胰腺导管腺癌（PDAC）是最恶性肿瘤之一，缺乏有效的治疗方案。癌症相关成纤维细胞（CAF）是肿瘤微环境的重要组成部分，与肿瘤进展、预后和治疗反应相关。本工作旨在探索新的CAFs相关靶点，以改善PDAC的治疗策略。下载PDAC单细胞测序数据（CRA001160，n = 35）并基于GSA数据库进行整合，将成纤维细胞分类为精细亚型。使用功能富集分析和共表达调控网络分析来鉴定不同成纤维细胞亚型的功能表型和生物学特性。使用伪时间分析构建成纤维细胞分化轨迹，以识别成纤维细胞的初始和终末分化亚型。比较有反应和无反应患者PDAC免疫治疗前后不同成纤维细胞亚型比例的变化，并基于GSA和GEO数据库确定成纤维细胞亚型与PDAC免疫治疗反应性之间的关系。利用分子生物学方法证实BNIP3对CAF缺氧和炎症的影响。将CAF与胰腺癌细胞共培养，检测其对胰腺癌迁移和侵袭的影响。单细胞数据分析将成纤维细胞分为6种亚型。分化轨迹表明BNIP3 Fibro亚型呈现终末分化，且随着分化时间的延长，与缺氧和炎症反应相关的基因表达量逐渐增加。 BNIP3 Fibro 亚型中特定的过表达基因与 PDAC 患者的总生存期和无疾病进展生存期显着相关。有趣的是，BNIP3 Fibro亚型的比例越大，PDAC患者对免疫治疗的反应越差，而CRTL治疗方案有效降低了BNIP3 Fibro亚型的比例。敲除BNIP3后，CAF的缺氧标志物和炎症因子受到抑制。 CAFs与胰腺癌细胞共培养可以增加胰腺癌的迁移和侵袭，但这可以通过敲除BNIP3来逆转。这项研究揭示了BNIP3 Fibro亚型与缺氧和炎症反应相关，这与胰腺癌的不良反应密切相关。 PDAC 患者的预后，并确定了预测 PDAC 免疫治疗反应的特征基因。© 2024。作者。

Pancreatic ductal adenocarcinoma (PDAC) is one of the most malignant tumors that lacks effective treatment options. Cancer-associated fibroblasts (CAFs), an important component of the tumor microenvironment, associated with tumor progression, prognosis, and treatment response. This work aimed to explore the novel CAFs-associated target to improve treatment strategies in PDAC.The PDAC single-cell sequencing data (CRA001160, n = 35) were downloaded and integrated based on GSA databases to classify fibroblasts into fine subtypes. Functional enrichment analysis and coexpression regulatory network analysis were used to identify the functional phenotypes and biological properties of the different fibroblast subtypes. Fibroblast differentiation trajectories were constructed using pseudochronological analysis to identify initial and terminally differentiated subtypes of fibroblasts. The changes in the proportions of different fibroblast subtypes before and after PDAC immunotherapy were compared in responsive and nonresponding patients, and the relationships between fibroblast subtypes and PDAC immunotherapy responsiveness were determined based on GSA and GEO database. Using molecular biology methods to confirm the effects of BNIP3 on hypoxia and inflammation in CAFs. CAFs were co cultured with pancreatic cancer cells to detect their effects on migration and invasion of pancreatic cancer.Single-cell data analysis divided fibroblasts into six subtypes. The differentiation trajectory suggested that BNIP3+ Fibro subtype exhibited terminal differentiation, and the expression of genes related to hypoxia and the inflammatory response increased gradually with differentiation time. The specific overexpressed genes in the BNIP3+ Fibro subtype were significantly associated with overall and disease progression-free survival in the patients with PDAC. Interestingly, the greater the proportion of the BNIP3+ Fibro subtype was, the worse the response of PDAC patients to immunotherapy, and the CRTL treatment regimen effectively reduced the proportion of the BNIP3+ Fibro subtype. After knocking out BNIP3, the hypoxia markers and inflammatory factors of CAFs were inhibited. Co-culture of CAFs with pancreatic cancer cells can increase the migration and invasion of pancreatic cancer, but this could be reversed by knocking out BNIP3.This study revealed the BNIP3+ Fibro subtype associated with hypoxia and inflammatory responses, which was closely related to the poor prognosis of patients with PDAC, and identified signature genes that predict the immunotherapy response in PDAC.© 2024. The Author(s).