Tracing hypoxic state and hypoxia memory in glioblastoma progression

缺氧与癌症预后较差有关，包括最常见和高度恶性的胶质母细胞瘤(GBM) 致命的原发脑瘤。低氧肿瘤细胞被认为更适合生存治疗和繁殖 然而，肿瘤复发，低氧细胞在基底膜扩张过程中的轨迹及其在复发中的作用 治疗后的基底膜仍然知之甚少。这项提案的主要目标是在基因上应用双重基因 具有谱系追踪能力的编码低氧敏感报告系统系统地表征低氧 基底膜进展过程中的肿瘤细胞。双重报告系统区分急性缺氧的肿瘤细胞 (用绿色荧光标记)来自有低氧暴露史的人(永久标记为红色 荧光)。他们的细胞命运将被追踪跨越时间和空间在GBM进展期间，在初级和 复发性肾小球基底膜模型。细胞研究将与单细胞水平的基因表达分析相结合，以 解开缺氧肿瘤细胞恶性潜能的分子基础。通过比较转录 在原发和复发的基底膜中，急性低氧和低氧暴露的细胞的特征，我们将测试 存在低氧记忆的假设，即低氧暴露的细胞保留了一组低氧诱导的 可能与生存、健康和致癌潜力有关的基因。结合的描述性部分 在我们的研究中，我们还设计了一个遗传平台，通过时间控制选择性地消融低氧人群 目的：研究它们在原发和复发的基底膜中对肿瘤进展的作用。此功能数据将 提供直接证据来检验缺氧有助于肿瘤进展的中心假设，以及更多 重要的是，肿瘤在治疗后会复发。总而言之，我们的创新研究将对这一联系获得新的见解 低氧和基底膜恶性之间的关系，并为发现新的预后工具和 治疗公司的目标是打击GBM。

Hypoxia is linked to worse prognosis for cancers, including glioblastoma (GBM), the most common and highly lethal primary brain tumor. Hypoxic tumor cells are thought to be better equipped to survive therapy and spawn tumor recurrence, however, the trajectory of hypoxic cells during GBM expansion and their roles in recurrent GBM after therapy remain poorly understood. The main goal of this proposal is to apply a dual genetically encoded hypoxia-sensitive reporter system with lineage tracing capability to systematically characterize hypoxic tumor cells during GBM progression. The dual reporter system distinguishes tumor cells that are acutely hypoxic (marked by green fluorescence) from those with a history of hypoxia exposure (permanently marked with red fluorescence). Their cell fate will be traced across time and space during GBM progression in both primary and recurrent GBM models. The cellular study will be coupled with gene expression analysis on single cell level to unravel the molecular underpinning of malignant potency of hypoxic tumor cells. By comparing transcription profiles of acutely hypoxic versus hypoxia-exposed cells in primary and recurrent GBM, we will test the hypothesis that there exists a hypoxia memory, i.e., hypoxia-exposed cells retaining a set of hypoxia-induced genes that may be linked to survival fitness and tumorigenic potential. In conjunction to the descriptive part of our study, we have also devised a genetic platform to selectively ablate hypoxic population with temporal control to study their contribution to tumor progression in both primary and recurrent GBM. This functional data will provide direct evidence to test the central hypothesis that hypoxia contributes to tumor progression, and more importantly, tumor relapse after treatment. In summary, our innovative study will gain new insights into the link between hypoxia and GBM malignancy, and set the stage for the discovery of new prognostic tools and therapeutics targets to combat GBM.