Isolation, characterization and translational development of glioma stem cells

项目摘要/摘要 实体瘤是癌基因和/或抑癌基因突变积累的结果。 这些突变是如何在一个细胞中产生并在一生中积累的仍然是一个谜。这很可能是一个 对形成肿瘤前细胞的早期事件的更好的理解可能对 对成熟肿瘤细胞的分析和对改进治疗发展的见解。我们已经完全发展起来了 突变型多形性胶质母细胞瘤基因工程小鼠模型的建立 肿瘤抑制基因(P53、PTEN和NF1)通常在人的GBM中发生突变。使用我们的联合 背景在发育生物学和神经科学中，我们已经追踪到这些肿瘤的起源是 成体干细胞/祖细胞群体。我们已经开发了工具来发现功能性GBM子类型，这些子类型是 预测基于起源的肿瘤细胞，而不是特定的驱动因素突变(阿尔坎塔拉，癌症细胞，2015)。 这些研究将扩展到确定细胞起源以及与基因和表型的关系。 此外，使用来自新的小鼠GBM亚型的基因表达签名，我们已经识别出 人类GBM对应签名表明相似的生物起源和人类的一种新策略 GBM分子分层。我们的数据为其他人类GBM亚型提供了证据，这些亚型也可能 与新的起源细胞有关。小鼠模型显示了内源性的GBM肿瘤细胞层级 在顶端放置一个癌症干细胞。我们正在进行的研究表明，每一种新的分层GBM 亚型由癌症干细胞的生长模式决定。我们将扩大和确认这些 观察。使用表型高通量小化合物筛选，我们已经鉴定出小分子 对原代低传代GBM来源细胞有纳摩尔毒性但对原代细胞无影响的分子 正常分裂的细胞，如小鼠胚胎成纤维细胞或新生星形胶质细胞。此外，铅 包括苯并咪唑类化合物及其衍生物在内的化合物对初生代 人GBM来源的肿瘤球体。这些化合物有望用于体内研究和识别新的 治疗发展的关键基底膜依赖途径。我们正在开发一种全面的GBM 患者衍生的异种移植程序，将用于进一步验证我们的小鼠模型发现并 鉴定、分离和中和人GBM肿瘤干细胞。

Project Summary/Abstract Solid tumors arise as the consequence of accumulation of oncogene and/or tumor suppressor mutations. How these mutations arise and accumulate in one cell over a lifetime remains a mystery. It is likely that an improved understanding of the early events that form a pre-tumorigenic cell could have implications for analysis of mature tumor cells and insights into improved therapy development. We have developed fully penetrant genetically engineered mouse models of glioblastoma multiforme (GBM) by mutation of three tumor suppressors commonly found mutated in human GBM (P53, PTEN, & NF1). Using our combined background in developmental biology and neuroscience, we have traced the origin of these tumors to the adult stem/progenitor cell population. We have developed tools to uncover functional GBM subtypes that are predicated on the tumor cell of origin rather than on specific driver mutations (Alcantara, Cancer Cell, 2015). These studies will be extended to identify cell of origin and relationship to genotype and phenotype. Moreover, using gene expression signatures from the novel mouse GBM subtypes, we have identified human GBM counterpart signatures that suggest similar biological origins and a novel strategy for human GBM molecular stratification. Our data provide evidence for additional human GBM subtypes that may also relate to novel cells of origin. The mouse models demonstrate an endogenous GBM tumor cell hierarchy placing a cancer stem cell at the apex. Our ongoing studies suggest that each of the new stratified GBM subtypes are governed by a cancer stem cell pattern of growth. We will expand and confirm these observations. Using a phenotypic high throughput small chemical compound screen we have identified small molecules that have nanomolar toxicity on primary low passage GBM derived cells but not on primary normally dividing cells such as mouse embryo fibroblasts or neonatal astrocytes. In addition, lead compounds including a benzimidazolium compound and its derivatives demonstrate toxicity on primary human GBM derived tumor spheres. These compounds hold promise for in vivo studies and identifying novel key GBM dependency pathways for therapeutic development. We are developing a comprehensive GBM patient derived xenograft program that will be employed to further validate our mouse model finding and to identify, isolate and neutralize human GBM cancer stem cells.