The Cancer Target Discovery and Development Network at UCSF

项目摘要 我们的总体战略是利用我们在开发过程中开发的新工具和方法， 第一个CTD^2资助期-更具体地说，开创和应用基于CRISPR的技术，以帮助 新型癌症靶点及其调节剂的发现和表征-使用创新的高通量 筛选方法我们的最终目标是发现微扰的最佳组合， 消除所有癌细胞，尽管它们的克隆异质性和环境背景。 一个目标是阐明新的分子靶点，以克服获得性耐药性。我们建立 在一个令人兴奋的系统，使我们能够定量基因型和表型细胞异质性为数百个 成千上万的单个癌细胞。我们提出了一系列治疗性小分子筛选， 与耐药性和来自TCGA，TARGET， CGCI、ICGC和相关举措。总的目标是确定合成基因组合所必需的， 临床耐药性，并与肿瘤间和肿瘤内异质性有关。 我们将开发和应用方法学来鉴定影响异型细胞的基因， 肿瘤中的相互作用。肿瘤进化是一个具有挑战性的研究领域，主要是由于细胞的复杂性， 类型和行为。在这个目标中，将进行高通量筛选以识别非细胞自主的 与肿瘤微环境相互作用相关的合成致死和合成存活相互作用。这些 研究将包括主要的T效应细胞/癌细胞相互作用，以确定新的治疗靶点和癌症 相关的巨噬细胞和成纤维细胞/癌细胞筛选以鉴定介导治疗抗性的基因。 这些系统是通过使用目前尚未公布的筛选平台，可能有助于 鉴定对癌症起始、维持和可能的转移重要的基因。由于我们将使用primary 和癌症组织，我们独特的平台将尽可能多地概括肿瘤的特征， 患者和解决癌症研究中的一个重要挑战。 我们已经开发出一种新的方法来建立哺乳动物细胞中的遗传上位相互作用， 扩大我们的努力，以产生特定的库，以映射上述目标的子集 卡位在这个目标中，我们将通过描述靶点在通路中的作用来解决靶点和机制 通过探索癌症定义分子的相互依赖性，使用新的靶点和筛选系统， 上面描述最终目标是揭示微扰的最佳组合， 消除所有癌细胞，尽管它们具有克隆异质性。

PROJECT SUMMARY Our general strategy is to take advantage of novel tools and methodologies that we have developed during our first CTD^2 funding period- more specifically pioneering and applying CRISPR based technologies to aid the discovery and characterization of novel cancer targets and their modulators– using innovative high throughput screening methods. Our end goal is to uncover optimal combinations of perturbagens with the potential to eliminate all cancer cells, despite their clonal heterogeneity and environmental context. One goal is to elucidate new molecular targets with the goal to overcome acquired drug resistance. We build upon an exciting system allowing us to quantitate genotypic and phenotypic cell heterogeneity for hundreds of thousands of single cancer cells. We propose a battery of therapeutic small molecule screens to identify candidate driver genes associated with drug resistance and with recurrent mutations from TCGA, TARGET, CGCI, ICGC and related initiatives. The overall goal is to identify synthetic gene combinations necessary for clinical resistance and related to inter- and intra-tumor heterogeneity. We will develop and apply methodologies for the identification of genes influencing heterotypic cell-cell interactions in tumors. Tumor evolution is a challenging area of research, largely due to the complexity of cell types and behaviors. In this aim, high-throughput screens will be performed to identify non-cell autonomous synthetic lethal and synthetic viable interactions relevant to tumor microenvironment interactions. These studies will include primary T-effector/cancer cell interactions to identify new therapeutic targets and cancer associated macrophage and fibroblast/cancer cell screens to identify genes mediating therapeutic resistance. These systems are made possible by using a currently unpublished screening platform that may help to identify genes important for cancer initiation, maintenance, and possibly metastasis. Since we will use primary and cancer tissue, our unique platform will recapitulate as much as possible the characteristics of tumors in patients and address an important challenge in cancer research. We have developed a novel means to establish genetic epistatic interactions in mammalian cells and will expand upon our efforts to generate specific libraries to map the subset of targets identified in the above screens. In this aim, we will address targets and mechanisms by delineating where targets act in the pathway by probing cancer-defining molecular interdependencies, using the novel targets and screening systems described above. The end goal is to uncover the optimal combination of perturbagens with the potential to eliminate all cancer cells, despite their clonal heterogeneity.