Department of General Surgery & Institute of Precision Diagnosis and Treatment of Digestive System Tumors, Carson International Cancer Center, Shenzhen University General Hospital, Shenzhen University, Shenzhen, Guangdong, 518055, China.
Guangdong Provincial Key Laboratory for Biomedical Measurements and Ultrasound Imaging, School of Biomedical Engineering, Shenzhen University Medical School, Xueyuan Road 1066, Shenzhen, 518060, China.
J Exp Clin Cancer Res. 2023 Aug 9;42(1):199. doi: 10.1186/s13046-023-02756-4.
Gemcitabine resistance (GR) is a significant clinical challenge in pancreatic adenocarcinoma (PAAD) treatment. Macrophages in the tumor immune-microenvironment are closely related to GR. Uncovering the macrophage-induced GR mechanism could help devise a novel strategy to improve gemcitabine treatment outcomes in PAAD. Therefore, preclinical models accurately replicating patient tumor properties are essential for cancer research and drug development. Patient-derived organoids (PDOs) represent a promising in vitro model for investigating tumor targets, accelerating drug development, and enabling personalized treatment strategies to improve patient outcomes.
To investigate the effects of macrophage stimulation on GR, co-cultures were set up using PDOs from three PAAD patients with macrophages. To identify signaling factors between macrophages and pancreatic cancer cells (PCCs), a 97-target cytokine array and the TCGA-GTEx database were utilized. The analysis revealed CCL5 and AREG as potential candidates. The role of CCL5 in inducing GR was further investigated using clinical data and tumor sections obtained from 48 PAAD patients over three years, inhibitors, and short hairpin RNA (shRNA). Furthermore, single-cell sequencing data from the GEO database were analyzed to explore the crosstalk between PCCs and macrophages. To overcome GR, inhibitors targeting the macrophage-CCL5-Sp1-AREG feedback loop were evaluated in cell lines, PDOs, and orthotopic mouse models of pancreatic carcinoma.
The macrophage-CCL5-Sp1-AREG feedback loop between macrophages and PCCs is responsible for GR. Macrophage-derived CCL5 activates the CCR5/AKT/Sp1/CD44 axis to confer stemness and chemoresistance to PCCs. PCC-derived AREG promotes CCL5 secretion in macrophages through the Hippo-YAP pathway. By targeting the feedback loop, mithramycin improves the outcome of gemcitabine treatment in PAAD. The results from the PDO model were corroborated with cell lines, mouse models, and clinical data.
Our study highlights that the PDO model is a superior choice for preclinical research and precision medicine. The macrophage-CCL5-Sp1-AREG feedback loop confers stemness to PCCs to facilitate gemcitabine resistance by activating the CCR5/AKT/SP1/CD44 pathway. The combination of gemcitabine and mithramycin shows potential as a therapeutic strategy for treating PAAD in cell lines, PDOs, and mouse models.
吉西他滨耐药（GR）是胰腺导管腺癌（PAAD）治疗中的一个重大临床挑战。肿瘤免疫微环境中的巨噬细胞与 GR 密切相关。揭示巨噬细胞诱导的 GR 机制有助于设计一种新的策略来改善 PAAD 中吉西他滨的治疗效果。因此，准确复制患者肿瘤特性的临床前模型对于癌症研究和药物开发至关重要。患者来源的类器官（PDO）代表了一种有前途的体外模型，可用于研究肿瘤靶点、加速药物开发，并实现个性化治疗策略以改善患者预后。
为了研究巨噬细胞刺激对 GR 的影响，使用来自三名 PAAD 患者的 PDO 与巨噬细胞共培养。为了鉴定巨噬细胞和胰腺癌细胞（PCC）之间的信号因子，使用了 97 个靶标细胞因子阵列和 TCGA-GTEx 数据库。分析显示 CCL5 和 AREG 是潜在的候选物。使用来自 48 名 PAAD 患者的临床数据和肿瘤切片、抑制剂和短发夹 RNA（shRNA）进一步研究了 CCL5 诱导 GR 的作用。此外，还分析了 GEO 数据库中的单细胞测序数据，以探讨 PCC 与巨噬细胞之间的串扰。为了克服 GR，评估了针对巨噬细胞-CCL5-Sp1-AREG 反馈环的抑制剂在细胞系、PDO 和胰腺癌细胞的原位小鼠模型中的效果。
巨噬细胞-CCL5-Sp1-AREG 反馈环在巨噬细胞和 PCC 之间导致 GR。巨噬细胞衍生的 CCL5 通过激活 CCR5/AKT/Sp1/CD44 轴赋予 PCC 干性和化疗耐药性。PCC 衍生的 AREG 通过 Hippo-YAP 通路促进巨噬细胞中 CCL5 的分泌。通过靶向反馈环，米托蒽醌改善了 PAAD 中吉西他滨治疗的效果。PDO 模型的结果得到了细胞系、小鼠模型和临床数据的证实。
我们的研究强调了 PDO 模型是临床前研究和精准医学的更好选择。巨噬细胞-CCL5-Sp1-AREG 反馈环通过激活 CCR5/AKT/SP1/CD44 通路赋予 PCC 干性，从而促进吉西他滨耐药。吉西他滨和米托蒽醌的联合应用显示出在细胞系、PDO 和小鼠模型中治疗 PAAD 的治疗潜力。
