利用患者来源的卵巢癌类器官进行体外药物测试

卵巢癌是最致命的妇科肿瘤。在晚期，化疗的反应率仅为60-70%，作为二线治疗时更低。尽管指南建议，但最有效的药物尚不明确，因此亟需制定优先化疗方案的策略。近年来，肿瘤类器官作为体外药物测试的新方法逐渐兴起，但在妇科肿瘤中，临床相关性有限。我们旨在建立患者来源的卵巢癌类器官（PDOs），评估其药物敏感性及与患者临床结局的关系。PDOs由手术切除的新鲜肿瘤组织制备，在基质胶和适宜生长因子中培养。通过相差显微镜和石蜡包埋的组织病理方法进行形态学和分子特征检测，免疫组化检测PAX8、TP53、WT1、CK7和CK20的表达，并与原发肿瘤组织及人类蛋白质图谱数据库进行比对。利用Titer-Glo® 3D细胞活力测定法进行体外药物敏感性测试，计算药物反应的曲线下面积（AUC），评估不同药物的反应差异。分析患者临床结局与药物测试结果的相关性。共建立了31个PDOs，其中28个可扩增，包括15个卵巢癌、11个子宫内膜癌及2个宫颈癌。PDOs保持了原发肿瘤的组织学特征。10个卵巢癌PDOs的药物测试显示不同个体对推荐药物的反应差异，即使在相同组织类型中也存在异质性。在4例对铂类药物敏感、耐药或难治的患者中，PDO的药物反应与临床表现高度相关。体外药物测试验证了卵巢癌类器官的可行性，并与临床反应密切相关，有助于精准化疗和个性化药物筛选，包括新药及旧药的再利用。

Ovarian cancer is the most lethal gynecological cancer. As the primary treatment, chemotherapy has a response rate of only 60-70% in advanced stages, and even lower as a second-line treatment. Despite guideline recommendations, which drugs will be most effective remains unclear. Thus, a strategy to prioritize chemotherapy options is urgently needed. Cancer organoids have recently emerged as a method for in vitro drug testing. However, limited clinical correlations have been assessed with test results from cancer organoids, particularly in gynecological cancers. We therefore aimed to generate patient-derived organoids (PDOs) of ovarian cancer, to assess their drug sensitivities and correlations with patient clinical outcomes.PDOs were generated from fresh tumors obtained during surgical resection, which was then cultured under matrix gel and appropriate growth factors. Morphological and molecular characterization of PDOs were assessed by phase contrast microscopy and paraffin-embedded histopathology. Expressions of PAX8, TP53, WT1, CK7, and CK20 were tested by immunohistochemical staining and compared with parental tumor tissues and the human protein atlas database. PDOs were subjected to in vitro drug testing to determine drug sensitivity using Titer-Glo® 3D Cell Viability Assay. PDO viability was measured, and area under the curve calculated, to compare responses to various compounds. Correlations were calculated between selected patients' clinical outcomes and in vitro drug testing results.We established 31 PDOs. Among them, 28 PDOs can be expanded, including 15, 11, and 2 from ovarian, endometrial, and cervical cancers, respectively. The PDOs preserved the histopathological profiles of their originating tumors. In vitro drug testing of 10 ovarian cancer PDOs revealed individual differential responses to recommended drugs, and interpersonal heterogeneity in drug sensitivity, even with the same histology type. Among four patients who were platinum sensitive, resistant, or refractory, PDO drug responses correlated well with their clinical courses.In vitro drug testing using ovarian cancer organoids is feasible and correlates well with patient clinical responses. These results may facilitate development of precision chemotherapy and personalized screening for repurposed or new drugs.