Department of Medicine, University of California, San Francisco; Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco; Department of Cellular and Molecular Pharmacology, University of California, San Francisco.
Department of Medicine, University of California, San Francisco; Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco.
J Vis Exp. 2021 Nov 22(177). doi: 10.3791/63039.
Novel 3D cancer organoid cultures derived from clinical patient specimens represent an important model system to evaluate intratumor heterogeneity and treatment response to targeted inhibitors in cancer. Pioneering work in gastrointestinal and pancreatic cancers has highlighted the promise of patient-derived organoids (PDOs) as a patient-proximate culture system, with an increasing number of models emerging. Similarly, work in other cancer types has focused on establishing organoid models and optimizing culture protocols. Notably, 3D cancer organoid models maintain the genetic complexity of original tumor specimens and thus translate tumor-derived sequencing data into treatment with genetically informed targeted therapies in an experimental setting. Further, PDOs might foster the evaluation of rational combination treatments to overcome resistance-associated adaptation of tumors in the future. The latter focuses on intense research efforts in non-small-cell lung cancer (NSCLC), as resistance development ultimately limits the treatment success of targeted inhibitors. An early assessment of therapeutically targetable mechanisms using NSCLC PDOs could help inform rational combination treatments. This manuscript describes a standardized protocol for the cell culture plate-based assessment of drug sensitivities to targeted inhibitors in NSCLC-derived 3D PDOs, with potential adaptability to combinational treatments and other treatment modalities.
从临床患者标本中衍生的新型 3D 癌症类器官培养物代表了一个重要的模型系统，可用于评估肿瘤内异质性和针对癌症的靶向抑制剂的治疗反应。在胃肠道和胰腺癌症方面的开创性工作强调了患者来源的类器官（PDO）作为一种接近患者的培养系统的潜力，越来越多的模型不断涌现。同样，其他癌症类型的工作也集中在建立类器官模型和优化培养方案上。值得注意的是，3D 癌症类器官模型保留了原始肿瘤标本的遗传复杂性，因此可以将肿瘤衍生的测序数据转化为在实验环境中使用具有遗传信息的靶向治疗。此外，PDO 可能有助于评估合理的联合治疗方法，以克服未来肿瘤耐药相关的适应性。后者侧重于非小细胞肺癌（NSCLC）的密集研究工作，因为耐药性的发展最终限制了靶向抑制剂的治疗成功。使用 NSCLC PDO 对治疗靶点机制进行早期评估，可以帮助为合理的联合治疗提供信息。本文描述了一种基于细胞培养板的标准化方案，用于评估 NSCLC 衍生的 3D PDO 中针对靶向抑制剂的药物敏感性，该方案具有针对联合治疗和其他治疗方式的潜在适应性。
