i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Rua Alfredo Allen 208, Porto, 4200-135, Portugal.
Cancer Drug Resistance Group, IPATIMUP - Institute of Molecular Pathology and Immunology, University of Porto, Rua Alfredo Allen 208, Porto, 4200-135, Portugal.
Cancer Chemother Pharmacol. 2024 Oct;94(4):585-597. doi: 10.1007/s00280-024-04712-1. Epub 2024 Sep 3.
Pancreatic ductal adenocarcinoma (PDAC) is among the most aggressive malignancies. Our previous work revealed Chitinase 3-like 1 (CHI3L1) involvement in PDAC resistance to gemcitabine, identifying it as a promising therapeutic target. Here, we aimed to identify putative CHI3L1 inhibitors and to investigate their chemosensitizing potential in PDAC.
Docking analysis for CHI3L1 identified promising CHI3L1 inhibitors, including darifenacin (muscarinic receptor antagonist). PDAC cell lines (BxPC-3, PANC-1) and primary PDAC cells were used to evaluate darifenacin's effects on cell growth (Sulforhodamine B, SRB), alone or in combination with gemcitabine or gemcitabine plus paclitaxel. Cytotoxicity against normal immortalized pancreatic ductal cells (HPNE) was assessed. Recombinant protein was used to confirm the impact of darifenacin on CHI3L1-induced PDAC cellular resistance to therapy (SRB assay). Darifenacin's effect on Akt activation was analysed by ELISA. The association between cholinergic receptor muscarinic 3 (CHRM3) expression and therapeutic response was evaluated by immunohistochemistry of paraffin-embedded tissues from surgical resections of a 68 patients' cohort.
In silico screening revealed the ability of darifenacin to target CHI3L1 with high efficiency. Darifenacin inhibited PDAC cell growth, with a GI of 26 and 13.6 µM in BxPC-3 and PANC-1 cells, respectively. These results were confirmed in primary PDAC-3 cells, while darifenacin showed no cytotoxicity against HPNE cells. Importantly, darifenacin sensitized PDAC cells to standard chemotherapies, reverted CHI3L1-induced PDAC cellular resistance to therapy, and decreased Akt phosphorylation. Additionally, high CHMR3 expression was associated with low therapeutic response to gemcitabine.
This work highlights the potential of darifenacin as a chemosensitizer for PDAC treatment.
胰腺导管腺癌（PDAC）是最具侵袭性的恶性肿瘤之一。我们之前的工作表明，几丁质酶 3 样 1（CHI3L1）参与了 PDAC 对吉西他滨的耐药性，将其确定为有前途的治疗靶点。在这里，我们旨在确定潜在的 CHI3L1 抑制剂，并研究其在 PDAC 中的化学增敏作用。
针对 CHI3L1 的对接分析确定了有前途的 CHI3L1 抑制剂，包括达非那新（毒蕈碱受体拮抗剂）。使用 PDAC 细胞系（BxPC-3、PANC-1）和原代 PDAC 细胞来评估达非那新对细胞生长（磺酰罗丹明 B，SRB）的影响，单独使用或与吉西他滨或吉西他滨加紫杉醇联合使用。评估对正常永生化胰腺导管细胞（HPNE）的细胞毒性。使用重组蛋白来确认达非那新对 CHI3L1 诱导的 PDAC 细胞对治疗的耐药性的影响（SRB 测定）。通过 ELISA 分析达非那新对 Akt 激活的影响。通过对 68 名患者手术切除组织的石蜡包埋组织进行免疫组织化学评估胆碱能受体毒蕈碱 3（CHRM3）表达与治疗反应之间的关联。
计算机筛选显示达非那新具有高效靶向 CHI3L1 的能力。达非那新抑制 PDAC 细胞生长，在 BxPC-3 和 PANC-1 细胞中的 GI 分别为 26 和 13.6µM。这些结果在原代 PDAC-3 细胞中得到了证实，而达非那新对 HPNE 细胞没有细胞毒性。重要的是，达非那新使 PDAC 细胞对标准化疗药物敏感，逆转了 CHI3L1 诱导的 PDAC 细胞对治疗的耐药性，并降低了 Akt 磷酸化。此外，高 CHRM3 表达与吉西他滨治疗反应低相关。
这项工作强调了达非那新作为 PDAC 治疗化学增敏剂的潜力。
