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Elucidating the role of PBRM1 in NPC via RNA-seq transcriptomic sequencing 通过RNA-seq转录组测序阐明PBRM1在鼻咽癌中的作用。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-12 DOI: 10.1016/j.slast.2025.100386
XingYu Yang , Qin Qiu , Yu Tang , WeiDi Sun , XiFang Wu , XiaoJiang Li , YanXin Ren

Purpose

The PBRM1 (PB1) gene, which encodes BAF180, a specific subunit of the PBAF SWI / SNF complex, is extensively studied in some other cancers, yet its effects and related mechanisms in NPC remains inadequately understood. Aerobic glycolysis is one of the hallmarks of cancer, and whether PBRM1 is involved in this metabolic metastasis in NPC remains unclear.

Methods

We established NPC cell lines with knockdown of PBRM1 and performed functional analysis to understand the impact of their production. Based on the RNA-seq data, we mainly analyzed the activity of the AKT-mTOR signaling pathway and examined the expression levels of some key glycolytic genes including HIF 1α, PFKP, ENO 1, PKM and LDHA. Using in vivo experiments, we verified the effect of PBRM1 on the proliferation of NPC.

Results

Our findings indicate that PBRM1 deficiency enhances proliferation, migration, and invasion in both CNE1 and CNE2 cells. Notably, PBRM1 downregulate activates the AKT-mTOR pathway, upregulating glycolytic enzymes and lactate production. Subcutaneous tumor formation assay in nude mice also showed that knockdown of PBRM1 promoted NPC cells growth.

Conclusion

This study illuminates PBRM1′s tumor suppressor role, highlighting the AKT-mTOR pathway and aerobic glycolysis as potential therapeutic targets in NPC.
目的:PBRM1 (PB1)基因编码PBAF SWI / SNF复合物的特异性亚基BAF180,在其他一些癌症中被广泛研究,但其在鼻咽癌中的作用和相关机制仍不充分了解。有氧糖酵解是癌症的标志之一,PBRM1是否参与鼻咽癌的代谢转移尚不清楚。方法:建立PBRM1基因敲低的鼻咽癌细胞系,并进行功能分析,了解其产生的影响。基于RNA-seq数据,我们主要分析了AKT-mTOR信号通路的活性,检测了HIF 1α、PFKP、ENO 1、PKM、LDHA等关键糖酵解基因的表达水平。通过体内实验,我们验证了PBRM1对鼻咽癌细胞增殖的影响。结果:我们的研究结果表明,PBRM1缺失增强了CNE1和CNE2细胞的增殖、迁移和侵袭。值得注意的是,PBRM1下调激活AKT-mTOR通路,上调糖酵解酶和乳酸生成。裸鼠皮下肿瘤形成实验也显示PBRM1的下调促进了鼻咽癌细胞的生长。结论:本研究阐明了PBRM1的肿瘤抑制作用,强调了AKT-mTOR途径和有氧糖酵解是鼻咽癌的潜在治疗靶点。
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引用次数: 0
A novel prognostic model in ovarian cancer based on the Nectin family and Necl-like molecules related transcriptomics 基于Nectin家族和necl样分子相关转录组学的卵巢癌预后新模型。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-12 DOI: 10.1016/j.slast.2025.100385
Yixian Liu , Xin Lan , Xiaoyi Zhang , Xinyi Ren , Hongmin Duan , Yanrong Dan , Dong Duan , Ganghua Lu
<div><h3>Background</h3><div>Ovarian cancer (OC) is the deadliest malignant tumor among gynecological tumors. The current treatment measures for OC remain not optimistic, so it is important to determine reliable prognostic biomarkers to prolong OC patients' survival.</div></div><div><h3>Methods</h3><div>The OC-related transcriptome data were downloaded from the University of California Santa Cruz (UCSC) and we obtained the Differentially Expressed Genes (DEGs) between OC and normal samples. The Nectins and Nectin-like (Necl) scores of OC patients were calculated by single-sample Gene Set Enrichment Analysis (ssGSEA), the correlation between these scores and the prognosis of OC patients was explored using the Kaplan-Meier survival curve. DEGs were overlapped with Nectins and Necls-related genes selected by Weighted Gene Co-expression Network Analysis (WGCNA) to obtain the differentially expressed Nectins and Necls-related genes (DENNGs). Next, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses were carried out on DENNGs. The Protein-Protein Interaction (PPI) network of DENNGs was constructed and hub genes were screened. In addition, univariate and multivariate Cox regression analysis were used to obtain the prognosis-related genes of OC patients and construct a prognostic model. Gene Set Variation Analysis (GSVA) was performed on the genes of high and low risk groups. SsGSEA algorithm was used to calculate the immune cell scores and the correlation between different immune cells and prognosis-related genes was explored.</div></div><div><h3>Results</h3><div>We obtained 583 DENNGs by crossing 6778 DEGs in OC and 584 Nectin- and Necl-related genes. The scores of Nectins and Necls in the OC group increased significantly, together with the poor prognosis. KEGG and GO analyses showed that DENNGs were mainly associated with cell proliferation, aging, canceration, and virus infection. Univariate and multivariate Cox analyses screened six prognosis-related genes (PTTG1, MELK, CENPF, PLK1, KIF20A, TOP2A) and modeled prognosis risks. Furthermore, a nomogram that integrated the risk model and patient age accurately predicted OC prognosis. The results of GSVA showed that TGF-β-mediated epithelial-mesenchymal transition, tumor cell invasion and metastasis were activated in the high-risk group, while pathways related to tumor cell invasiveness, such as hedgehog (Hh) mediated by hypoxia, were activated in the low-risk group. Finally, we found five immune cells (activated CD4+ <em>T</em> cells, central memory CD4+ <em>T</em> cells, central memory CD8+ <em>T</em> cells, T follicular helper cells (tTFH), and type II helper T cells(Th2)) different between OC and normal samples, and prognosis-related genes were positively correlated with activated CD4+ cells and Th2 of immune cells but negatively correlated with tTFH.</div></div><div><h3>Conclusion</h3><div>We identified six prognosis-related genes and constructed a prognostic mo
背景:卵巢癌是妇科肿瘤中最致命的恶性肿瘤。目前OC的治疗措施仍然不容乐观,因此确定可靠的预后生物标志物对于延长OC患者的生存期至关重要。方法:从加州大学圣克鲁兹分校下载OC相关转录组数据,获得OC与正常样本之间的差异表达基因(differential Expressed Genes, DEGs)。通过单样本基因集富集分析(ssGSEA)计算OC患者的nectin和nectin样蛋白(Necl)评分,并利用Kaplan-Meier生存曲线探讨这些评分与OC患者预后的相关性。deg与加权基因共表达网络分析(Weighted Gene Co-expression Network Analysis, WGCNA)选择的Nectins和necls相关基因重叠,得到差异表达的Nectins和necls相关基因(differential expression genes, denng)。接下来,对denng进行京都基因与基因组百科全书(KEGG)和基因本体(GO)富集分析。构建了denng蛋白-蛋白相互作用(PPI)网络,筛选了枢纽基因。此外,采用单因素和多因素Cox回归分析,获得OC患者预后相关基因,构建预后模型。对高、低危组基因进行基因集变异分析(GSVA)。采用SsGSEA算法计算免疫细胞评分,探讨不同免疫细胞与预后相关基因的相关性。结果:通过6778个OC基因和584个Nectin及necl相关基因杂交,获得583个denng。OC组Nectins和Necls评分明显升高,预后较差。KEGG和GO分析显示,dengs主要与细胞增殖、衰老、癌变和病毒感染有关。单因素和多因素Cox分析筛选了6个预后相关基因(PTTG1、MELK、CENPF、PLK1、KIF20A、TOP2A),并对预后风险进行建模。此外,结合风险模型和患者年龄的nomogram预测了OC的预后。GSVA结果显示,高危组TGF-β介导的上皮-间质转化、肿瘤细胞侵袭转移被激活,低危组缺氧介导的hedgehog (Hh)等与肿瘤细胞侵袭相关的通路被激活。最后,我们发现OC与正常样本的5种免疫细胞(活化CD4+ T细胞、中枢记忆CD4+ T细胞、中枢记忆CD8+ T细胞、T滤泡辅助细胞(tTFH)和II型辅助T细胞(Th2))存在差异,且预后相关基因与免疫细胞的活化CD4+细胞和Th2呈正相关,与tTFH呈负相关。结论:我们鉴定了6个预后相关基因,构建了预后模型,为OC患者的临床预后预测和治疗提供了理论依据。
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引用次数: 0
High-throughput dissection of inter-organ genetic networks: A multi-omic systems biology approach 器官间遗传网络的高通量解剖:多组学系统生物学方法。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-11 DOI: 10.1016/j.slast.2025.100376
Rana Alabdan , Hechmi Shili , Ghada Moh. Samir Elhessewi , Mukhtar Ghaleb , Eman M Alanazi , Nouf Helal Alharbi , Rowida Mohammed Alharbi , Asma A. Alhashmi
The existing multi-omic analyses are frequently confined to individual tissues, and the regulatory picture of the systemic regulator of complex physiology and disease is hidden. To fill this gap, we have created a unified systems biology model of the high-throughput dissection of inter-organ genetic networks. Our model incorporates transcriptomic, epigenomic and proteomic analysis of five major organs (liver, kidney, heart, lung, brain) using the Multi-Omics Factor Analysis (MOFA+) tool, specifically, cross-tissue coordination. We characterized 27 evidence-heavy cross-tissue modules (FDR < 0.05) that are major hubs such as *HNF4Aenda NRF2cheng8loadmasterregulatingconstitutionembryonicstemcellularinfoncogenes recognize them. One notable observation was liver-kidney metabolic axis, significant cross-talks in hepatocyte organoids are confirmed with CRISPR knockdown, which suppresses the expression of transporters expressed by the kidney. Our work offers a scalable validated framework that goes beyond organ-centric perspectives, which can be used as a potent tool of systemic disease modelling and precision medicine.
现有的多组学分析往往局限于单个组织,而隐藏了复杂生理和疾病的系统性调节机制。为了填补这一空白,我们创建了一个统一的高通量解剖器官间遗传网络的系统生物学模型。我们的模型结合了五个主要器官(肝、肾、心、肺、脑)的转录组学、表观基因组学和蛋白质组学分析,使用多组学因子分析(MOFA+)工具,特别是跨组织协调。我们鉴定了27个证据丰富的跨组织模块(FDR < 0.05),这些模块是主要的枢纽,如*HNF4Aenda nrf2cheng8loadmaster调控结构,胚胎系统细胞信息癌基因识别它们。一个值得注意的观察是肝-肾代谢轴,肝细胞类器官的显著交叉对话被CRISPR敲低证实,这抑制了肾脏表达的转运蛋白的表达。我们的工作提供了一个可扩展的验证框架,超越了以器官为中心的观点,它可以作为系统性疾病建模和精准医学的有力工具。
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引用次数: 0
Spatial transcriptomic modeling of vascular remodeling in aortic aneurysm using integrated single-cell RNA sequencing analysis 基于单细胞RNA测序分析的主动脉瘤血管重构的空间转录组学建模。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-06 DOI: 10.1016/j.slast.2025.100377
Shaodong Xiong , Ludong Liang , Tao Liu

Background

The phenotypic regulation of vascular smooth muscle cells (VSMCs) is a critical characteristic of aortic aneurysm formation, although its spatial and transcriptional dynamics remain incompletely understood.

Methods

We developed a computational workflow integrating single-cell RNA sequencing (scRNA-seq) with pseudo-spatial transcriptomic inference to model vascular remodeling at cellular resolution. Using the publicly available SCP1361 dataset from the Broad Institute's Single Cell Portal (15,698 high-quality cells from ascending aorta of normal and high- fat diet mice), we employed Seurat v4.3.0 for quality control and clustering, Tangram v1.0 for pseudo-spatial projection onto synthetic tissue scaffolds, and Monocle3 for trajectory inference. Statistical analyses included chi-square tests for cell proportion differences, Wilcoxon rank-sum tests for differential expression, and Moran's I for spatial variability (α = 0.05, adjusted for multiple testing).

Results

We identified 25 transcriptionally distinct cell populations including 5 VSMC subtypes and 4 fibroblast subtypes showing region-specific localization patterns. High-fat diet significantly increased synthetic VSMC_2 populations (+13.1 %, p = 0.008) and monocyte infiltration (+16.3 %, p = 0.042) while decreasing contractile VSMC_3 (-8.2 %, p = 0.041). Pseudo-spatial reconstruction revealed anatomically compartmentalized cell states with contractile VSMCs localizing to the media and synthetic/inflammatory phenotypes enriching in adventitial regions. Trajectory analysis identified 847 genes with pseudotemporal dynamics (q < 0.05) associated with VSMC phenotypic transitions.

Conclusions

This framework demonstrates how publicly available scRNA-seq data can be leveraged for hypothesis-generating spatial modeling of vascular disease. The approach reveals cell-type-specific transcriptional programs and phenotypic transitions that warrant experimental validation through immunohistochemistry and true spatial transcriptomics.
背景:血管平滑肌细胞(VSMCs)的表型调控是动脉瘤形成的一个关键特征,尽管其空间和转录动力学尚不完全清楚。方法:我们开发了一个计算工作流,将单细胞RNA测序(scRNA-seq)与伪空间转录组推断结合起来,在细胞分辨率上模拟血管重构。使用来自Broad研究所单细胞门户网站的公开可用的SCP1361数据集(来自正常和高脂肪饮食小鼠升主动脉的15698个高质量细胞),我们使用Seurat v4.3.0进行质量控制和聚类,Tangram v1.0用于伪空间投影到合成组织支架上,Monocle3用于轨迹推断。统计分析包括细胞比例差异的卡方检验,差异表达的Wilcoxon秩和检验,空间变异性的Moran's I检验(α = 0.05,经多重检验调整)。结果:我们鉴定了25个转录不同的细胞群,包括5个VSMC亚型和4个显示区域特异性定位模式的成纤维细胞亚型。高脂饲料显著增加了合成VSMC_2 (+13.1%, p=0.008)和单核细胞浸润(+16.3%,p=0.042),显著降低了收缩VSMC_3 (-8.2%, p=0.041)。伪空间重建显示解剖上的细胞状态区隔化,收缩的VSMCs定位于介质,合成/炎症表型在外膜区丰富。轨迹分析确定了847个具有伪时间动力学的基因(结论:该框架证明了如何利用公开可用的scRNA-seq数据进行血管疾病的假设生成空间建模。该方法揭示了细胞类型特异性转录程序和表型转变,需要通过免疫组织化学和真正的空间转录组学进行实验验证。
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引用次数: 0
Guide to liquid volume measurements: A review of methods and technologies 液体体积测量指南:方法和技术综述。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-05 DOI: 10.1016/j.slast.2025.100382
Nathaniel G. Hentz , John Thomas Bradshaw
The need to measure volume is an important component of any liquid-based assay, because the final result is often based upon the concentration of some analyte within the assay. Volume measurements can be used to determine the actual amount of sample throughout an assay. They can also be used to calibrate, routinely verify performance of, or optimize liquid classes for liquid handling platforms or other automated or manual pipetting devices. This review surveys current liquid volume measurement methods and technologies available for use in life science laboratories. Volumes considered in this review range from picoliters to a few milliliters, which are typical for life science assays. While the need for volume measurement extends across many different industries, the focus here is on applications in biopharmaceutical and clinical life science laboratories. This review evaluates key measurable attributes, including the addressable volume range, as well as the attainable precision and accuracy of the method or technology. Several important features are also considered such as the ability to measure actual samples, commercial availability, regulatory compliance, sample type, workflow integration, suitability for in-line use, and relative ease of use.
测量体积的需要是任何液基分析的重要组成部分,因为最终结果通常基于分析中某些分析物的浓度。体积测量可用于确定整个分析过程中样品的实际量。它们还可用于校准,常规验证液体处理平台或其他自动或手动移液装置的性能或优化液体类别。本文综述了目前生命科学实验室中可用的液体体积测量方法和技术。本综述中考虑的体积范围从皮升到几毫升,这是典型的生命科学分析。虽然体积测量的需求扩展到许多不同的行业,但这里的重点是生物制药和临床生命科学实验室的应用。这篇综述评估了关键的可测量属性,包括可寻址的体积范围,以及方法或技术的可实现的精度和准确性。还考虑了几个重要的特性,例如测量实际样本的能力、商业可用性、法规遵从性、样本类型、工作流集成、联机使用的适用性以及相对易用性。
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引用次数: 0
Low-Cost CNC-Based media dispensing system for biotechnology laboratories 低成本的基于cnc的生物技术实验室介质分配系统。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-05 DOI: 10.1016/j.slast.2025.100384
Ping Wu , Matthew Moore , Peter Cieszewski, Kayla Beam, Sivamani Elumalai, Stephen B. Rigoulot
This report outlines the development and implementation of a Computer Numerical Control (CNC) based Automated Media Dispensing System (AMDS) in a plant biotechnology research laboratory. The AMDS, a dual-channel liquid dispensing system, underwent comprehensive testing and refinement over a two-year period. Designed to serve the production demand for a commercial transformation pipeline, the system represents an initial automation step to replace manual media dispensing operations. Performance evaluation demonstrates that this iteration surpasses manual dispensing metrics, achieving a process capability index (Cpk) of 2.28 compared to 1.43 for manual operations, while maintaining comparable time per tray efficiency. The system delivers significant ergonomic benefits by reducing prolonged standing periods associated with manual dispensing tasks. Building upon established automated dispensing principles, the custom-built AMDS achieves core functionality while reducing the barrier to entry, with an estimated material cost of approximately one-fiftieth that of more comprehensive market solutions. The system's modular architecture facilitates future modifications and adaptations, positioning it as a valuable tool for addressing evolving research needs in plant biotechnology while enabling broader adoption of automated dispensing technology.
本报告概述了在植物生物技术研究实验室中基于计算机数控(CNC)的自动介质分配系统(AMDS)的开发和实施。AMDS是一种双通道液体分配系统,经过了两年的全面测试和改进。该系统旨在满足商业改造管道的生产需求,代表了取代人工介质分配操作的初始自动化步骤。性能评估表明,这一迭代超越了手动点胶指标,实现了2.28的过程能力指数(Cpk),而手动操作为1.43,同时保持了相当的每托盘时间效率。该系统通过减少与人工分配任务相关的长时间站立时间,提供了显著的人体工程学效益。基于既定的自动化点胶原理,定制的AMDS实现了核心功能,同时降低了进入门槛,估计材料成本约为更全面的市场解决方案的五分之一。该系统的模块化架构便于未来的修改和调整,将其定位为解决植物生物技术不断发展的研究需求的有价值的工具,同时使自动化分配技术得到更广泛的采用。
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引用次数: 0
An explainable hybrid CNN-LSTM framework for accurate sequence-based classification of RNA N6-methyladenosine (m6A) modification 基于序列的RNA n6 -甲基腺苷(m6A)修饰精确分类的可解释的CNN-LSTM混合框架
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-05 DOI: 10.1016/j.slast.2025.100383
Kainat Ali Rehman , Muhammad Sohail Khan , Faiza Tila , Inayat Khan , Jawad Khan , Mohammad Shabaz , Ahlam Almusharraf , Mohammad Tabrez Quasim
N6-methyladenosine (m6A) is one of the most prevalent and functionally significant RNA modifications in eukaryotic transcriptomes, playing critical roles in post-transcriptional gene regulation. Accurate identification of m6A sites remains a considerable challenge due to complex sequence dependencies and limited labeled data. To address these issues, this paper proposed a hybrid Convolutional Neural Network and Long Short-Term Memory (CNN-LSTM) framework integrated with Shapley additive explanations (SHAP) for feature selection and interpretability. The proposed scheme aims to improve both predictive performance and biological insight by identifying and emphasizing the most informative features in RNA sequences. Firstly, multiple biologically relevant feature extraction methods are employed to encode RNA sequences into numerical representations. Secondly, SHAP is used to quantify each feature's contribution, enabling the selection of the most impactful subset while enhancing model interpretability. Thirdly, the selected data is reshaped and standardized to ensure Conv1D compatibility and capture local sequence motifs effectively. Fourthly, CNN layers extract high-level spatial features, which are then fed into LSTM layers to model long-range dependencies within the sequences. Finally, the output is processed through fully connected layers with a sigmoid activation function to perform binary prediction. The experimental results indicate that the CNN-LSTM model, combined with SHAP-based feature selection, outperforms traditional classifiers and standalone deep learning models. The proposed CNN-LSTM framework, evaluated using 10-fold stratified cross-validation, achieved an accuracy of 87.39 %, sensitivity of 83.25 %, specificity of 91.52 %, and an MCC of 0.7534. The results of the proposed model demonstrate its strong ability to accurately classify RNA m6A modification sites, highlighting its potential for large-scale transcriptome-wide epitranscriptomics analysis. The proposed CNN-LSTM model was rigorously compared with several traditional machine learning classifiers and state-of-the-art deep learning approaches, including DT, SVM, KNN, AdaBoost Classifier, Gaussian NB, and DNN.
n6 -甲基腺苷(m6A)是真核生物转录组中最普遍和功能最重要的RNA修饰之一,在转录后基因调控中起着关键作用。由于复杂的序列依赖性和有限的标记数据,准确识别m6A位点仍然是相当大的挑战。为了解决这些问题,本文提出了一个混合卷积神经网络和长短期记忆(CNN-LSTM)框架,并结合Shapley加性解释(SHAP)进行特征选择和可解释性。该方案旨在通过识别和强调RNA序列中最具信息量的特征来提高预测性能和生物学洞察力。首先,采用多种生物学相关特征提取方法将RNA序列编码为数值表示。其次,使用SHAP来量化每个特征的贡献,在增强模型可解释性的同时选择最具影响力的子集。再次,对选取的数据进行重构和标准化,以保证Conv1D的兼容性,有效捕获局部序列基元。第四,CNN层提取高级空间特征,然后将其馈送到LSTM层中,以模拟序列内的长期依赖关系。最后,用sigmoid激活函数对输出进行全连接层处理,进行二值预测。实验结果表明,CNN-LSTM模型结合基于shap的特征选择,优于传统的分类器和独立的深度学习模型。采用10倍分层交叉验证对提出的CNN-LSTM框架进行评估,准确率为87.39%,灵敏度为83.25%,特异性为91.52%,MCC为0.7534。该模型的结果表明,其具有准确分类RNA m6A修饰位点的强大能力,突出了其在大规模转录组全表转录组学分析方面的潜力。提出的CNN-LSTM模型与几种传统的机器学习分类器和最先进的深度学习方法进行了严格的比较,包括DT、SVM、KNN、AdaBoost Classifier、高斯NB和DNN。
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引用次数: 0
Deciphering OCT4A-dose-dependent transcriptional profiles associated with tumorigenic potential in somatic cancer cells 解读体细胞中与致瘤潜能相关的oct4a剂量依赖性转录谱。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-04 DOI: 10.1016/j.slast.2025.100381
Min Tang , Rui Liang , Zhenyu Wu , Cheng Chen , Bo He , Ning Zhou , Sujuan Wang , Xinqiang Xiao , Guangdi Li , Yongfang Jiang , Guozhong Gong , Yanwen Zhou

Aims

The transcription factor OCT4A, a well-established master pluripotency factor, exerts regulatory effects on cell fate determination that are closely associated with its protein levels. This study aims to uncover the downstream gene profile features relevant to tumorigenic potential mediated by OCT4A under varying protein abundance in somatic cancer cells (SCCs).

Materials and methods

CRISPR-Cas9-mediated knockout and doxycycline-inducible OCT4A expression systems were established in cervical (HeLa) and hepatocellular (HepG2, Huh7) cancer cells. Single-cell sequencing, spatial transcriptomic and survival analysis data were used to elucidate the expression pattern of OCT4 in somatic cancer tissues and its prognostic relevance. The plate colony formation assay was performed to assess the tumorigenic capacity of SCCs, and Bulk RNA sequencing coupled with weighted gene co-expression network analysis (WGCNA) identified dose-relevant downstream pathways. Functional enrichment, survival modeling, and RT-qPCR validation were used to construct OCT4A-dose-dependent transcriptional regulatory networks.

Key findings

OCT4 transcript, is heterogeneously present and confined to a small subset of tumor cells within somatic cancer tissues, with a significantly higher proportion of OCT4-positive cells in tumor tissues compared to paired paraneoplastic tissues and is significantly correlated with poor prognosis in SCCs. Endogenous low-level OCT4A positively regulates tumorigenic capacity predominantly through targeting non-coding genes, whereas high-level OCT4A suppresses tumorigenic capacity primarily via protein-coding genes in SCCs. A prognostic model based on high-level OCT4A-regulated protein-coding genes was associated with favorable clinical outcomes, aligning with in vitro phenotypic results.

Significance

Our findings further confirm in SCCs that the functional pleiotropy of OCT4A is closely linked to its protein abundance, and further systematically elucidate the molecular signatures of OCT4A-regulated downstream gene networks associated with tumorigenic phenotypes at differential protein levels, providing novel insights for its translational exploitation in both oncological intervention and regenerative medicine strategies.
目的:转录因子OCT4A是一种公认的主控多能因子,其对细胞命运的调控作用与其蛋白水平密切相关。本研究旨在揭示体细胞(SCCs)中不同蛋白丰度下OCT4A介导的致瘤潜能相关的下游基因谱特征。材料和方法:在宫颈(HeLa)和肝细胞(HepG2, Huh7)癌细胞中建立crispr - cas9介导的敲除和强力霉素诱导的OCT4A表达系统。利用单细胞测序、空间转录组学和生存分析数据来阐明OCT4在体细胞癌组织中的表达模式及其与预后的相关性。通过平板集落形成实验评估SCCs的致瘤能力,Bulk RNA测序结合加权基因共表达网络分析(WGCNA)确定了剂量相关的下游途径。功能富集、生存模型和RT-qPCR验证用于构建oct4a剂量依赖性转录调控网络。关键发现:OCT4转录物异质性存在,局限于体细胞癌组织内的一小部分肿瘤细胞,与配对的副肿瘤组织相比,肿瘤组织中OCT4阳性细胞的比例明显更高,并且与SCCs的不良预后显著相关。内源性低水平OCT4A主要通过靶向非编码基因积极调节致瘤能力,而高水平OCT4A主要通过SCCs中的蛋白质编码基因抑制致瘤能力。基于高水平oct4a调节蛋白编码基因的预后模型与良好的临床结果相关,与体外表型结果一致。意义:我们的研究结果进一步证实了OCT4A的功能多效性与其蛋白丰度密切相关,并进一步系统阐明了OCT4A调控的下游基因网络在不同蛋白水平下与致瘤表型相关的分子特征,为OCT4A在肿瘤干预和再生医学策略中的转化利用提供了新的见解。
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引用次数: 0
Viral peptides remodel the tumor immune microenvironment of cold tumors with RNA-seq insights 病毒肽重塑肿瘤免疫微环境与RNA-Seq的见解。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-04 DOI: 10.1016/j.slast.2025.100379
Zhengqi Peng , Qian Chen , Pengkhun Nov , Mohsin Farid Sulehri , Yujing Liu , Hexi Zhu , Wulikaixi Yagufu , Yiteng Chen , Haipeng Tang , Linlang Guo , Mengchuan Wang

Background

Cold tumors, characterized by low cytotoxic T cell infiltration, limit immunotherapy efficacy. Current strategies often depend on tumor-specific antigens or non-specific immune activation, restricting translational potential. Here, we present a peptide-based platform that leverages intratumoral expression of influenza-derived peptides to redirect virus-specific CD8⁺ T cells and remodel the tumor immune microenvironment in an antigen-independent manner.

Methods

In murine 4T1 and CT26 tumor models, the immunodominant influenza A virus peptide NP147–155 was delivered via intratumoral peptide injection or stable lentiviral expression. Immune remodeling was assessed using flow cytometry, cytokine profiling, and transcriptomic profiling. Differential gene expression and pathway analyses identified mechanisms underlying cold-to-hot TME conversion.

Results

Intratumoral NP147–155 expression robustly activated virus-specific CD8⁺ T cells and enhanced IFN-γ secretion. Transcriptomic profiling revealed upregulation of genes associated with cytotoxicity, chemokine signaling, antigen presentation, and the NKG2D-NKG2DL pathway, consistent with increased T cell infiltration and conversion of cold tumors into hot tumors. Tumor growth was significantly suppressed, and durable anti-tumor immunity was established, including in MHC class I-deficient tumors.

Conclusion

This study demonstrates a peptide-based platform to remodel cold tumor immune microenvironments. Integration of transcriptomic profiling provides mechanistic insights and establishes a robust workflow for evaluating immunomodulatory technologies. This tumor-agnostic approach may enhance anti-tumor immunity and improve immunotherapy efficacy in cold tumors.
背景:冷肿瘤以低细胞毒性T细胞浸润为特征,限制了免疫治疗的效果。目前的策略通常依赖于肿瘤特异性抗原或非特异性免疫激活,限制了翻译潜力。在这里,我们提出了一个基于肽的平台,利用流感衍生肽的肿瘤内表达来重定向病毒特异性CD8+ T细胞,并以抗原独立的方式重塑肿瘤免疫微环境。方法:在小鼠4T1和CT26肿瘤模型中,通过瘤内肽注射法或稳定慢病毒表达法给药免疫优势甲型流感病毒肽NP147-155。利用流式细胞术、细胞因子谱和转录组谱评估免疫重塑。差异基因表达和途径分析确定了冷到热TME转化的机制。结果:瘤内NP147-155的表达强烈激活了病毒特异性CD8 + T细胞,增强了IFN-γ的分泌。转录组学分析显示,与细胞毒性、趋化因子信号、抗原呈递和NKG2D-NKG2DL通路相关的基因上调,与T细胞浸润增加和冷肿瘤向热肿瘤转化一致。肿瘤生长被显著抑制,持久的抗肿瘤免疫被建立,包括MHC i类缺陷肿瘤。结论:本研究提供了一个基于肽的平台来重塑冷肿瘤免疫微环境。转录组分析的整合提供了机制的见解,并建立了一个稳健的工作流程,以评估免疫调节技术。这种肿瘤不可知的方法可以增强抗肿瘤免疫,提高冷肿瘤的免疫治疗效果。
{"title":"Viral peptides remodel the tumor immune microenvironment of cold tumors with RNA-seq insights","authors":"Zhengqi Peng ,&nbsp;Qian Chen ,&nbsp;Pengkhun Nov ,&nbsp;Mohsin Farid Sulehri ,&nbsp;Yujing Liu ,&nbsp;Hexi Zhu ,&nbsp;Wulikaixi Yagufu ,&nbsp;Yiteng Chen ,&nbsp;Haipeng Tang ,&nbsp;Linlang Guo ,&nbsp;Mengchuan Wang","doi":"10.1016/j.slast.2025.100379","DOIUrl":"10.1016/j.slast.2025.100379","url":null,"abstract":"<div><h3>Background</h3><div>Cold tumors, characterized by low cytotoxic T cell infiltration, limit immunotherapy efficacy. Current strategies often depend on tumor-specific antigens or non-specific immune activation, restricting translational potential. Here, we present a peptide-based platform that leverages intratumoral expression of influenza-derived peptides to redirect virus-specific CD8⁺ T cells and remodel the tumor immune microenvironment in an antigen-independent manner.</div></div><div><h3>Methods</h3><div>In murine 4T1 and CT26 tumor models, the immunodominant influenza A virus peptide NP<sub>147–155</sub> was delivered via intratumoral peptide injection or stable lentiviral expression. Immune remodeling was assessed using flow cytometry, cytokine profiling, and transcriptomic profiling. Differential gene expression and pathway analyses identified mechanisms underlying cold-to-hot TME conversion.</div></div><div><h3>Results</h3><div>Intratumoral NP<sub>147–155</sub> expression robustly activated virus-specific CD8⁺ T cells and enhanced IFN-γ secretion. Transcriptomic profiling revealed upregulation of genes associated with cytotoxicity, chemokine signaling, antigen presentation, and the NKG2D-NKG2DL pathway, consistent with increased T cell infiltration and conversion of cold tumors into hot tumors. Tumor growth was significantly suppressed, and durable anti-tumor immunity was established, including in MHC class I-deficient tumors.</div></div><div><h3>Conclusion</h3><div>This study demonstrates a peptide-based platform to remodel cold tumor immune microenvironments. Integration of transcriptomic profiling provides mechanistic insights and establishes a robust workflow for evaluating immunomodulatory technologies. This tumor-agnostic approach may enhance anti-tumor immunity and improve immunotherapy efficacy in cold tumors.</div></div>","PeriodicalId":54248,"journal":{"name":"SLAS Technology","volume":"36 ","pages":"Article 100379"},"PeriodicalIF":3.7,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145696199","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Implementation of a modular digital laboratory infrastructure for SiLA 2 based devices 为基于sila2的设备实现模块化数字实验室基础设施。
IF 3.7 4区 医学 Q3 BIOCHEMICAL RESEARCH METHODS Pub Date : 2025-12-04 DOI: 10.1016/j.slast.2025.100380
Ferdinand Lange, Tessa Habich, Sascha Beutel
Laboratory digitalization is a complex field that includes many aspects to consider, such as device control, data management, and data evaluation. This article presents a potential laboratory digitalization process, including the use of open-source software and hardware components from laboratory equipment to data evaluation. It shows what is required and how it can be used. In this example, a SiLA-based continuous chromatography system is used as a model device to demonstrate the possible purification of GFP from crude cell lysate. The current version of the Gateway Module is a translation unit for driver protocols used for device control. It uses SiLA-based drivers within a Docker environment to communicate with the server infrastructure. The Gateway Module software was managed and rolled out with the mkbox-project. Finally, the recorded laboratory data are stored in a data management system and made available for later analysis. The article also discusses how software and hardware are maintained in the background, as well as the update strategy used.
实验室数字化是一个复杂的领域,包括许多方面需要考虑,如设备控制、数据管理和数据评估。本文介绍了一个潜在的实验室数字化过程,包括使用开源软件和硬件组件,从实验室设备到数据评估。它显示了需要什么以及如何使用它。在这个例子中,一个基于sila的连续色谱系统被用作一个模型装置来演示从粗细胞裂解物中纯化绿色荧光蛋白的可能性。网关模块的当前版本是用于设备控制的驱动协议的转换单元。它在Docker环境中使用基于sila的驱动程序与服务器基础设施进行通信。网关模块软件是通过mkbox项目管理和推出的。最后,记录的实验室数据存储在数据管理系统中,供以后分析使用。本文还讨论了如何在后台维护软件和硬件,以及所使用的更新策略。
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引用次数: 0
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SLAS Technology
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