Abstract Topic Esophageal Cancer: Adjuvant and Neo-Adjuvant Therapies Background Despite improved interventions with radical esophagectomy accompanied by perioperative systematic therapy, recurrence remains frequent in locally advanced esophageal squamous cell carcinoma (ESCC). A clinically important subset experiences early recurrence (ER), which is often followed by rapid deterioration and limited opportunities for effective salvage therapy. Identifying patients at high risk of ER and understanding the dominant failure patterns may inform postoperative surveillance intensity and risk-adapted adjuvant strategies. We aimed to determine clinicopathologic predictors of ER and to characterize recurrence patterns after curative resection. Methods We retrospectively analyzed a prospectively maintained cohort of patients with locally advanced ESCC treated with neoadjuvant chemotherapy (nCT) or chemoradiotherapy (nCRT) followed by curative esophagectomy. ER was defined as any recurrence within 12 months after surgery, non-ER comprised patients recurring later or remaining recurrence-free. Clinicopathologic characteristics were compared between groups. Factors associated with ER were ssessed using Cox regression, and recurrence patterns were compared using categorical models. Results Among 517 patients, 238 (46.0%) developed recurrence, and 126/238 (52.9%) occurred within 12 months. Univariable predictors of ER included comorbidity (HR=1.356; p=0.049), pathologic regression (HR=0.506; p=0.006 for complete; HR=0.572; p=0.001 for major), invasion depth (HR=0.543; p=0.001), lymph-node invasion (HR=2.380; p<0.001), and tumor deposits (HR=2.368; p<0.001). Female sex showed borderline significance (HR=0.584; p=0.053). Multivariable analysis identified major pathologic regression (HR=0.741; p=0.089), lymph-node invasion (HR=2.000; p<0.001), and tumor deposits (HR=1.656; p=0.021) as independent predictors for ER. ER was more often distant than locoregional (RR=1.70; p=0.07). Non-ER patients had markedly better overall survival (HR=0.102; p<0.001), most evident within 24 months. Conclusion Early recurrence within 12 months after neoadjuvant therapy and curative esophagectomy is common in locally advanced ESCC and is independently associated with treatment response, lymph-node involvement, and tumor deposits. ER shows a tendency toward distant failure and confers substantially worse early postoperative survival. supporting intensified surveillance and risk-adapted postoperative strategies.

Disease-free survival (DFS) is increasingly proposed as a surrogate endpoint for overall survival (OS) in oncology trials, yet its validity in esophageal squamous cell carcinoma (ESCC) remains unconfirmed. This study evaluated the surrogacy of DFS for OS in patients with locally advanced ESCC receiving neoadjuvant therapy. Individual patient data from a prospectively maintained cohort of patients who underwent neoadjuvant therapy followed by radical esophagectomy (N = 578) were analyzed. Individual-level surrogacy was assessed by comparing long-term OS with expected survival in a demographically matched general population using standardized mortality ratios (SMR) and landmark analysis. Trial-level surrogacy was evaluated via weighted linear regression of treatment effects on DFS and OS across published clinical trials. Patients remaining recurrence-free at 3 years achieved a 5-year OS rate of 93.0

Background In esophageal cancer, the ypN0 status after induction therapy could be categorized into two primary groups: "natural N0" (cN0/ypN0) and "down-staged N0" (cN+/ypN0). The assessment of cN status is typically based on clinical imagination or pathological regression. However, there is no standardized method for evaluating cN/ypN status. This study aims to investigate the prognosis of patients with cN+/ypN0 using both assessment methods through a cohort study and meta-analysis. Methods A prospectively maintained database encompassing esophageal cancer patients undergoing induction therapy followed by radical esophagectomy was comprehensively reviewed. The prognostic significance of cN+/ypN0 across two evaluation methods was quantified. Additionally, a meta-analysis using data from previous studies was conducted. Results 578 patients were identified from the cohort analysis, with 342 classified as ypN0 and 236 as ypN+. When evaluated with clinical imagination, patients with cN+/ypN0 had survival outcomes comparable to those with natural N0 but significantly better than those with ypN+ (p < 0.001). Using pathological nodal regression, cN+/ypN0 patients showed superior overall survival compared to ypN+ patients (p = 0.0043), although their disease-free survival was notably inferior to that of natural N0 patients (p = 0.0088). A meta-analysis of 20 previous studies confirmed the prognostic value of cN+/ypN0 status in both clinical imagination and pathological regression. Conclusions For esophageal cancer patients receiving neoadjuvant, cN+/ypN0 status, assessed through both clinical imagination and pathological regression, serves as a significant prognostic factor. It holds precedence over ypN+ yet falls short of the natural N0. The pre-treatment categorizations warrant recognition as a novel and pertinent staging metric.

BACKGROUND Transforming growth factor-β (TGF-β) superfamily plays an important role in tumor progression and metastasis. Activin A receptor type 1C (ACVR1C) is a TGF-β type I receptor that is involved in tumorigenesis through binding to different ligands. AIM To evaluate the correlation between single nucleotide polymorphisms (SNPs) of ACVR1C and susceptibility to esophageal squamous cell carcinoma (ESCC) in Chinese Han population. METHODS In this hospital-based cohort study, 1043 ESCC patients and 1143 healthy controls were enrolled. Five SNPs (rs4664229, rs4556933, rs77886248, rs77263459, rs6734630) of ACVR1C were assessed by the ligation detection reaction method. Hardy-Weinberg equilibrium test, genetic model analysis, stratified analysis, linkage disequilibrium test, and haplotype analysis were conducted. RESULTS Participants carrying ACVR1C rs4556933 GA mutant had significantly decreased risk of ESCC, and those with rs77886248 TA mutant were related with higher risk, especially in older male smokers. In the haplotype analysis, ACVR1C Trs4664229Ars4556933Trs77886248Crs77263459Ars6734630 increased risk of ESCC, while Trs4664229Grs4556933Trs77886248Crs77263459Ars6734630 was associated with lower susceptibility to ESCC. CONCLUSION ACVR1C rs4556933 and rs77886248 SNPs were associated with the susceptibility to ESCC, which could provide a potential target for early diagnosis and treatment of ESCC in Chinese Han population.

Tuberculosis, a major global health threat, necessitates understanding the pharmacological mechanisms of current drugs to combat multidrug resistance. In addition to alterations at the proteome level, the dynamic changes occurring at various levels of post-translational modifications (PTMs) following pharmacological intervention remain unclear. In our current study, we employed a quantitative proteomic approach to systematically analyze the dynamic molecular alterations at both the proteome and PTM levels in response to clinical drugs, including ethambutol, bedaquiline, moxifloxacin, and streptomycin. Our findings revealed enriched bioprocesses beyond known functions, phosphorylation-level changes in kinases and phosphatases, and increased acetylation levels with all four drugs. Overexpression of CobB in Mycobacterium smegmatis significantly increased its susceptibility to ethambutol, indicating enhanced drug sensitivity. Our study provides integrated multiomics resources for understanding the dynamic molecular characteristics and drug resistance associated with clinical drug interventions and proposes novel therapeutic strategies targeting the PTM levels.

Introduction LAMA1, also known as laminin subunit α1, is a member of the laminin family, which is widely reported to be a key basement membrane molecule that affects various biological activities and is associated with many kinds of diseases. We aimed to investigate the association between LAMA1single-nucleotide polymorphisms and the occurrence and progression of esophageal squamous cell carcinoma in the Chinese population. Method 2,186 participants were collected retrospectively between October 2008 and January 2017, including 1,043 ESCC patients and 1,143 noncancer patients. A 2 mL blood sample was obtained intravenously for the LDR for SNP analysis. The 6 SNP loci of LAMA1 were selected and examined. We analyzed the association of several genetic models of 6 LAMA1 SNP loci, sex, age, smoking and drinking status, and the occurrence of esophageal squamous cell carcinoma. Results In the rs62081531 G > A locus, genotype GA was a protective factor for ESCC compared with GG (OR: 0.830, P=0.046), especially among the younger and nondrinkers. At rs607230 T > C, genotype TC was linked with a lower risk of ESCC compared with TT. (OR: 0.613, P=0.034). Haplotype Frequencies revealed that Ars62081531Grs621993Ars539713Trs566655Ars73938538Crs607230 (OR: 0.803, P=0.028) and Grs62081531Grs621993Ars539713Trs566655Crs73938538Crs607230 (OR: 0.679, P=0.010) were strongly associated with lower susceptibility of ESCC. Conclusion The LAMA1 rs62081531, rs539713, rs566655, and rs607230 polymorphisms were demonstrated to be related to susceptibility to ESCC in the Chinese population. LAMA1 SNPs may have a significant impact on the occurrence of esophageal cancer and may serve as potential diagnostic biomarkers.

BACKGROUND:Pathological response is a critical factor in predicting long-term survival of patients with esophageal cancer after preoperative therapy. However, the validity of using pathological response as a surrogate for overall survival (OS) for esophageal cancer has not yet been established. In this study, a literature-based meta-analysis was conducted to evaluate pathological response as a proxy endpoint for survival in esophageal cancer.METHODS:Three databases were systematically searched to identify relevant studies investigating neoadjuvant treatment for esophageal cancer. The correlation between pathological complete response (pCR) and OS were assessed using a weighted multiple regression analysis at the trial level, and the coefficient of determination (R2) was calculated. The research design and histological subtypes were considered in the performance of subgroup analysis.RESULTS:In this meta-analysis, a total of 40 trials, comprising 43 comparisons and 55,344 patients were qualified. The surrogacy between pCR and OS was moderate (R2 = 0.238 in direct comparison, R2 = 0.500 for pCR reciprocals, R2 = 0.541 in log settings). pCR could not serve as an ideal surrogate endpoint in randomized controlled trials (RCTs) (R2 = 0.511 in direct comparison, R2 = 0.460 for pCR reciprocals, R2 = 0.523 in log settings). A strong correlation was observed in studies comparing neoadjuvant chemoradiotherapy and neoadjuvant chemotherapy (R2 = 0.595 in direct comparison, R2 = 0.840 for pCR reciprocals, R2 = 0.800 in log settings).CONCLUSIONS:A lack of surrogacy of pathological response for long-term survival at trial level is established in this study. Hence, caution should be exercised when using pCR as the primary endpoint in neoadjuvant studies for esophageal cancer.

Background: Deep learning methods have demonstrated great potential for processing high-resolution images. The U-Net model, in particular, has shown proficiency in the segmentation of biomedical images. However, limited research has examined the application of deep learning to esophageal squamous cell carcinoma (ESCC) segmentation. Therefore, this study aimed to develop deep learning segmentation systems specifically for ESCC. Methods: A Visual Geometry Group (VGG)-based U-Net neural network architecture was utilized to develop the segmentation models. A pathological image cohort of surgical specimens was used for model training and internal validation, with two additional endoscopic biopsy section cohort for external validation. Model efficacy was evaluated across several metrics including Intersection over Union (IOU), accuracy, positive predict value (PPV), true positive rate (TPR), specificity, dice similarity coefficient (DSC), area under the receiver operating characteristic curve (AUC), and F1-Score. Results: Surgical samples from ten patients were analyzed retrospectively, with each biopsy section cohort encompassing five patients. Transfer learning models based on U-Net weights yielded optimal results. For mucosa segmentation, the in internal validation achieved 93.81% IOU, with other parameters exceeding 96% (96.96% accuracy, 96.45% PPV, 96.65% TPR, 98.41% specificity, 96.81% DSC, 96.11% AUC, and 96.55% F1-Score). The tumor segmentation model attained an IOU of 91.95%, along with other parameters surpassing 95% (95.90% accuracy, 95.62% PPV, 95.71% TPR, 97.88% specificity, 95.81% DSC, 94.92% AUC, and 95.67% F1-Score). In the external validation for tumor segmentation model, IOU was 59.86% for validation database 1 (72.74% for accuracy, 76.03% for PPV, 77.17% for TPR, 83.80% for specificity, 74.89% for DSC, 71.83% for AUC, and 76.60% for F1-Score), and 50.88% for validation cohort 2 (68.03% for accuracy, 59.02% for PPV, 66.87% for TPR, 78.48% for specificity, 67.44% for DSC, 64.68% for AUC, and 62.70% for F1-Score). Conclusions: The models exhibited satisfactory results, paving the way for their potential deployment on standard computers and integration with other artificial intelligence models in clinical practice in the future. However, limited to the size of study, the generalizability of models is impaired in the external validation, larger pathological section cohort would be needed in future development to ensure robustness and generalization.

Overall survival (OS) benefits of neoadjuvant immunotherapy remain elusive in locally advanced esophageal squamous cell carcinomas (ESCC). Here, we reported the results of a phase 1b trial of neoadjuvant PD-L1 blockade with adebrelimab in resectable ESCC. Patients received two neoadjuvant doses of adebrelimab followed by surgery. The primary endpoints were safety and feasibility; secondary endpoints included pathologic complete response (pCR) and OS. Our data showed the primary endpoints of safety and feasibility had been met. Common treatment-related adverse events were anorexia (32%) and fatigue (16%), without grade 3 or more adverse events. Of the 30 patients enrolled in the trial, 25 underwent successful resection without surgery delay and 24% had major pathologic responses including a pCR rate of 8%. The 2-year OS was 92%. Responsive patients had an immune-enriched tumor microenvironment phenotype, whereas nonresponsive patients had greater infiltration of cancer-associated fibroblasts at baseline. Clonotypic dynamics of pre-existing intratumoral T cells was a hallmark of responsive patients. These findings provide a rational for neoadjuvant anti-PD-L1 monotherapy as a therapeutic strategy for patients with resectable ESCC. ClinicalTrials.gov identifier: NCT04215471 .

Background and Objective:Lymph nodes constitute an integral component of the secondary lymphoid organs, housing a diverse population of macrophages. Macrophages exhibit heterogeneity in terms of localization, phenotype and ontogeny. Recent evidence has established that subcapsular sinus macrophages (SCSMs) are the initial cells exposed to antigens from afferent lymph vessels, playing a crucial role in the host immune response against invading pathogens and tumor cells. In order to summarize the role and mechanisms of SCSM in tumor immunity, this study systematically reviews research on SCSMs in tumor immunity.Methods:A systematic search was conducted in PubMed and Web of Science to identify articles investigating clinical significance and mechanisms of SCSMs. Study eligibility was independently evaluated by two authors based on the assessment of titles, abstracts and full-texts.Key Content and Findings:The narrative review included a total of 17 studies. Previous research consistently showed that a high level of SCSM in patients with various carcinomas is associated with a favorable long-term prognosis. SCSM acts as the front-line defender in antitumor activity, engaging in intricate communication with other immune cells. Moreover, SCSM could directly and indirectly modulate tumor immunity, and the integrity of SCSM layer is interrupted in disease status. Several studies explored the feasibility of targeting SCSM to activate immunity against tumors. However, the direct molecular interactions and alternation in signal pathway in the tumor immunity of SCSM are less well established in previous researches.Conclusions:This narrative review underscores the critical role of SCSM in tumor immunity. Future studies should focus on the deeper mechanism underlying SCSMs and explore their clinical applications.

Background:The σ1A subunit of the adaptor protein 1 (AP1S1) participates in various intracellular transport pathways, especially the maintenance of copper homeostasis, which is pivotal in carcinogenesis. It is therefore rational to presume that AP1S1 might also be involved in carcinogenesis. In this hospital-based case-control study, we investigated the genetic susceptibility to ESCC in relation to SNPs of AP1S1 among Chinese population. Methods:A database containing a total of 1303 controls and 1043 ESCC patients were retrospectively studied. The AP1S1 SNPs were analyzed based on ligation detection reaction (LDR) method. Then, the relationship between ESCC and SNPs of AP1S1 was determined with a significant crude P<0.05. Then the logistic regression analysis was used for the calculation for adjusted P in the demographic stratification comparison if a significant difference was observed in the previous step. Results:AP1S1 rs77387752 C>T genotype TT was an independent risk factor for ESCC, while rs4729666 C>T genotype TC and rs35208462 C>T genotype TC were associated with a lower risk for ESCC, especially in co-dominant model and allelic test for younger, male subjects who are not alcohol-drinkers nor cigarette smokers. Conclusion:AP1S1 rs77387752, rs4729666 and rs35208462 polymorphisms are associated with susceptibility to ESCC in Chinese individuals. AP1S1 SNPs may exert an important role in esophageal carcinogenesis and could serve as potential diagnostic biomarkers.

Background: Esophagectomy offers the chance of cure for esophageal cancer, however, the optimal circumferential extent of surgery remains uncertain. En bloc esophagectomy (EBE) and total mesoesophagectomy (TME) have yielded inconsistent results. Therefore, the purpose of this study was to evaluate the surgical and oncological effects of EBE and TME on esophageal cancer patients. Methods: Four databases including PubMed, Cochrane Library, Web of Science, and Embase were searched through to March 1st, 2022, and the references of eligible studies were further evaluated. Randomized controlled trials comparing the efficacy of EBE and TME were included, and the risk of biases for included studies was assessed with the Cochrane risk of bias tool by two reviewers independently. The outcomes were recorded as mean difference, risk ratio, odds ratio, and hazard ratio with its corresponding 95% confidence interval. Results: Overall, a total of 14 randomized controlled trials involving 3,106 subjects were included. Compared with standard resection, higher blood loss [mean difference =56.29 (14.80, 97.77), P=0.008], more dissected lymph nodes [mean difference =14.39 (9.79, 19.00), P<0.001], and superior long-term outcomes for early [overall survival: hazard ratio =0.31 (0.10, 0.96), P=0.04; disease-free survival: hazard ratio =0.71 (0.41, 1.21), P=0.21] and advanced-stage esophageal cancer patients [overall survival: hazard ratio =0.47 (0.33, 0.66), P<0.001; disease-free survival: hazard ratio =0.62 (0.38, 0.99), P=0.05] were observed in the EBE group, while TME showed less blood loss [mean difference =-74.03 (-96.69, -51.38), P<0.001], shorter operation time [mean difference =-32.37 (-65.12, 0.37), P=0.05], and better overall survival [hazard ratio =0.74 (0.55, 0.98), P=0.04]. Conclusions: EBE is highly technically demanding and is associated with comparable surgical trauma and better long-term outcomes comparted to the standard esophagectomy. TME has a better long-term prognosis without improving operative bleeding and operation time. Further prospective studies are required to verify the efficacy of EBE and TME.

The σ1A subunit of the adaptor protein 1 (AP1S1) participates in intracellular transport pathways, which is pivotal in carcinogenesis. It is therefore rational to presume that AP1S1 might also be involved in carcinogenesis. In this hospital-based case–control study, we investigated the genetic susceptibility to esophageal squamous cell carcinoma (ESCC) in relation to Single Nucleotide Polymorphisms (SNPs) of AP1S1 among Chinese population and made a preparation for the subsequent esophageal SNP risk model building. A database containing a total of 1143 controls and 1043 ESCC patients were retrospectively studied. The AP1S1 SNPs were analyzed based on ligation detection reaction (LDR) method. Then the relationship between ESCC and SNPs of AP1S1 was determined with a significant crude P < 0.05. Then the logistic regression analysis was used for the calculation for adjusted P in the demographic stratification comparison if a significant difference was observed in the previous step. AP1S1 rs77387752 C > T mutated homozygous TT was an independent risk factor for ESCC, while SNP rs4729666 C > T and rs35208462 C > T mutated heterozygote TC were associated with a lower risk for ESCC, especially in co-dominant model and allelic test for younger, male subjects who are not alcohol-drinkers nor cigarette smokers. These three AP1S1 SNP sites could be incorporated into subsequent SNPs risk models for ESCC in specific populations. AP1S1 rs77387752, rs4729666 and rs35208462 polymorphisms are associated with susceptibility to ESCC in Chinese individuals. AP1S1 SNPs may exert an important role in esophageal carcinogenesis and could serve as potential diagnostic biomarkers. These SNP sites might be added into the building of risk model and then the early diagnosis of ESCC. Besides, further mechanical exploration is required. Table. Logistic regression analyses of associations between AP1S1 SNPs rs77387752 C > T, rs4729666 C > T, rs35208462 C > T and risk of ESCC.

Abstract Background Metabolic reprogramming and redox homeostasis contribute to esophageal squamous cell carcinoma (ESCC). CDC‐like kinase 4 (CLK4) is a dual‐specificity kinase that can phosphorylate substrates’ tyrosine or serine/threonine residue. However, the role and mechanism of CLK4 in ESCC remain unknown. Methods CLK4 expression was analysed using publicly available datasets and confirmed in ESCC tissues and cell lines. The biological roles of CLK4 were studied with gain and loss‐of‐function experiments. Mass spectrometry was employed to examine the effects of CLK4 on metabolic profiling. In vitro kinase assay, co‐immunoprecipitation, glutathione S‐transferase pulldown, chromatin immunoprecipitation and luciferase reporter were used to elucidate the relationship among CLK4, microphthalmia‐associated transcription factor (MITF), COP1 and ZRANB1. Results CLK4 down‐regulation was observed in ESCC cell lines and clinical samples and associated with the methylation of its promoter. Low levels of CLK4 promoted ESCC development by affecting the purine synthesis pathway and nicotinamide adenine dinucleotide phosphate (NADPH)/nicotinamide adenine dinucleotide phosphate (NADP+) ratio. Interestingly, CLK4 inhibited ESCC development by blocking MITF‐enhanced de novo purine synthesis and redox balance. Mechanistically, wild type CLK4 (WT‐CLK4) but not kinase‐dead CLK4‐K189R mutant phosphorylated MITF at Y360. This modification promoted its interaction with E3 ligase COP1 and its K63‐linked ubiquitination at K308/K372, leading to sequestosome 1 recognition and autophagic degradation. However, the deubiquitinase ZRANB1 rescued MITF ubiquitination and degradation. In turn, MITF bound to E‐ rather than M‐boxes in CLK4 promoter and transcriptionally down‐regulated its expression in ESCC. Clinically, the negative correlations were observed between CLK4, MITF, and purine metabolic markers, which predicts a poor clinical outcome of ESCC patients. Notably, CLK4 itself was a redox‐sensitive kinase, and its methionine oxidation at M307 impaired kinase activity, enhanced mitochondria length and inhibited lipid peroxidation, contributing to ESCC. Conclusions Our data highlight the potential role of CLK4 in modulating redox status and nucleotide metabolism, suggesting potential therapeutic targets in ESCC treatment.

Background Gut microbiota (GMB) alteration has been reported to influence the Alzheimer’s disease (AD) pathogenesis through immune, endocrine, and metabolic pathways. This study aims to investigate metabolic output of the dysbiosis of GMB in AD pathogenesis. In this study, the fecal microbiota and metabolome from 21 AD participants and 44 cognitively normal control participants were measured. Untargeted GMB taxa was analyzed through 16S ribosomal RNA gene profiling based on next-generation sequencing and fecal metabolites were quantified by using ultrahigh performance liquid chromatography-mass spectrometry (UPLC-MS). Results Our analysis revealed that AD was characterized by 15 altered gut bacterial genera, of which 46.7% (7/15 general) was significantly associated with a series of metabolite markers. The predicted metabolic profile of altered gut microbial composition included steroid hormone biosynthesis, N-Acyl amino acid metabolism and piperidine metabolism. Moreover, a combination of 2 gut bacterial genera ( Faecalibacterium and Pseudomonas ) and 4 metabolites (N-Docosahexaenoyl GABA, 19-Oxoandrost-4-ene-3,17-dione, Trigofoenoside F and 22-Angeloylbarringtogenol C) was able to discriminate AD from NC with AUC of 0.955 in these 65 subjects. Conclusions These findings demonstrate that gut microbial alterations and related metabolic output changes may be associated with pathogenesis of AD, and suggest that fecal markers might be used as a non-invasive examination to assist screening and diagnosis of AD.