BNIP3+与缺氧和炎症相关的成纤维细胞预测胰腺导管腺癌的预后和免疫疗法反应

胰腺导管腺癌（PDAC）是缺乏有效治疗选择的最恶性肿瘤之一。癌症相关的成纤维细胞（CAF）是肿瘤微环境的重要组成部分，与肿瘤进展，预后和治疗反应有关。这项工作旨在探索与CAFS相关的新型目标，以改善PDAC中的治疗策略。PDAC单细胞测序数据（CRA001160，n = 35）的下载并基于GSA数据库进行了集成，以将成纤维细胞分类为细胞亚型。功能富集分析和共表达调节网络分析用于识别不同成纤维细胞亚型的功能表型和生物学特性。使用假体学分析构建成纤维细胞分化轨迹，以鉴定成纤维细胞的初始和末端分化的亚型。在反应性和无反应患者中比较了PDAC免疫疗法前后不同成纤维细胞亚型的比例变化，并根据GSA和GEO数据库确定了成纤维细胞亚型与PDAC免疫疗法反应性之间的关系。使用分子生物学方法来确认BNIP3对CAF中缺氧和炎症的影响。将CAF与胰腺癌细胞一起培养，以检测其对胰腺癌迁移和侵袭的影响。单核数据分析将成纤维细胞分为六个亚型。分化轨迹表明，BNIP3+纤维亚型表现出终末分化，并且与缺氧相关的基因表达和炎症反应随着分化时间而逐渐增加。 BNIP3+纤维亚型中的特异性过表达基因与PDAC患者的总体和无疾病进展生存率显着相关。有趣的是，BNIP3+纤维亚型的比例越大，PDAC患者对免疫疗法的反应越多，并且CRTL治疗方案有效地降低了BNIP3+纤维亚型的比例。拆除BNIP3后，抑制了CAF的缺氧标记和炎症因子。 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.

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.