Core 2: Stanford Breast Metastasis Center Organoid Core

核心 2：斯坦福乳腺转移中心类器官核心

基本信息

批准号：
10704695
负责人：
CALVIN J KUO
金额：
$ 18.94万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-14 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10704695
关键词：
3-Dimensional Address Air Breast Breast Cancer Model Breast Cancer Patient Breast cancer metastasis CRISPR interference CRISPR-mediated transcriptional activation CRISPR/Cas technology Cancer Biology Cancer Relapse Carcinoma Cell Communication Cell Line Cells Characteristics Clinical Clinical Pathology Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Communication Consent Cryopreservation Development Disease Drug Modelings Drug Targeting Drug resistance Epithelial Cells Epithelium Experimental Models Feedback Genes Genetic Genetic Engineering Genomics Goals Heterogeneity Histologic Histology Human Immune Immune Evasion Immunological Models Impairment Information Dissemination Liquid substance Macrophage Malignant Epithelial Cell Malignant Neoplasms Mediating Metastatic breast cancer Methods Modeling Mus Mycoplasma Neoplasm Metastasis Occupational activity of managing finances Organoids Pathology Patients Pattern Phagocytosis Pharmaceutical Preparations Physiological Population Positioning Attribute Primary Neoplasm Production Property Proteomics Protocols documentation Qualifying Quality Control Research Research Personnel Resources Services Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Standardization Study models System Technology Testing Tissues Tumor Tissue Work anticancer research biobank breast cancer genomics cancer subtypes cell type digital functional genomics genome sequencing glycosylation in vitro Model malignant breast neoplasm mammary molecular subtypes nano-string next generation novel patient derived xenograft model reconstitution response single-cell RNA sequencing synergism therapeutic target therapy resistant tool transcriptome tumor tumor microenvironment tumor-immune system interactions whole genome

项目摘要

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.

摘要/项目摘要由于缺乏有效反映乳腺癌的实验模型，乳腺癌研究受到严重阻碍。生理疾病需要提供对转移性疾病的全面了解。传统的细胞系和 PDX 模型极大地促进了我们对乳腺癌生物学的理解，但派生效率低下，没有完全捕捉乳腺癌亚型之间的异质性，并且很少模仿临床观察。我们实验室和其他实验室在 3D 类器官模型方面取得的进展表明，患者可以有效地生成衍生类器官（PDO），保留原始肿瘤的组织学和遗传特征，密切模仿患者的药物反应，并且可以修改以用于基因组功能筛选。最近， Kuo 实验室通过开发气液界面 (ALI) 培养系统开发了下一代 PDO，该系统能够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。在这里，类器官核心提供重要的患者衍生类器官模型，用于识别进化动力学和斯坦福乳腺转移中心转移性乳腺癌的微环境决定因素调查人员。由乳腺癌基因组学专家 Christina Curtis 博士和类器官先驱 Calvin Kuo 博士共同领导，斯坦福乳腺转移中心类器官核心具有独特的定位并有资格解决以下问题：本 RFA 强调了未满足的需求：转移研究网络，通过提供新颖的实验模型研究转移扩散模式、细胞微环境串扰和药物反应。在这里，类器官核心将为患者提供来自原发性和转移性乳腺癌的浸没类器官组织来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，以及使用 CRISPR/CAS9 和巨噬细胞介导的基因组功能筛选项目 3 中的吞噬作用。新型人类乳腺癌 ALI 类器官模型将得到广泛表征与项目 1 和 2 合作的原始组织相比。此外，ALI 培养条件将与项目 3 一起针对功能性肿瘤免疫串扰研究进行了优化。成功完成后，新颖的类器官模型将为定义转移驱动基因和识别转移驱动基因提供关键的实验工具。克服耐药性和免疫逃避的治疗目标。