Isolation, characterization and translational development of glioma stem cells

项目总结/摘要 实体瘤是癌基因和/或肿瘤抑制突变积累的结果。 这些突变是如何在一个细胞中产生和积累的，这仍然是一个谜。很可能是一个 对形成前致瘤细胞的早期事件的更好理解可能会对 成熟肿瘤细胞的分析和对改进治疗开发的见解。我们已经充分发展 通过三个突变的多形性胶质母细胞瘤（GBM）的渗透性基因工程小鼠模型 在人GBM中通常发现突变的肿瘤抑制因子（P53、PTEN和NF 1）。使用我们的组合 在发育生物学和神经科学的背景下，我们追踪了这些肿瘤的起源， 成体干/祖细胞群。我们已经开发了工具来发现功能性GBM亚型， 这取决于肿瘤细胞的起源，而不是特定的驱动突变（Alcantara，Cancer Cell，2015）。 这些研究将扩展到确定细胞的起源以及与基因型和表型的关系。 此外，使用来自新的小鼠GBM亚型的基因表达特征，我们已经鉴定了 人类GBM对应物签名表明相似的生物起源和人类GBM的新策略 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 与起源的新细胞有关。小鼠模型显示了内源性GBM肿瘤细胞分级 将癌症干细胞放置在顶端。我们正在进行的研究表明，每个新的分层GBM 亚型由癌症干细胞生长模式控制。我们将扩大和确认这些 意见。使用表型高通量小化学化合物筛选，我们已经鉴定了小 对原代低传代GBM衍生细胞具有纳摩尔毒性但对原代GBM衍生细胞不具有纳摩尔毒性的分子 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 包括苯并咪唑鎓化合物及其衍生物的化合物显示出对初级 人GBM衍生的肿瘤球。这些化合物有望用于体内研究和鉴定新的 治疗开发的关键GBM依赖途径。我们正在开发一个全面的GBM 患者来源的异种移植程序，将用于进一步验证我们的小鼠模型发现， 鉴定、分离和中和人GBM癌干细胞。

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.