTissue-resident CD8(+)T cells (CD103(+)CD8(+)T cells) are the essential effector cell population of anti-tumor immune response in tissue regional immunity. And we have reported that IL-33 can promote the proliferation and effector function of tissue-resident CD103(+)CD8(+)T cells. As of now, the immunolocalization and the prognostic values of tissue-resident CD8(+)T cells in human hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC) still remain to be illustrated.MethodsIn our present study, we used the tissue microarrays of HCC and ICC, the multicolor immunohistochemistry (mIHC), and imaging analysis to characterize the tissue-resident CD8(+)T cells in HCC and ICC tissues. The prognostic values and clinical associations were also analyzed. We also studied the biological functions and the cell-cell communication between tumor-infiltrating CD103(+)CD8(+)T cells and other cell types in HCC and ICC based on the published single-cell RNA sequencing (scRNA-seq) data.ResultsOur work unveiled the expressions of CD8 and CD103 and immunolocalization of tissue-resident CD8(+)T cells in human HCC and ICC. Elevated CD8(+)T cells indicated a better overall survival (OS) rate, implying that tumor-infiltrating CD8(+)T cells in HCC and ICC could serve as an independent prognostic factor. Moreover, the number of CD103(+)CD8(+)T cells was increased in HCC and ICC tissues compared with adjacent normal tissues. HCC patients defined as CD8(high)CD103(high) had a better OS, and the CD8(low)CD103(low) group tended to have a poorer prognosis in ICC. Evaluation of the CD103(+)CD8(+)T-cell ratio in CD8(+)T cells could also be a prognostic predictor for HCC and ICC patients. A higher ratio of CD103(+)CD8(+)T cells over total CD8(+)T cells in HCC tissues was negatively and significantly associated with the advanced pathological stage. The percentage of higher numbers of CD103(+)CD8(+)T cells in ICC tissues was negatively and significantly associated with the advanced pathological stage. In contrast, the higher ratio of CD103(+)CD8(+)T cells over total CD8(+)T cells in ICC tissues was negatively and significantly associated with the advanced pathological stage. In addition, single-cell transcriptomics revealed that CD103(+)CD8(+)T cells were enriched in genes associated with T-cell activation, proliferation, cytokine function, and T-cell exhaustion.ConclusionThe CD103(+) tumor-specific T cells signified an important prognostic marker with improved OS, and the evaluation of the tissue-resident CD103(+)CD8(+)T cells might be helpful in assessing the on-treatment response of liver cancer.

As a practical local therapeutic approach to destroy tumor tissue, thermal ablation can activate tumor-specific T cells via enhancing tumor antigen presentation to the immune system. In the present study, we investigated changes in infiltrating immune cells in tumor tissues from the non-radiofrequency ablation (RFA) side by analyzing single-cell RNA sequencing (scRNA-seq) data of tumor-bearing mice compared with control tumors. We showed that ablation treatment could increase the proportion of CD8+T cells and the interaction between macrophages and T cells was altered. Another thermal ablation treatment, microwave ablation (MWA), increased the enrichment of signaling pathways for chemotaxis and chemokine response and was associated with the chemokine CXCL10. In addition, the immune checkpoint PD-1 was especially up-regulated in the infiltrating T cells of tumors on the non-ablation side after thermal ablation treatment. Combination therapy of ablation and PD-1 blockade had a synergistic anti-tumor effect. Furthermore, we found that the CXCL10/CXCR3 axis contributed to the therapeutic efficacy of ablation combined with anti-PD-1 therapy, and activation of the CXCL10/CXCR3 signaling pathway might improve the synergistic effect of this combination treatment against solid tumors.

Background Methyltransferase-like 3 (METTL3) expression could be found in various normal and cancerous tissues. As of now, the clinical significance of METTL3 expression in human pancreatic cancer (PC) tissues still remains to be understood. Our present study aims to investigate the prognostic value and clinical implications of METTL3 expression in PC tissues. Methods The TCGA, GTEx, and GEO public databases were used to study the mRNA expression level of the m 6 A family members and its relationship among PC tissues and normal pancreatic tissue. The immunohistochemistry was used to analyze the difference of METTL3 expression between cancer tissues and adjacent normal tissues. The prognostic value was evaluated by using the Log-rank survival analysis and Cox model analysis. PAAD samples from TCGA and GEO databases were used to perform the immune infiltration analysis and gene set enrichment analysis based on the genes that were highly correlated with METTL3. Results Based on the analysis of TCGA, GTEx, and GEO public database, we found that the m 6 A family members showed a higher correlation in PC tissues compared to normal pancreatic tissues, and the mRNA expression level of the m 6 A family members showed a significant difference between PC tissues and adjacent normal tissues. Moreover, scRNA-seq data indicated that METTL3 showed a higher expression level in malignant epithelial cells. Our immunohistochemistry results also confirmed that the intensity of METTL3 immunostaining in PC tissues was significantly higher than that in adjacent normal tissues ( P = 0.015). The overall survival (OS) of PC patients with high expression of METTL3 protein were significantly poorer than those with low expression of METTL3 protein (HR = 1.788, 95% CI 1.071–2.984, P = 0.026). Further analysis of PC data from the database showed that METTL3 expression was associated with a variety of tumor-infiltrating immune cells and was involved in m 6 A modification and metabolism in PC tissues. Conclusion Increased METTL3 expression at the protein level could be found in PC tissues, suggesting that the METTL3 expression was involved in the progression of PC and could serve as an important marker for prognostic prediction of this malignancy.

本研究用慢病毒转染法构建人内源性反转录病毒-H长末端重复关联蛋白质 2(human endogenous retrovirus subfamily H long terminal repeat associating protein 2,HHLA2)稳定上调表达的人胰腺癌细胞株,探讨HHLA2过表达对胰腺癌细胞生物学功能的影响.

Background The immune checkpoint inhibitors (ICIs) combined with other therapeutic strategies have shown exciting results in various malignancies, and ICIs have now become the gold standard for current cancer treatment. In several preclinical and clinical investigations, ablation coupled with immunotherapy has proved to be quite effective. Our previous studies have shown that ablation coupled with ICI is a potential anti-cancer regimen for colorectal cancer liver metastases (CRLM). Furthermore, we have reported that following microwave ablation (MWA), the expression of LAG3 is up-regulated in tumor microenvironment (TME), indicating that LAG3 is implicated in the regulation of immunosuppressive immune response, and combination therapy of MWA and LAG3 blockade can serve as a promising therapeutic strategy against cancer. Methods The expression of LAG3 was investigated in this study utilizing a preclinical mouse model treated with MWA. Moreover, we monitored the tumor development and survival in mice to assess the anti-cancer effects of MWA alone or in combination with LAG3 blockade. Flow cytometry was also used to phenotype the tumor-infiltrating lymphocytes (TILs) and CD8 + T cell effector molecules. We finally analyzed the single-cell RNA sequencing (scRNA-seq) data of infiltrating CD45 + immune cells in the tumors from the MWA alone and MWA combined with LAG3 blockade groups. Results After MWA, the expression of LAG3 was up-regulated on sub-populations of TILs, and introducing LAG3 blockade to MWA postponed tumor development and extended survival in the MC38 tumor model. Flow cytometry and scRNA-seq revealed that LAG3 blockade in combination with MWA markedly boosted the proliferation and the function of CD8 + TILs, leading to altered myeloid cells in the TME. Conclusion Combination therapy of LAG3 blockade and MWA was a unique therapeutic regimen for some solid tumors, and such combination therapy might reprogram the TME to an anti-tumor manner.

BACKGROUND:Pancreatic cancer (PC) is one of the most malignant solid tumors and its 5-year survival rate remains poor. Although immunotherapy has achieved certain therapeutic efficacy in some clinical trials, such treatment still shows low responses and overall remission rate. Therefore, it is urgently necessary to dissect the tumor microenvironment and optimize the immunotherapeutic strategies against this malignancy.METHODS:Using the multi-color immunohistochemistry assay, we investigated the expressions of B7-H3, B7-H4, HHLA2, CD8, and CD68 in 63 cases of PC tissues in a tissue microarray. Moreover, we analyzed immunolocalization features, clinical associations and prognostic values of these molecules.RESULTS:The expressions of B7-H3, B7-H4, and HHLA2 could be detected in cytokeratin staining positive (CK+) cancer epithelial cells, CD68+tumor-associated macrophages (TAMs), and even other cells defined as CK-CD8-CD68-. Higher expression of B7-H3 in tumor cells could predict a better survival of the PC patients. A positive correlation was found between the expressions of B7-H3 and HHLA2 in tumor cells, while there was a negative correlation between the expressions of B7-H4 and HHLA2 in tumor cells. A positive correlation was found between the expressions of B7-H3 and B7-H4 or HHLA2 in CD68+TAMs, but not B7-H4 and HHLA2. Tumor-infiltrating CD8+T cells in combination with CD68+TAMs could serve as an important predictor for the postoperative prognosis of PC patients. Higher expression of B7-H3, or HHLA2 in CD68+TAMs could serve as an important predictor for poorer prognosis of PC patients. Patients with B7-H3lowB7-H4low, B7-H3lowHHLA2low, or B7-H4lowHHLA2low on CD68+TAMs could have a better postoperative prognosis compared with the other sub-populations in the combinational analysis.CONCLUSIONS:Taken together, our study indicated variable expressions and prognostic values of B7-H3, B7-H4, and HHLA2, in human PC tissues, and demonstrated that these co-stimulator molecules expressed by CD68+TAMs could be used as important bio-markers for the prognostic prediction of PC patients. Moreover, these results supported that the evaluation of these markers could be used as essential candidate targets for immunotherapy against PC.