Hu Qingwen, Chen Jiali, Liu Yang, Chen Haiqing, Lai Haotian, Jiang Lai, Zhou Xuancheng, Zhang Shengke, Huang Jinbang, Chi Hao, Li Bo, Zhong Xiaolin
Clinical Medical College, Southwest Medical University, Luzhou, China.
Department of General Surgery (Hepatopancreatobiliary Surgery), The Affiliated Hospital of Southwest Medical University, Luzhou, China.
Front Immunol. 2025 May 15;16:1594879. doi: 10.3389/fimmu.2025.1594879. eCollection 2025.
Pancreatic cancer (PC) is a highly aggressive pancreatic malignant tumor with poor prognosis due to its complex tumor microenvironment (TME) and metastatic potential. Fibroblasts play an important role in tumor progression and metastasis by remodeling the extracellular matrix (ECM) and influencing the immune response. This study explored migrasome-associated fibroblasts, especially TSPAN4 and ITGA5, as key regulators of pancreatic cancer metastasis, aiming to provide new ideas for therapeutic strategies targeting TME.
We employed single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics to analyze pancreatic cancer tissues. Data from the GEO and TCGA databases were integrated and processed using batch correction techniques. Single-cell data were analyzed with Seurat and Monocle for dimensionality reduction and pseudotime trajectory analysis. Cell communication was assessed using CellCall and CellChat. Spatial transcriptomic analysis was conducted with RCTD and MISTy tools to investigate cell interactions within the TME. Additionally, gene enrichment, deconvolution, and prognostic analyses were performed, alongside experimental validation through siRNA transfection, qRT-PCR, and various functional assays to investigate the role of TSPAN4 in metastasis.
Our results underscore the critical role of TSPAN4 fibroblasts in pancreatic cancer. These fibroblasts were predominantly located at the tumor periphery and exhibited elevated migrasome gene expression, which was associated with enhanced ECM remodeling and immune suppression. Intercellular communication analysis revealed that TSPAN4 fibroblasts engaged in extensive interactions with immune cells, such as macrophages and endothelial cells, facilitating metastasis and immune evasion. Moreover, the high expression of immune checkpoint markers indicated their involvement in modulating the immune response.
TSPAN4 fibroblasts are key regulators of pancreatic cancer progression, contributing to metastasis, immune suppression, and ECM remodeling. Targeting these fibroblasts represents a promising therapeutic strategy to improve clinical outcomes and enhance the effectiveness of immunotherapies in pancreatic cancer.
胰腺癌（PC）是一种侵袭性很强的胰腺恶性肿瘤，由于其复杂的肿瘤微环境（TME）和转移潜能，预后较差。成纤维细胞通过重塑细胞外基质（ECM）和影响免疫反应，在肿瘤进展和转移中发挥重要作用。本研究探索了与迁移体相关的成纤维细胞，特别是四跨膜蛋白4（TSPAN4）和整合素α5（ITGA5），作为胰腺癌转移的关键调节因子，旨在为靶向TME的治疗策略提供新思路。
我们采用单细胞RNA测序（scRNA-seq）和空间转录组学分析胰腺癌组织。利用批次校正技术整合和处理来自基因表达综合数据库（GEO）和癌症基因组图谱（TCGA）数据库的数据。使用Seurat和Monocle对单细胞数据进行分析，以进行降维和伪时间轨迹分析。使用CellCall和CellChat评估细胞通讯。使用RCTD和MISTy工具进行空间转录组分析，以研究TME内的细胞相互作用。此外，还进行了基因富集、反卷积和预后分析，并通过小干扰RNA（siRNA）转染、定量逆转录聚合酶链反应（qRT-PCR）和各种功能试验进行实验验证，以研究TSPAN4在转移中的作用。
我们的结果强调了TSPAN4成纤维细胞在胰腺癌中的关键作用。这些成纤维细胞主要位于肿瘤周边，迁移体基因表达升高，这与增强的ECM重塑和免疫抑制相关。细胞间通讯分析显示，TSPAN4成纤维细胞与免疫细胞（如巨噬细胞和内皮细胞）进行广泛相互作用，促进转移和免疫逃逸。此外，免疫检查点标志物的高表达表明它们参与调节免疫反应。
TSPAN4成纤维细胞是胰腺癌进展的关键调节因子，有助于转移、免疫抑制和ECM重塑。靶向这些成纤维细胞代表了一种有前景的治疗策略，可改善临床结局并提高胰腺癌免疫治疗的有效性。
