Guerrero Quiles Conrado, Fahy Siobhan, Bartak Matej, Gonzalez Abalos Julia, Powell Emilia, Lodhi T, Reed Rachel, Reeves Kim, Baker Alex, James Nicholas D, Hall Emma, Huddart Robert A, Porta Nuria, Hoskin Peter, West Catharine, Biolatti Luisa V, Choudhury Ananya
Division of Cancer Sciences, The University of Manchester, The Christie Hospital National Health Service (NHS) Foundation Trust, Manchester, United Kingdom.
Institute Curie, Immunity and Cancer Unit (U932), Paris, France.
Front Oncol. 2025 Jul 28;15:1616943. doi: 10.3389/fonc.2025.1616943. eCollection 2025.
Muscle-invasive bladder cancer (MIBC) is a prevalent disease that can be treated with radiotherapy, but has a poor prognosis. Radiation-induced extracellular matrix (ECM) remodelling and fibrosis can induce tumour resistance and recurrence, but have not been studied in MIBC. Here, we aimed to characterise the impact of radiation on the ECM composition of MIBC. Three MIBC cell lines (T24, UMUC3, J82) were treated with fractionated radiation. We used proteomics to analyse the ECM composition produced by surviving cancer cells and immunofluorescence to investigate changes in the morphology and number of ECM fibres. We evaluated the RNA expression of identified ECM proteins (FN1, COL5A1, COL1A1, TNF6AIP6, FLG) in one cystectomy (TCGA-BLCA, n=397) and two radiotherapy (BC2001, n=313; BCON, n=151) cohorts. There were 613 proteins affected by radiation (p<0.05, fold change >2 or <-2), 68 of which were ECM-associated proteins. There was a general increase in proteases and protease regulators but heterogeneity across cell lines. Enrichment analysis showed ECM organisation was the primary pathway affected. Immunofluorescence confirmed radiation affected ECM structure, generally, reducing the number, length and width of fibres. Five ECM genes of interest were identified (, , , , ), constituting an ECM signature. High , mRNA levels and ECM signature scores were independent poor prognostic markers, while mRNA expression independently predicted radiotherapy benefit in a meta-analysis (n=861). We found high expression levels predicted hypoxia-modifying treatment benefit. Prognostic significance of , and the ECM signature was dependent on patients harbouring -mutations. Radiation alters the composition and structure of the ECM produced by MIBC. As a proof-of-concept, we showed that radiation-affected ECM genes are independent prognostic and predictive markers of radiotherapy benefit in MIBC. Future studies should validate these radiation-induced ECM changes in clinical samples, and explore the role of in radioresistance.
肌肉浸润性膀胱癌（MIBC）是一种常见疾病，可采用放射治疗，但预后较差。辐射诱导的细胞外基质（ECM）重塑和纤维化可诱导肿瘤耐药性和复发，但在MIBC中尚未得到研究。在此，我们旨在描述辐射对MIBC细胞外基质组成的影响。对三种MIBC细胞系（T24、UMUC3、J82）进行分次辐射处理。我们使用蛋白质组学分析存活癌细胞产生的细胞外基质组成，并通过免疫荧光研究细胞外基质纤维的形态和数量变化。我们评估了一个膀胱切除术队列（TCGA - BLCA，n = 397）和两个放射治疗队列（BC2001，n = 313；BCON，n = 151）中已鉴定的细胞外基质蛋白（FN1、COL5A1、COL1A1、TNF6AIP6、FLG）的RNA表达。有613种蛋白质受辐射影响（p < 0.05，倍数变化>2或< -2），其中68种是与细胞外基质相关的蛋白质。蛋白酶和蛋白酶调节剂总体上有所增加，但各细胞系存在异质性。富集分析表明细胞外基质组织是受影响的主要途径。免疫荧光证实辐射影响细胞外基质结构，一般会减少纤维的数量、长度和宽度。鉴定出五个感兴趣的细胞外基质基因（此处原文缺失具体基因名称），构成一个细胞外基质特征。高（此处原文缺失具体基因名称）、（此处原文缺失具体基因名称）mRNA水平和细胞外基质特征评分是独立的不良预后标志物，而（此处原文缺失具体基因名称）mRNA表达在一项荟萃分析（n = 861）中独立预测放射治疗获益。我们发现高（此处原文缺失具体基因名称）表达水平预测低氧修饰治疗获益。（此处原文缺失具体基因名称）、（此处原文缺失具体基因名称）和细胞外基质特征的预后意义取决于携带（此处原文缺失具体基因突变名称）突变的患者。辐射改变了MIBC产生的细胞外基质的组成和结构。作为概念验证，我们表明受辐射影响的细胞外基质基因是MIBC放射治疗获益的独立预后和预测标志物。未来的研究应在临床样本中验证这些辐射诱导的细胞外基质变化，并探索（此处原文缺失具体基因名称）在放射抗性中的作用。
