PURPOSE:Esophageal squamous cell carcinoma (ESCC) is frequently refractory to radiation therapy, in part because of inefficient DNA damage repair and an immunosuppressive tumor microenvironment. We investigated whether ALYREF (Aly) coordinates radioresistance and immune evasion in ESCC. METHODS AND MATERIALS:Aly expression and its clinical associations were analyzed in the Cancer Genome Atlas (TCGA) cohorts and validated by immunohistochemistry in an independent ESCC cohort. Aly was genetically silenced or overexpressed in ESCC cell lines, followed by irradiation and subsequent assessment of clonogenic survival, reactive oxygen species (ROS) production, and apoptosis. Multi-omics analyses and mechanistic studies (protein interaction modeling, immunofluorescence, and DNA damage readouts) were performed to define pathways linking Aly to radioresponse and immune contexture. RESULTS:Aly was overexpressed in ESCC and associated with adverse clinical outcomes. Aly depletion suppressed ESCC growth and enhanced radiosensitivity, characterized by increased radiation-induced reactive oxygen species and apoptosis, as well as reduced clonogenic survival. Aly's loss was accompanied by changes consistent with impaired resolution of DNA double-strand break repair and activation of inflammatory signaling. Mechanistically, Aly engaged REV7-associated nuclear programs and Hsp70-linked cytoprotective signaling, thereby supporting DNA damage tolerance and survival after irradiation, and was correlated with an immunosuppressive contexture. CONCLUSIONS:Aly functions as a dual regulator of ESCC radioresistance by integrating DNA repair capacity and immune suppression. Targeting Aly may represent a strategy to radiosensitize ESCC and potentially improve responses to combined radiation therapy-immunotherapy.

Colorectal cancer (CRC) has high global incidence and mortality, yet effective diagnostic tools for detecting the adenoma to adenocarcinoma transition are lacking, making identification of adenomas with malignant transformation potential essential for early diagnosis and intervention. This study aimed to identify key biomarkers involved in the adenoma-carcinoma sequence. Based on this discovery, we developed an integrated radionuclide probe for diagnosis and therapy. Through a combination of retrospective clinical analysis, proteomics, and cellular experiments, we identified a significant upregulation of heat shock protein beta-1 (HSPB1) during the adenoma-carcinoma transition, which is closely associated with tumor proliferation, invasiveness, and metastatic potential. Based on this finding, we constructed HSPB1-targeted radiolabeled probes-177Lu-DPTP. In vitro and in vivo, including early CRC patient-derived xenograft (PDX) and low rectal cancer models, 177Lu-DPTP demonstrates high targeting specificity and favorable pharmacokinetics, selectively accumulates in HSPB1-high tumors, and produces effective imaging and therapeutic outcomes. It enabled noninvasive diagnosis via molecular imaging while significantly inhibiting tumor growth through targeted radiotherapy, without notable toxicity. In conclusion, our study highlights the pivotal role of HSPB1 in colorectal adenoma-carcinoma progression and demonstrates the potential of HSPB1-targeted probes for early diagnosis and precision therapy of CRC, offering a promising strategy for improved screening and treatment outcomes.

Neutrophils exhibit significant plasticity within the tumor microenvironment (TME), such as promoting immune suppression, tumor growth, and metastasis. Although various nano drug delivery systems targeting neutrophils have been developed based on their tumor chemotaxis, few studies have focused on designing nano-vaccines that specifically target and activate neutrophils to exert anti-tumor effects, mainly due to the lack of specific targets. To address these issues, we genetically engineered tumor cell membranes to first overexpress the CD300LD protein, which was subsequently coated onto the surface of liposomes. Additionally, we encoded and delivered the cytokine IL-36γ, which can effectively activate neutrophils, thus constructing an mRNA-based biomimetic nanovaccine, termed CMNPs (cell-membrane coated mRNA nanovaccine platform), which is designed to elicit potent anti-tumor immunity by targeted activation of neutrophils. Through analysis of single-cell public databases, we discovered that neutrophils in the TME express CD300LD receptors at significantly higher levels compared to other immune cells. Consequently, we further transduced tumor cells with lentivirus to induce overexpression of the CD300LD protein and utilized cell membrane biomimetic technology to modify the surface of CMNPs. The modified CMNPs can specifically target neutrophils via the extracellular domain (ECD) of the CD300LD protein. Upon targeted delivery of CMNPs to neutrophils in the TME, the internally encoded mRNA, fused with albumin and IL-36γ, is transcribed and translated. Since albumin prolongs the half-life of the cytokine, IL-36γ continuously activates neutrophils via the IL-36R receptor on their surface, enhancing their tumoricidal functions and activating other immune cells. Experimental results confirmed that CMNPs effectively activate neutrophils, leading to robust anti-tumor immune responses and tumor growth inhibition. Compared to the control group, CMNPs increased mouse survival rates to 85%. Collectively, the CMNPs vaccine demonstrates broad therapeutic efficacy and remarkable functional diversity, not only improving the effectiveness of immunotherapy but also providing a novel strategy for neutrophil-targeted tumor immunotherapy.

Background: Treatment options for unresectable locally advanced esophageal squamous cell carcinoma (ESCC) that cannot tolerate concurrent chemoradiotherapy (CCRT) are exceedingly limited. This study aimed to evaluate the efficacy and safety of tislelizumab, a programmed cell death protein 1 (PD-1) inhibitor, in combination with definitive radiotherapy as a novel chemotherapy-free first-line treatment for patients with unresectable, locally advanced ESCC who are ineligible for standard CCRT. Methods: This prospective, single-arm, phase II clinical trial (ChiCTR2100053182) recruited treatment-naïve patients diagnosed with unresectable, locally advanced ESCC at Binzhou Medical University Hospital. Eligibility was limited to patients considered ineligible for standard CCRT due to medical contraindications or patient preference. The investigational regimen consisted of tislelizumab (200 mg intravenously) administered on a three-week cycle, initiated concurrently with the first fraction of radiotherapy. The primary endpoint was progression-free survival (PFS). Additionally, the protocol included a prespecified exploratory analysis of tumor immune microenvironment markers using high-throughput multiplex immunofluorescence. Results:From May 2021 to October 2022, 32 eligible patients were enrolled. The mPFS was 23.5 months (95% CI 18.2–not reached), the 1-, 2-, and 3-year PFS rates were 75.0%, 57.1% and 42.9% respectively. The median overall survival (mOS) was not reached, with a 3-year OS rate of 53.6%. The regimen demonstrated substantial efficacy, achieving an objective response rate (ORR) of 82.1% and a pathological complete response (pCR) rate of 64.3%. Treatment was well-tolerated; the most frequent grade ≥3 treatment-related adverse event (TRAE) was lymphopenia (53.6%). Exploratory biomarker analysis revealed that high densities of T-cell exhaustion markers (e.g., intratumoral PD-1+ cells) were significantly associated with poorer PFS and OS, whereas high expression of stromal CD68+PD-L1+ cells was a strong favorable prognostic factor. Conclusion: This phase II study demonstrates that the combination of tislelizumab and definitive radiotherapy yields promising efficacy and manageable toxicity in patients with unresectable, locally advanced ESCC who are ineligible for standard CCRT. Moreover, the observed associations between immune cell signatures and treatment outcomes highlight the potential for biomarker-driven patient selection in future trials.

Introduction:Terminal uridyl transferase 4 (TUT4), a nucleotide transferase that modifies miRNA sequences, plays a critical role in regulating miRNA target interactions and function. However, its involvement in radiation-induced esophageal injury remains poorly understood. Methods:To investigate this, we performed computational analysis of RNA-seq data from irradiated esophageal tissues of wild-type and TUT4-knockout (TUT4-/-) mice, identifying 53 differentially expressed mRNAs (DEmRNAs), of which 30 were upregulated and 23 downregulated. Results:Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that these DEmRNAs were significantly associated with biological processes including lipid metabolism, fatty acid metabolism, proteolysis, and broader metabolic functions. Notably, DEmRNAs in TUT4-/- esophageal tissues showed marked enrichment in the renin-angiotensin system and peroxisome proliferator-activated receptor signaling pathways, implicating their potential roles in the pathogenesis of radiation-induced esophageal injury. In addition, we identified a regulatory axis in which a long non-coding RNA competes with miR-182 to modulate the competing endogenous RNA network governing TUT4 target genes. Collectively, our transcriptomic analysis offers novel mechanistic insights into how TUT4 may confer protection against radiation-induced damage in esophageal tissues.

In recent years, with the continuous development of single-cell sequencing (sc-seq) technology, Sc-seq can conduct high-throughput and high-resolution transcriptome analysis at the single-cell level, revealing gene expression differences and molecular characteristics among individual cells. This provides more precise guidance for determining tumor molecular subtypes and formulating treatment strategies. The neoadjuvant therapy (NAT) for esophageal cancer, including chemotherapy, radiotherapy, molecular targeted therapy, immunotherapy, and comprehensive treatment, has achieved significant progress. However, there remain considerable challenges in enhancing the quality of life for esophageal cancer patients, extending their survival period, and reducing the rate of late-stage recurrence. This article explores the Sc-seq technology, delineating its advantages and disadvantages. It further examines the application progress of this technology in the tumor microenvironment (TME) of esophageal cancer, as well as its application for various neoadjuvant treatment regimens. The article offers a novel perspective on the effectiveness of NAT and provides a basis for the neoadjuvant treatment of esophageal tumors.

Background:Radiotherapy and Chimeric Antigen Receptor(CAR)-T therapy may exhibit a synergistic effect, suggesting that incorporating radiotherapy into CAR-T could improve the prognosis for patients with refractory/relapsed diffuse large B-cell lymphoma (R/R DCBCL). A lack of standardized treatment protocols and relevant guidelines in bridging radiotherapy(BRT) prior to CAR-T therapy still exists. Consequently, we retrospectively analyzed the outcomes of R/R DLBCL patients treated with BRT prior to CAR-T therapy or not, aiming to evaluate the efficacy and satety of BRT as well as the impact of radiotherapy dose on prognosis. Methods:Between December 2017 and January 2025, 80 patients diagnosed with R/R DLBCL were treated with CAR-T. Thirty-five of them received BRT during leukapheresis and lymphodepletion. The primary endpoint of this study was progression-free survival(PFS), and secondary endpoints included overall survival(OS), disease-specific survival(DSS), in-field PFS, best objective response rate(ORR), and complete response rate(CRR). PFS and OS of CAR-T were compared between BRT group and no BRT group. In the subgroup of radiotherapy patients, PFS, OS and in-field PFS were compared between the low-Equivalent dose to 2 Gy per fraction(EQD2) subgroup and the high-EQD2 subgroup. Results:BRT group showed obviously longer PFS and OS than no BRT group(p = 0.001, p = 0.043). In addition, BRT did not increase the incidence of CAR-T toxicities during follow-up (median:35.27 months). Comprehensive BRT subgroup improved prognosis in PFS(p = 0.015) and OS(p = 0.029) when compared with focal BRT subgroup, no significant effect on DSS was noted(p = 0.109). High-EQD2 subgroup did not significantly improve PFS(p = 0.181) and OS(p = 0.665) except for local control(p = 0.079) especially in patients with high tumor burden(p = 0.005). There is no impact on prognosis between early salvage radiotherapy(SRT) and salvage chemotherapy(SCT) cohorts in patients with PR response to CAR-T therapy. Conclusions:Our analysis demonstrated that BRT is an effective and safe approach for patients with R/R DLBCL preparing for CAR T-cell therapy, which indicated that BRT may enhance the anti-tumor effect of CAR T-cells.

The advancement of single-cell sequencing technology has promoted the generation of a large amount of single-cell transcriptional profiles, providing unprecedented opportunities to identify drug-resistant cell subpopulations within a tumor. However, few studies have focused on drug response prediction at single-cell level, and their performance remains suboptimal. This paper proposed scAdaDrug, a novel multi-source domain adaptation model powered by adaptive importance-aware representation learning to predict drug response of individual cells. We used a shared encoder to extract domain-invariant features related to drug response from multiple source domains by utilizing adversarial domain adaptation. Particularly, we introduced a plug-and-play module to generate importance-aware and mutually independent weights, which could adaptively modulate the latent representation of each sample in element-wise manner between source and target domains. Extensive experimental results showed that our model achieved state-of-the-art performance in predicting drug response on multiple independent datasets, including single-cell datasets derived from both cell lines and patient-derived xenografts (PDX) models, as well as clinical tumor patient cohorts. Moreover, the ablation experiments demonstrated our model effectively captured the underlying patterns determining drug response from multiple source domains.

Despite advances in non-small cell lung cancer (NSCLC) therapies, resistance remains a major challenge. Ferroptosis, a form of regulated cell death, plays a key role in cancer progression and treatment response. However, the mechanisms governing ferroptosis in NSCLC are not fully understood. The Hippo pathway, which regulates cell proliferation, has recently been implicated in ferroptosis regulation. In this study, we identify Mitotic Spindle Positioning (MISP) as a critical inhibitor of ferroptosis in NSCLC. MISP is upregulated in NSCLC tissues, and its loss sensitizes cells to ferroptosis, reducing cell proliferation in vitro and in vivo. Mechanistically, MISP binds to the SARAH domain of MST1/2 kinases, inhibiting their homodimerization and autophosphorylation, leading to sustained activation of YAP, a transcriptional coactivator in the Hippo pathway. YAP activation increases SLC7A11 expression, which protects cells from ferroptosis. We also identify a mutant MISP-R390/391A that disrupts MISP-MST1/2 binding, further illustrating the MST1/2-dependent inhibition of Hippo signaling. Notably, MISP is a target of YAP, creating a feedback loop that amplifies YAP signaling. Our findings suggest a novel MISP-YAP axis regulating ferroptosis, positioning MISP as a potential therapeutic target for NSCLC, especially in cases with dysregulated YAP.

Background:A lot of studies have shown a close relationship between cuproptosis and cancer. The main purpose of this study is to analyze the impact of cuproptosis on cervical cancer (CC). Methods:Using The Cancer Genome Atlas (TCGA) public database, we analyzed the genetic correlation, expression, and prognostic value of 25 cuproptosis-related genes (CRGs) in CC. A least absolute shrinkage and selection operator (LASSO) risk regression model was constructed to compare the changes in associated pathways, prognosis, immune infiltration, and antibody programmed cell death-ligand 1 (anti-PD-L1) treatment response of the high- and low-risk groups. In addition, we collected CC tissue samples before and after radiotherapy for ribonucleic acid (RNA) sequencing, and analyzed the relationship between CRGs and radiotherapy. Results:The results showed CRGs were differentially expressed and were associated with multiple metabolic pathways. High expression of COX7B, PIH1D2, NDUFA1, NDUFA2 and NDUFB1 indicated a better prognosis. CRGs signature could predict prognosis (P<0.001) and affect immune infiltration. The prognosis was better in the low-risk group, while the high-risk group was more correlated with PD-L1. SLC25A5 downregulated expression (P=0.001) and SLC6A3 upregulated (P=0.02) after radiotherapy. SLC25A5 was related to the degree of differentiation of CC; the worse the differentiation, the higher the expression. Conclusions:CRGs may further affect patient prognosis and response to immunotherapy by influencing metabolic pathways and immune infiltration. Radiation could alter the expression of CRGs, which may have potential research value in evaluating the efficacy of radiotherapy.

The artificial intelligence-powered computational pathology has led to significant improvements in the speed and precision of tumor diagnosis, while also exhibiting substantial potential to infer genetic mutations and gene expression levels. However, current studies remain limited in predicting molecular subtypes and clinical outcomes in breast cancer. In this paper, we proposed a weakly supervised contrastive learning framework to address this challenge. Our framework first performed contrastive learning pretraining on a large number of unlabeled patches tiled from whole slide images (WSIs) to extract patch-level features. The gated attention mechanism was leveraged to aggregate patch-level features to produce slide feature that was then applied to various downstream tasks. To confirm the effectiveness of the proposed method, three public cohorts and one external independent cohort of breast cancer have been used to conducted evaluation experiments. The predictive powers of our model to infer gene expression, molecular subtypes, recurrence events and drug responses were validated across cohorts. In addition, the learned patch-level attention scores enabled us to generate heatmaps that were highly consistent with pathologist annotations and spatial transcriptomic data. These findings demonstrated that our model effectively established the high-order genotype-phenotype associations, thereby potentially extend the application of digital pathology in clinical practice.

Episil is a bioadhesive barrier-forming liquid gel that can relieve mucositis caused by radiotherapy (RT) and effectively relieve pain. The purpose of this trial is to compare the efficacy and safety of Episil in improving acute radiation dermatitis (ARD) in patients with breast cancer. This study included patients who met the criteria for postoperative RT for breast cancer. The primary end point was the grade of RD during treatment. A total of 102 patients were included in this study. The patients were grouped in a 2:1 ratio using the randomized number table method: 67 patients received Episil combined with conventional skin care (the Episil group), whereas the remaining 35 patients served as the control group and received conventional skin care only (the control group). According to the grading criteria of the Radiation Therapy Oncology Group (RTOG), the skin reaction rate and severity were significantly better in the Episil group than the control group (24.62%, 72.31%, 3.08, 0, 0 vs. 0, 85.71%, 14.29%, 0, 0, 0) across grades 0 to 4 (P < 0.05). The itchiness score exhibited s significant reduction in the Episil group as compared with the control group (P < 0.05). The results of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) showed that the overall health (z = -5.855, P < 0.001) and overall quality of life (z = -6.583, P < 0.001) were better in the Episil group than the control group after RT. Overall, in patients with breast cancer receiving RT, the topical application of Episil may significantly reduce the grading of ARD, alleviate patient symptoms, and improve the patient's overall quality of life.

Ductal carcinoma in situ and invasive ductal carcinoma represent two stages of breast cancer progression. A multitude of studies have shown that genomic instability increases during tumor development, as manifested by higher mutation and copy number variation rates. The advent of single-cell and spatial transcriptomics has enabled the investigation of the subtle differences in cellular states during the tumor progression at single-cell level, thereby providing more nuanced understanding of the intercellular interactions within the solid tumor. However, the evolutionary trajectory of tumor cells and the establishment of the immunosuppressive microenvironment during breast cancer progression remain unclear. In this study, we performed an exploratory analysis of the single-cell sequencing dataset of 13 ductal carcinoma in situ and invasive ductal carcinoma samples. We revealed that tumor cells became more malignant and aggressive during their progression, and T cells transited to an exhausted state. The tumor cells expressed various coinhibitory ligands that interacted with the receptors of immune cells to create an immunosuppressive tumor microenvironment. Furthermore, spatial transcriptomics data confirmed the spatial colocalization of tumor and immune cells, as well as the expression of the coinhibitory ligand-receptor pairs. Our analysis provides insights into the cellular and molecular mechanism underlying the formation of the immunosuppressive landscape during two typical stages of breast cancer progression. The Journal of Immunology, 2024, 213: 1392-1401.

IntroductionLung adenocarcinoma, a prevalent and lethal malignancy globally, is characterized by significant tumor heterogeneity and a complex tumor immune microenvironment during its histologic pattern progression. Understanding the intricate interplay between tumor and immune cells is of paramount importance as it could potentially pave the way for the development of effective therapeutic strategies for lung adenocarcinoma.MethodsIn this study, we run comparative analysis of the single-cell transcriptomic data derived from tumor tissues exhibiting four distinct histologic patterns, lepidic, papillary, acinar and solid, in lung adenocarcinoma. Furthermore, we conducted immunofluorescence assay and spatial transcriptomic sequencing to validated the spatial co-localization of typical co-inhibitory factors.Results and DiscussionOur analysis unveiled several co-inhibitory receptor-ligand interactions, including PD1-PDL1, PVR-TIGIT and TIGIT-NECTIN2, that potentially exert a pivotal role in recruiting immunosuppressive cells such as M2 macrophages and Tregs into LUAD tumor, thereby establishing immunosuppressive microenvironment and inducing T cells to exhaustion state. Furthermore, The expression level of these co-inhibitory factors, such as NECTIN2 and PVR, were strongly correlated with low immune infiltration, unfavorable patient clinical outcomes and limited efficacy of immunotherapy. We believe this study provides valuable insights into the heterogeneity of molecular, cellular interactions leading to immunosuppressive microenvironment during the histological progression of lung adenocarcinoma. The findings could facilitate the development of novel immunotherapy for lung cancer.

ObjectiveThis study endeavored to explore the optimal treatment strategy and conduct a prognostic analysis for patients diagnosed with pT4M0 (pathologic stage T4) colon adenocarcinoma (COAD).Methods and materialsA total of 8,843 patients diagnosed with pT4M0 COAD between January 2010 and December 2015 were included in this study from the Surveillance, Epidemiology, and End Results (SEER) database. These patients were randomly divided into a training set and an internal validation set using a 7:3 ratio. Variables that demonstrated statistical significance (P<0.05) in univariate COX regression analysis or held clinical significance were incorporated into the multivariate COX regression model. Subsequently, this model was utilized to formulate a nomogram. The predictive accuracy and discriminability of the nomogram were assessed using the C-index, area under the curve (AUC), and calibration curves. Decision curve analysis (DCA) was conducted to confirm the clinical validity of the model.ResultsIn the entire SEER cohort, the 3-year overall survival (OS) rate (74.22% vs. 63.20%, P<0.001) and the 3-year cancer-specific survival (CSS) rate (76.25% vs. 66.98%, P<0.001) in the surgery combined with postoperative adjuvant therapy (S+ADT) group surpassed those in the surgery (S) group. Multivariate COX regression analysis of the training set unveiled correlations between age, race, N stage, serum CEA (carcinoembryonic antigen), differentiation, number of resected lymph nodes, and treatment modalities with OS and CSS. Nomograms for OS and CSS were meticulously crafted based on these variables, achieving C-indexes of 0.692 and 0.690 in the training set, respectively. The robust predictive ability of the nomogram was further affirmed through receiver operating characteristic (ROC) and calibration curves in both the training and validation sets.ConclusionIn individuals diagnosed with pT4M0 COAD, the integration of surgery with adjuvant chemoradiotherapy demonstrated a substantial extension of long-term survival. The nomogram, which incorporated key factors such as age, race, differentiation, N stage, serum CEA level, tumor size, and the number of resected lymph nodes, stood as a dependable tool for predicting OS and CSS rates. This predictive model held promise in aiding clinicians by identifying high-risk patients and facilitating the development of personalized treatment plans.

PURPOSE:Radiation-induced pneumonitis (RIP) seriously limits the application of radiation therapy in the treatment of thoracic tumors, and its etiology and pathogenesis remain elusive. This study aimed to elucidate the role of ubiquitin-specific peptidase 11 (USP11) in the progression of RIP and the associated underlying mechanisms. METHODS AND MATERIALS:Changes in cytokines and infiltrated immune cells were detected by enzyme-linked immunosorbent assays and immunohistochemistry after exposure to 20 Gy x-ray with whole-thorax irradiation. The effects of USP11 expression on endothelial cell proliferation and apoptosis were analyzed by costaining of CD31/Ki67 and CD31/caspase-3 in vivo, and the production of cytokines and reactive oxygen species was confirmed by reverse-transcription polymerase chain reaction and flow cytometry in vitro. Comprehensive proteome and ubiquitinome analyses were used for USP11 substrate screening after radiation. Results were verified by Western blotting and coimmunoprecipitation experiments. Recombinant adeno-associated virus lung vectors expressing OTUD5 were used for localized overexpression of OTUD5 in mouse pulmonary tissue, and immunohistochemistry was conducted to analyze cytokine expression. RESULTS:The progression of RIP was significantly alleviated by reduced expression of proinflammatory cytokines in both Usp11-knockout (Usp11-/-) mice and in mice treated with the USP11 inhibitor mitoxantrone. Likewise, the absence of USP11 resulted in decreased permeability of pulmonary vessels and neutrophils and macrophage infiltration. The proliferation rates of endothelial cells were prominently increased in the Usp11-/- lung, whereas apoptosis in Usp11-/- lungs decreased after irradiation compared with that observed in Usp11+/+ lungs. Conversely, USP11 overexpression increased proinflammatory cytokine expression and reactive oxygen species production in endothelial cells after radiation. Comprehensive proteome and ubiquitinome analyses indicated that USP11 overexpression upregulates the expression of several deubiquitinating enzymes, including USP22, USP33, and OTUD5. We demonstrate that USP11 deubiquitinates OTUD5 and implicates the OTUD5-STING signaling pathway in the progression of the inflammatory response in endothelial cells. CONCLUSIONS:USP11 exacerbates RIP by triggering an inflammatory response in endothelial cells both in vitro and in vivo, and the OTUD5-STING pathway is involved in the USP11-dependent promotion of RIP. This study provides experimental support for the development of precision intervention strategies targeting USP11 to mitigate RIP.

Tumor-associated macrophages (TAMs) are known to promote tumor growth, invasion, metastasis, and protumor angiogenesis, but the role of TAMs in evading radiotherapy in esophagus cancer remains unclear. In this study, we first induced TAMs from human monocytes (THP-1) and identified using immunofluorescence and Western blotting assays. We then co-cultured them with human esophageal cancer cell lines. CCK-8, colony formation, Transwell, scratch test, and TUNEL assays showed that TAMs could promote proliferation, survival rate, invasion, migration, and radioresistance and could inhibit apoptosis of the esophageal squamous carcinoma cell lines KYSE-150 and TE-1 before and after radiotherapy both in vivo and in vitro. Using LV-VEGFA-RNAi lentiviral vectors, we also found that TAMs could increase the expression of VEGFA and that inhibition of VEGFA could inhibit the biological function caused by TAMs. Finally, a Western blotting assay was used to evaluate the expression of various factors underlying the mechanism of TAMs. VEGFA, MAPK, P-MAPK, BCL-2, and Snail proteins were found to be overexpressed in co-cultured groups, whereas after VEGFA inhibition, MAPK, P-MAPK, BCL-2, and Snail proteins were found to be significantly downregulated in the radiotherapy group. These study results offer important information regarding the mechanism of radioresistance in esophageal cancer.

PURPOSE:To analyze the gene mutation, immune infiltration and tumor growth of primary tumor and distant tumor under different treatment modes.MATERIALS AND METHODS:Twenty B16 murine melanoma cells were injected subcutaneously into the of both sides of the thigh, simulating a primary tumor and a secondary tumor impacted by the abscopal effect, respectively. They were divided into blank control group, immunotherapy group, radiotherapy group, and radiotherapy combined immunotherapy group. During this period, tumor volume was measured, and RNA sequencing was performed on tumor samples after the test. R software was used to analyze differentially expressed genes, functional enrichment, and immune infiltration.RESULTS:We found that any treatment mode could cause changes in differentially expressed genes, especially the combination treatment. The different therapeutic effects might be caused by gene expression. In addition, the proportions of infiltrating immune cells in the irradiated and abscopal tumors were different. In the combination treatment group, T-cell infiltration in the irradiated site was the most obvious. In the immunotherapy group, CD8+ T-cell infiltration in the abscopal tumor site was obvious, but immunotherapy alone might have a poor prognosis. Whether the irradiated or abscopal tumor was evaluated, radiotherapy combined with anti-programmed cell death protein 1 (anti-PD-1) therapy produced the most obvious tumor control and might have a positive impact on prognosis.CONCLUSION:Combination therapy not only improves the immune microenvironment but may also have a positive impact on prognosis.

Abstract Background Radiation-induced esophageal injury remains a limitation of radiotherapy for lung and esophageal cancer patients. Esophageal epithelial cells are extremely sensitive to irradiation; nevertheless, factors involved in the radiosensitivity of esophageal epithelial cells are still unknown. Terminal uridyl transferase 4 (TUT4) can modify the sequence of miRNAs, which affects their regulation of miRNA targets and function. Methods In this study, we used transcriptome sequencing technology to identify mRNAs that were differentially expressed before and after radiotherapy in esophageal epithelial cells. Results We further explored the mRNA expression profiles between wild-type and TUT4 knockout esophageal epithelial tissue. Volcano and heatmap plot unsupervised hierarchical clustering analyses were performed to classify the samples. Enrichment analysis of Gene Ontology (GO) functional annotations and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways was performed. Annotation of differential genes from metabolism, genetic information processing, environmental information processing, cellular processes and organismal systems human diseases was conducted. Conclusions The aberrantly expressed genes were significantly enriched in irradiation-related biological processes, such as lipid metabolic process, fatty acid metabolic process, proteolysis, and metabolic process. Moreover, we explored the distribution of the transcription factor family and its target genes among the differentially expressed genes. In addition, we found that PPAR pathway and the renin-angiotensin pathway might serve as therapeutic targets in TUT4-related radiation-induced esophageal injury.

Abstract Background Male breast cancer (MBC) is a rare condition, and the effectiveness of radiotherapy in treating MBC patients remains uncertain. This study aimed to investigate the role of adjuvant radiotherapy (RT) in the management of MBC. Methods MBC patients from the Surveillance Epidemiology and End Results (SEER) database were included in the study and were divided into RT and no-RT groups. A 1:1 propensity score matching (PSM) method was employed to balance baseline characteristics. Kaplan-Meier curves were used to evaluate the impact of RT on overall survival (OS) and breast cancer-specific survival (BCSS). Cox analyses were conducted to identify factors associated with survival. Subgroup analysis was performed to identify subgroups of MBC patients who might benefit from RT. Results In the matched cohort, the 5-year OS and BCSS rates were higher in the RT group compared to the no-RT group (p = 0.023, p = 0.035). Univariate and multivariate analysis demonstrated significant differences in both OS and BCSS associated with RT (p = 0.024, p = 0.037, p = 0.025, p = 0.028). Forest plots revealed a greater OS benefit in patients with T1 stage, age ≥ 60 years, estrogen receptor positivity (ER+), absence of distant metastasis (M0), married status, and with local or regional metastases. Furthermore, a greater BCSS benefit was observed in patients aged ≥ 60 years, ER+, progesterone receptor negativity (PR-), M0, married status, and with regional metastases who received RT for MBC. Conclusion RT in MBC patients is associated with improved survival and is recommended for patients aged ≥ 60 years with ER+, PR-, M0, married status, and regional metastases.