Xu Haiyan, Xue Shengbai, Sun Yang, Ma Jingyu, Li Shumin, Wang Yanling, Mao Tiebo, Ge Weiyu, Yue Ming, Shentu Daiyuan, Lu Wenxin, Wang Yongchao, Hu Jiong, Cui Jiujie, Zhang Xiaofei, Cai Li, Wang Yu, Wang Liwei
State Key Laboratory of Systems Medicine for Cancer of Oncology Department and Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China
State Key Laboratory of Systems Medicine for Cancer of Oncology Department and Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
J Immunother Cancer. 2025 Jan 6;13(1):e010029. doi: 10.1136/jitc-2024-010029.
To date, a growing body of evidence suggests that unfolded protein response (UPR) sensors play a vital role in carcinogenesis. However, it remains unclear whether they are involved in pancreatic ductal adenocarcinoma (PDAC) and how they relate to clinical outcomes. This study aims to investigate the biological function and mechanism of how a novel UPR sensor, CREB3L1 works in PDAC and further evaluate its clinical application prospect.
We tested UPR signaling including CREB3L1 in Thapsigargin-treated PDAC cells. Subsequently, we defined CREB3L1 expression and further analyzed its expression with clinical characteristics in PDAC. Then, we established gene-modified cells to determine whether CREB3L1 functions in cell proliferation and migration capacity. Besides, we constructed subcutaneously and orthotopically transplanted mice models to verify their progrowing function and pulmonary metastasis models to prove their proinvasion role. What's more, RNAseq, qPCR, Western blotting, immunohistochemistry and multicolor flow cytometry experiments were used to explore the mechanism of how CREB3L1 worked in PDAC. Lastly, CREB3L1 expression correlation with PDAC immunotherapy outcome and immune cell signatures were explored in the patients with advanced PDAC who received PD-1 antibody therapy.
We first confirmed CREB3L1 could be induced by endoplasmic reticulum stressor and found its aberrant activation was associated with poorer overall survival in PDAC patients indicating the protumor function of the new UPR sensor. Functionally, we confirmed CREB3L1 contributing to PDAC malignant progression including growth and metastasis by multiple in in vitro and in vivo models. Mechanistically, CREB3L1 upregulated COL3A1 and promoted dense stroma formation for facilitating PDAC and knocking down COL3A1 disrupted CREB3L1 protumor function. Furthermore, CREB3L1-induced TAM polarization toward an M2 phenotype and reduced the infiltration of CD8 T cells. Clinically, CREB3L1 correlated with immune cell signatures as well as immune checkpoint blockade (ICB) treatment response and outcome that CREB3L1aberrant activation indicated poorer efficacy and worse prognosis than the low in PDAC which might empower clinical decision.
Collectively, this study revealed CREB3L1 facilitated PDAC progression, shaped an immune exclude tumor microenvironment and distinguished therapy response and outcome of ICB therapy indicating CREB3L1 could be a promising novel molecular target and biomarker for PDAC treatment.
迄今为止，越来越多的证据表明未折叠蛋白反应（UPR）传感器在肿瘤发生中起着至关重要的作用。然而，它们是否参与胰腺导管腺癌（PDAC）以及它们与临床结局的关系仍不清楚。本研究旨在探讨一种新型UPR传感器CREB3L1在PDAC中的生物学功能和作用机制，并进一步评估其临床应用前景。
我们检测了毒胡萝卜素处理的PDAC细胞中的UPR信号，包括CREB3L1。随后，我们确定了CREB3L1的表达，并进一步分析了其在PDAC中的表达与临床特征的关系。然后，我们建立了基因修饰细胞，以确定CREB3L1是否在细胞增殖和迁移能力中发挥作用。此外，我们构建了皮下和原位移植小鼠模型，以验证其促生长功能，并构建了肺转移模型，以证明其促侵袭作用。更重要的是，我们使用RNA测序、qPCR、蛋白质免疫印迹、免疫组织化学和多色流式细胞术实验来探索CREB3L1在PDAC中发挥作用的机制。最后，在接受PD-1抗体治疗的晚期PDAC患者中，探讨了CREB3L1表达与PDAC免疫治疗结果及免疫细胞特征的相关性。
我们首先证实CREB3L1可被内质网应激诱导，并发现其异常激活与PDAC患者较差的总生存期相关，表明这种新的UPR传感器具有促肿瘤功能。在功能上，我们通过多种体外和体内模型证实CREB3L1促进了PDAC的恶性进展，包括生长和转移。在机制上，CREB3L1上调COL3A1并促进致密基质形成以促进PDAC，敲低COL3A1破坏了CREB3L1的促肿瘤功能。此外，CREB3L1诱导肿瘤相关巨噬细胞向M2表型极化，并减少CD8 T细胞的浸润。在临床上，CREB3L1与免疫细胞特征以及免疫检查点阻断（ICB）治疗反应和结果相关，CREB3L1的异常激活表明其疗效比低表达者差，预后更差，这可能有助于临床决策。
总体而言，本研究揭示了CREB促进了PDAC的进展，塑造了免疫排斥性肿瘤微环境，并区分了ICB治疗的反应和结果，表明CREB3L1可能是PDAC治疗中一个有前景的新型分子靶点和生物标志物。
