Project 2: Dissection of clonal architecture and evolution in solid tumors

标题 项目2：实体瘤克隆结构和进化的剖析 摘要 越来越多的证据表明，实体瘤的耐药性和治疗失败的原因是 抗病亚系的自然选择。肿瘤内异质性与癌症 亚克隆多样性是肿瘤药物相对于其他药物的高失败率的关键驱动力。 这些专业将药物应用于稳定的体细胞基因组，而不是不稳定的基因组， 人类癌症在这个建议中，我们关注的是一种最难治愈的遗传异质性肿瘤。 （人类胶质母细胞瘤），以预测和验证标志着启动的驱动程序改变的景观，创始人 进化和个体患者内的治疗适应。我们将开发和应用新技术， 高通量转录和恶性神经胶质瘤组织内单个细胞的基因组分析。我们 目前的系统能够单细胞mRNA捕获、cDNA条形码化和芯片上扩增， 用于直接转化成标准的合并的测序文库的扩增子。我们将采用相同的设备， 大规模并行芯片上捕获来自单个细胞的基因组DNA用于全基因组扩增， 外显子组捕获接下来，我们将在原位小鼠中对单细胞胶质瘤模型进行功能验证， 体外和体内的人体系统。这一建议的成功结果将是提供一个综合的 计算实验管道，将能够预测未来的任何固体的进化运动， 在一组确定的选择性压力的存在下治疗肿瘤。这一信息将具有非常重要的意义 以破译不断演变的肿瘤依赖性，并提供最准确的治疗预测。

TITLE Project 2: Dissection of clonal architecture and evolution in solid tumors ABSTRACT Increasing evidence in solid tumors suggests that drug resistance and therapeutic failure results from natural selection of resistant subclones during the disease course. Intra-tumor heterogeneity and cancer subclonal diversity is the key driving force of the high failure rate of oncology drugs relative to other medical specialties where drugs are applied to stable somatic genomes rather than the unstable genomes found in human cancer. In this proposal, we focus on one of the most incurable and genetically heterogeneous tumors (human glioblastoma), to predict and validate the landscape of driver alterations that mark initiation, founder evolution, and therapy adaptation within individual patients. We will develop and apply novel technologies for high-throughput transcription and genomic analysis of individual cells within malignant glioma tissues. Our current system is capable of single cell mRNA capture, cDNA barcoding, and on-chip amplification, generating amplicons for direct conversion into a standard, pooled sequencing library. We will adapt the same device for massively parallel, on-chip capture of genomic DNA from individual cells for whole genome amplification and exome capture. Next, we will functionally validate the single-cell glioma models in orthotopic mouse and human systems in vitro and in vivo. The successful outcome of this proposal will be to deliver an integrated computational-experimental pipeline that will be able to predict the forthcoming evolutionary moves of any solid tumor in the presence of a defined set of selective pressures. This information will be of invaluable significance to decipher evolving tumor dependencies and provide the most accurate therapeutic predictions.