使用患者衍生的卵巢癌器官进行体外药物测试

卵巢癌是最致命的妇科癌。作为主要治疗方法，化学疗法在晚期阶段的缓解率仅为60-70％，甚至作为二线治疗率较低。尽管有指南建议，哪种药物将最有效。因此，迫切需要一种优先考虑化学疗法选择的策略。最近，癌症器官已成为体外药物测试的一种方法。然而，通过癌症类器官，特别是在妇科癌症中的测试结果评估了有限的临床相关性。因此，我们的目的是产生卵巢癌的患者衍生的器官（PDOS），以评估其药物敏感性和与患者临床结果的相关性。PDOS是由手术切除期间获得的新鲜肿瘤产生的，然后在基质凝胶和适当的生长因子下进行培养。通过相对比显微镜和石蜡包埋的组织病理学评估PDO的形态和分子表征。通过免疫组织化学染色测试了PAX8，TP53，WT1，CK7和CK20的表达，并与亲本肿瘤组织和人类蛋白质ATLAS数据库进行了比较。对PDO进行了体外药物测试，以使用Titer-Glo®3D细胞活力测定法确定药物敏感性。测量了PDO生存力，并计算出曲线下的面积，以比较对各种化合物的反应。在选定患者的临床结果和体外药物测试结果之间计算了相关性。我们确定了31个PDO。其中，可以扩展28个PDO，包括卵巢，子宫内膜和宫颈癌的15、11和2个PDO。 PDO保留了其起源肿瘤的组织病理学特征。 10个卵巢癌PDOS的体外药物测试揭示了对推荐药物的单个差异反应，即使具有相同的组织学类型，也具有人际异质性。在四个具有铂敏感，抗性或难治性的患者中，PDO药物反应与其临床课程非常相关。在使用卵巢癌的体外药物测试中是可行的，并且与患者的临床反应很好地相关。这些结果可能有助于开发精确的化学疗法和对新药的个性化筛查。

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.