[摘 要] 目的：构建胆管癌（CCA）患者来源癌组织类器官（PDO）模型，并评估其在检测顺铂与吉西他滨单药及联合用药敏感 性中的应用价值。方法：收集4例经病理确诊、术前未行系统治疗的胆管癌患者的肿瘤组织，采用基质胶三维培养法构建PDO 模型，并通过苏木精-伊红（H-E）染色法、免疫组化（IHC）法及全外显子组测序（WES）验证其与原发肿瘤的组织学形态及遗传特 征一致性。对稳定传代的PDO采用三磷酸腺苷（ATP）法进行药物敏感性检测，拟合剂量-反应曲线计算IC50 ；并通过Chou-Talalay 方法计算联合指数（CI）评估协同作用。结果：成功构建3例可稳定传代超过7代的CCA的PDO模型，H-E和IHC结果显示PDO 模型与原代肿瘤组织高度一致，WES显示配对PDO保留了其来源组织约78.4%的遗传变异。3例PDO对顺铂的IC50 分别为 （277.50 ± 4.056）、 （9.129 0 ± 1.012）及（115.50 ± 3.034） μmol/L；对吉西他滨的IC50 分别为（0.581 8 ± 0.020）、 （0.012 1 ± 0.008）及 （0.048 9 ± 0.004） μmol/L，显示显著个体差异。联合用药实验中，PDO#1表现为显著协同作用（CI = 0.116~0.573）；PDO#2在低剂 量出现拮抗，部分中等剂量呈协同；PDO#3在低剂量出现拮抗，在中剂量呈协同，而在高剂量再次出现拮抗（最高组CI ≈ 1.99），表 明该药物组合的协同作用仅限定于一个剂量范围内。结论：本研究构建的CCA的PDO模型能再现原代肿瘤组织的形态学及遗 传学特征，有效模拟患者肿瘤的药物反应异质性，为临床个性化治疗方案的选择和优化提供了高效、可靠的检测平台。

[Abstract] Objective: To construct patient-derived organoid (PDO) models of cholangiocarcinoma (CCA) and evaluate their application values in assessing the drug sensitivity to cisplatin and gemcitabine, both as single agents and in combination. Methods: TTumor tissues were collected from four patients with pathologically confirmed cholangiocarcinoma who had not received any prior systematic treatment. PDO models were constructed using matrigel 3D cultures. The consistency of histological morphology and genetic characteristics between PDO and primary tumors was validated by hematoxylin-eosin (H-E) staining, immunohistochemistry (IHC) and whole-exome sequencing (WES). Drug sensitivity of stably passaged PDO was detected using the adenosine triphosphate (ATP) assay, and dose-response curves were used to calculate the half-maximal inhibitory concentration (IC50 ). Synergistic effects were evaluated by calculating the combination index (CI) through the Chou-Talalay method. Results: PDO models for three CCA strains capable of stable subculturing for over seven generations were successfully constructed. H-E staining and IHC results demonstrated high consistency between the PDO models and the primary tumors. WES analysis revealed that the paired PDO retained approximately 78.4% of the genetic variations present in the source tissues. The IC50 values of the three PDO for cisplatin were (277.50 ± 4.056), (9.129 0 ± 1.012), and (115.50 ± 3.034) μmol/L, respectively, while the IC50 values for gemcitabine were (0.581 8 ± 0.020), (0.012 1 ± 0.008), and (0.048 9 ± 0.004) μmol/L, indicating significant inter-individual differences. In the combination therapy experiment, PDO#1 exhibited a significant synergistic effect (CI = 0.116-0.573); PDO#2 showed antagonism at low doses and synergy at moderate doses; PDO#3 displayed antagonism at low doses, synergy at moderate doses, and a return to antagonism at high doses (CI ≈ 1.99 in the highest concentration group). These results indicated that the synergistic effect of the drug combination was strictly confined to a specific dose range. Conclusion: The CCA PDO model faithfully reproduces the morphological and genetic features of the original tumor, and effectively simulates the heterogeneous drug response patterns in patients' tumors, which provides an efficient and reliable platform for selecting and optimizing personalized clinical treatment strategies.

[基金项目] 福建省自然科学基金资助项目（No. 2022J01742）