Acute heart failure (AHF) is a life-threatening clinical syndrome due to impaired cardiac function. Ferroptosis has emerged as a contributor to cytotoxicity in cardiomyocytes. However, the functional interplay between USP8 and ferroptosis during AHF has not been investigated. H9C2 rat cardiomyocytes were treated with doxorubicin (Dox) to establish an experimental model. Cell cytotoxicity was evaluated by measuring cell viability, LDH release, and cell death. Ferroptosis was assessed by analyzing Fe2+, lipid ROS, MDA, and GSH levels in treated cells. Immunoprecipitation (IP), Co-IP, and protein stabilization assays were performed to validate the USP8/murine double minute 4 (MDM4) interaction and the regulation of USP8 in MDM4. Expression of mRNA and protein was quantified by quantitative PCR and immunoblot analyses, respectively. USP8 and MDM4 were downregulated in Dox-exposed H9C2 cardiomyocytes. USP8 overexpression alleviated Dox-triggered cytotoxicity and cell death in H9C2 cardiomyocytes. Moreover, USP8 overexpression mitigated H9C2 cardiomyocyte ferroptosis induced by Dox. Mechanistically, USP8 stabilized MDM4 via deubiquitination. Inhibition of MDM4 counteracted the ability of USP8 overexpression to attenuate Dox-triggered cell death and ferroptosis in H9C2 cardiomyocytes. Our findings indicate that USP8 overexpression protects H9C2 cardiomyocytes from Dox-induced ferroptosis by stabilizing MDM4 via deubiquitination.

The efficacy of prophylactic cranial irradiation (PCI) is still controversial in small cell lung cancer (SCLC) patients with pT1-2N0M0 disease after complete resection. The majority of previous studies haven’t discerned the potential benefit of PCI in this subgroup of patients, probably due to the paucity of these patients and distinct treating modalities. The aim of this study is to demonstrate the potential medical benefit of PCI in these patients. A retrospective study was conducted to evaluate the potential benefit of PCI in pT1-2N0M0 SCLC patients after complete resection. We retrospectively reviewed 112 pT1-2N0M0 SCLCs after complete resection and adjuvant chemotherapy between January 2013 and January 2022. Survival and Cox regression analysis were conducted to elucidate the potential medical benefit of PCI in these patients. The median overall survival (OS) has not been reached. The 2-year, 5-year, 8-year OS rate was 93.7

Abstract Background: Inetetamab is a neotype HER2-targeted monoclonal antibody with amino acids modified Fc segment which optimizes the antibody-dependent cellular cytotoxicity effect. However, robust evidence evaluating the combination of inetetamab combined with pertuzumab, paclitaxel and carboplatin (TCbIP) for neoadjuvant therapy is still lacking. This study aimed to evaluate the efficacy and safety of TCbIP as a neoadjuvant therapy for patients with locally advanced HER2-positive breast cancer. We now present the updated results of this trial. Methods: This phase II trial included female patients with histologically confirmed stage IIA to IIIC HER2-positive primary invasive breast cancer. Eligible patients received TCbIP treatment every three weeks for a maximum of six cycles followed by surgery. The primary endpoint was pathologic complete response (pCR, ypT0/is ypN0) rate. Key secondary endpoints included near pCR (npCR, residual breast disease < 1cm) rate, objective response rate (ORR) and safety. Results: From November 2021 to July 2023, 40 patients were enrolled in the trial. One patient received one cycle of the study treatment but was lost to follow-up without surgery, and five patients received one cycle of the study treatment and were still undergoing treatment, leaving 34 patients in the intention-to-treat (ITT) population. Among these 34 patients (82.4% in stage III), 22 patients completed the study treatment and surgery (per-protocol [PP] population) and 12 patients were still undergoing neoadjuvant treatment. The ORR was 91.2% (31/34) in the ITT population and 86.4% (19/22) in the PP population. Among the 22 patients in the PP population, 12 patients (54.5%) achieved pCR, and18 patients (81.8%) achieved npCR. The pCR rates for patients with hormone receptor (HR) negative and positive tumors were 80.0% (8/10) and 33.3% (4/12), respectively. The most common grade 3 adverse event was neutropenia (17.6%). No significant reduction in the left ventricular ejection fraction was observed in any patient. Conclusions: Neoadjuvant therapy with TCbIP demonstrated promising efficacy and manageable toxicity in patients with HER2-positive locally advanced breast cancer. Registration number: NCT05749016 (www.clinicaltrials.gov). Citation Format: Yue Chai, Jiaxuan Liu, Mingxia Jiang, Maiyue He, Xin Wang, Yipeng Wang, Xue Yang, Jing Wang, Binghe Xu, Qiao Li. Updated Results of a Phase II Study: Neoadjuvant Inetetamab Combined with Pertuzumab, Paclitaxel and Carboplatin for Locally Advanced HER2-Positive Breast Cancer [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO2-18-12.

Abstract Background Inetetamab is the first domestically developed innovative anti-HER2 monoclonal antibody in China, proven effective and safe in HER2-positive advanced breast cancer. However, its efficacy and safety in neoadjuvant treatment of HER2-positive locally advanced breast cancer (LABC) remain to be validated. Methods This prospective cohort study aimed to evaluate the efficacy and safety of inetetamab combined with pertuzumab, taxanes, and carboplatin (TCbIP) in neoadjuvant therapy for HER2-positive LABC, comparing it to data from patients treated with the TCbHP regimen (trastuzumab combined with pertuzumab, taxanes, and carboplatin) using propensity score matching (PSM). The primary endpoint was total pathological complete response (tpCR). Adverse events (AEs), objective response rate (ORR), and near-pCR were key secondary endpoints. Results Forty-four patients with clinical stage IIA-IIIC HER2-positive LABC were prospectively enrolled and treated with the TCbIP regimen. The tpCR rate among 28 patients who completed surgery was 60.7%, comparable to and slightly higher than the TCbHP group in PSM (60.7% vs. 53.6%, P = 0.510). The ORR was 96.4%, and the DCR reached 100.0%. The most common ≥ grade 3 AE was neutropenia (21.4% vs. 11.9%, P = 0.350). No significant reduction in left ventricular ejection fraction was observed, and no patient withdrew from treatment due to AEs. Conclusion Neoadjuvant therapy with TCbIP showed good efficacy and safety in patients with HER2-positive LABC and might be another promising option for neoadjuvant treatment. Trial registration NCT05749016 (registration date: Nov 01, 2021).

e12609 Background: Inetetamab is an Fc segment-modified innovative anti-HER2 monoclonal antibody. It has been proven to be effective and safe in HER2-positive advanced breast cancer. However, its efficacy and safety in the neoadjuvant treatment of HER2-positive locally advanced breast cancer (LABC) remains to be validated. This study aimed to evaluate the efficacy and safety of Inetetamab-based neoadjuvant therapy (TCbIP). Methods: In this prospective cohort study, we investigated the efficacy and safety of TCbIP in neoadjuvant therapy for HER2-positive LABC, and compared with TCbHP regimen (trastuzumab combined with pertuzumab, paclitaxel, and carboplatin) with propensity score matching (PSM). Thirty-four patients (clinical stage IIA-IIIC, HER2-positive LABC) were prospectively enrolled in this study and treated with the TCbIP regimen. The primary endpoint was total pathological complete response (tpCR). Adverse events (AEs), objective response rate (ORR), and near-pCR were key secondary endpoints. Results: The pCR rate of 28 patients who finished surgery was 60.7%, and was comparable and slightly higher (60.7% vs. 53.6%, P = 0.510) than the TCbHP group in PSM. The ORR was 96.4% and DCR reached 100.0%. The most common ≥grade 3 AE was neutropenia (21.4% vs. 11.9%, P = 0.350). No significant reduction in left ventricular ejection fraction was observed, and no patient withdrew from treatment due to AEs. Conclusions: Neoadjuvant therapy with TCbIP resulted in good efficacy and safety in patients with HER2-positive LABC, and might be another promising option for neoadjuvant treatment. Clinical trial information: NCT05749016 .

Abstract Background Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer lacking specific targeted therapeutic options. The deubiquitinizing enzyme OTUD6B is emerging as a potential player in TNBC progression. Objective The study aimed to ascertain the role of OTUD6B in TNBC progression and to exploit its therapeutic potential using a novel delivery system. Methods OTUD6B expression in TNBC was studied in relation to patient survival outcomes. We explored the potential of OTUD6B siRNA for TNBC treatment and its combined delivery with the chemotherapeutic agent, DOX, using a synthesized DNA tetrahedral nanoparticle (Td). The uptake and efficacy of the siOTUD6B/DOX@Td combination in TNBC cells were then evaluated, alongside an investigation into the underlying molecular mechanisms. Results There was a significant upregulation of OTUD6B in TNBC, inversely correlated with patient survival. The siOTUD6B/DOX@Td composite demonstrated efficient cellular uptake by TNBC cells, leading to the gene silencing of OTUD6B and controlled DOX release. This dual treatment approach enhanced apoptosis rates, increased DOX sensitivity, and inhibited TNBC cell growth, migration, and metastasis. In vivo results further confirmed the inhibition of tumor growth and metastasis without harming primary organs. Mechanistically, OTUD6B influenced TNBC progression through MDM2 stabilization and FOXO3 degradation. Conclusion Our findings highlight the pivotal role of OTUD6B in TNBC progression and underscore the potential of DNA tetrahedral nanoparticles as drug delivery systems. The combined siOTUD6B/DOX@Td strategy offers a promising therapeutic approach for TNBC and demonstrates the broader applicability of DNA nanotechnology in biomedicine.

Background Small cell lung cancer (SCLC) is a highly malignant cancer characterized by metastasis and an extremely poor prognosis. Although combined chemoimmunotherapy improves the prognosis of extensive-stage (ES)-SCLC, the survival benefits remain limited. Furthermore, no reliable biomarker is available so far to predict the treatment outcomes for chemoimmunotherapy.Methods This retrospective study included patients with ES-SCLC treated with first-line combined atezolizumab or durvalumab with standard chemotherapy between Janauray 1, 2019 and October 1, 2022 at five medical centers in China as the chemoimmunotherapy group. The patients were divided into one training cohort and two independent external validation cohorts. Additionally, we created a control group of ES-SCLC who was treated with first-line standard chemotherapy alone. The Radiomics Score was derived using machine learning algorithms based on the radiomics features extracted in the regions of interest delineated on the chest CT obtained before treatment. Cox proportional hazards regression analysis was performed to identify clinical features associated with therapeutic efficacy. The log-rank test, time-dependent receiver operating characteristic curve, and Concordance Index (C-index) were used to assess the effectiveness of the models.Results A total of 341 patients (mean age, 62±8.7 years) were included in our study. After a median follow-up time of 12.1 months, the median progression-free survival (mPFS) was 7.1 (95% CI 6.6 to 7.7) months, whereas the median overall survival (mOS) was not reached. The TNM stage, Eastern Cooperative Oncology Group performance status, and Lung Immune Prognostic Index showed significant correlations with PFS. We proposed a predictive model based on eight radiomics features to determine the risk of chemoimmunotherapy resistance among patients with SCLC (validation set 1: mPFS, 12.0 m vs 5.0 m, C-index=0.634; validation set 2: mPFS, 10.8 m vs 6.1 m, C-index=0.617). By incorporating the clinical features associated with PFS into the radiomics model, the predictive efficacy was substantially improved. Consequently, the low-progression-risk group exhibited a significantly longer mPFS than the high-progression-risk group in both validation set 1 (mPFS, 12.8 m vs 4.5 m, HR=0.40, p=0.028) and validation set 2 (mPFS, 9.2 m vs 4.6 m, HR=0.30, p=0.012). External validation set 1 and set 2 yielded the highest 6-month area under the curve and C-index of 0.852 and 0.820, respectively. Importantly, the integrated prediction model also exhibited considerable differentiation power for survival outcomes. The HR for OS derived from the low-progression-risk and high-progression-risk groups was 0.28 (95% CI 0.17 to 0.48) in all patients and 0.20 (95% CI 0.08 to 0.54) in validation set. By contrast, no significant differences were observed in PFS and OS, between high-progression-risk patients receiving chemoimmunotherapy and the chemotherapy cohort (mPFS, 5.5 m vs 5.9 m, HR=0.90, p=0.547; mOS, 14.5 m vs 13.7 m, HR=0.97, p=0.910).Conclusions The integrated clinical and radiomics model can predict the treatment outcomes in patients with ES-SCLC receiving chemoimmunotherapy, rendering a convenient and low-cost prognostic model for decision-making regarding patient management.

Objective: Head and neck squamous cell carcinoma (HNSCC) is a common heterogeneous cancer with complex carcinogenic factors. However, the current TNM staging criteria to judge its severity to formulate treatment plans and evaluate the prognosis are particularly weak. Therefore, a robust diagnostic model capable of accurately diagnosing and predicting HNSCC should be established.Methods: Gene expression and clinical data were retrieved from The Cancer Genome Atlas and Gene Expression Omnibus databases. Key prognostic genes associated with HNSCC were screened with the weighted gene co-expression network analysis and least absolute shrinkage and selection operator (LASSO) Cox regression model analysis. We used the timeROC and survival R packages to conduct time-dependent receiver operating characteristic curve analyses and calculated the area under the curve at different time points of model prediction. Patients in the training and validation groups were divided into high- and low-risk subgroups, and Kaplan-Meier (K-M) survival curves were plotted for all subgroups. Subsequently, LASSO and support vector machine algorithms were used to screen genes to construct diagnostic model. Furthermore, we used the Wilcoxon signed-rank test to compare the half-maximal inhibitory concentrations of common chemotherapy drugs among patients in different risk groups. Finally, the expression levels of eight genes were measured using quantitative real-time polymerase chain reaction and immunohistochemistry.Results: Ten genes (SSB, PFKP, NAT10, PCDH9, SHANK2, PAX8, CELSR3, DCLRE1C, MAP2K7, and ODF4) with prognostic potential were identified, and a risk score was derived accordingly. Patients were divided into high- and low-risk groups based on the median risk score. The K-M survival curves confirmed that patients with high scores had significantly worse overall survival. Receiver operating characteristic curves proved that the prognostic signature had good sensitivity and specificity for predicting the prognosis of patients with HNSCC. Univariate and multivariate Cox regression analyses confirmed that the gene signature was an independent prognostic risk factor for HNSCC. Diagnostic model was built by identifying eight genes (SSB, PFKP, NAT10, PCDH9, CELSR3, DCLRE1C, MAP2K7, and ODF4). The high-risk group showed higher sensitivity to various common chemotherapeutic drugs. DCLRE1C expression was higher in normal tissues than in HNSCC tissues.Conclusion: Our study identified the important role of tumor-driver genes in HNSCC and their potential clinical diagnostic and prognostic values to facilitate individualized management of patients with HNSCC.

Objectives: To examine the associations between physical activity and tinnitus development and physical activity and tinnitus severity in a large representative sample of US adults. Design: Data were obtained from 3826 eligible participants (20 to 69 years) in the National Health and Nutrition Examination Survey between 2015 and 2016. Physical activity was assessed using a Global Physical Activity Questionnaire. We used multivariable logistic regression to test the associations of physical activity (without physical activity, with physical activity) and amount of physical activity (min/week, in quartiles) with tinnitus symptoms. Adults with depressive symptoms were excluded, and the models were controlled for relevant sociodemographic, lifestyle, and health-related covariates. A restricted cubic spline was used to explore the dose-response relationship between the amount of physical activity and tinnitus. Results: Overall, 12.8% of the population who engaged in physical activity reported tinnitus, compared with 18.5% of the population who did not (p = 0.005). Subgroup analysis based on the amount of physical activity showed that participants who performed physical activity (150 to 300, 310 to 540, and 550 to 4800 min/week) had lower risks of tinnitus than those with no physical activity (odds ratio = 0.72, 0.56, and 0.62, respectively), after adjusting for covariates. However, no correlation was observed between physical activity and tinnitus severity in the present study. The dose-response analysis showed a nonlinear relationship (P for nonlinearity = 0.04) between the amount of physical activity and the risk of tinnitus. Conclusions: Physical activity may be associated with a reduced risk of tinnitus. Further research using a longitudinal design is required to confirm these findings and clarify the direction of causation.

Purpose: To achieve biofunctional annotation of imaging features and establish a prognostic risk prediction model for colorectal cancer.Experimental Design: This study used the Dazhou Central Hospital of Sichuan province and The Cancer Genome Atlas (TCGA) to collect the colorectal cancer data, which was divided into three parts. Firstly, Radiomics dataset was used to obtain the prognostic imaging features through univariate Cox and Lasso-Cox analysis. Next, Radiogenomics dataset and TCGA dataset were used for robustness module search based on Weighted correlation network analysis (WGCNA) and Z-Summary. The pathway information significantly correlated with prognostic imaging features was used to realize biofunctional annotation of imaging features. Finally, genes in pathways significantly associated with prognostic imaging features were extracted. The gene expression was transformed into an artificial image object as the input of the classification task of prognostic risk level based on ResNet18 deep learning model.Results: GSVA pathway enrichment of genes in modules significantly associated with prognosis-related imaging features revealed that 124 pathways were significantly associated with 23 (23/26) imaging features (|R| > 0.4 & P < 0.01). Most of these pathways (73/124) related to immunity. In the prognostic risk level prediction task, our model achieved an accuracy of 0.84 and 0.81 for the training set and test set, respectively, with area under curve of 0.93 and 0.83.Conclusions: This study successfully realized biofunctional annotation of 23 imaging features and innovatively transformed transcriptome data into artificial image objects to establish a ResNet18-based prognostic risk level prediction model.Funding Information: This work was supported by the National Natural Science Foundation of China (No. 21873034), the Key Projects fund of Science & Technology Department of Sichuan Province (2022YFS0588), Innovative Scientific Research Project of Medical in Sichuan Province (Q20073), the Scientific Research Fund of Technology Bureau in Dazhou (21ZDYF0029, 2021CDDZ-26 ).Declaration of Interests: They decleared no conflict of interest.Ethics Approval Statement: The institutional review board at Dazhou Central Hospital of Sichuan Province approved the study (No. 2020030).

Nonsmall cell lung cancer (NSCLC) accounts for the majority of lung cancers. Studies have revealed the regulatory role of lncRNAs in cancer pathogenesis and their potential use as diagnostic and prognostic biomarkers. The epidermal growth factor receptor antisense RNA 1 (EGFR-AS1) has been reported to be upregulated in NSCLC tissues, while its detailed mechanism in lung cancer needs to be explored. DNA damage-regulated autophagy modulator 1 (DRAM1) has been known to act as a tumor suppressor in NSCLC, and miR-524-5p has been reported to be a biomarker in idiopathic pulmonary fibrosis and different lung disorders. Our investigation revealed that EGFR-AS1 is highly expressed in lung cancer tissues, and its knockdown inhibited lung cancer cell invasion and viability and reduced tumor growth in vivo. We also found that EGFR-AS1 targets miR-524-5p, and there was a negative correlation between their expressions in lung cancer tissues. Simultaneously, miR-524-5p has been found to promote DRAM1 expression. In addition, the inhibition of miR-524-5p diminished DRAM1 protein expression and promoted lung cancer cell invasion. Our study has revealed that EGFR-AS1 contributes to the pathogenesis of NSCLC by inhibiting autophagic-lysosomal degradation via targeting the miR-524-5p/DRAM1 axis. This finding elucidated for the first time the role of EGFR-AS1 in lung cancer progression and the positive regulatory function of miR-524-5p in regulating DRAM1 protein and suppressing lung cancer progression. This novel mechanism provided a better insight into the pathogenesis of lung cancer and presented a better strategy for the treatment of lung cancer.

Background: Laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors of the head and neck, and it has shown increasing incidence and mortality. The mechanistic target of rapamycin complex 1 (mTORC1) is frequently dysregulated in LSCC, but its underlying mechanisms remain unclear.Methods: Establishment of a novel LSCC cell line using primary LSCC tumor tissues with dysregulated mTORC1 activity and then stable knockdown of Raptor (an mTORC1 specific component) in this cell line. Transcriptomic sequencing, quantitative real-time PCR, western blot analysis, and immunofluorescence assays were used to identify the crucial downstream effector of mTORC1. A series of experiments were conducted to investigate the functions and underlying mechanisms of the mTORC1 target gene in LSCC progression. Clinical LSCC samples were used to evaluate the association of mTORC1 and its downstream targets with clinicopathological features and patient prognosis. Finally, the influence on cisplatin (CDDP) sensitivity upon depletion of the mTORC1 target gene was assessed using a cell culture system, a cell line-derived xenograft (CDX) model, and a patient-derived xenograft (PDX) model.Results: We successfully established a novel LSCC cell line with hyperactivated mTORC1 activity and then identified integrin subunit alpha 5 (ITGA5) as a novel functional downstream effector of mTORC1 in the progression of LSCC. Elevated ITGA5 promotes LSCC progression through augmentation of ephrin-B2 (EFNB2). Clinical data analysis indicated that the activation of the mTORC1-ITGA5-EFNB2 signaling pathway is associated with malignant progression and poor prognosis of LSCC patients. Inhibition of ITGA5 significantly sensitized LSCC cells to CDDP.Conclusions: Our findings highlight a novel molecular mechanism for the tumorigenesis driven by deregulated mTORC1 signaling in LSCC, suggesting that the ITGA5-EFNB2 axis may be a therapeutic target for the treatment of mTORC1-related LSCC.

As the major effector of cellular adaptive immune response, CD8+ T cells play crucial roles in the host's defense against tumor and infection. Two recent studies revealed that basic leucine zipper ATF-like transcriptional factor (BATF) not only blocks the exhaustion of CD8+ engineered chimeric antigen receptor (CAR)-T cells, but also regulates the differentiation of stem-like CD8+ T precursor cells into effector cells during chronic infection [1, 2]. These results emphasized the important roles of transcriptional regulation in promoting T cell effector functions and preventing T cell exhaustion as well, suggesting the potential application of motivating transcriptional regulators such as BATF to improve the efficacy of adaptive T cell immunotherapy of cancer and chronic infection. In response to antigenic peptide stimulation, naive T cells proliferate and differentiate to effector T cells to eliminate malignant cells and invading pathogens, part of which further transform to memory T cells to provide long-term immune protection [3]. When the stimulation persists in the tumor microenvironment (TME) and chronic infections, CD8+ T cells can gradually enter an exhausted state characterized by high expression of inhibitory receptors as well as the overall epigenetic and metabolic profile changes, leading to the failure of clearing tumor cells and pathogens [4]. Therefore, discovering the key transcriptional regulators to T cell exhaustion will provide the potential target to improve the therapeutic effect of cancer immunotherapy by blocking or reversing the exhaustion of effector T cells. Transcription factor nuclear factor of activated T cells (NFAT) has been recognized as the crucial regulator of effector responses of T cells by interacting with its partner activator protein 1 (AP-1) [5]. Based on a preliminary screening of transcription factors that promote NFAT-AP-1 activity, Seo et al. [1] constructed CAR-T cells overexpressing three candidates (BATF, JUN and MAF basic leucine zipper transcription factor F [MAFF]) and then transferred them to mice inoculated with B16F0-hCD19 tumors. As a result, they found that CAR-T cells overexpressed with BATF, a cellular transcription factor that participates in the regulation of various immune cells differentiation, showed the most significant antitumor abilities with the enhanced effector functions and increased infiltration in the TME. Transcriptome results confirmed that BATF-overexpressing CAR-T cells manifest with increased tumor infiltration, intratumor expansion, effector functions and decreased exhaustion tendency. In addition, BATF-transduced CAR-T cells can survive several weeks after tumor clearance and acquire similar phenotype of central memory T cells, rendering mice more resistant to the secondary occurrence of tumor. Further assays for transposase-accessible chromatin using sequencing (ATAC-seq) showed decreased accessibility of exhaustion-related chromatin regions in BATF overexpressing cells, which was consistent with the reduced exhaustion tendency of CAR-T cells by cytometry by time of flight (CyTOF) analysis. Next, Seo and colleagues affirmed that BATF exert the regulatory function by binding with interferon regulatory factor 4 (IRF4), which is essential for the survival and expansion of BATF-transduced CAR-T cells in the TME. Surprisingly, chromatin immunoprecipitation-sequencing (ChIP-seq) analysis results showed that in BATF-overexpressing CAR-T cells, the reduced IRF4 can redistribute and preferentially bind at higher-affinity AP-1-IRF composite element (AICE) sites of chromatin to promote the expression of T cell effector-related genes. Conclusively, overexpression of BATF blocked CAR-T cell exhaustion to enhance CAR-T cells proliferation and cytotoxic capacity, with the underlying mechanisms by forming BATF-IRF4-DNA complexes to alter chromatin accessibility of target genes, indicating the potential application of BATF for improving the effectiveness of CAR-T cells in treating solid tumors. In another independent study, Chen et al. [2] sorted virus-specific splenic GP33-41+CD8+ T cells, a pool of heterogeneous exhausted T cells from chronic lymphocytic choriomeningitis virus (LCMV) infection. On the basis of the distribution and activity of transcription factors and the corresponding binding targets, they further divided the above-mentioned cell population into three major CD8+ T cell subsets: Ly108+ progenitor (TPRO) with the stem-like signature, CX3CR1+ cytolytic (TEFF) and CX3CR1−Ly108− exhausted (TEXH) CD8+ T subsets. Then, they focused on the epigenetic characteristics of three subsets by analyzing the chromatin accessibility as well as the activity of promoter and enhancer regions in each subtype using multiple sequencing tools including single-cell RNA-sequencing (RNA-seq), cleavage under targets and tagmentation sequencing (CUT & Tag-seq), and ATAC-seq. Notably, the authors also discovered BATF is mostly enriched within the CX3CR1+ effector T subset, which shows a potential role of BATF in promoting TEFF cell differentiation and avoiding TEXH cell formation. Further analysis showed that BATF mainly bound to the active enhancer regions during the differentiation of TPRO and TEFF cells, therefore, promoting the expression of key effector genes. While in BATF-deficient virus-specific CD8+ T cells, the proportion and cytotoxicity of TEFF subsets both significantly reduced, which was accompanied by the remarkably decreased active enhancer regions of effector genes. Collectively, Chen and colleagues demonstrated that BATF promotes the differentiation from TPRO cells to TEFF cells and suppresses the generation of TEXH cells upon the sustained viral infection. However, there are still several questions that need to be addressed. Although both studies described that BATF regulated the expression of key effector genes accompanied with altered chromatin accessibility, neither study elucidated whether and how BATF epigenetically affected the structure of exhaustion-related chromatin regions. Therefore, whether there exists a regulatory network among epigenetic changes induced by BATF and other T cell exhaustion characteristics such as metabolic changes, and whether BATF-IRF4 interaction has the crosstalk with other transcription regulatory factors need further exploration [6]. In addition, previous studies have reported that BATF can be induced by CD4+ T cell-derived IL-21 signaling pathway. Since CD4+ T cells mostly exhibit dysfunctional in the TME and chronic infection, which might promote CD8+ T exhaustion due to decreased IL-21 production [7]. Therefore, whether restoration of CD4+ T function and supplementation of IL-21 have similar effects as BATF overexpression, or whether there are other BATF regulators in these pathological conditions still require further clarification. Currently, immune checkpoint blockade (ICB) therapy has been widely used to treat patients with different malignancies [8], which are based on monoclonal antibodies targeting inhibitory receptors to change T cell exhaustion state and restore its effector functions. Thus, it is worth investigating whether there is a synergistic effect by combining BATF overexpression with checkpoint inhibitors. Besides, the authors did not show the long-term effect of BATF overexpression in human CAR-T cells as well as the unintended adverse effects such as cytokine release syndrome and neurologic toxicity. Altogether, both studies demonstrated that BATF can block the CD8+ T cell exhaustion pathway and facilitate the effector phenotype of CD8+ T cells, suggesting the great potential of BATF in preventing CD8+ T cell exhaustion (Figure 1). In immunotherapy for solid tumors and chronic viral infection with Epstein-Barr virus, hepatitis B virus or human papillomavirus, which is significantly related to the carcinogenesis, CD8+ T cell exhaustion has been recognized as the main obstacle of adaptive immune response [9]. Therefore, inhibiting CD8+ T cell exhaustion to restore immune system function could have important implications for the treatment of virus-related diseases and tumors. According to these two papers, BATF plays an important role in the regulation of CD8+ T cell exhaustion, implicating its potentiality in clinical application. It is impressive that both articles performed comprehensive bioinformatics including single-cell transcriptomics and genome-wide epigenetics to analyze the individual cell characteristics from a microscopic perspective and discovered the shift of regulatory networks among different CD8+ T cell subsets. The discovery of BATF-overexpressing CAR-T cells displaying outstanding anti-tumor effects provides new insights into the potential use of transcriptional suppressors of T cell exhaustion for specifically improving CAR-T cancer immunotherapy effectiveness. The transcription factor BATF epigenetically regulating the exhaustion and differentiation of CD8+ T cells in a solid tumor model case and chronic infection case. (A) In a CAR-T therapy of solid tumor model, BATF binds with IRF4 at AICE site to reshape the chromatin landscape, therefore, inducing chromatin accessibility changes to promote the mRNA expression of effector genes and suppress the mRNA expression of exhaustion genes. Consequently, BATF-transduced CAR-T cells can show enhanced anti-tumor effect with increased TME infiltration, proliferation capacity and cytokine production, as well as the decreased exhaustion tendency. (B) In a chronic viral infection model, transcription factor BATF promotes Ly108+ progenitor T (TPRO) cell subset to differentiate into CX3CR1+ cytolytic (TEFF) cells by improving chromatin accessibility in the enhancer regions and blocks the formation of CX3CR1−Ly108− exhausted (TEXH) cells in the meantime. Abbreviations: BATF, basic leucine zipper ATF-like transcriptional factor; AICE, AP-1-IRF composite element; Bhlhe40, basic helix-loop-helix family member e40; CAR, Chimeric antigen receptor; Ccl5, C-C motif chemokine ligand 5; Elk4, ETS-like transcription factor 4; Eomes, Eomesodermin; Foxo1, Forkhead box o1; E2f4, E2f transcription factor 4; Gzmb, Granzyme B; Icos, Inducible T cell costimulatory; Ifnar1, Interferon alpha and beta receptor subunit 1; IRF4, Interferon regulatory factor 4; Klf2/3, Kruppel-like factor 2/3; Lag3, Lymphocyte activation gene 3 protein; Pd-1, Programmed cell death-1; Nr4a2, Nuclear receptor subfamily 4 group A member 2; Runx1, RUNX family transcription factor 1; Runx3, RUNX family transcription factor 3; Tcf-1, T-cell factor 1; Tim3, T cell immunoglobulin and mucin domain 3; TIL, Tumor-infiltrating lymphocyte; Tox, Thymocyte selection-associated high mobility group box factor. Not applicable. Not applicable. Not applicable. The authors declare that they have no competing interests. This work was supported by the grant from the National Key R&D Program of China (2018YFA0507400) and National Natural Science Foundation of China (81788101). X.Y. and B.W. drafted the manuscript. X.C. supervised and revised the manuscript. All authors read and approved the final manuscript.

Acute promyelocytic leukemia (APL) is characterized by a specific chromosome translocation involving RARα and its fusion partners. For decades, the advent of all- trans retinoic acid (ATRA) synergized with arsenic trioxide (As 2 O 3 ) has turned most APL from highly fatal to highly curable. TBLR1-RARα (TR) is the tenth fusion gene of APL identified in our previous study, with its oncogenic role in the pathogenesis of APL not wholly unraveled. In this study, we found the expression of TR in mouse hematopoietic progenitors induces blockade of differentiation with enhanced proliferative capacity in vitro. A novel murine transplantable leukemia model was then established by expressing TR fusion gene in lineage-negative bone marrow mononuclear cells. Characteristics of primary TR mice revealed a rapid onset of aggressive leukemia with bleeding diathesis, which recapitulates human APL more accurately than other models. Despite the in vitro sensitivity to ATRA-induced cell differentiation, neither ATRA monotherapy nor combination with As 2 O 3 confers survival benefit to TR mice, consistent with poor clinical outcome of APL patients with TR fusion gene. Based on histone deacetylation phenotypes implied by bioinformatic analysis, HDAC inhibitors demonstrated significant survival superiority in the survival of TR mice, yielding insights into clinical efficacy against rare types of APL.

Acute myeloid leukemia (AML) refers to a heterogeneous group of hematopoietic malignancies. The well-known European Leukemia Network (ELN) stratifies AML patients into three risk groups, based primarily on the detection of cytogenetic abnormalities. However, the prognosis of cytogenetically normal AML (CN-AML), which is the largest AML subset, can be hard to define. Moreover, the clinical outcomes associated with this subgroup are diverse. In this study, using transcriptome profiles collected from CN-AML patients in the BeatAML cohort, we constructed a robust prognostic Cox model named NEST (Nine-gEne SignaTure). The validity of NEST was confirmed in four external independent cohorts. Moreover, the risk score predicted by the NEST model remained an independent prognostic factor in multivariate analyses. Further analysis revealed that the NEST model was suitable for bone marrow mononuclear cell (BMMC) samples but not peripheral blood mononuclear cell (PBMC) samples, which indirectly indicated subtle differences between BMMCs and PBMCs. Our data demonstrated the robustness and accuracy of the NEST model and implied the importance of the immune dysfunction in the leukemogenesis that occurs in CN-AML, which shed new light on the further exploration of molecular mechanisms and treatment guidance for CN-AML.

Objectives: Progesterone receptor membrane component-1 (PGRMC1) in breast cancer tissue has been suggested to predict a worse prognosis. The aim of this study was to assess for the first time whether PGRMC1 expressed in cancer tissue is associated with PGRMC1 blood concentrations and whether both are correlated with clinical tumour characteristics known to predict a worse outcome. Methods: In total, 201 patients with invasive breast cancer and 65 with benign breast disease (control group) were recruited. PGRMC1 blood concentrations were measured by a recently developed ELISA, PGRMC1 in breast cancer tissue was assessed by immunohistochemistry, and the correlation between the two was calculated. Receiver-operating characteristic (ROC) curve analysis was used to assess area under the curve (AUC). Furthermore, PGRMC1 was correlated with tumour characteristics such as tumour diameter, tumour grade and metastatic status, and with known blood tumour markers. Results: AUC for the breast cancer group was 0.713, which was significantly higher than in the control group (p < 0.01). Blood PGRMC1 concentrations had a strong (positive) correlation with tissue PGRMC1 expression (p < 0.01) but were not associated with serum tumour markers CEA, CA125, CA153 and TPS. Tissue PGRMC1, ER and cancer stage were positively associated with blood PGRMC1 (p < 0.05). Conclusions: As PGRMC1 expression levels in cancer tissue were significantly correlated with PGRMC1 in blood, and because concentrations in blood were also positively associated with breast tumour characteristics known to predict a worse prognosis, PGRMC1 may be valuable as a new tumour marker and may be superior to known tumour markers such as CEA, CA125, CA153 and TPS.

Objective To study the effects of Prunella vulgaris polysaccharide (PVP) on human breast carcinoma-associated fibroblasts (CAFs). Methods Cell viability was detected by 3-[4,5-dimethylthiazol-2-yl]-2,5-(3-carboxymethoxyphenyl)-2-4-sulfophenyl)-2H-tetrazolium (MTS) assay. Wound healing experiment and transwell migration assay were used to investigate the anti-migration effects. Flow cytometry was applied to detect cell apoptosis and cell cycle distribution. Reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were used to detect the expression of basic fibroblast growth factor (bFGF) in CAFs. Culture SKBr-3 with CAFs conditioned medium (CAFs-CM) to evaluate the indirect function on the proliferation of breast cancer SKBr-3 cells. Results PVP inhibited the viability of CAFs by inducing apoptosis (P <0.01) and arresting cell cycle (P <0.01). It also inhibited the migration of CAFs (P <0.01). bFGF promoted CAFs proliferation (P <0.01) and migration (P <0.01), protected CAFs from apoptosis (P <0.05) and reduced G(0) phase to 49.06% (P <0.01). However, these effects of bFGF on CAFs could be abrogated by PVP. Culturing SKBr-3 with CAFs-CM, PVP could inhibit the viability of breast cancer SKBr-3 cells indirectly. Moreover, PVP reduced the mRNA expression (P <0.01) and protein secretion of bFGF (P <0.01) in CAFs. Conclusion PVP could exert an anti-cancer effect on breast CAFs by inhibiting bFGF expression, thus inhibiting the growth of breast cancer SKBr-3 cells indirectly.

Previously, several protein-coding tumor suppressors localized at 1p36 have been reported. In the present work, we focus on functional long non-coding RNAs (lncRNAs) embedded in this locus. Small interfering RNA was used to identify lncRNA candidates with growth-suppressive activities in breast cancer. The mechanism involved was also explored. LINC01355 were downregulated in breast cancer cells relative to non-malignant breast epithelial cells. Overexpression of LINC01355 significantly inhibited proliferation, colony formation, and tumorigenesis of breast cancer cells. LINC01355 arrested breast cancer cells at the G0/G1 phase by repressing CCND1. Moreover, LINC01355 interacted with and stabilized FOXO3 protein, leading to transcriptional repression of CCND1. Importantly, LINC01355-mediated suppression of breast cancer growth was reversed by knockdown of FOXO3 or overexpression of CCND1. Clinically, LINC01355 was downregulated in breast cancer specimens and correlated with more aggressive features. There was a negative correlation between LINC01355 and CCND1 expression in breast cancer samples. LINC01355 acts as a tumor suppressor in breast cancer, which is ascribed to enhancement of FOXO3-mediated transcriptional repression of CCND1. Re-expression of LINC01355 may provide a potential therapeutic strategy to block breast cancer growth and progression.

Department of Breast Surgical Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, People’s Republic of China; Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, Harvard Medical School, Harvard University, Boston, MA 02114-3117, USA; Department of Cancer Prevention, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, People’s Republic of China; Department of Laboratory Medicine, Mayo Clinic, Rochester, MN 55902, USA; Department of Breast Surgery, Yantai Yuhuangding Hospital, Yantai 264000, People’s Republic of China; Department of Pathology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, People’s Republic of China

Colorectal cancer is a complex multistage process following the adenoma-carcinoma sequence. Additional research on the basis of molecular dysregulations, particularly in the precancerous stage, may provide insight into the realization of potential biomarkers and therapeutic targets for the disease. In the present study, the expression profile of human multistage colorectal mucosa tissues, including healthy, adenoma and adenocarcinoma samples, was downloaded. Genes that were consistently differentially expressed in precancerous tissues and cancer samples were collected. Based on a merged biological network, the biggest connected component composed of these identified genes and their one-step neighbors were retrieved to conduct random walk with restart algorithm, in order to identify genes significantly affected during carcinogenesis. Therefore, 35 genes significantly affected by carcinogenic dysregulation were successfully identified. Survival and Cox analysis indicated that the expression of these genes was an independent prognostic factor confirmed by six cohorts. In summary, based on the transcription profile of multi-stage carcinogenesis and bioinformatics analysis, 35 genes significantly associated with patient survival were successfully identified, which may serve as promising therapeutic targets for the disease.