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

项目摘要 尽管手术、放疗和化疗是标准的治疗方法，但胶质母细胞瘤不可避免地会复发 会带来致命的后果。基底节细胞弥漫性地渗入大脑，通过手术切除完全切除。 不可能。手术后残留在浸润性脑内的肿瘤细胞和非肿瘤细胞的混合物 脱髓形成了辅助治疗干预和复发的生物学背景。此外， 放射治疗虽然有益于延长生存期，但可能会损害靶向治疗的疗效和 诱导治疗耐药。这项研究的目标是确定神经胶质瘤的分子特征。 对复发和治疗耐药负责。将纳入四个创新功能：1)a) 对基底膜及其周围脑组织进行放射定位取样的方法学；2)新方法 从MRI定位的活检组织中获取和分析大规模单细胞RNA-Seq数据3)Ribotag小鼠 实验上跟踪与进展和复发相关的特定细胞类型的改变。 在这个建议中，肿瘤的异质性是从中枢神经系统谱系的角度来探讨的，特别是 在肿瘤及其周围的细胞类型和细胞表型的变化的混合 渗入的脑组织。AIM1将确定浸润性边缘的基底膜的细胞和分子组成 为了证明浸润性边缘的胶质瘤细胞和反应性胶质细胞不同于肿瘤核心的细胞和反应性胶质细胞。 我们还将确定在最初的基底膜切除之间发生的细胞和分子组成的变化。 和治疗后肿瘤复发。在目标2中，我们将使用RiboTag小鼠神经胶质瘤模型来 确定辐射是否诱导间充质转化，从而对靶向治疗产生抵抗。通过 了解基底膜和浸润性脑组织的细胞和分子组成，并比较 在胶质瘤进展过程中和放射治疗后发生的细胞类型特异性改变的模式， 这项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 对目前的治疗方法有反应。

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.