Project 3: Systematic characterization of factors controlling breast cancer progression and resistance

家长资助项目3概要：控制乳腺癌因素的系统表征 癌症进展和耐药性 两个核心挑战限制了癌症治疗的有效性并使转移成为可能： 对靶向药物的抗性，以及对癌细胞的识别和破坏的抗性的发展， 免疫系统.传统的体外癌症模型在规模上受到限制，并且通常缺乏关键特性 肿瘤微环境。我们最近开发了一种可扩展的癌症球体系统， 在3D培养中首次进行全基因组CRISPR筛选;该系统中的表型更好地反映了体内肿瘤 (Han例如，2020年）。此外，我们开发了一种磁分离策略，以快速识别调节剂， 巨噬细胞的吞噬作用（Haney等人，2018年），并成功地将这一战略扩展到研究 巨噬细胞-肿瘤细胞相互作用。在这里，我们建议使用这些系统来确定监管机构， 转移性乳腺癌对靶向治疗和内分泌治疗的耐药性以及巨噬细胞杀伤。 目的1：确定高复发风险ER+乳腺癌细胞生长和耐药性的关键驱动因素 使用CRISPRi/a屏幕的生产线和PDO 目的2：系统鉴定和表征限制乳腺巨噬细胞吞噬作用的因素 癌细胞

Summary of the Parent Grant Project 3: Systematic characterization of factors controlling breast cancer progression and resistance Two central challenges limit effectiveness of cancer therapies and enable metastasis: development of intrinsic resistance to targeted drugs, and development of resistance to recognition and destruction of cancer cells by the immune system. Traditional in vitro cancer models have been limited in scale and often lack key properties of the tumor microenvironment. We recently developed a scalable cancer spheroid system that enabled the first genome-wide CRISPR screens in 3D culture; phenotypes in this system much better reflect in vivo tumors (Han et al., 2020). In addition, we developed a magnetic separation strategy to rapidly identify regulators of phagocytosis by macrophages (Haney et al., 2018), and have successfully extended this strategy to study macrophage-tumor cell interactions. Here we propose to use these systems to identify regulators of resistance to targeted and endocrine therapies and macrophage killing in metastatic breast cancer. Aim 1: Identify critical drivers of growth and drug resistance in high relapse risk ER+ breast cancer cell lines and PDOs using CRISPRi/a screen Aim 2: Systematic identification and characterization of factors limiting macrophage phagocytosis of breast cancer cells