Single cell analysis of the infiltrative margins of glioblastoma and post-treatment recurrence

项目摘要 尽管标准治疗包括手术、放疗和化疗，胶质母细胞瘤（GBM）仍不可避免地复发 带来致命的后果GBM弥漫性浸润大脑，通过手术切除完全清除 不可能的手术后残留在浸润脑内的肿瘤细胞和非肿瘤细胞的混合物 减积形成辅助治疗干预和复发的生物学背景。此外，委员会认为， 放射治疗虽然有利于延长生存期，但可能损害靶向治疗的疗效， 诱导治疗抵抗。本研究的目的是确定胶质瘤的分子特征 负责复发和治疗抗性。该系统将包含四个创新功能：1）a GBM和周围脑组织的放射学定位取样方法; 2） 从mri定位的活组织检查中获取并分析大规模单细胞RNA-Seq数据3）Ribotag小鼠 GBM模型，以实验性地追踪与进展和复发相关的细胞类型特异性改变。 在这个建议中，肿瘤异质性是从中枢神经系统谱系的角度来探讨的，特别是寻找 细胞类型的混合物和细胞表型的改变， 侵入脑组织Aim 1将决定浸润边缘GBM的细胞和分子组成 以证明浸润边缘的胶质瘤细胞和反应性胶质细胞与肿瘤中心的不同。 我们还将确定在初次GBM切除术后， 和治疗后肿瘤复发。在目标2中，我们将使用RiboTag小鼠前神经胶质瘤模型， 确定辐射是否诱导间充质转化，从而对靶向治疗产生抗性。通过 了解GBM和浸润脑组织的细胞和分子组成， 在神经胶质瘤进展期间和放射治疗后发生的细胞类型特异性改变的模式， 这一建议将揭示胶质瘤治疗的假定靶点，并确定特定的胶质瘤表型 对目前的治疗有反应。

Project Summary Despite standard treatment with surgery, radiation and chemotherapy, glioblastomas (GBMs) inevitably recur with fatal consequences. GBMs diffusely infiltrate the brain, making complete removal by surgical resection impossible. The mixtures of neoplastic and non-neoplastic cells that remain in the infiltrated brain after surgical debulking form the biological context for both adjuvant therapeutic intervention and recurrence. Furthermore, radiation therapy, while beneficial for extending survival, may compromise the efficacy of targeted therapy and induce treatment resistance. The goal of this study is to identify the molecular signatures in gliomas responsible for recurrence and treatment resistance. Four innovative features will be incorporated: 1) a methodology for radiographically -localized sampling of GBM and surrounding brain tissue; 2) new methods to acquire and analyze Large-Scale Single Cell RNA-Seq data from mri-localized biopsies 3) a Ribotag mouse model of GBM to experimentally track cell type-specific alterations associated with progression and recurrence. In this proposal, tumor heterogeneity is approached from the perspective of CNS lineages, specifically looking at the mixture of cell-types and the alterations in cell phenotypes that populate the tumor and surrounding infiltrated brain tissue. Aim1 will determine the cellular and molecular composition of GBM at infiltrative margins to demonstrate that glioma cells and reactive glia in the infiltrating margins differ from those in the tumor core. We will also determine changes in cellular and molecular composition that occur between initial GBM resection and post-treatment tumor recurrence. In Aim 2 we will use the RiboTag mouse model of proneural glioma to determine if radiation induces mesenchymal transformation with a resultant resistance to targeted therapy. By understanding the cellular and molecular composition of GBM and infiltrated brain tissue and comparing the patterns of cell type-specific alterations that occur during glioma progression and following radiation therapy, this proposal will reveal putative targets for glioma treatment and determine how specific glioma phenotypes respond to current therapies.