Core 2: Stanford Breast Metastasis Center Organoid Core

摘要/项目摘要 乳腺癌研究因缺乏有效反映乳腺癌的实验模型而受到严重阻碍。 生理性疾病，提供转移性疾病的全面了解所必需的。常规 细胞系和PDX模型极大地促进了我们对乳腺癌生物学的理解，但 这些结果不能完全捕获乳腺癌亚型之间的异质性， 临床观察。我们实验室和其他实验室的3D类器官模型的进展表明，患者- 衍生的类器官（PDO）可以有效地产生，保留原始肿瘤的组织学和遗传学特征， 紧密模拟患者药物反应，并可用于基因组功能筛选。近日 Kuo实验室通过开发气液界面（ALI）培养系统开发了下一代PDO， 肿瘤上皮与内源性免疫基质的整体培养（Cell，2018）。ALI类器官的出现 培养为研究肿瘤微环境提供了关键的实验模型。这里，类器官核心 提供了用于鉴定进化动力学的基本的患者来源的类器官模型， 斯坦福大学乳腺转移中心转移性乳腺癌的微环境决定因素 investigators. 由乳腺癌基因组学专家克里斯蒂娜·柯蒂斯博士和类器官先驱卡尔文·郭博士共同领导， 斯坦福大学乳腺转移中心类器官核心是唯一定位和资格，以解决 RFA中强调的未满足的需求：转移研究网络，通过提供新的实验模型， 研究转移性播散模式、细胞微环境串扰和药物反应。这里 类器官核心将提供来自原发性和转移性乳腺癌的患者来源的浸没类器官， 研究靶向耐药性并确定乳腺癌亚克隆的特定转移模式 在项目1中，沿着使用CRISPR/CAS 9和巨噬细胞介导的基因组功能筛选， 项目3中的吞噬作用。新型人乳腺癌ALI类器官模型将被广泛表征 与与项目1和2合作的原始组织相比。此外，ALI培养条件将是 针对项目3的功能性肿瘤免疫串扰研究进行了优化。成功完成后，小说 类器官模型将为定义转移驱动基因和识别 治疗靶点，以克服耐药性和免疫逃避。

Abstract/Project Summary Breast cancer research is substantially hampered by lack of experimental models that efficiently reflect physiological disease necessary to provide a comprehensive understanding of metastatic disease. Conventional cell lines and PDX models have contributed greatly to our understanding of breast cancer biology, but are inefficiently derived, do not fully capture the heterogeneity across breast cancer subtypes and rarely mimic clinical observations. Advancements in 3D organoid models from our lab and others have shown that patient- derived organoids (PDOs) can be generated efficiently, retain histologic and genetic features of originating tumor, closely mimic patient drug response and are amendable to scale for genomic functional screens. Recently, the Kuo lab developed next generation PDOs by developing an Air-Liquid Interface (ALI) culture system that enables culture of tumor epithelium en bloc with endogenous immune stroma (Cell, 2018). The advent of ALI organoid culture provides critical experimental models for studying the tumor microenvironment. Here, the Organoid Core provides essential patient derived organoid models for identifying the evolutionary dynamics and microenvironmental determinants of metastatic breast cancer for Stanford Breast Metastasis Center investigators. Co-led by breast cancer genomics expert Dr. Christina Curtis and organoid pioneer Dr. Calvin Kuo, the Stanford Breast Metastasis Center Organoid Core is uniquely positioned and qualified to address the unmet needs highlighted in this RFA: Metastasis Research Network, by providing novel experimental models to study metastatic dissemination patterns, cellular microenvironment crosstalk, and drug response. Here, the organoid core will provide patient-derived breast cancer submerged organoids from primary and metastatic tissue to study targeted drug resistance and to define specific metastatic patterns of breast cancer sub-clones in Project 1, along with genomic functional screens using CRISPR/CAS9 and macrophage-mediated phagocytosis in Project 3. Novel human breast cancer ALI organoid models will be extensively characterized as compared to originating tissue in collaboration with Projects 1 and 2. Further, ALI culture conditions will be optimized for functional tumor-immune crosstalk studies with Project 3. Upon successful completion, novel organoid models will provide critical experimental tools for defining metastatic driver genes and identifying therapeutic targets to overcome drug resistance and immune evasion.